Journal of Thrombosis and Haemostasis, 11 (Suppl. 1): 46–66
12253
INVITED REVIEW
Evolutionary origins of the blood vascular system and
endothelium
R . M O N A H A N - E A R L E Y , * † A . M . D V O R A K * † and W . C . A I R D * ‡ §
*The Center for Vascular Biology Research, Beth Israel Deaconess Medical Center; †Department of Pathology, Beth Israel Deaconess Medical
Center; ‡Department of Medicine, Beth Israel Deaconess Medical Center, Boston, MA; and §Mount Desert Island Biological Laboratory,
Salisbury Cove, ME, USA
To cite this article: Monahan-Earley R, Dvorak AM, Aird WC. Evolutionary origins of the blood vascular system and endothelium. J Thromb
Haemost 2013; 11 (Suppl. 1): 46–66.
Summary. Every biological trait requires both a proxi-
mate and evolutionary explanation. The field of vascular
biology is focused primarily on proximate mechanisms in
health and disease. Comparatively little attention has
been given to the evolutionary basis of the cardiovascular
system. Here, we employ a comparative approach to
review the phylogenetic history of the blood vascular sys-
tem and endothelium. In addition to drawing on the pub-
lished literature, we provide primary ultrastructural data
related to the lobster, earthworm, amphioxus, and hag-
fish. Existing evidence suggests that the blood vascular
system first appeared in an ancestor of the triploblasts
over 600 million years ago, as a means to overcome the
time-distance constraints of diffusion. The endothelium
evolved in an ancestral vertebrate some 540–510 million
years ago to optimize flow dynamics and barrier function,
and/or to localize immune and coagulation functions.
Finally, we emphasize that endothelial heterogeneity
evolved as a core feature of the endothelium from the
outset, reflecting its role in meeting the diverse needs of
body tissues.
Keywords: biological evolution, blood vessels, endothe-
lium, heart, phylogeny.
Introduction
Every biological trait requires both a proximate and evo-
lutionary explanation (reviewed in [1]). Proximate expla-
nations (‘how’) employ traditional tools, including
biochemistry, molecular biology, and cell biology to
Correspondence: William C. Aird, Beth Israel Deaconess Medical
Center, Molecular and Vascular Medicine; RN-227, 330 Brookline
Ave., Boston, MA 02215, USA.
Tel.: +1 617 667 1031; fax: +1 617 667 1035.
E-mail: waird@bidmc.harvard.edu
DOI: 10.1111/jth.
describe the anatomy, physiology, and ontogeny of a trait
at the level of a modern-day organism. Evolutionary
explanations (‘when and why’) employ a combination of
the fossil record, and comparative morphology and DNA
sequences to describe the phylogenetic history of a trait
and the fitness advantage that the trait provides at the
level of a population or species [2].
The field of cardiovascular biology is largely concerned
with proximate mechanisms. How does the heart generate
force? How do blood vessels form during development?
How do blood vessels and their endothelial lining func-
tion in health and how do they become dysfunctional in
disease? By contrast, little attention is paid to the evolu-
tionary mechanisms of the cardiovascular system. When
and why did the cardiovascular system evolve in the first
place? Why are certain blood circulatory systems open,
while others are closed? Why are some systems lined by
endothelium, whereas others have no cell lining? These
are important questions because they provide insights into
the design constraints, path dependence, trade-offs, and
selective pressures that underlie human physiology and
vulnerability to disease.
The goal of this review is to explore the evolutionary
origins of the blood vascular system and endothelium.
There is no trace of the cardiovascular system in the
fossil record. Molecular phylogenetic analyses have
yielded interesting, although limited insights into evolu-
tionarily conserved mechanisms of heart development
and tube formation. By contrast, comparative biology
provides a rich source of information that can be used
to infer and reconstruct the evolutionary history of the
cardiovascular system. We will begin with an overview
of body plans in extant animals. Next, we will discuss
concrete examples (and introduce primary ultrastructural
data) from a spectrum of representative species, with a
particular emphasis on invertebrates. Finally, we will use
these data to draw conclusions about the evolutionary
origins of the blood vascular system. A glossary of
terms and concepts used in the current review is
included in the Appendix S2.
© 2013 International Society on Thrombosis and Haemostasis

Body plans
All animals have evolved to survive and reproduce, and
to do so they share common tasks that include the cap-
ture, ingestion, absorption, and distribution of food/nutri-
ents; the acquisition and distribution of oxygen for
cellular respiration; and the excretion of metabolic wastes
and undigested materials [3]. Different species use differ-
ent strategies to achieve these goals. Such strategies
diverge most prominently with changes in body size and
complexity. However, the number of identifiable themes
or body plans (including the structural design of cardio-
vascular systems) is limited by developmental/genetic con-
straints and the laws of chemistry and physics. A
consideration of these designs provides important insights
into the evolutionary history of the blood vascular system
and endothelium.
A word on taxonomy
Eukaryotes consist of two kingdoms, the Protozoa and
Animalia (Metazoa). For our purposes, Metazoa, which
represent multicellular eukaryotes, can be further classi-
fied in one of two ways. The first classification employs
molecular and/or morphological data to describe the evo-
lutionary relationships among major metazoan lineages.
The results, which may be represented in the form of phy-
logenetic trees (an example is shown in Fig. 1), can be
used to infer evolutionary histories. The deep branches on
the animal tree of life remain controversial. Although the
details of the metazoan phylogenic tree continue to be
revised as new evidence emerges, several important
themes emerge. The origin of the Metazoa dates back to
approximately 770–850 million years. The most primitive
living phylum of animals is the Porifera (sponges), fol-
lowed by Cnidaria (corals and jellyfish) and Ctenophora
(comb jellies). Only two germ layers (endoderm and ecto-
derm) develop in these latter phyla. Hence, they are called
diploblastic animals. Between 600 and 700 million years
ago, a new body plan emerged that demonstrated bilat-
eral symmetry and a third germ layer (mesoderm).1 These
animals, referred to as triploblasts, gave rise to two sepa-
rate lineages: the protostomes and deuterostomes.2 The
deuterostome lineage gave rise to the chordates (including
cephalochordates, urochordates, and vertebrates) as well
as hemichordates (acorn worms) and echinoderms (e.g.
sea urchins and starfish). Protostomes are further divided
into two groups: the Ecdysozoans (including the arthro-
1
Bilaterality is associated with cephalization (anterior–posterior
head-to-tail body axis); a dorsoventral back-to-front axis; and a divi-
sion of the body into left and right sides.
2
The deuterostomes and protostomes are considered superphyla of
Metazoa. They differ primarily in their embryonic origins. For exam-
ple, in protostomes, the blastopore becomes the mouth, whereas in
deuterostomes, it becomes the anus.
© 2013 International Society on Thrombosis and Haemostasis
Evolutionary origins of the blood vascular system 47
pods and nematodes) and the Lophotrochozoans (includ-
ing the mollusks and annelids).3 As we shall see, these
various branch points mark important transitions in the
development of vascular systems.
A second, simpler classification separates Metazoa into
vertebrates (those animals that possess a backbone/verte-
bral column consisting of bone or cartilage) and inverte-
brates (those that do not possess such a structure).
Vertebrates, which constitute the subphylum Vertebra of
the phylum Chordata, comprise only 3% of all living ani-
mal species. Invertebrates, which span over 30 phyla, con-
stitute the remainder of metazoan species. Invertebrates
are highly paraphyletic4 and thus display an enormous
diversity of body plans. By contrast, all vertebrates are
derived from a common ancestor and are thus con-
structed on a single common body plan.5 Although the
invertebrate–vertebrate dichotomy carries little taxonomic
meaning, it allows for a distinction between the proto-
typic vertebrate cardiovascular system and a broad spec-
trum of vascular phenotypes in invertebrates, many of
which arose independently in response to common selec-
tive pressures.
Size matters
Larger body size is a major trend in animal evolution.
Changes in size mandate changes in structural design. All
unicellular and multicellular animals depend on diffusion
to supply oxygen and nutrients, and to remove carbon
dioxide. Diffusion, while energetically inexpensive, is a
very slow process and works only over small distances
(diffusion path < 1 mm).6 A change in body size (whether
for a single cell or a multicellular organism) dispropor-
tionately changes the ratio of surface area to volume.
Specifically, as a solid 3-dimensional body enlarges, its
surface area increases in proportion to the radius squared
(r2), whereas its volume increases more rapidly (r3). At
some point, the cell will reach a size where its surface
area cannot meet the needs of its volume. Single cells
3
These two groups were initially proposed based on molecular
phylogenetic analyses of the protostomes (particularly similarities in
18S rRNA genes).
4
A paraphyletic group is a group whose member species are all
descendants of a common ancestry, but that does not contain all the
species descended from that ancestor.
5
For example, all vertebrates have an axial skeleton with a verte-
bral column, brain complexity, an anterior mouth, a posterior anus,
a digestive tract, a liver, kidneys, a ventrally located heart (two
hearts in the case of hagfish), a closed circulatory system with an
endothelial lining, a neural crest, and acquired immunity.
6
According to Fick’s first law, diffusion is proportional to the con-
centration gradient and surface area and inversely proportional to
the distance. dS/dt = D 9 A 9 dC/dx, where dS/dt is the rate of
transport, A is the area through which diffusion occurs, dC/dx is the
concentration gradient, and D is the diffusion coefficient of the sub-
stance [5].
48 R. Monahan-Earley et al

Mollusca (e.g. clams, squid)

Lophotro-
chozoa Annelida (e.g. earthworms)

Phoronida
Brachiopoda
Nemertea (“ribbon worm”)

Platyhelminthes (“flatworm”)
Polyzoa
Gnathifera Protostomes
Triploblasts

Chaetognatha

Tardigrada
Ecdysozoa Nematoda
Nematomorpha
Scalidophora
Onychophora (“velvet worm”)
Protostome
Arthropoda (e.g. lobsters, insects)
deuterostome
ancestor
Echinodermata (e.g. star fish, sea urchins)
Hemichordata (e.g. acorn worms)
Chordata Deuterostomes
Deutero-
Cephalochordates (e.g. amphioxus)
stomia
Urochordates (e.g. sea squirt)
Agnathans (e.g. hagfish, lamprey)
Gnathostomes (e.g. jawed fish)
Diploblasts

Cnidaria (e.g. corals, jellyfish)

Porifera (“sponges”)
Ctenophora (“comb jellies”)

Fig. 1. Metazoan phylogenetic tree. Adapted and modified from Juliano et al. [4].

optimize their surface area-to-volume ratio by developing
folded surfaces, or a flattened or thread-like shape.
Another strategy to increase size is to incorporate many
cells into a single organism. Simple multicellular organ-
isms (diploblastic animals and some of the early triplo-
blastic animals, such as flatworms) obtain oxygen by
diffusion alone. They do so by minimizing metabolic
demands, by assuming a body geometry that maximizes
the surface area, by localizing most of their cells at the
environment/body interface and/or by pumping external
environmental water to their internal surfaces. However,
these strategies have inherent design constraints that place
an upper limit on body size. To achieve further 3-dimen-
sional increases in size, it is necessary to employ internal
transport and exchange systems (i.e. circulatory systems)
to provide bulk flow delivery of substances (e.g. gases,
nutrients, wastes) to and from each cell in the body.
Circulatory systems
A circulatory system is any system of moving fluids that
reduces the functional diffusion distance that nutrients,
gases, and metabolic waste products must traverse regard-
less of its embryological origin or its design [3]. Circula-
tory systems are highly varied. In diploblasts, they
involve circulation of seawater into a body cavity that is
open to the environment. In triploblasts, however, circu-
latory fluid is an internal, extracellular, aqueous medium
produced by the animal, and distributed either through
body cavities or through integrated networks of vessels,
sinuses, and pumping organs. Circulatory fluids can be
moved by ciliary function or muscle action. Directional
flow is achieved by means of coordinated peristaltic waves
of contraction and/or the presence of one-way valves.
There are two internal circulatory systems: coelomic and
blood vascular. Most triploblastic animals possess both a
coelomic circulatory system and a blood vascular system.7
Coelomic circulatory systems Some small and flat triplo-
blastic animals (e.g. flatworms) have no system of internal
fluid support (i.e. they lack a coelom or blood vascular
system). They are called acoelomates (Fig. 2). The space
7
There are some exceptions such as higher leeches and the nemer-
tines, in which a coelom exists without a blood vascular system.
Conversely, animals with an open circulation (e.g. mollusks and ar-
thropods) have a blood vascular system, but a significantly regressed
coelomic system.

© 2013 International Society on Thrombosis and Haemostasis


between their ectoderm and endoderm tissue layers is
filled with a meshwork of mesodermal cells called paren-
chyma. These creatures obtain all of their oxygen and
food by simple diffusion across the skin and gut and
throughout the intercellular medium.8 If there are fluid-
filled clefts in this meshwork, the animal is termed a pseu-
docoelomate.9 However, most triploblastic animals have a
fluid-filled body cavity between the outer body wall (ecto-
derm) and the digestive tube (endoderm), termed the coe-
lom. The coelom is lined by mesoderm-derived epithelium
(termed mesothelium), with the apical surface of the meso-
thelial cells facing toward the lumen.10 The cavity is filled
with coelomic fluid, which in some cases contains cells
(termed coelomocytes). Coelomic cavities have never
developed a pumping system. Instead, fluid transport is
produced by cilia on the surface of the mesothelial cells,
by contraction of mesothelial cells (which have acquired a
myoepithelial phenotype and are thus termed myoepitheli-
al cells), or by contraction of body wall muscle. The coe-
lom is usually subdivided into multiple compartments by
septa and mesenteries (Fig. 2).11 In addition to providing
convective flow of gases, nutrients, and wastes, the
appearance of the coelom allowed organs to move freely,
rather than being embedded in solid mesoderm tissue.
The coelom also provided space for organ development.
Functionally, a coelom can absorb shock or provide a
hydrostatic skeleton essential for certain types of move-
ment.12 Because they tend to be compartmentalized, coe-
lomic cavities function in the local circulation of fluid. In
contrast, blood vascular systems have evolved in the vast
majority of coelomates to provide bulk fluid transport
throughout the body of segmented animals.
Blood vascular systems The blood vascular system con-
sists of blood-filled spaces (vessels, sinuses, hemocoels,
and/or pumping organs) within the connective tissue com-
partment, which is continuous around and between all tis-
sue layers in the body [6]. In invertebrates, the spaces are
lined only by matrix. Vertebrates have evolved a secondary
cell lining, termed endothelium. Fluid transport is produced
8
Large flatworms have highly branched guts that carry out the
functional equivalent of internal transport. Reliance on the gut for
diffusion of gases and nutrients limits the degree to which the gut
may be regionally specialized, for example for digestion, and absorp-
tion.
9
The pseudocoelom, also referred to as a blastocoelom, is not
formed from mesoderm and is not lined by mesothelium. It may rep-
resent remnants of the embryonic blastocoel [3, p. 48–9].
10
Mesothelial cells are also called myoepithelial cells, peritoneal
cells, and cardiac cells.
11
Examples in mammals include the pericardial cavity, pleural
space, and peritoneal cavity.
12
In fact, the coelom may have evolved initially to provide a
hydrostatic skeleton for early soft-bodied, bilaterally symmetrical
animals that assumed a crawling or burrowing lifestyle.
© 2013 International Society on Thrombosis and Haemostasis
Evolutionary origins of the blood vascular system 49
by contraction of specialized mesothelial cells (myoepitheli-
al cells) and/or muscular pumps.13 The blood vascular sys-
tem is used for the transport of substances (e.g. nutrients,
oxygen, carbon dioxide), hydraulic force generation (e.g.
head–foot protrusion in mollusks and penile erection in
vertebrates), regulation of heat (e.g. via countercurrent
flow), ultrafiltration (e.g. in the kidney), defense (e.g.
through delivery of clotting factors, immune factors/cells),
and whole-body integration (e.g. hormonal regulation).
Blood vascular systems follow one of two principal designs:
open or closed. In keeping with their paraphyletic origins,
invertebrates display diverse phenotypes ranging from open
to closed systems. In contrast, all vertebrates have a closed
cardiovascular system. As we shall discuss, the division
between closed and open systems is not always clear-cut.
Open circulatory systems occur in arthropods (e.g.
insects and crustaceans) and non-cephalopod mollusks
(e.g. clams, snails and slugs). In these animals, the meso-
derm forms coelomic cavities during embryogenesis.
However, the cavities and their cell lining regress in the
adult. Some populations of mesodermal cells reaggregate
and form the blood vessels (e.g. the dorsal vessel in
insects). The system is considered open because the blood
(called hemolymph) empties from a contractile heart and
major supply vessels into the body cavity (termed a hemo-
coel), where it directly bathes the organs. In other words,
the hemocoel is bordered not by mesoderm-derived meso-
thelial cells, but rather by the basal surface of the tissue
cells themselves.14 Thus, once blood empties from the
lumen of the distributing vessels, there is no distinction
between hemolymph and interstitial fluid/extracellular
fluid. Hemolymph returns to the heart either through
ostia in the ventricle (arthropods) or via the atrium (mol-
lusks).15 Compared with the closed circulation in lower
invertebrates (e.g. annelids), the open circulation boasts a
more efficient pump. Whereas annelids rely on peristaltic
blood vessels to propel blood, animals with an open cir-
culation have evolved true hearts (containing muscle stria-
tions and Z-bands), which display automatic and
synchronized beating. However, compared with more
13
The following types of pumps or hearts may be distinguished: (i)
myoepithelial cell-containing peristaltic pulsating vessels which pro-
pel blood by peristalsis (e.g. annelids and amphioxus); (ii) cardio-
myocyte-containing tubular hearts (which represent an adaptation of
an original peristaltic design) whose beating is synchronous or near-
synchronous (e.g. insects); and (iii) cardiomyocyte-containing cham-
bered hearts whose beating is synchronous or near-synchronous (e.g.
mollusks and vertebrates). The latter two designs are controlled
myogenically or neurogenically [7].
14
The hemocoel is not a coelom. It may be viewed as a persistent
blastoceolic remnant, which has been secondarily derived during evo-
lution [3, p. 476].
15
Mollusks and vertebrates are the only animals that possess a
chambered heart with at least one ventricle and atrium. The occur-
rence of chambered hearts in these two phylogenetically distant
groups is an example of evolutionary convergence.
50 R. Monahan-Earley et al

Key: L R
Planes of
longitudinal Longitudinal
Cross-section Notes
sections V section

Acoelomate Acoelomate
Epidermis Parenchyma Mesoderm forms somewhat solid mass of tissue, called
parenchyma
(ectoderm) (mesoderm)
No system of internal fluid support
Mouth Gut
Oxygen exchange/delivery occurs by diffusion across
the skin and gut
e.g., flatworms
Gut (endoderm)

Epidermis (ectoderm) Pseudocoelomate

Pseudocoelomate
Body wall (mesoderm) Fluid-filled clefts in mesoderm form a body
cavity, termed a pseudocoelom
Pseudocoelom Pseudo- e.g., nematodes
coelom
Mouth Gut
Eucoelomate
Fluid-filled body cavity forms within mesoderm,
Gut (endoderm) termed the coelom
Coelom is lined by mesoderm-derived epi-
thelium termed mesothelium
Epidermis (ectoderm) The apical surface of the mesothelial cells
Eucoelomate Body wall (mesoderm) faces towards the lumen
Basal
Mesothelium (mesoderm) The cavity is filled with coelomic fluid and in some cases cells
lamina
No pumping system
Fluid transport is produced by cilia on the surface of the
Coelom Dorsal mesentery mesothelial cells, by contraction of mesothelial cells or by
contraction of body wall muscle
Mouth Gut Gut (endoderm)
Coelom provides convective flow of gases, nutrients and
Coelom wastes, allows organs to move freely, provides space for
Ventral mesentery organ development, absorbs shock and provides a hydro-
static skeleton essential for certain types of movement
e.g., mollusks, annelids, arthropods, chordates

Epidermis (ectoderm)
Eucoelomate + Blood vascular system
Body wall (mesoderm) Eucoelomate + Blood vascular system
Mesothelium (mesoderm) In most animals, the coelom is subdivided into several
Closed System Blood vessel compartments by septa and mesenteries
(Note: Longitudinal Direction of flow in Owing to compartmentalization, fluid circulation in the
Ventral dorsal blood vessel coelomic cavities is localised
blood section at level
of ventral blood vessel) Septum Blood vessels form to bypass the septal bulkheads
vessel
(wall of Blood transport is produced by contraction of specialized
in mes-
coelom) mesothelial cells (termed myoepithelial cells) and/or muscular
entery
Coelom pumps
Gut
The blood vascular systems is used for transport of substances,
hydraulic force generation, regulation of heat, ultrafiltration,
protection, immune factors/cells and whole-body integration

Closed System
Blood vessel in septum Direction of flow in
Blood remains inside distinct channels or chambers, physically
ventral blood vessel separated from the intercellular fluid, body cells and coelom
Invertebrate Vertebrate In invertebrates, blood vessels consist of a network of spaces
Microfilaments
in the extracellular matrix, lined by the basal lamina/basement
Smooth membrane of surrounding epithelial cells
ECs muscle cell
Lumen (No ECs) In vertebrates, blood vessels are lined by endothelium and
in Basal lamina (–) invested with smooth muscle cells or pericytes
Mesothelial cells lumen and ECs lining Blood transport is produced by the pumping action of the
Basal with cilia blood vessels blood vessels (most invertebrates) or a chambered heart
(cephalopods, vertebrates)
lamina
e.g., annelids, cephalopods, chordates

Epidermis (ectoderm)
Open System Open System
Heart Body wall (mesoderm)
Coelomic cavities and their cell lining regress in the adult,
leaving a body cavity, termed a hemocoel (a type of acquired
HEART Unidirectional flow pseudocoelom)
Hemocoel is bordered not by mesoderm-derived mesothelial
cells, but rather by the basal surface of tissue cells
Mouth Gut
Blood is propelled from a contractile heart (+/– supply vessels)
Hemocoel into the hemocoel, where it directly bathes the organs
e.g., mollusks (except cephalopods), arthropods
Gut (endoderm)

Fig. 2. Schematic of different circulatory systems.

© 2013 International Society on Thrombosis and Haemostasis


advanced closed systems (e.g. in cephalopods and verte-
brates), open circulatory systems have a larger blood vol-
ume and lower flow rates/pressures. Indeed, the velocity
and blood pressure drop abruptly once the blood leaves
the heart and vessels and enters the hemocoel. As a final
point of comparison, open systems provide flow to organs
in series, such that tissues lying further downstream of
the heart receive less oxygen compared with more proxi-
mal organs. By contrast, the capillary networks of closed
circulatory systems are arranged in parallel allowing for
equal (and regulated) distribution among tissues.
In insects, the open circulation is not responsible for
delivering oxygen. Oxygen delivery is carried out by an
elaborate, highly branched tracheal system, which facili-
tates diffusion to each and every cell of the body. As a
result, insects have a larger capacity for aerobic metabo-
lism compared with other open circulation invertebrates.
Closed circulatory systems occur in a wide variety of
invertebrates including annelids, cephalopods (e.g. octo-
pus and squid), and non-vertebrate chordates, as well as
in vertebrates. In closed systems, the blood remains inside
distinct channels or chambers, where it is physically sepa-
rated from the intercellular fluid, body cells, and coe-
lom.16 Closed systems consist of collecting and
distributing vessels, usually with a central meeting site in
a propulsive pump. Exchange with the interstitial fluid
and body cells takes place in special areas such as capil-
lary beds or plexi, where the walls are thin to optimize
diffusion. The closed system of invertebrates consists of a
network of spaces in the extracellular matrix between epi-
thelial cells where the coelom contacts coelom (e.g. mes-
entery or septa) or where coelom contacts endoderm or
ectoderm (Fig. 2). Thus, the wall of these vessels consists
of basement membranes derived from different types of
epithelia, and the vessels are outlined by the basal sur-
faces of these cells. In some cases (e.g. annelids), blood is
propelled by the pumping action of the blood vessels,
which in turn is mediated by the contraction of special-
ized, differentiated coelomic mesothelial cells (myoepithe-
lial cells),17 or by muscular blood vessels that contract in
peristaltic waves. In other cases (e.g. in cephalopods),
16
Some authors have used ‘closed’ to describe systems lined by
endothelium [8] (e.g. see McMahon B.R. Comparative evolution and
design in non-vertebrate cardiovascular systems. In: Sedmera and
Wang [9]). Our definition does not require the presence of a cell lin-
ing.
17
These latter cells, which contain contractile myofibrils, may
remain in the coelomic cavity or they may delaminate and constitute
a differentiated contractile cell layer between the hemal cavity and
coelomic epithelium (giving rise to a multilayered blood vessel wall)
[10,11]. In addition to giving rise to contractile myoepithelial cells,
coelomic epithelial cells are believed to differentiate into hemocytes
during development [12]. Hemocytes are diverse in structure and
function, but participate in functions similar to vertebrate blood
cells.
© 2013 International Society on Thrombosis and Haemostasis
Evolutionary origins of the blood vascular system 51
chambered hearts have evolved to promote fluid move-
ment.
In vertebrates, the closed vascular system consists of a
series of closed vessels with an endothelial lining, invested
with smooth muscle cells or pericytes. Vertebrate blood
vessels, while contractile, do not propel blood. Rather,
transport is mediated by a central muscular pump. Most
vertebrates have a lymphatic system, which collects and
recycles interstitial fluid back to the circulation. There are
variations on the common vertebrate plan, many of which
are related to different requirements of living in water or
on land (reviewed in [13,14]). For example, fish have a
single circulatory system, consisting of an undivided heart
with a single atrium and single ventricle in series with
oxygen-exchanging gills. By contrast, adult birds and
mammals have a double circulatory system in which the
heart is completely divided into right and left sides, result-
ing in separation of deoxygenated and oxygenated blood.
The circulatory systems of lungfish, amphibians, and rep-
tiles demonstrate a broad array of intermediate designs,
each of which is characterized by partial separation of the
air-breathing organ and the systemic circulation, and thus
the potential for left–right and right–left shunts.
The idea that cardiovascular systems are either closed
or open is an oversimplification [9]. For example, in many
mollusks and crustaceans, blood is ejected from the heart
into a highly ramified network of branching vessels before
emptying into the hemocoel. This is illustrated in the lob-
ster (see section below), and even more prominently in
the sea snail, Haliotis, where the abdominal arteries end
up in a complex meshwork of small capillary-like vessels
[15].18 Moreover, studies in drosophila have shown that
blood flows along preferred routes in the open hemocoel
despite the absence of discrete return channels [16]. Con-
versely, the closed system of vertebrates contains vascular
beds, such as the sinusoids of the liver, spleen, and bone
marrow, where there is direct contact between blood and
the interstitial space. In hemochorial placentation (e.g. in
primates), the maternal spiral arteries become open-
ended, and blood is released into a placental labyrinth
where it bathes the chorionic villi and is drained by the
spiral veins. Such an arrangement is highly reminiscent of
an open system.
Endothelium
Invertebrate vessels are always lined by extracellular
matrix. Only vertebrates possess a true endothelial lining,
defined as a layer of epithelial cells expressing basoapi-
cal polarity (with the apical surface facing the lumen),
18
There is no structural equivalent of a vertebrate artery in inverte-
brates. The term ‘artery’ is often used to describe distributing vessels
that supply blood downstream of a pump. In other words, an artery
is defined by its efferent relationship to the pump and not by its wall
structure.
52 R. Monahan-Earley et al
intercellular junctions, and anchoring to basement mem-
brane. The blood vessels of some invertebrates, including
cephalopods, annelids, and amphioxus (discussed next)
have cells clinging to the luminal surface, internal to the
basement membrane. These cells have sometimes been
referred to as ‘endothelial cells’ [8,17–19]. However, they
form an incomplete lining, they lack intercellular junc-
tions typical of vertebrate endothelial cells, and they
rarely appear attached to the underlying basal lamina. A
more appropriate term for this cell type is an amoebocyte.
It likely represents a type of circulating hemocyte, which
may or not be an evolutionary precursor of the endothe-
lial cell.
Gas exchange
The circulatory system has co-evolved with gas exchange
mechanisms. Some invertebrates obtain their oxygen by
simple diffusion through the skin. Integumentary gas
exchange is characteristic of small soft-bodied animals
with high surface/volume ratios. Gas exchange in the
majority of marine and many freshwater invertebrates
occurs via gills. Terrestrial invertebrates employ special-
ized invaginated structures, including book lungs (spiders)
or trachea (insects). Many invertebrates have metal ion-
containing respiratory pigments to increase the oxygen-
carrying capacity of their blood. For example, hemoglo-
bin is found in many annelids, as well as some crusta-
ceans, insects, and mollusks, whereas hemocyanins are
the most commonly occurring respiratory pigments in
mollusks and arthropods. Invertebrate respiratory pig-
ments are usually carried in solution in blood, but they
are occasionally packaged in blood cells. Vertebrates
breathe through their gills, skin, and/or lungs. All verte-
brates with the exception of Antarctic icefishes (Chann-
ichthyidae) have hemoglobin, which is packaged in red
blood cells.
Case studies
In this section, we will consider the blood vascular sys-
tems of four different animals: 3 invertebrates and one
basal vertebrate. These examples will be used to highlight
features of cardiovascular design in the animal kingdom.
We begin with an example of an open circulation (lob-
ster). We then turn to three examples of a closed circula-
tion (earthworm, amphioxus, and hagfish). The reader is
referred to the Appendix S1 for a description of the
methods employed and for additional figures.
Case study 1: The lobster (Homarus americanus)
Lobsters belong to the phylum Arthropoda, subphylum
Crustacea. Like other arthropods, they have an open cir-
culation (Fig. 3A, Fig. S1). They have a well-developed
single-chambered heart suspended in a pericardial sinus
in the dorsal thorax by several pairs of alary ligaments
(Fig. 3B). These elastic ligaments are stretched during
systole. In diastole, they expand the heart, which results
in passive filling with hemolymph from the pericardial
sinus through three pairs of muscular valved ostia [21].
The heart is comprised of striated cardiac muscle cells
and is conspicuous for its absence of an endocardial lin-
ing (Fig. 3C, Fig. S2). The heart empties into 7 arteries,
which then branch multiple times along their length into
thin-walled microvessels before emptying into tissue lacu-
nae (i.e. the hemocoel) (Fig. 3A,F) [22]. Muscular car-
dio-arterial valves are present at the origins of all but
the dorsal abdominal artery. Blood is collected into a
series of interconnected sinuses and passes through
either the gills or the branchiostegal sinus before being
delivered back to the pericardial sinus surrounding the
heart [22].
Arteries leaving the heart are trilaminar vessels consist-
ing of an inner acellular tunic interna, a tunica intermedia
which contains fibroblast-like cells, and an outer tunica
externa (Fig. 3D, Fig. S3) [23,24]. Of the seven arteries,
only the dorsal abdominal artery has occasional muscle
fibers in its wall [23,25]. Electron microscopy reveals the
presence of an internal lamina containing microfibrils and
a medial lamina composed of a dense network of microfi-
brils (Fig. 3E, Fig. S4) [26]. All invertebrates lack elastin.
Nonetheless, lobster arteries demonstrate non-linear elas-
ticity (typical of vertebrate arteries) [23]. This property
may be related to the reorientation of the microfibrils
within the vessel wall [27].
Previous studies in lobster have demonstrated heart
rates of 61–83 beats per minute [23,28] and systolic pres-
sures of approximately 20 mmHg [23]. Although all but
one of the arteries lack muscle, they contract with expo-
sure to cardioactive drugs, suggesting that they are not
simply passive supply tubes but rather are used to regu-
late arterial resistance [23]. Oxygen exchange in lobsters
takes place in gills. Oxygen transport is facilitated by the
copper-containing respiratory pigment, hemocyanin [29],
which is found in solution in the hemolymph. Lobster
blood contains hemocytes (Fig. 3C, Fig. S2). Between 3
and 11 morphologically distinct types of hemocytes have
been described [30]. These cells are likely involved in
innate immunity and coagulation.
In summary, the lobster has evolved highly efficient
cardiovascular and respiratory systems to meet its meta-
bolic requirements. Indeed, the very fact that mollusks
and arthropods represent the largest and most diverse
phyla in the animal kingdom is testament to the enor-
mous success of the open circulatory design.
Case study 2: The earthworm (Lumbricus terrestris)
The earthworm belongs to the phylum, Annelida. Like
most triploblastic animals, the earthworm has a well-
developed coelom and a blood vascular system. The coe-
© 2013 International Society on Thrombosis and Haemostasis
 Evolutionary origins of the blood vascular system 53

Anterior
Hepatic a.
lateral a.
Anterior
Cor frontale Heart Sternal a.
aorta D
A Posterior
aorta
Abdominal L
segmental a.
Antennal
a.

Ventral
thoracic a. Ventral
abdominal a.

B E

lumen

C F

h cm *

Fig. 3. Blood vascular system of the lobster. (A) Schematic of the blood vascular system of the lobster. a, artery. (B) Open view of the dorsal
thorax showing the single-chambered heart suspended in the pericardial sinus by alary ligaments. (C) Electron micrograph of the heart showing
a cardiomyocyte (cm) filled with cross sections of thick and thin microfibrils (consistent with myosin and actin) (*) and mitochondria. The
inner surface of the heart chamber is covered by a thin basal lamina. There is no endocardial lining. A hemocyte (h) is shown in the heart
chamber. 9 4,500 (D) A 1-lm transverse histological section of the dorsal abdominal artery stained with Giemsa. (E) Electron micrograph of
the dorsal abdominal artery shows the wall consisting of extracellular matrix composed of a dense meshwork of collagen fibrils. There is no
endothelial lining. 912,900 (F) Photomicrograph of a swimmeret of a lobster that has been injected with Evans blue dye in the dorsal abdomi-
nal artery. Note the branching network of vessels ending in plumes of extravasated dye in the interstitial space. Panel A is adapted from
McLaughlin [20].

lom is highly compartmentalized, with distinct left and
right cavities housed in each body segment. The coelomic
fluid provides an important hydrostatic skeleton that
enables locomotion.19 The blood vascular system is
closed. There is no heart. Rather, circulation is carried
out by contractile blood vessels. There are three main
longitudinal vessels: the dorsal vessel, the ventral vessel,
19
The action of the body wall muscles on the coelomic fluids pro-
vides the hydraulic changes associated with locomotion.
and the subneural vessel (Fig. 4A, Fig. S5). The largest
and most conspicuous vessel in the earthworm is the dor-
sal vessel, which lies on the dorsal surface of the alimen-
tary tract and traverses the length of the animal (Fig. 4B,
Fig. S6) [31]. The dorsal vessel collects blood from other
vessels and drives it forwards through contractile peristal-
tic waves that originate at the posterior end of the animal
and move forward [31,32]. Anteriorly the dorsal vessel
ceases to be a collecting vessel and gives rise to five pairs
of large commissures that connect to the ventral vessel.

© 2013 International Society on Thrombosis and Haemostasis

54 R. Monahan-Earley et al

Epidermis
A B
Circumesophageal Muscle
Lateral
esophageal
vessel
vessels (hearts)

Dorsal vessel
*
Dorsosubneural
vessel

Gut

Ventral vessel
Head Subneural vessel

C D
Coelom Myoepithelial cells
ep mu

lumen

lumen
*

lumen Amoebocyte

Fig. 4. Blood vascular system of the earthworm. (A) Schematic of the blood vascular system of the earthworm. In this case, all vessels are col-
ored red because there is no specialized respiratory organ where blood is oxygenated and because there is no central heart that divides vessels
into afferent channels (veins) and efferent channels (arteries). (B) Transverse histological section through the body of the earthworm shows the
dorsal vessel (arrow) on the dorsal side of the gut. The section was stained with H&E. *, coelom. (C) A 1-lm histological section stained with
Giemsa shows two small blood vessels (arrows) in the body wall. Ep, epidermis; mu, circular muscle layer. (D) Electron micrograph of a small
blood vessel surrounded by a continuous layer of myoepithelial cells. The lumen is filled with hemoglobin particles. 9 4,300 (E) Higher power
electron micrograph shows a blood vessel lined by several myoepithelial cells. The cells are connected by specialized lateral borders. They con-
tain numerous thick myofilaments (arrows) consistent with myosin that are oriented circumferentially around the vessel. A well-formed basal
lamina (*) separates the myoepithelial cells from the lumen of the blood vessel. The fact that the hemoglobin particles are retained in the lumen
indicates that the basal lamina forms an effective barrier. 9 20,500 (F) A similar electron micrograph to E, but shows the presence of an amoe-
bocyte in the blood vessel lumen adjacent to the myoepithelial lining. 9 14,800. Panel A is adapted from Brusca and Brusca [3].

© 2013 International Society on Thrombosis and Haemostasis


These commissures contain lateral ‘hearts’ (pseudohearts),
whose contractions are asynchronous with those of the
dorsal vessel. The ventral vessel, which is suspended in
the mesentery beneath the gut, is the main distributing
channel, sending blood forwards anterior to the hearts,
and posteriorly behind the hearts. The ventral vessel has
no peristaltic activity. In each segment, the ventral vessel
gives rise to segmental vessels and capillaries (Fig. 4C,
Fig. S7 show capillaries in the body wall), through which
blood is carried back to the dorsal vessel. Blood also
flows posteriorly through the subneural vessel, which lies
ventral to the nerve cord.
A previous study in L. terrestris demonstrated a resting
dorsal vessel pulse rate of 11  2.1 beats per minute [31].
The giant earthworm (Glossoscolex giganteus) reaches
sizes of 500–600 g (the body weight of a typical earth-
worm used in our studies is 3-4 g) and may be as long as
120 cm. Frequency of peristalsis in the dorsal vessel of
the giant earthworm was measured to be 8 per minute,
with an average systolic and diastolic pressure of 24 and
14 cm H2O, respectively, in resting healthy specimens
[32]. Pressures in the ventral vessel in response to lateral
heart contractions can exceed 100 cmHg [32]. There is
evidence that blood flow is physiologically regulated in
annelids. For example, dorsal vessel pulse rates are
increased in L. terrestris after forced exercise [31]. More-
over, the load or degree of filling of the dorsal vessel
determines the frequency as well as the force of contrac-
tion of the peristaltic waves [31,32]. Finally, a previous
study in L. variegatus demonstrated a role for biogenic
amines in regulating the dorsal vessel pulse rate [33].
In electron microscopy, blood vessels are readily identi-
fied by the presence of electron-dense hemoglobin parti-
cles within the lumen (Fig. 4D, Fig. S8). The vessels are
outlined by the basal lamina/basement membrane of
mesodermal cells, mesodermal and endodermal cells (of
the gut), or mesodermal and ectodermal cells (of the
skin). The thickness of this extracellular layer (basement
membrane) varies across the vascular tree [34]. Mesoder-
mal cells have varying numbers of microfilaments
(Fig. 4E, Fig. S9) [34,35]. Microfilament-rich mesodermal
cells (myoepithelial cells) are presumed to have a contrac-
tile function. Scattered amoebocytes are found on the
inner surface of the lumen (Fig. 4F, Fig. S10; these cells
can also be seen on 1-lm Giemsa-stained sections of
blood vessels, as shown in Fig. S11) [34].20 The amoebo-
cytes contain electron-dense granules and vesicles [34]. In
tracer experiments in which colloidal gold or horseradish
peroxidase was injected into annelid blood vessels, most
of the vesicles in the amoebocytes were filled with tracer,
suggesting that they are endosomes [34]. Amoebocytes are
distinct from endothelial cells. They do not appear to be
20
These cells have been variously referred to as vasothelial myo-
blasts, amoebocytes, blood cells, hemocytes, or endothelial cells [35,
and references therein].
© 2013 International Society on Thrombosis and Haemostasis
Evolutionary origins of the blood vascular system 55
firmly attached to the basement membrane. They cover
only a small fraction of the surface of the blood vessel.
There is no evidence for junctional complexes between
neighboring cells. Finally, the large number of dense
granules is atypical for endothelial cells. In summary,
blood vessels of the earthworm are lined by basement
membrane and lack an endothelial layer.
Earthworms have no specialized respiratory organs.21
Rather, gas exchange takes place across the general body
surface. The earthworm has an extensive intradermal cap-
illary network, which provides a high surface area for
such exchange. The blood contains a high molecular
weight (3.6 MDa) hemoglobin dissolved in the plasma
[36,37].22 The oxygen content in blood samples from the
dorsal vessel of giant earthworms has been reported to be
between 0.7% and 9.8% [39].
In summary, the earthworm provides an example of an
invertebrate with a closed blood circulatory system.
Unlike its vertebrate counterpart, the closed circulation of
annelids lacks a true heart and consists of blood vessels
that are delimited by the basal surface of epithelial cells,
some of which exhibit a contractile function.
Case study 3: Amphioxus (Branchiostoma lanceolatum)
Amphioxus belongs to the phylum Chordata, subphylum
Cephalochordata, which is the most basal subphylum of
the chordates [40,41].23 They have extensive coelomic cavi-
ties and a closed circulation. The blood does not contain
circulating cells or hemoglobin (or any other respiratory
pigment). As a result, the blood vessels are not readily iden-
tifiable. However, previous tracer studies have yielded a
roadmap of the circulatory system in amphioxus (Fig. 5A,
Fig. S12) [18,42,43]. Amphioxus do not have a heart.
Instead, circulation is accomplished by contractile vessels,
including the endostylar artery, subintestinal vein, and
hepatic vein. As in vertebrates, blood moves forward ven-
trally and backwards dorsally. The endostylar artery (also
called the ventral aorta) is ventral and carries blood in a
cranial direction. The endostylar artery sends vessels to the
gills, which comprise more than 80 pairs of gill slits with
adjacent slits separated by gill bars (Fig. 5B, Figs S12–S15)
21
In fact, the giant earthworm represents the largest living terres-
trial animal without specialized respiratory organs.
22
A recent study demonstrated the use of ultra-pure earthworm Hb
as a means to improve oxygen delivery in hamsters with severe ane-
mia [38].
23
Like vertebrates, amphioxus have a hollow dorsal nerve cord, a
notochord, segmented muscles, and pharyngeal gill slits. In contrast
to vertebrates, the notochord persists in adults, and it is the only
skeletal structure in its body. Like other chordates, they possess a
series of V-shaped muscle blocks (myomeres or myotomes).
Although fish-like in shape, they lack eyes, paired fins, ears, and
jaws. They are always half buried in benthic substrate, and filter feed
on small particles of plankton.
56 R. Monahan-Earley et al

A B
NC
Gills
Dorsal Common
aorta cardinal v.
Posterior
+
Anterior
cardinal v.
Branchial a. P

Endostylar a. Mesenteric a.
Liver
Hepatic v. Sinus Subintestinal v. Hepatic
plexus
venosus cecum

C E
ae
skr

lumen a
a
lumen
Icc

F
lumen
g

D
G
lumen

lumen a

Fig. 5. Blood vascular system of amphioxus. (A) Schematic of the blood vascular system of amphioxus. a, artery; v, vein. (B) Transverse histo-
logical section through the pharynx shows the paired dorsal aortae (arrows) lying on either side of and ventral to the notochord (NC). The
hepatic vein is on the dorsal surface of the liver (also referred to as the hepatic cecum). The section was stained with H&E. P, pharynx. (C) A
1-lm histological section stained with Giemsa shows a discontinuous coverage of dorsal aorta with amoebocytes (arrows). (D) Electron micro-
graph of the contractile hepatic vein shows the lumen delimited by parts of three myoepithelial cells, which contain myofilaments (*) in their
basal portion. a, amoebocyte. 99,200 (E) Electron micrograph of a skeletal vessel in the gill surrounded by skeletal rod (skr). The skeletal rod,
in turn is surrounded by atrial epithelial cells (ae) and lateral ciliated cells (lcc). Occasional chromaffin-like cells (with membrane-delimited elec-
tron-dense granules) are scattered between the atrial epithelial cells. Note the presence of amoebocytes (a) on the luminal surface of the blood
vessel wall.91,750. (F) Electron micrograph shows an amoebocyte ‘clinging’ to the luminal surface of the dorsal aorta. The cell contains a
well-developed Golgi apparatus (g), numerous vesicles, and granules containing tubular structures (arrows). Note that the amoebocyte has no
underling basal lamina. Rather, it is applied to the connective tissue matrix. 911,500. (G) Higher magnification of an amoebocyte from dorsal
aorta shows two tubule-filled granules. 955,000.

[44]. The gills are used for filter feeding, not for oxygen ondary only have two such vessels: the visceral and skeletal.
exchange [45]. The gill bars are of two types: primary and Blood from the gills moves to the dorsal aorta. In the ante-
secondary. The primary gill bars have three longitudinal rior region of the pharynx, the dorsal aorta is paired and
vessels: the visceral, the skeletal, and the coelomic. The sec- lies immediately under the notochord. It is fused posteri-

© 2013 International Society on Thrombosis and Haemostasis


orly. The dorsal aorta sends branches (segmental dorsal
and ventral arteries) to supply the myomeres. It also sends
short mesenteric arteries, which open directly into the
plexus of vessels surrounding the intestine [42]. Blood in
the intestinal plexus is drained into the subintestinal vein
followed in turn by the afferent liver vessel, the liver plexus,
and the efferent liver vessel (also called the hepatic vein).
The hepatic vein runs caudally along the dorsal surface of
the liver (a simple blind diverticulum of the intestine) to
ultimately join the common cardinal vein at the sinus veno-
sus to form the endostylar artery (Figs S15 and S16). Previ-
ous tracer studies have shown that the various afferent and
efferent vessels of amphioxus are connected by extensive
plexi of tiny vessels [42].
Blood vessels in amphioxus are not lined by endothe-
lium, but rather by the basal surfaces of epithelial cells
and extracellular matrix. For example, the dorsal aorta is
delimited by the basal surface of coelomic cells and by
connective tissue around the sheath of the notochord and
the hyperpharyngeal groove (Fig. 5B,C, Fig. S18). Blood
vessels in the gills are lined by varying types of epithelial
cells, including coelomic cells, atrial cells, lateral ciliated
cells, lateral pharyngeal cells, and medial pharyngeal cells.
In the gut, the blood vessel lumen forms between the
basal surfaces of coelomic cells and intestinal epithelial
cells. A previous tracer study demonstrated that the basal
lamina in the visceral vessel of the gill is more permeable
to horseradish peroxidase compared with endostylar artery
[19], suggesting vascular bed-specific differences in permeabil-
ity properties.
In contractile vessels (e.g. the hepatic vein and endo-
stylar artery), the surrounding mesoderm consists of
myoepithelial cells. These cells contain contractile fila-
ments in the basal part, next to the basal lamina [18].
Many filaments end in patches of higher density
(Fig. 5D, Fig. S18). Isolated masses of connective tissue
are distributed around the luminal margin of the hepatic
vein near attachment of the vein to the cecum. These
finger-like extensions gain entrance into the lumen and
form a web-like supporting structure for the vessel wall
[18].
As in the earthworm, scattered amoebocytes are found
clinging to the inner surfaces of amphioxus blood vessels
(Fig. 5E,F, Figs S19 and S20). These cells, which are
rounded or flattened, are rarely in contact with one
another and when they are, there are no junctional special-
izations. The cells contain a well-formed Golgi apparatus.
They are packed with polyribosomes and rough endoplas-
mic reticulum. They also possess many vesicles. In previ-
ous tracer studies, ferritin and horseradish peroxidase were
taken up by coated and smooth-surfaced vesicles within
the amoebocytes, suggesting that these latter cells possess
endocytotic capability [19]. The most conspicuous finding
(which is not observed in the earthworm amoebocyte) is
the presence of tubules in membrane-bound organelles
(granules), which occur at varying orientations (Fig. 5G,
© 2013 International Society on Thrombosis and Haemostasis
Evolutionary origins of the blood vascular system 57
Fig. S21). The size of the tubules appears larger than what
is observed in Weibel–Palade bodies and microtubules in
the cytoskeleton. It has been proposed that these tubules
are precursors of cytoplasmic microtubules involved in cell
motility, and that the amoebocytes function by ‘policing’
the luminal surface of the boundary layer, removing resi-
dues of filtration, and facilitating exchange of material
between blood and surrounding tissues [18]. Others have
suggested that these cells are responsible for depositing
and/or clearing the lumen of extracellular matrix as blood
vessels develop in the amphioxus larvae [46,47].
In summary, despite its close evolutionary relationship
with vertebrates, amphioxus seem to possess a relatively
‘primitive’ closed circulatory system, conspicuous for its
absence of a central heart and endothelial lining, as well
as lack of circulating blood cells and respiratory pigment.
Hagfish (Myxine glutinosa)
Modern vertebrates are classified into two major groups:
the gnathostomes (jawed vertebrates) and the agnathans
(jawless vertebrates). The agnathans are further classified
into two groups, myxinoids (hagfish) and petromyzonids
(lamprey). Most evidence suggests that the hagfish
diverged before lamprey and is thus the oldest living ver-
tebrate. Shared traits in hagfish and jawed vertebrates
indicate that the trait was present in the common ances-
tor of all craniates.
Hagfish have a closed circulation (Fig. 6A, Fig. S22).
Similar to the case in other fishes, the branchial (i.e. gill)
circulation is found in series with the systemic circulation.
The gills are unique among vertebrates in that they are
internalized and organized as pouches, typically six on
each side in M. glutinosa (Fig. 6B). The hagfish circula-
tion also possesses a series of blood sinuses, which are in
direct communication with systemic vessels [50]. The most
prominent of these is the large subcutaneous vascular
sinus located between skeletal muscle and the skin,
stretching from the tentacles of the snout to the caudal
fin fold. Hagfish can hold up to 30% of their blood vol-
ume within the sinus system [50]. It is believed that the
caudal and cardinal ‘hearts’ (composed of skeletal muscle)
function to reintroduce sinus blood into the systemic cir-
culation. The hagfish heart exhibits a Frank–Starling
mechanism [51]. Thus, blood that is redistributed from
sinus to central compartments may serve to increase pre-
load and cardiac output.
Hagfish maintain the lowest arterial blood pressures
(dorsal aorta blood pressure 5.8–9.8 mmHg) and highest
relative blood volumes (18%) of any vertebrate [52–54].
Cardiac output in M. glutinosa is 8–9 mL min 1 kg 1
[55], which approaches the values seen in some teleosts.
Similar to other fish, hagfish have a double-chambered
heart containing a single atrium and a single ventricle.
The heart consists of an avascular mesh of muscle cells (a
so-called spongy heart). There is no coronary circulation.
58 R. Monahan-Earley et al

Rather, deoxygenated blood in the ventricle circulates heart lacks cardioregulatory nerves. The heart rate ranges
through spaces (sinusoidal channels or lacunae) in the from 20 to 30 beats per minute [55]. Hagfish possess a
myocardial wall. Unlike other vertebrates, the hagfish second cardiomyocyte-containing chambered heart, the
A Skin Subcutaneous sinus B
Neural tube
Aorta Notochord
Head Posterior
cardinal vein Somite
Digestive tract
Lumen
Va
Slime gland
Cross-
section 2 cm

Subcutaneous sinus

Muscle

Dorsal aorta
Cardinal Slime glands
hearts Kidney
Gill pouches Caudal
hearts
Branchial Portal
Ventral aorta Liver Gut
heart heart
Internal jugular vein Posterior cardinal vein Caudal vein

C D

lumen
lumen
EC

*
ECM
*

cm

Podocyte F
lumen

EC1
EC1

EC2
lumen Space of Disse

Hepatocyte

© 2013 International Society on Thrombosis and Haemostasis

 Evolutionary origins of the blood vascular system 59

Fig. 6. Blood vascular system of the hagfish. (A) Schematic of the blood vascular system of hagfish. Top, Transverse section through mid-
region shows several features that are typical of other vertebrates, including the arrangement of myomeres, neural tube, and aorta. Features
that are unique to hagfish include the large subcutaneous sinus between skin and skeletal muscle, the retention of the notochord in the adult,
and the presence of slime glands on the ventrolateral surface. Bottom, schematic of hagfish circulation. The gills are in series with the systemic
circulation. Blood enters the subcutaneous sinus via skeletal muscle capillaries and reenters the systemic circulation via accessory hearts (the
caudal and cardinal hearts). The portal heart pumps blood from the intestinal vasculature into the systemic heart via the common portal vein.
(B) Photomicrograph of ventral aorta and two (of a total of 12) gills in an animal that has been injected with Evans blue dye through the
heart. (C) Electron micrograph of the heart shows electron-lucent endothelial cells (EC) overlying a thick extracellular matrix containing a
chromaffin-like cell, and a cardiomyocyte (cm) with electron-lucent cytoplasm, and well-preserved mitochondria and muscle filaments. 915,100.
(D) Electron micrograph of the dermis shows a microvessel in cross-section containing two nucleated red blood cells. The blood vessel is lined
by a continuous layer of endothelial cells. A melanocyte is seen on the left side of the vessel. 92,500. (E) Electron micrograph of a kidney glo-
merulus shows podocyte foot processes abutting a well-formed basal lamina. On the other side of the basal lamina is an endothelial cell (EC)
facing the lumen of a glomerular capillary. The endothelial cell contains many vesicles, vacuoles and tubular structures. 99,100. (F) Electron
micrograph of a liver sinusoid shows a large gap in a single endothelial cell (E1, double-headed arrow), well-preserved attachments between
two endothelial cells (EC1 and EC2), and a continuum of proteinaceous material from the lumen to extravascular space (Space of Disse). EC2
is a second endothelial cell. 932,400. Panel A is reprinted with permission from Cheruvu et al. [48]. Panels C, D and F are reprinted with per-
mission from Yano et al. [49].

portal heart, which beats asynchronously with the sys-

temic heart, as it propels blood from the gut to the liver
via the common portal vein [55].
We and others have previously shown that hagfish
blood vessels are lined by a single layer of endothelial cells
[49,56,57]. All endothelial cells have vesicles along both
the blood and tissue fronts of the cell. The lateral borders
of endothelial cells are typical for those seen in mammals.
Occasional Weibel–Palade bodies are observed, suggesting
that hagfish express von Willebrand factor. Similar to
other vertebrates, endothelial phenotypes vary from one
vascular bed to another. For example, endothelial cells lin-
ing the heart sinuses are attenuated, electron lucent, and
contain many vesicular structures (Fig. 6C, Fig. S23).
Dermal microvessels display a continuous endothelium,
with lateral plasma membrane borders containing special-
izations typical of other vertebrates (Fig. 6D, Fig. S24).
The kidney glomerulus contains endothelial cells with few
fenestrae, and many vesicle and vacuoles of varying sizes
(Fig. 6E, Fig. S25). As a final example, ultrastructural
studies of the liver demonstrate a discontinuous endothe-
lium with many gaps (Fig. 6F, Fig. S26).
In addition to vascular bed-specific differences in ultra-
structure, the endothelium of hagfish also demonstrates
molecular and functional heterogeneity. For example, lec-
tin staining of various tissues revealed significant differ-
ences in lectin binding to the endothelium [49]. Moreover,
in intravital microscopy studies of the dermal microvascu-
lature, histamine was shown to induce neutrophil adhe-
sion in capillaries and postcapillary venules, but not
arterioles [49]. As a final example, arteries from the mes-
entery and skeletal muscle demonstrated site-specific
mechanisms of endothelium-dependent vasomotor relaxa-
tion [58].
In summary, our findings in hagfish confirm that the
closed system of even the most basal vertebrate is lined
by endothelium and that phenotypic heterogeneity of the
endothelium is a conserved property of this cell type.
Evolutionary implications
When approaching the evolutionary origins of the blood
vascular system and the endothelium, we must consider
three important questions. First, when did these systems
evolve? Second, why did they evolve? In other words,
what survival advantage do these structures confer at the
level of species? Finally, how did the blood vessels and
their endothelial linings develop ontogenetically (as evolu-
tionary novelties) in the first place? In this section, we will
consider each question in turn.
When did the blood vascular system and endothelium first
evolve?
As we survey the present landscape of body plans, we
find a wide variety of blood vascular systems (Fig. 7).
Some are closed, others are open. Some use hearts to pro-
pel blood, whereas others employ pulsatile blood vessels.
Only a minority are lined by endothelium. When and
how did all of these diverse structures evolve? The answer
is likely through a combination of homology and conver-
gence.24 The last common ancestor of vertebrates and
annelids, or of vertebrates and mollusks was the ancestor
of the protostome–deuterostome ancestor, which lived
between 600 and 700 million years ago [59]. Although the
fossil record is scarce, it is widely believed that this pre-
cursor animal was a segmented bilaterian (triploblastic
coelomate) [9]. If we are to accept that the blood vascular
system evolved as a means to bypass the bulkheads of a
segmented animal (see next section), then the first such
system likely arose during this time. Flow would have
been mediated by peristaltic vessels, perhaps like those
described in the annelid. Blood probably percolated
through spaces in the extracellular matrix, and thus, the
24
The reader is referred to an excellent discussion of these concepts
in Xavier-Neto et al. [7].

© 2013 International Society on Thrombosis and Haemostasis

60 R. Monahan-Earley et al

Closed
Closed
Blood

Open
None

Open
None
vascular Closed circulation
system type

Single-cham-

Three-cham-
bered heart
Multi-cham-
bered heart

bered heart

Two-cham-
bered heart

Four-cham-
bered heart
Contractile

Contractile
vessels

vessels
Major

None

None
propulsive
organ

(noncephalopod)
(cephalopod)

Amphibians
Arthropods

Amphioxus

Mammals
Bony fish
Mollusks

Mollusks

Annelids
Cnidaria

Reptiles
Porifera

Hagfish

Birds
T Y
um EI
eli N
o th GE
d O
En ER
T
HE
EC

Last common ancestor

of vertebrates
First endothelium
(540-510 mya)

Protostome deuterostome ancestor

First blood vascular system
(600-700 mya)

Fig. 7. Phylogenetic perspective of the blood vascular system. Blood vascular system types and major propulsive organs are shown for repre-
sentative extant phyla. The phylogenetic tree is schematic only, and the timelines are not drawn to scale. EC, endothelial cell; mya, million
years ago.

Why did the blood vascular system and endothelium

system was by definition closed, albeit primitive. This sce-
nario supports homology of all blood vascular systems.
Over the past 600–700 million years of evolution, the
blood vascular system has undergone significant modifica-
tions, in response to selective pressures experienced by
individual phyla. In some cases, the blood vascular sys-
tem has regressed (e.g. in flatworms and nematodes). In
other cases, the primitive system transitioned to an open
circulation. The open system reverted back to a closed
design in an ancestral cephalopod. This is an example of
convergent evolution, whereby an analogous structure
(the closed circulation in cephalopods) arose indepen-
dently of the closed circulation in other invertebrates (e.g.
worms) and in vertebrates. Finally, the endothelium is
present in all vertebrates, yet distinctly absent in inverte-
brates. Although cells appear to cling to the inner surface
of some invertebrate closed vessels, these lack the classical
morphological features of endothelial cells. Therefore, the
endothelium appeared in an ancestral vertebrate following
the divergence from the urochordates and cephalochor-
dates between 540 and 510 million years ago. Our find-
ings in hagfish indicate that endothelial heterogeneity
appeared during the same narrow window of time [49].
evolve?
Circulatory systems most certainly evolved to overcome
the time and distance constraints of diffusion, thus per-
mitting increased body size and metabolic rates, as well
as increased levels of integration and organization in
Metazoa. Although the primitive coelom provided bulk
flow delivery of substances, these structures never devel-
oped an efficient pumping mechanism. More importantly,
they became increasingly localized to distinct compart-
ments in the body. Ruppert and Carle hypothesized that
the blood vascular system evolved as a means to provide
bulk fluid transport along the entire length of a seg-
mented animal, in effect bypassing the septal bulkheads
[6]. The earliest design may have involved the formation
of spaces in between the basal laminae of endodermal
and coelomic epithelia, allowing for improved distribution
of nutrients absorbed by the intestine [60]. Myoepithelial
differentiation of the visceral layer of coelomic epithelium
would have provided the necessary pumping action to
drive nutrients toward the viscera [10]. Subsequent vascu-
larization of the skin would have facilitated gas exchange
at the body surface.

© 2013 International Society on Thrombosis and Haemostasis


The movement of fluid in blood vascular systems was
originally mediated by the peristaltic motion of certain
blood vessels, similar to what occurs in the earthworm
and amphioxus. However, peristaltic pumps lack effective
coordination between the fluid that is entering the con-
tractile region and the fluid that is leaving it. Despite
some tweaks to the peristaltic design, such as the evolu-
tion of one-way valves and partial coordination in con-
tractions, the loss of fluid energy associated with
backflow, distension of wall segments ahead of the
stream, and pump reversals would have constrained body
size and metabolic activity [7]. This would have provided
a selective pressure for the appearance of true hearts, in
which inflow and outflow are tightly coupled via
multiple chambers, efficient electrical connection between
myocytes (hence, simultaneous contraction) and one-way
valves [7].
If the closed circulatory system was the ancestral condi-
tion, then why did the open circulatory system evolve in
the precursors of mollusks and arthropods? One explana-
tion for this change in the circulatory blueprint is that the
rigid exoskeleton of these animals, which evolved for pro-
tection against predation and physical injury, eliminated
the need for a coelomic hydrostatic skeleton [6]. The sub-
sequent regression of the coelom removed barriers to bulk
transport and therefore the need for a closed system of
vessels. At the same time, the exoskeleton impaired any
contribution of body wall musculature to the movement
of blood. This may have provided selective pressure for
the evolution of a well-developed muscular heart, which
is a characteristic feature of most animals with an open
circulation.25
Cephalopods are monophyletic with other mollusks, yet
they have secondarily evolved a closed circulation. More-
over, they have a far more efficient heart compared with
other members of the phylum [10,15].26 It seems likely
that these modifications were selected for to enable an
active lifestyle, which includes predatory behavior, swim-
ming, and jet propulsion. The same association between
closed circulatory system, highly efficient hearts, and
active lifestyles is seen in the vertebrates. There are sev-
eral reasons why a closed system might better serve the
needs of a highly active animal. First, it provides for a
more efficient (i.e. fractal-like) distribution system, which
can be tailored to the unique architecture of the various
25
This is a nice example of an evolutionary trade-off. In this case,
the exoskeleton evolved to provide protection from predators and
physical support. However, body size could no longer increase
(hence, the evolution of molting), the body wall muscles could no
longer contribute to blood flow (hence, the evolution of the heart),
and it was no longer possible to breathe through the skin (hence, the
evolution of gills).
26
The cephalopod heart typically consists of a ventricle and two
atria. The myocardium contains multiple layers of striated muscle
cells, which in turn are covered by a coelomic epithelium.
© 2013 International Society on Thrombosis and Haemostasis
Evolutionary origins of the blood vascular system 61
tissues and organs.27 Second, closed circulatory systems
can be subject to regulation so that flow is directed pref-
erentially to one organ or another. Third, the closed sys-
tem enables carriers to remain inside the vessels and
release their cargo where needed. This allows for homeo-
static integration of diverse tissues in the body. Finally,
as alluded to earlier, the closed system provides for paral-
lel flow (equal opportunity) to all organs of the body.
The transition from aquatic to terrestrial life required
modifications of several body systems, including those
involved with respiration, feeding, locomotion, water bal-
ance, and reproduction. The formation of invaginated gas
exchangers (i.e. lungs) reflected the need to minimize
water loss across the respiratory surface [14].28 Ancillary
skin breathing (i.e. cutaneous gas exchange) was possible
so long as the epidermis was highly vascularized and thin.
Animals with thin skin have high evaporative loss. There-
fore, skin breathing is confined to aquatic or semi-aquatic
tetrapods, particularly amphibians. Reptiles, which
evolved thick impermeable skin to prevent water loss and
provide physical protection, were the first vertebrates to
rely entirely on lungs for gas exchange. A limitation of
the single circulation in fish is that gills receive a greater
pressure head than the systemic circulation. As higher
pressures evolved in response to increased metabolic
demands and gravitation, there was a need to depressur-
ize the gas exchanger. This was accomplished by the sepa-
ration of systemic and pulmonary circulations.
In water, buoyancy provides virtual weightlessness,
whereas on land, animals are subjected to gravitational
force. As the density of water and blood is similar, gradi-
ents of hydrostatic pressure in vertical blood columns are
counter-balanced by pressure gradients in surrounding
water [61]. Thus, transmural pressure throughout the vas-
culature of aquatic animals remains relatively constant. In
contrast, on land, gravity causes increased pressure in the
lower part of a fluid column, resulting in increased trans-
mural pressure, pooling of blood in dependent vascula-
ture, reduced venous return, and decreased arterial
pressures. This effect is particularly important in tall ani-
mals or in animals with postural behavior (e.g. arboreal
or climbing snakes). Countermeasures to gravitational
stress have evolved in tetrapods, including higher arterial
pressure, baroreceptor reflex, and low-compliance tissues
(e.g. stiff skin) [61].
27
It is often said that the closed system ‘allowed’ for higher pres-
sures. However, in reality, the closed system required high pressures
to overcome the systemic resistance associated with a complex distri-
bution system.
28
The original selection pressure for lungs was aquatic hypoxia,
resulting in the appearance of air-breathing fish lungfish. In fish, the
heart receives (and is therefore perfused by) deoxygenated mixed
venous blood. It has been hypothesized that the lung (in conjunction
with the coronary circulation) evolved as an adaptation to myocar-
dial hypoxia [13].
62 R. Monahan-Earley et al
What selective forces were responsible for the appear-
ance of the endothelium in the ancestral vertebrate? One
suggestion is that that the smooth lining provided by the
endothelium minimizes energy required to move blood.29
The endothelium mediates vasomotor control in the earli-
est extant vertebrate. Thus, the appearance of the endo-
thelium may have provided a critical new level of
vasoregulation, allowing for more highly pressurized sys-
tems. Also, the barrier function provided by the endothe-
lium would have prevented the loss of plasma proteins
necessary to maintain equilibrium between osmotic and
hydrostatic pressures [6]. Higher pressures required not
only higher osmotic pressures, but also the formation of
the lymphatic system to drain net surplus of interstitial
fluid. The ancestral vertebrate also evolved novel immune
and coagulation mechanisms. Thus, the co-evolution of
the endothelium during this narrow window of time may
be explained in part by the role of endothelial cells in
localizing these activities to areas of need [11]. Endothe-
lial cells have been shown to play a pivotal role in the
early development of organs, such as the pancreas and
liver [62]. Such bidirectional dialog between the endothe-
lium and other tissues may have been instrumental in
shaping the evolution of this cell layer.
The fitness advantage of endothelial cell heterogeneity
may be explained on several levels [63]. First, phenotypic
heterogeneity reflects the remarkably pleiotropic role of
the endothelium in meeting the varying needs of diverse
tissue types. Second, some site-specific properties reflect a
local self-preserving adaptation of the endothelial cells to
their particular microenvironment. Third, phenotypic
plasticity provides the endothelium with flexibility to tem-
porally adjust to a wide range of physiological and patho-
physiological influences, including pathogens and toxins.
How did the blood vascular system and endothelium
evolve?
The coelomic and blood vascular systems (as well as
excretory systems) arose within the mesoderm of triplo-
blastic animals. Indeed, the appearance of the mesoderm
provided new building material for animal construction
and allowed for the evolution of increasingly complex
and large animals. At some early evolutionary stage, per-
haps in the ancestral triploblastic bilaterian condition, a
subpopulation of mesodermal cells differentiated into a
mesothelium whose apical side faced the coelom and
29
Laminar flow is required to minimize the energy needed to move
blood through these complex vascular systems. Laminar flow
through a cylindrical tube can be predicted based on vessel diameter,
mean blood velocity, and blood density and viscosity (Reynolds’s
number). However, if there are sudden variations in vessel diameter
or irregularities in the vessel walls, turbulent flow can result. In tur-
bulent flow, a significantly greater pressure is required to move a
fluid through the vessels as compared to laminar flow.
whose basal side faced clefts (i.e. blood vessels) between
the mesothelial walls [12]. Some of these mesothelial cells
acquired myofilaments and the capacity to contract
[11,12]. If the earliest blood vessels formed between these
myoepithelial cells and gut epithelial cells, then there must
have been an intimate coordination between mesodermal
and endodermal lineages. In vertebrates, the mesothelium
no longer contributes to the vasculature. Instead, endo-
thelial cells are derived from distinct mesodermal zones
arising from the primitive streak, including the lateral
plate mesoderm, cardiac mesoderm, and paraxial meso-
derm. Other components of the vascular wall, including
smooth muscle cells, are derived from mesoderm or the
neural crest (an ectoderm derivative found exclusively in
vertebrates). Thus, at some point following the divergence
of the cephalochordates, the mesoderm of the ancestral
vertebrate acquired novel specialization, which allowed
for the formation of endothelium.
There is an interesting body of the literature debating
the degree to which the drosophila and vertebrate hearts
are homologous [7,12,64,65]. An important concept is
that homology can apply to different levels of organiza-
tion (e.g. genetic, molecular, cellular, tissue structure,
function) [3,7].30 For example, the drosophila and verte-
brate hearts are hardly homologous in three-dimensional
structure (at least in the adult stages). However, the dro-
sophila and vertebrates share orthologous genes involved
in making a heart, including tinman/Nkx2–5. Indeed, it
has been argued that the homologies lie at the level of
gene regulatory pathways that make a muscle cell rather
than at the level of the organ (i.e. the three-dimensional
pump) [7]. It has been proposed that the heart evolved
through modification and expansion of an ancestral net-
work of regulatory genes encoding a core set of cardiac
transcription factors, including NK2, myocyte enhancer
factor 2 (MEF2), GATA, Tbx, and Hand, virtually all of
which are present in triploblasts [47,66]. The ancestral cir-
cuit was probably responsible for the contractile mesothe-
lial cell that lined the earliest blood vessels and ultimately
acquired a contractile phenotype to propel the blood.
Complexity of the heart increased during evolution owing
to the expansion of the number of ancestral regulatory
genes, modification of the timing, and pattern of their
expression, as well as their regulatory interactions with
each other and with other regulatory inputs.
What is the origin of the endothelial cell? One possibil-
ity is that it arose from the mesothelial cell. Alternatively,
endothelial cells are derived from amoebocytes, through
the acquisition of an epithelial phenotype [11,67]. A previ-
ous study demonstrated that amoebocytes in amphioxus
30
Brusca offers a nice example of the importance of the level of
analysis being considered: The wings of bats and birds are homolo-
gous as tetrapod forelimbs, but they are not homologous as ‘wings’
because wings evolved independently in these two groups (i.e. the
wings of bats and birds do not share a common ancestral wing) [3].
© 2013 International Society on Thrombosis and Haemostasis

larvae express Pdvegfr (a single member of the platelet-
derived growth factor receptor [PDGFR]/vascular endo-
thelial growth factor receptor [VEGFR] subfamily) [47].
The authors proposed that vertebrate endothelial cells
may have derived from ancestral-free hemal Pdvegfr+
cells [47]. Consistent with an amoebocyte origin of endo-
thelial cells is the observation that endothelial cells arise
in close association with hematopoietic cells. For exam-
ple, they share a common progenitor cell (the hemangio-
blast), and some hematopoietic cells are derived from
hemogenic endothelium. It also noteworthy that hemo-
cytes in Drosophila express VEGFR [68].
As in the case of the heart, the endothelium likely arose
through the mutation and selection of a small number of
pre-existing regulatory genes involved in mesoderm func-
tion. It is, of course, highly improbable that selection has
coordinated hundreds of different DNA mutations to
yield such a broad array of endothelial cell phenotypes
[63]. Instead, the endothelium has been selected for its
plasticity and exploratory behavior [69]. As a result,
organs can acquire changes without depending on co-evo-
lution in the endothelium.
The ancestral vertebrate developed the means to assem-
ble endothelial cells into tubes, to recruit pericytes and sta-
bilize blood vessels, and to specify different subtypes of
vessels, including arteries and veins. The extent to which
these processes ‘borrowed’ from established building plans
is unclear. It is notable that VEGF-like factors have been
identified in a number of invertebrates, including drosoph-
ila [68] and squid [70]. Moreover, a blueprint for tubulo-
genesis exists in multiple systems, including the tracheal
system of insects [71,72]. The formation of blood vessels in
the closed circulatory system of invertebrates has been
referred to as ‘myoepithelial angiogenesis’ [60]. The process
may involve hemocyte-mediated clearance of extracellular
matrix to form a patent lumen [46,67], or the separation of
apposing mesothelial basement membranes, perhaps via
cell–cell repulsion [67]. Studies in Drosophila have impli-
cated a role for Robo-Slit in heart tube formation [67].
Evolutionary medicine
In the Introduction, we emphasized the importance of both
proximate and evolutionary explanations when approach-
ing human physiology and pathophysiology. Evolutionary
medicine is the application of evolutionary principles to an
understanding of human health and disease [1,73,74]. Evo-
lutionary biology teaches us that no trait is perfect. Every
trait can be made better, but by making it better something
else would be made worse. Indeed, a central tenet of evolu-
tionary medicine is that the human body is a jerry-rigged
bundle of trade-offs and that an understanding of the cost–
benefits of these trade-offs will provide novel insights into
our vulnerability to disease. For example, consider the
closed circulatory system. Such a design permits more effi-
cient bulk flow delivery to the tissues of the body. However,
© 2013 International Society on Thrombosis and Haemostasis
Evolutionary origins of the blood vascular system 63
a closed vasculature is by definition branched and curved.
Blood flow is disturbed at branch points and curvatures.
Microdomains of flow disturbance render the endothelium
dysfunctional and vulnerable to atherosclerosis. This is a
classic example of an evolutionary tradeoff, where the
advantages of closing the circulation are balanced against
the disadvantage of increased susceptibility to atheroscle-
rotic lesion formation. Another important evolutionary
principle when considering human health and disease is the
notion that the present-day blood vascular system evolved
to maximize fitness in a far earlier era, perhaps tens of
thousands of years ago, which is the time it takes for natu-
ral selection to filter the gene pool. Our evolved state has
not had time to adapt to changes in longevity, physical
activity, diet, and recreational drug use (especially smok-
ing). The resulting gene-environment mismatch has led to
an epidemic of hypertension, hypercholesterolemia, diabe-
tes, and atherosclerosis. A goal of preventive medicine is to
recalibrate the balance by adapting the environment to the
genetic blueprint.
Conclusions
Clinicians and clinician-scientists tend to view the cardio-
vascular system through the narrow lens of proximate
causation. In this review, we have attempted to place the
blood vascular system in a broader temporal context,
with the goal of emphasizing the selection pressures that
have led to its emergence. An important take home mes-
sage is that cardiovascular systems are highly diverse in
their structure and function. The design of a given system
is exquisitely matched to the needs of the animal. It is not
helpful to think of one system being ‘superior’ to another.
Mollusks have no more need for a human heart, than
humans have for a mollusk heart. All extant animals have
survived the rigor of natural selection and are by defini-
tion highly successful. An appreciation of the particular
adaptations that underlie these many successes provides
important insights into basic principles of oxygen delivery
and homeostasis in all animals, including humans.
Addendum
R. Monahan-Earley designed and performed experiments
and analyzed the data. A. M. Dvorak designed and per-
formed experiments and analyzed the data. W. C. Aird
designed and performed experiments, analyzed the data
and wrote the manuscript.
Acknowledgements
We thank S. Moskowitz for his artwork, E. Morgan and
K. Yano for their technical help, and D. Groutsis and
James Aird for their help in procuring specimens. This
work was supported by National Institutes of Health
grant HL076540.
64 R. Monahan-Earley et al
Disclosure of Conflicts of Interest
The authors state that they have no conflict of interest.
Supporting Information
Additional Supporting Information may be found in the
online version of this article:
Appendix S1. Methods
Appendix S2. Glossary of terms
Fig. S1. Schematic of lobster circulation.
Fig. S2. Electron micrograph of the lobster heart.
Fig. S3. Transverse 1-lm Giemsa-stained histological sec-
tion of the dorsal abdominal artery from lobster.
Fig. S4. Electron micrograph of the lobster aorta.
Fig. S5. Schematic of earthworm circulation.
Fig. S6. Transverse 5-lm H&E-stained histological sec-
tion through the body of the earthworm.
Fig. S7. Transverse 1-lm Giemsa-stained histological sec-
tion of the body wall of the earthworm.
Fig. S8. Electron micrograph of a small earthworm blood
vessel.
Fig. S9. Electron micrograph of an earthworm blood
vessel.
Fig. S10. Electron micrograph of an earthworm blood
vessel with an amoebocyte adhering to the inner side of
the lumen.
Fig. S11. Transverse 1-lm Giemsa-stained histological
section of a blood vessel in the earthworm.
Fig. S12. Schematic of amphioxus circulation.
Fig. S13. Transverse 5-lm H&E-stained histological sec-
tion through the pharynx of amphioxus.
Fig. S14. Transverse 1-lm Giemsa-stained histological
section through the gills of amphioxus.
Fig. S15. Transverse 1-lm Giemsa-stained histological
section through the mid-body region of amphioxus.
Fig. S16. Transverse 1-lm Giemsa-stained histological
section of the hepatic vein in amphioxus.
Fig. S17. Transverse 1-lm Giemsa-stained histological
section of the dorsal aorta in amphioxus.
Fig. S18. Electron micrograph of the contractile hepatic
vein from amphioxus.
Fig. S19. Electron micrograph of a secondary gill bar in
amphioxus.
Fig. S20. Electron micrograph of an amoebocyte adhering
to the inner wall of the dorsal aorta in amphioxus.
Fig. S21. Electron micrograph of two tubule-filled gran-
ules in an amoebocyte from amphioxus.
Fig. S22. Schematic of hagfish circulation.
Fig. S23. Electron micrograph of a heart sinus in hagfish.
Fig. S24. Electron micrograph of a dermal capillary in
hagfish.
Fig. S25. Electron micrograph of a a glomerular capillary
in hagfish.
Fig. S26. Electron micrograph of an hepatic sinusoid in
hagfish.
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