JOURNAL OF MORPHOLOGY 275:247–257 (2014)
Structure of the Female Gonoduct of the Viviparous
Teleost Poecilia reticulata (Poeciliidae) During
Nongestation and Gestation Stages
Juan Carlos Campuzano-Caballero and Mari Carmen Uribe*
Departamento de Biologıa Comparada, Facultad de Ciencias, Universidad Nacional Autonoma de Mexico,
Ciudad Universitaria, Mexico DF 04510
ABSTRACT Female teleosts do not have oviducts
because M€ ullerian ducts do not develop. Instead, the
caudal region of the ovary, the gonoduct, connects to
the exterior. Because of the lack of oviducts in vivipa-
rous teleosts, the embryos develop in the ovary, as an
intraovarian gestation, unique in vertebrates. This is
the first study to address the histology of the gonoduct
in a viviparous teleost. The gonoduct of Poecilia reticu-
lata was analyzed during previtellogenesis, vitellogene-
sis, and gestation. The gonoduct lacks germinal cells.
From deep to superficial, the wall has simple cuboidal
or columnar epithelium, loose connective tissue, longi-
tudinal layer of smooth muscle, and visceral perito-
neum. Cells of the immune system occur in the lumen
and in the mucosa. The gonoduct was divided in three
regions: 1) cephalic, 2) middle, and 3) caudal. At the
initial part of each region, thin mucosal folds extend
into the lumen. The cephalic region forms a tubular
structure with light and irregular folds. The middle
region has a wider lumen and is more irregular due to
ventral invaginations and irregular and short mucosal
folds; beneath the epithelium there are melano-
macrophage centers. The caudal region is delimited
from the middle region by folds; however, they are thin-
ner than these of the other regions. Ventral invagina-
tions form exocrine glands, and the smooth muscle is
thicker than in the other regions. During gestation,
cells of the immune system are abundant; melano-
macrophage centers become larger and the glands
exhibit desquamated cells. These observations suggest
roles of the gonoduct in reducing the diameter of the
lumen; receiving sperm during vitellogenesis; produc-
ing secretions, more abundant during vitellogenesis;
and in immunological activity throughout the reproduc-
tive cycle. The ciliated epithelium and the thick muscle
of the caudal region may be involved during
birth. J. Morphol. 275:247–257, 2014. V C 2013 Wiley
Periodicals, Inc.
KEY WORDS: gonoduct; ovary; viviparous fishes; Poeci-
lia reticulata; histology
INTRODUCTION
During embryogenesis of female vertebrates,
excluding teleosts, the M€ullerian or parameso-
nephric ducts differentiate, becoming oviducts.
According to Mendoza (1943), Turner (1947),
Wourms (1981), Wake (1985, 1989), Wourms et al.
C 2013 WILEY PERIODICALS, INC.
V
(1988),and Wourms and Lombardi (1992), the term
oviduct in vertebrates is restricted to the ducts
derived from the M€ ullerian ducts. As a result,
female teleosts do not have oviducts because the
lack of development of M€ ullerian ducts during
embryogenesis. Therefore, and exclusively in
female teleosts, the posterior part of the ovary is
called the gonoduct (Wourms, 1981; Wake, 1985).
The gonoduct connects the ovary to the exterior,
and opens at the genital pore. The lumen of the
gonoduct is continuous with the lumen of the ger-
minal part of the ovary (Wake, 1985; Lahnsteiner
et al., 1997).
The gonoduct is characterized by the absence of
germinal cells. This feature differentiates the mor-
phology and physiology of the gonoduct from the
germinal part of the ovary, where germinal cells
are included, and it is the site where oogenesis
occurs (Turner, 1947; Wake, 1985; Wourms et al.,
1988; Greven, 2005, 2010; Uribe et al., 2009,
2010).
Because teleosts lack oviducts, viviparity, includ-
ing that in the Poeciliidae, differs from all other
vertebrates due to the intraovarian gestation
(Turner, 1947; Wourms, 1981, 2005; Wourms et al.,
1988; Grier et al., 2005; Uribe et al., 2009, 2010).
There are no studies that are exclusively dedi-
cated to morphological analysis and description of
the gonoduct in any teleost, neither oviparous nor
viviparous species. Only several studies on ovarian
structure have documented some morphological
and physiological characteristics of the gonoduct.
They mentioned the following aspects: a) the
*Correspondence to: Dra. Mari Carmen Uribe; Departamento de
Biologıa Comparada, Facultad de Ciencias, Circuito Exterior,
Ciudad Universitaria, Universidad Nacional Aut
onoma de M exico,
Insurgentes Sur 3000, Delegacion Coyoac
an, D. F. M
exico 04510,
Mexico. E-mail: mari3uribe3@gmail.com
Received 19 May 2013; Revised 10 July 2013;
Accepted 16 August 2013.
Published online 18 December 2013 in
Wiley Online Library (wileyonlinelibrary.com).
DOI 10.1002/jmor.20200
248 J.C. CAMPUZANO-CABALLERO AND M.C. URIBE
gonoduct is a single structure, even in species that
have double or partially fused ovaries (Turner, 1947;
Wake, 1985; Wourms et al. 1988; Greven, 2005,
2010); b) the gonoduct is the growth of somatic
elements of the caudal ovarian wall (Turner, 1947;
Wourms, 1981; Wake, 1985; Wourms et al., 1988;
Lahnsteiner et al., 1997; Potter and Kramer, 2000;
Greven, 2010); and c) in viviparous teleosts, the
gonoduct is involved in internal fertilization, receiv-
ing the spermatozoa during insemination (Wourms,
1981; Koya et al., 1998; Potter and Kramer, 2000;
Kobayashi and Iwamatsu, 2002; Evans et al., 2003;
Burns and Weitzman, 2005; Greven, 2005).
The guppy, Poecilia reticulata, is one of the most
frequently studied viviparous fishes and has been
selected for numerous studies related to its repro-
duction (Jollie and Jollie, 1964; Jalabert and Bill-
ard, 1969; Lambert, 1970; Pilastro and Bisazza,
1999; Evans et al., 2003; Urriola Herna ndez et al.,
2004; Gasparini et al., 2012). The gonoduct of this
species, however, has not been analyzed. The goal
of this study is to describe the histological struc-
ture of the gonoduct of the viviparous teleost
P. reticulata during previtellogenesis, vitellogene-
sis, and gestation stages.
MATERIAL AND METHODS
Adult females P. reticulata (N 5 15), ranging in total stand-
ard length between 4.5–5.4 cm were obtained from the Aquar-
ium of the Facultad de Ciencias, Universidad Nacional
Autonoma de Mexico. The specimens were selected during three
reproductive stages: previtellogenesis (N 5 5), vitellogenesis
(N 5 5), and gestation (N 5 5). Guppies were anesthetized and
decapitated. The abdominal cavity was opened by a lateral inci-
sion, and the ovary, including the gonoduct, was quickly
excised. Ovaries were fixed in alcoholic Bouin’s (8–15 h). After
fixation, the ovaries were washed in water, dehydrated in a
series of graded alcohols (70, 80, 96, and 100%), cleared in
xylene and embedded in paraffin. Tissues were serially sec-
tioned at 6 mm in longitudinal plane and processed with peri-
odic acid-Schiff’s reagent and stained with hematoxylin-eosin
(H-E) and Masson’s trichrome (Aguilar-Morales et al., 1996).
Selected morphometrics were obtained such as: length of the
gonoduct of each specimen, epithelium cell height (n 5 10 per
specimen), diameter of melano-macrophage centers (n 5 3 per
specimen), and glandular cell height (n 5 10 per specimen).
Measurements were recorded using a Carl Zeiss Axion Vision
Imaging System. Digital photomicrographs were taken using
an Olympus digital camera model C5050Z coupled to an Olym-
pus CX31 microscope.
RESULTS
The ovary of P. reticulata is a single organ of
the cystovarian type (Figs. 1A,B), having a central
lumen. The gonoduct is the caudal part of the
ovary and communicates to the exterior by the
genital pore (Figs. 1A–C). Consequently, the gono-
duct separates the germinal part of the ovary,
where development of oocytes and gestation occur,
from the exterior. The gonoduct lacks germinal tis-
sue, but its lumen is continuous with the germinal
part of the ovary (Figs. 1A,B). The gonoduct, as is
Journal of Morphology
the rest of the ovary, is dorsal to the gut
(Figs. 1A–C). The length of the gonoduct is similar
during the three reproductive stages: in previtello-
genesis, it was 2.64 6 0.21 mm; during vitellogene-
sis, it was 2.71 6 0.42 mm; and during gestation, it
was 2.76 6 0.62 mm.
According to their morphological features, three
regions of the gonoduct were defined: 1) cephalic
region, 2) middle region, and 3) caudal region
(Figs. 1C,D). At the initial portion of each region,
there are folds of the mucosa that extend into the
gonoducal lumen. These folds form structures sim-
ilar to a cervix. Throughout the three regions of
the gonoduct, the wall contains luminal columnar
epithelium; loose and vascularized connective
tissue; longitudinal layer of smooth muscle and
visceral peritoneum at the periphery.
Cephalic Region
Observed during previtellogenesis (Figs. 2A,B),
vitellogenesis (Figs. 2C,D), and gestation (Figs.
2E,F). It is adjacent to the germinal part of the
ovary. At the anterior part of this region, there are
large and thin mucosal folds that extend into the
lumen (Figs. 2A,C,E). The rest of the cephalic
region has a wider lumen and forms a tubular
structure, with a lightly irregular margin (Figs.
2A,C,E). The mucosa of the gonoducal wall and
the folds are lined by columnar epithelium with
ciliated and nonciliated cells. The height of the
epithelial cells varies during the reproductive
cycle: during nongestation their average height is
8.38 6 1.12 mm (Fig. 2B); this increases during
vitellogenesis (Fig. 2D) to 11.51 6 2.31 mm, but
during gestation (Fig. 2F) this decreases slightly
to 10.84 6 1.21 mm. Subjacent to the epithelium,
the folds contain loose and vascularized connective
tissue (Figs. 2B,D,F). The wall has similar histo-
logical components as was mentioned in the struc-
ture of the folds. Additionally, at the periphery,
there are a longitudinal layer of smooth muscle
and visceral peritoneum (Figs. 2B,F). The connec-
tive tissue of this region is looser during gestation
(Fig. 2F), suggesting that it retains more fluid.
Middle Region
Observed during previtellogenesis (Figs. 1A, 3A,B),
vitellogenesis (Figs. 1B,C, 3C), and gestation
(Figs. 3D–F) stages. Similar to that observed in
the cephalic region, at the initial part of this
region, there are large mucosa folds which
extend into the lumen (Figs. 1C). At this level,
throughout the year the lumen is clearly
reduced. The lumen of this region is more irregu-
lar than that of the cephalic region due to the
presence of several, irregular and short folds of
the mucosa (Fig. 1C). At the ventral side, there
is a deep invagination with large and irregular
folds (Fig. 1C). The epithelium is columnar with
 STRUCTURE OF THE GONODUCT OF POECILIA RETICULATA 249

Fig. 1. Ovary of P. reticulata. Panoramic views of the single, sacular ovary during previtellogenesis (A) and vitellogenesis (B, C). (A,
B) Germinal part of the ovary and gonoduct. The germinal part of the ovary contains follicles in different stages of development; in con-
trast, the gonoduct lacks germinal cells. The germinal part of the ovary and the gonoduct are situated dorsally to the gut. The lumen of
the germinal part of the ovary is continuous with the lumen of the gonoduct. The gonoduct communicates to the exterior by the genital
pore. (C) Detail of a serial section of the ovary of Fig. 1B, where the three regions of the gonoduct: cephalic region, middle region, and
caudal region are depicted. Mucosal folds occur at the beginning of each region. At the ventral side of the middle region, there is a deep
invagination having large, thick, and irregular folds. Outside the genital pore, there are abundant cells of the immune system. (A) H-E,
Bar 5 200 mm. (B) H-E, Bar 5 400 mm. (C) PAS, Bar 5 200 mm. Caudal region, CaR; cephalic region, CeR; deep invagination of the
ventral side, *; follicles, F; genital pore, GP; germinal part of the ovary, Ge; gonoduct, Go; gut, Gu; lumen, L; middle region, MiR; mucosal
folds, f.

ciliated and nonciliated cells, similar to those
observed in the cephalic region. The connective
tissue is loose and vascularized. Cells of the
immune system, such as lymphocytes, macro-
phages, and neutrophils, are seen dispersed in
the connective tissue.
Melano-macrophage centers are irregularly
located in the connective tissue, adjacent to the
epithelium (Figs. 3A–D) of the middle region.
They are round or oval aggregates that contain
several types of cells such as macrophages,
lymphocytes, and melanocytes (Figs. 3A,D).
The melanocytes are irregularly distributed among
the immunological cells (Figs. 3B,C,E,F).
During previtellogenesis, the melano-macrophage
centers (Figs. 3A,B) have a mean diameter of

Journal of Morphology
250 J.C. CAMPUZANO-CABALLERO AND M.C. URIBE

Fig. 2. Cephalic region of the gonoduct of P. reticulata, during previtellogenesis (A, B), vitellogenesis (C, D), and gestation (E, F). (A, C,
E) The germinal part of the ovary contains vitellogenic follicles, and embryos during gestation. Thick mucosal folds, similar to a cervix,
extend into the gonoducal lumen at the initial portion of this region. (B, D, F) The histological components of the gonoduct wall consists of
cuboidal, luminal epithelium; vascularized loose connective tissue, which is looser during gestation as is observed in Fig. 2F, longitudinal
smooth muscle and visceral peritoneal epithelium at the periphery. The mucosal fold observed in Fig. 2D has the identical histological com-
ponents as does the gonoducal wall. (A–F) H-E. (A) Bar 5 60 mm. (B) Bar 5 20 mm. (C) Bar 5 40 mm. (D) Bar 5 10 mm. (E) Bar 5 200 mm. (F)
Bar 510 mm. Connective tissue, Ct;, connective tissue during gestation, Ct1; embryos, Em; epithelium, E; follicles, F; lumen, L; mucosal
folds, f; peritoneal epithelium, P; smooth muscle, M.

46.25 6 4.34 mm, during vitellogenesis (Fig. 3C),
they have a mean diameter of 51.61 6 3.21 mm.,
compared with these stages, during gestation are
larger (Figs. 3D–F), attaining a mean diameter of
84.52 6 12.68 mm.
Caudal Region
Observed during previtellogenesis (Fig. 1A),
vitellogenesis (Figs. 1B,C, 4A), and gestation
(Fig. 7A) stages. In similar fashion to that men-
tioned for the other regions, in the anterior part
of this region there are thin folds of the mucosa
that project into the lumen (Figs. 4A,B). Adja-
cent to the thin folds, there are two thick evagi-
nations or outpocketings of the mucosa of the
gonoducal wall, observed in the three reproduc-
tive stages. One of them is located at the ventral
side, and the other one is located at the dorsal
side (Fig. 4A). The ventral evagination is larger
and more anterior than that of the dorsal side
(Fig. 4A). Both evaginations reduce noticeably
the lumen in this part of the gonoduct. The epi-
thelium bordering the folds (Fig. 4B), and the
gonoducal wall (Figs. 4C,D) is columnar with
ciliated and nonciliated cells, also similar to

Journal of Morphology
 STRUCTURE OF THE GONODUCT OF POECILIA RETICULATA 251

Fig. 3. Melano-macrophage centers in the middle region of the gonoduct of P. reticulata. (A, B) During previtellogenesis. (C) Dur-
ing vitellogenesis. (D–F) During gestation. Melano-macrophage centers are adjacent to the luminal epithelium and surrounded by
connective tissue. They contain several types of cells such as macrophages, including lymphocytes and melanocytes. During gesta-
tion, the melano-macrophage centers are larger than those observed during previtellogenesis and vitellogenesis. At the periphery of
the gonoduct, there is longitudinal smooth muscle and visceral peritoneal epithelium. (A) PAS, Bar 5 20 mm. (B) PAS, Bar 5 60 mm.
(C) H-E. Bar 5 20 mm. (D) H-E, Bar 5 200 mm. (E) H-E, Bar 5 20 mm. (F) Masson’s trichrome, Bar 5 20 mm. Connective tissue, Ct;
luminal epithelium, E; lymphocytes, ly; macrophages, ma; melanocytes, me; melano-macrophage centers, Mc; peritoneal epithelium,
P; smooth muscle, M.

those observed in the cephalic and middle
regions. The connective tissue is loose and vas-
cularized (Fig. 4D), especially during gestation.
This suggests that it retains more fluid, similar
to those observed in the cephalic region during
gestation.
Ventrally, at the posterior side of the thick eva-
gination of the gonoducal wall, there are several
exocrine glands (Figs. 5A,B). The glands are lim-
ited by tall columnar epithelium (Figs. 5C–E).
The glandular cells contain a basal nucleus and
fine granular cytoplasm (Figs. 5C–E). Some glan-
dular cells include vacuoles of different sizes
in their cytoplasm suggesting the presence of
lipid secretory material (Figs. 5D,E). During pre-
vitellogenesis (Fig. 5A), the glandular cells have
a height of 15.78 6 1.7 mm; they are larger dur-
ing vitellogenesis (Figs. 5B–E) when they attain
22.84 6 2.31 mm in height, and gestation
(Figs. 6A–E) when they are 21.50 6 2.11 mm.
During gestation, the glands have portions with
irregular shape; some of the glandular cells des-
quamate into the gonoducal lumen and are
mixed with abundant cells of the immune system
(Figs. 6B–E).
During vitellogenesis, abundant spermatozoa
were observed in this region (Fig. 7A). Some sper-
matozoa were seen near the genital pore (Figs. 1C,
7A). Others were dispersed in the gonoducal
lumen (Figs. 7B,C). Still others had their heads

Journal of Morphology
252 J.C. CAMPUZANO-CABALLERO AND M.C. URIBE

Fig. 4. Caudal region of the gonoduct of P. reticulata during vitellogenesis. (A) Thin mucosal folds in the initial part of this region where
the lumen is reduced. Two thick evaginations of the mucosa, one of them at the ventral side and the other one at the dorsal side reduce the
gonoducal lumen. Adjacent and caudally to the ventral evagination there are exocrine glands. The gonoduct communicates with the exte-
rior at the genital pore. There are cells of the immune system in the lumen. (B) Detail of the thin mucosa folds expanded to the gonoducal
lumen in the initial part of this region; they contain columnar luminal epithelium and loose connective tissue. (C, D) The mucosa consists
of luminal columnar epithelium with ciliated and nonciliated cells and loose connective tissue with blood vessels. (A–D) Masson’s tri-
chrome. (A) Bar 5 60 mm. (B) Bar 5 20 mm. (C) Bar 5 20 mm. (D) Bar 5 10 mm. Blood vessels, v; cells of the immune system (arrow head),
connective tissue, Ct; dorsal side evagination, De; genital pore, GP; glands, G; lumen, L; luminal epithelium, E; mucosal folds, f; ventral
side evagination, Ve.

embedded in the apical ends of the epithelial cells
(Fig. 7D). Abundant cells of the immune system,
such as macrophages, neutrophils, and lympho-
cytes, were seen among the spermatozoa (Fig. 7B).
Near the caudal edge of the gonoduct, through
the genital pore, abundant cells of the immune
system were seen dispersed in the lumen and out-
side of the genital pore (Fig. 7A). These cells
include lymphocytes, neutrophils, and macro-
phages (Fig. 7E). At this farthest part of the gono-
duct, the epithelium becomes stratified (Fig. 7F)
and is formed by four or five layers of irregularly

Journal of Morphology
 STRUCTURE OF THE GONODUCT OF POECILIA RETICULATA 253

Fig. 5. Glands in the caudal region of the gonoduct of P. reticulata during previtellogenesis (A) and during vitellogenesis (B–E).
(A, B) At the ventral side of this region, several invaginations form glands. There are cells of the immune system in the lumen.
(C–E) The epithelium of the glands is columnar; the nuclei of the glandular cells are located on the basal side. Some cells of the
immune system, such as lymphocytes, are seen through the epithelium. Spherical vacuoles of different sizes are also observed in the
glandular cells. (A) H-E, Bar 5 20 mm. (B) H-E, Bar 5 20 mm. (C) H-E, Bar 5 10 mm. (D) Masson’s trichrome, Bar 5 10 mm. (E) Masson’-
s trichrome, Bar 5 10 mm. Cells of the immune system (arrow head), epithelium, E; glands, G; lumen, L; lymphocytes, ly; nuclei, n;
vacuoles, va.

shaped cells. The apical epithelial cells are oval or
squamous.
DISCUSSION
The ovary of P. reticulata observed in this study is
a single, elongated structure, of cystovarian type
and oriented longitudinally, coinciding with previ-
ous descriptions in this species by Jollie and Jollie
(1964) and Lambert (1970). The caudal part of the
ovary forms the gonoduct that lacks germinal cells.
The gonoduct of P. reticulata was divided in three
regions according to the presence of mucosa folds at
the initial part of each region: cephalic, middle, and
caudal regions. The morphological features of the
gonoduct of P. reticulata described in this study
indicates relevant functions of this part of the ovary
during the reproductive cycle, they are:
a. The gonoduct is the structure through which
embryos pass during birth, facilitated by the

Journal of Morphology
254 J.C. CAMPUZANO-CABALLERO AND M.C. URIBE

Fig. 6. Glands in the caudal region of the gonoduct of P. reticulata during gestation. (A) The germinal part of the ovary contains embryos
and follicles with large, yolked oocytes in development. The glands of the gonoduct are dorsal to the gut. (B–E) The glands have portions
where the epithelium has an irregular shape. Some of the glandular cells are desquamated and mixed with cells of the immune system.
Connective tissue and longitudinal smooth muscle are observed. (A–E) H-E. (A) Bar 5 200 mm. (B) Bar 5 20 mm. (C) Bar 5 20 mm. (D)
Bar 5 20 mm. (E) Bar 5 20 mm. Cells of the immune system (arrow head), connective tissue, Ct; embryos, Em; epithelium, E; follicles, F;
glands, G; gut, Gu; smooth muscle, M.

ciliated epithelial cells and the longitudinal
layer of smooth muscle that surrounds the gono-
ducal wall. Lahnsteiner et al. (1997) docu-
mented the presence of ciliated and nonciliated
epithelial cells, restricted to the gonoduct, in
the ovary of the oviparous species Alburnus
alburnus (Cyprinidae), the ciliated cells favoring
the transport of eggs throughout their passage
by the gonoduct, and the nonciliated cells hav-
ing a secretory activity. The viviparous condi-
tion of P. reticulata suggests that functions of
the gonoduct also include the transport of
sperm during insemination and the passage of
embryos during birth.
b. The gonoduct of P. reticulata forms partial bar-
riers that are defined particularly by the reduc-
tion of the ovarian lumen at the beginning of
the cephalic, middle, and caudal regions by the
presence of large folds of the mucosa; and by
the development of ventral and dorsal thick
evaginations in the caudal region.
c. The presence of abundant cells such as macro-
phages, neutrophils, and lymphocytes that were
observed in the gonoduct of P. reticulata indi-
cates immune functions, coinciding with the
descriptions by Tazumi et al. (2004), who
observed these three cell types, in the germinal
part of the ovary of the viviparous teleost,

Journal of Morphology
 STRUCTURE OF THE GONODUCT OF POECILIA RETICULATA 255

Fig. 7. Spermatozoa in the caudal region of the gonoduct of P. reticulata during vitellogenesis. (A) There are abundant spermatozoa near
the genital pore and extending into the gonoduct. (B) Detail of the previous figure where the presence of abundant spermatozoa in the
lumen and near the luminal epithelium is evident. (C, D) Some spermatozoa are in the gonoducal lumen while the heads of others appear
to be embedded in the apical ends of the epithelial cells. (E) Cells at the genital pore and in the gonoducal lumen where there are lympho-
cytes, macrophages, and neutrophils. (F) View of the lumen at the caudal end of the gonoduct. (A–F) H-E. (A) Bar 5 100 mm. (B) Bar 5 20
mm. (C) Bar 5 20 mm. (D) Bar 5 10 mm. (E) Bar 5 10 mm. (F) Bar 5 10 mm. Cells of the immune system (arrow head), genital pore, GP; gono-
ducal lumen, L; lymphocytes, ly; macrophages, ma; neutrophils, ne; spermatozoa, z.

Neoditrema ransonneti. These cells were seen in
the gonoduct of P. reticulata in the connective
tissue subjacent to the luminal epithelium and
in the gonoducal lumen similar to those
observed in the viviparous teleosts Ilyodon
whitei and Goodea atripinnis (Uribe et al.,
2005, 2009).
The presence of melano-macrophage centers in
the gonoduct of P. reticulata, during the three
reproductive stages described in this study,
suggests the involvement of the gonoduct in
the protection of the ovary and embryos by
immunological functions, against microbes,
phagocytosis of foreign particles, especially,
during gestation when these centers increase
their size. Melano-macrophage centers have
been observed previously in teleosts in differ-
ent organs such as kidney, spleen, and liver
(Agius, 1980; Agius and Agbede, 1984; Wolke,
1992; Agius and Roberts, 2003). Similar to
those seen in the gonoduct of P. reticulata, they
consist of lymphocyte, melanocyte, and macro-
phage aggregations. Kranz (1989), Bucke et al.
(1992), Wolke (1992), and Leknes (2007)
concluded that the functions of melano-
macrophage centers in fishes are multiple and

Journal of Morphology
256 J.C. CAMPUZANO-CABALLERO AND M.C. URIBE
complex, involved in immunological sequestra-
tion and destruction of products of cellular
degradation or foreign materials, including
infectious agents. Coinciding with these state-
ments, Agius and Roberts (2003) and Vigliano
et al. (2006), suggested an evolutionary rela-
tionship between the melano-macrophage cen-
ters of fish and the germinal centers of lymph
nodes of birds and mammals.
d. Exocrine glands in the gonoduct of P. reticulata
are located on the ventral side of the caudal
region. The glandular cells have common mor-
phological features such as being columnar cells
with basal nuclei and fine granular cytoplasm
that indicates secretory activity. Several authors
have documented secretory activity in the ovary
of viviparous teleosts, but all of them refer their
observations to the luminal epithelial cells in
the germinal part of the ovary (Mendoza, 1941;
Igarashi, 1961; Lombardi and Wourms, 1985;
Koya et al., 1998; Potter and Kramer, 2000).
However, there is no description of glandular
development, as was observed in this study in
the gonoduct of P. reticulata. The gland cells of
P. reticulata are more developed during vitello-
genesis. This feature suggests a role during
insemination such as providing a proper envi-
ronment for the passage of the spermatozoa.
These glands probably add a complex secretion
to the gonoduct fluid since they show different
vacuolar inclusions. Furthermore, during gesta-
tion, the glands observed in P. reticulata showed
desquamated gland cells accompanied with cells
of the immune system. Further analysis is
needed to understand the function of these gon-
oducal glands, the nature of their secretions,
the dynamics of the fluid environment, and the
mechanisms that control the changes of the
glandular components during the reproductive
cycle of P. reticulata.
e. The gonoduct of P. reticulata observed in this
study has morphological similarities to the ovi-
ducts of other vertebrates that also have inter-
nal fertilization and gestation. These
similarities include ciliated and nonciliated epi-
thelial cells that facilitate the transport of both
substances and cells, the spermatozoa and eggs
(Howarth, 1974; Johnson and Foley, 1974; Lom-
bardi, 1998), exocrine glands, and smooth mus-
cle that contracts at specific times during the
reproductive cycle, in particular, during oviposi-
tion or during birth of embryos.
As observed in this study, spermatozoa occur in
the caudal region of the gonoduct of P. reticulata
during vitellogenesis. Coinciding with our observa-
tion, Greven (2005) also observed, in P. reticulata,
spermatozoa in the caudal region of the gonoduct
during insemination. Spermatozoa are introduced
into the gonoduct after which they move to the
Journal of Morphology
germinal part of the ovary. In the present study,
the spermatozoa were seen in the gonoduct among
macrophages, suggesting that these cells phagocy-
tize additional spermatozoa, coinciding with the
descriptions of spermatozoa in the lumen of the
germinal part of the ovary documented by Tazumi
et al. (2004) in the viviparous teleost Neoditrema
ransonneti, and by Koya et al. (1997) in the ovipa-
rous teleost Alcichthys alcicornis. Future research
on the role that the gonoduct plays in modulating
sperm function and how these modifications affect
fertilization is needed.
ACKNOWLEDGMENTS
The authors thank Harry J. Grier for his valua-
ble comments that improved this manuscript,
Humberto Olvera-Quezada, Marıa Estela Perez-
Cruz, and Ignacio Andres Morales-Salas for valua-
ble help taking care of fishes in captivity, Adriana
Garcıa-Alarcon for help in histological processing,
Gabino De la Rosa Cruz and Ana Isabel Bieler
Antolin for the kind technical assistance with digi-
tal photography; and Jose Antonio Hern andez
Gomez for the excellent digital preparation of the
figures.
LITERATURE CITED
Agius C. 1980. Phylogenetic development of melano-
macrophage centres in fish. J Zool 191:11–31.
Agius C, Agbede SA. 1984. Electron microscopical studies on
the genesis of lipofuscin, melanin and haemosiderin in the
haemopoietic tissues of fish. J Fish Biol 24:471–488.
Agius C, Roberts RJ. 2003. Melano-macrophage centers and
their role in fish pathology. J Fish Dis 26:499–509.
Aguilar-Morales M, Couti~ no-Bello B, Salinas-Rosales P. 1996.
Manual general de t ecnicas histologicas y citoquımicas.
M exico: Las Prensas de Ciencias, Facultad de Ciencias,
UNAM. p 106.
Bucke D, Vethaak AD, Lang T. 1992. Quantitative assessment
of melanomacrophage centres (MMCs) in dab Limanda
limanda along pollution transect in the German Bight. Mar
Ecol Prog Ser 91:193–196.
Burns JR, Weitzman SH. 2005. Insemination in Ostariophysan
fishes. In: Uribe MC, Grier HJ, editors. Viviparous Fishes.
Homestead, FL: New Life Publications. pp 105–132.
Evans JP, Pilastro A, Ramnarine IW. 2003. Sperm transfer
through forced matings and its evolutionary implications in
natural guppy (Poecilia reticulata) populations. Biol J Linn
Soc 78:605–612.
Gasparini C, Andreatta G, Pilastro A. 2012. Ovarian fluid of
receptive females enhances sperm velocity. Naturwissenschaf-
ten 99:417–420.
Greven H. 2005. Structural and behavioral traits associated
with sperm transfer in Poeciliinae. In: Uribe MC, Grier HJ,
editors. Viviparous Fishes. Homestead, FL: New Life Publica-
tions. pp 145–163.
Greven H. 2010. What do we know about reproduction of inter-
nally fertilizing halfbeaks Zenarchopteridae)? In: Uribe MC,
Grier HJ, editors. Viviparous Fishes II. Homestead, FL. USA:
New Life Publications. pp 121–141.
Grier HJ, Uribe MC, Parenti LR, De la Rosa-Cruz G. 2005.
Fecundity, the germinal epithelium, and folliculogenesis in
viviparous fishes. In: Uribe MC, Grier HJ, editors. Viviparous
Fishes. Homestead, FL: New Life Publications. pp 191–126.
 STRUCTURE OF THE GONODUCT OF POECILIA RETICULATA
Howarth BJ. 1974. Sperm storage: As a function of the female
reproductive tract. In: Johnson AD, Foley CW, editors.
The Oviduct and its Functions. New York, London: Academic
Press. pp 237–270.
Igarashi T. 1961. Histological and cytological changes in the ovary
of a viviparous teleost, Neoditrema ransonneti Steindachner,
during gestation. Bull Fac Fish Hokkaido Univ 12:181–188.
Jalabert B, Billard R. 1969. Etude ultrastructurale du site de
conservation des spermatozoides dans l’ovaire de Poecilia
reticulata (Poisson Teleosteen). Ann Biol Anim Biochim
Biophys 9:273–280.
Johnson AD, Foley CW. 1974. The Oviduct and Its Functions.
New York, London: Academic Press. pp 369.
Jollie WP, Jollie LG. 1964. The fine structure of the ovarian follicle
of the ovoviviparous poeciliid fish, Lebistes reticulates. II. Forma-
tion of follicular pseudoplacenta. J Morphol 114:503–526.
Kobayashi H, Iwamatsu T. 2002. Fine structure of the storage
micropocket of spermatozoa in the ovary of the guppy Poecilia
reticulata. Zool Sci 19:545–555.
Koya Y, Munehara H, Takano K. 1997. Sperm storage and deg-
radation in the ovary of a marine copulating sculpin, Alcich-
thys alcicornis (Teleostei: Scorpaeniformes): Role of
intercellular junctions between inner ovarian epithelial cells.
J Morphol 233:153–163.
Koya Y, Itazu T, Inoue M. 1998. Annual reproductive cycle based
on histological changes in the ovary of the female mosquitofish,
Gambusia affinis in central Japan. Ichthyol Res 45:241–248.
Kranz H. 1989. Changes in splenic melano-macrophage centers
of dab Limanda limanda during and after infection with
ulcer disease. Dis Aquat Organ 6:167–173.
Lahnsteiner F, Weismann T, Patzner RA. 1997. Structure and
function of the ovarian cavity and oviduct and composition of
the ovarian fluid in the bleak, Alburnus alburnus (Teleostei,
Cyprinidae). Tissue Cell 29:305–314.
Lambert JGD. 1970. The ovary of the guppy Poecilia reticulata.
The granulosa cells as sites of steroid biosynthesis. Gen
Comp Endocrinol 21:287–304.
Leknes IL. 2007. Melano-macrophage centers and endocytic cells
in kidney and spleen of pearl gouramy and platyfish (Anabanti-
dae, Poeciliidae: Teleostei). Acta Histochem 109:164–168.
Lombardi J. 1998. Comparative Vertebrate Reproduction. Boston,
Dordrecht, London: Kluwer Academic Publisher. pp 1–469.
Lombardi J, Wourms JP. 1985. The trophotaenia placenta of a
viviparous goodeid fish. I. Ultrastructure of the internal ovar-
ian epithelium, the maternal component. J Morphol 184:
277–292.
Mendoza G. 1941. The reproductive cycle of the viviparous tele-
ost, Neotoca bilineata, a member of the family goodeidae: III.
The germ cell cycle. Biol Bull 81:70–79.
Mendoza G. 1943. The reproductive cycle of the viviparous tele-
ost Neotoca bilineata, a member of the family Goodeidae. IV.
The germinal tissue. Biol Bull 84:87–97.
Pilastro A, Bisazza A. 1999. Insemination efficiency of two
alternative male mating tactics in the guppy (Poecilia reticu-
lata). Proc R Soc London B Biol 266:1887–1891.
257
Potter H, Kramer CR. 2000. Ultrastructural observations on
sperm storage in the ovary of the platyfish, Xiphophorus
maculatus (Teleostei: Poeciliidae): The role of the duct epithe-
lium. J Morphol 245:110–129.
Tazumi Y, Nakamura O, Watanabe T. 2004. Intraovarian cavity
leucocytes of viviparous fish, Neoditrema ransonneti
(Perciformes, Embiotocidae). Zool Sci 21:739–746.
Turner CL. 1947. Viviparity in teleost fishes. Sci Mon 65:
508–518.
Uribe MC, De la Rosa-Cruz G, Garcia-Alarcon A. 2005.
The ovary of viviparous teleosts. Morphological differences
between the ovaries of Goodea atripinnis and Ilyodon whitei.
In: Uribe MC, Grier HJ, editors. Viviparous Fishes.
Homestead, FL: New Life Publications. pp 217–235.
Uribe MC, Grier HJ, De la Rosa Cruz G, Garcıa Alarc on A.
2009. Modifications in ovarian and testicular morphology
associated with viviparity in teleosts. In: Jamieson B, editor.
Reproductive Biology and Phylogeny of Fish (Agnatha and
Osteichthyes).Enfield, N H, Plymouth, U K: Science Publish-
ers, Inc. pp 85–117.
Uribe MC, Aguilar-Morales M, De la Rosa-Cruz G, Garcıa-
Alarcon A, Campuzano-Caballero JC, Guerrero-Est evez SM.
2010. Ovarian structure and embryonic traits associated with
viviparity in poeciliids and goodeids. In: Uribe MC, Grier HJ,
editors. Viviparous Fishes II. Homestead, FL: New Life Publi-
cations. pp 211–229.
Urriola Hern andez M, Cabrera Pe~ na J, Protti Quesada M.
2004. Fecundidad, fertilidad e ındice gonadosom atico de
Poecilia reticulata (Pisces:Poeciliidae) en Heredia, Costa Rica.
Rev Biol Trop 52:945–950.
Vigliano FA, Berm udez R, Quiroga MI, Nieto JM. 2006.
Evidence for melano-macrophage centres of teleost as evolu-
tionary precursors of germinal centres of higher vertebrates:
An immunohistochemical study. Fish Shellfish Immunol 21:
467–471.
Wake WH. 1985. Oviduct structure and function in
non-mammalian vertebrates. Forts Zool 30:427–435.
Wake WH. 1989. Phylogenesis of direct development and
viviparity in vertebrates. In: Wake DB, Roth G, editors.
Complex Organismal Functions: Integration and Evolution in
Vertebrates. Chichester: Wiley. pp 235–250.
Wolke RE. 1992. Piscine macrophage aggregates: A review.
Annu Rev Fish Dis 2:91–108.
Wourms JP. 1981. Viviparity: The maternal-fetal relationship in
fishes. Am Zool 21:473–515.
Wourms JP. 2005. Functional morphology, development, and
evolution of trophotaeniae. In: Uribe MC, Grier HJ, editors.
Viviparous Fishes. Homestead, FL: New Life Publications.
pp 217–242.
Wourms JP, Lombardi J. 1992. Reflections on the evolution of
piscine viviparity. Am Zool 32:276–293.
Wourms JP, Grove BD, Lombardi J. 1988. The maternal-
embryonic relationships in viviparous fishes. In: Hoar WS,
Randall DJ, editors. Fish Physiology, Vol. XI. New York:
Academic Press. pp 1–134.
Journal of Morphology