Term Paper

Proteomics of Human Aqueous Humor

Prof.dr.Sabina Semiz

Table of Contents
Abstract

1. INTRODUCTION 3
2. AQUEOUS HUMOR

2.1 Aqueous vs.Vitreous Humor 5

2.2 Formation and Flow of Human Aqueous Humor 6

3. PROTEOMICS OF HUMAN AQUEOUS HUMOR

3.1 Classification of Proteins found in Human Aqueous Humor

10

3.2 Proteins Previously Identified in Human Aqueous Humor

11

3.2.1

Vitamin D-binding protein (GC) and Pigment epithelium derived factor

(SERPINF1)

13

3.2.2

Transforming growth factor beta 2 (TGFB2) 14

3.2.3

Components of the complement pathway

3.2.4

Osteopontin and Vasorin

14

15

3.3 Proteins identified in human aqueous humor for the first time
3.3.1

Sorbitol dehydrogenase (SORD)

3.3.2

Filensin, Phakinin and Platelet derived growth factors

16

3.4 Enzymes identified in human aqueous humor

4. DISCUSSION

18

5. CONCLUSION

20

6. References

15

16

16

21
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Abstract
The eye is a double organ which is responsible for receiving information from the
outside world and transforming it into electrical impulses, so they could get to the brain. Its
principal function is to create vision. To be able to do that, it needs proper functioning of all of
it's parts. Besides the well know parts of the eye, such as cornea or lens, the eye contains
many others which have an important function. One of those is the aqueous humor, a
transparent fluid in the front chamber of the eye. The regulation and secretion of aqueous
humor and it's flow, are physiologically significant processes for the eye. Besides that, the
composition of the aquous humor is also important. This specific article is dealing with the
proteomics of the aqueous humor, different proteins and enzymes disscovered in previous
studies and those disscovered here for the first time. Incredible 763 proteins were found to
circulate through the aqueous humor, 386 identified for the first time in this part of the eye.
The results of this study will serve as a great baseline for other studies, whether on the
aqueous humor itself or diseases in relation with it.

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1. INTRODUCTION
After the brain, eyes are the most complex organ. One eye is composed of over 2
milion of working parts, but only 1/6 of it is exposed and visible from the outside. The eye is
a small organ, which does not change it's dimensions from birth until death, and it is there to
convert light into electrical impulses. (Lamb, 2011)
The human eye is a double organ which is working on a principle similar to cameras. The
transparent front of the eye is breaking the beams of light, projecting a smaller and inverted
image on the photosensitive retina where the specialized nerve cells perform conversion into
electrical nerve impulses. (Lamb, 2011)
The eye is the most important human sense because it receives 80% of all information from
the environment, allows conscious perception of light, recognition of colors and depth
perception. (McBride, 2010) In the area of the eye are a number of morphological and
functional systems: optical, accommodative, the system of external eye muscles,
photoreceptive, nervous system, the system for creating and circulating aqueous humor,
cardiovascular, eye protective apparatus and system for maintaining the size and shape of the
eye. The main role in keeping the shape of the eye and provision of nutrients has the aqueous
humor, the central theme of this work. In this study, Krishna R.Murthy et al. (2015) took
aqueous humor samples from different patients, processed further to different fractionation
strategies and did an analysis by mass spectrometry. The results were more than satisfactory
with 386 novel proteins identified in the human aqueous humor.
2. AQUEOUS HUMOR

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The eyeball consists of three envelopes. The external, fibrous envelope at the front is
transparent and we call it cornea. In the back of the eye there is the non-transparent sclera.
The cornea has an optical and supporting role, and the sclera does only support. Beneath the
sclera is the middle eye envelope or uvea. Under the eye envelopes are the lens, vitreous and
aqueous humor. (McCaa, 1982)
The eyeball itself can be divided into two psegments, which are both containing fluids. The
front segment, or front chamber, is localized from the cornea up to the lens, and the back
segment, or back chamber, from the back edges of the lens to the retina. The front chamber is
filled with a fluid that we know as human aqueous humor that feeds the internal tissue of the
eye, while the back chamber carries a gel-like substance that we call the vitreous humor.
These fluids are there to help the eyeball keep its shape. A bigger amount, 4/5 of human
aqueous humor is in the front chamber and 1/5 in the back chamber of the eye. The total
amount of human aqueous humor in one eye is 0.30 ml. The human aqueous humor is
secreted from blood of ciliary fringes by different mechanisms like ultrafiltration, active
secretion and dialysis. (McCaa, 1982) Parts of the eye are shown in Figure 1.

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Figure 1. Parts of the Eye (Picture taken from:

https://en.wikipedia.org/wiki/Aqueous_humour#/media/File:Schematic_diagram_of_the_hum
an_eye_en.svg)
What are actually the most important roles of the human aqueous humor? Aqueous humor has
a function in balancing intraocular pressure, refraction, oxydation and it is important in
nutrition. Besides that, the human aqueous humor takes away wastes, products of
inflammation and metabolism overall. It has also a role in maintaining the temperature
difference of the colder cornea and warmer planks. (Murthy et al., 2015)
Primary aqueous humor, which is the one made in the back chamber circulating to get into the
front chamber, is made out of 90% water, and 5-16 mg / 100 ml protein (suggesting that it is
poor in proteins). There are also other molecules besides proteins in the aqueous humor, like
ascorbic acid who's concentration is higher in aqueous humor than in plasma. The same
situation is with amino acids. Both of them, ascorbic acid and amino acids, are regulary
transported from blood into the human aqueous humor. Talking about cations, sodium is the
most important in plasma as well as in aqueous humor. The calcium and phosphorus
concentrations are similar in blood and human aqueous humor. (De Berardinis et al., 1965)
The secondary aqueous humor is the one which reached the front chamber and it also reached
an increase in protein concentration. This happens because of the disturbed hematoocular
barrier and proteins can easily pass the barrier. Mostl globulins, albumins, and
gammaglobulins antibodies are found in the human aqueous humor. Besides that, the
secondary aqueous humor has increased concentrations of substances which are usually
deficit in it, like urea or glucose, and substances that are in excess like hyaluronic acid, lactic
acid and ascorbic acid). (Fredo et al. 1990)
2.1 Aqueous vs. Vitreous Humor
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One may find it hard to distinguish between aqueous and vitreous humor in the eye.
The difference is simply made. Vitreous humor is a gel-like fluid which is placed in the back
chamber of the eye and maintains mostly its shape. It consists mostly of water, but it contains
also collagen and hyaluronic acid which are making it gelatinous. The vitreous humor also
absorbs shocks and keeps the eye parts, retina and back wall well connected. While getting
older, the vitreous humor losses it's gel consistency and becomes more fluid, which makes the
retina separate from the back wall and cause posterior vitreous detachment in older people.
Aqueous humor is more water-like and located in the front chamber of the eye, between iris
and cornea. It keeps the shape of this part of the eye, and brings nourishment to the eye.
(Healthline, 2015)
2.2 Formation and Flow of Human Aqueous Humor
Making aqueous humor happens in the ciliary body by different and compley
processes. It is made from the blood which is flowing through the cilia extensions. From this
part, the aqueous humor is secreted in the back chamber, from which it flows besides the lens
and enters the front chamber. In the part of the chamber angle (area of the front chamber
which is placed in the part of the cornea joining the planks) we find the so called trabecular
meshwork, aqueous veins (venae aquosae) and Schlems channel. This is the main way
through which the human aqueous humor exits the anterior chamber. (see Figure 2) (Johnson,
2010)
The relation between formation and swelling of the human aqueous humor on the pressure in
the eye which is called intraocular pressure. The disruption of this relation and balance, an
increased or decreased formation, or increased and decreased swelling of the human aqueous
humor, will make an fall or rise in the intraocular pressure, what can mean a risk to the health

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of the eye. One of the risks is eye glaucoma, which is the consequence of increased
intraocular pressure. Because of that, the maintenance of intraocular pressure is crucial and
for the prevention and regulation of this and other dangerous diseases. (Brubaker, 1991)

Figure 2. Human Aqueous Humor pathway (Picture taken from:

http://medicforyou.blogspot.ba/2015/09/what-is-glaucoma-glaucome-glucoma.html)

3. PROTEOMICS OF HUMAN AQUEOUS HUMOR

Human aqueous humor (hAH) is a fluid consisting of a mixture of cytokines,
electrolytes, growth factors, organic solutes and some proteins that make the metabolic
requirements to the tissues in the front segment. (Chowdhury et al.,2010) The protein
component of the secondary human aqueous humor is minimal, and makes between 120 and
500 ng/L of protein. (Cole, 1947)
The human aqueous humor has antioxidant functions and studies have shown an immune
response role during infection and inflammation. Still, the proteins which are responsible for
most of this functions are unknown. The protein composition of the human aqueous humor

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has been studied earlier. But, because of limitations in technology and methods, a good and
definitive description of human aqueous humor proteins must be provided in future.
(Richardson et al.,2009)
Rohde et al. (1998) did a research and used a proteomic approach to analyze human aqueous
humor proteins. In this early studies, because of technological restrictions, the proteins could
be identified only based on molecular weight and so have to be considered tentative. The
study in this time suggested that the most appropriate techniques for these kind of
experimentation include coupling of 1 and 2-dimensional electrophoresis with mass
spectrometric of the sample proteins. The researchers usedsed membrane preconcentrationCE-MS (mPC-CE-MS) and analised 13 patient samples of human aqueous humor (AH).
Measured molecular weights (Mr) as already stated, were used to identify different proteins
after searching in the SWISS-PROT database. Proteins which were identified in this time are
apolipoprotein A1 (Mr 28078.6), beta-2 microglobulin (Mr 11731.2) and serum albumin (Mr
66400). ( Rohde et al., 1998)
Funding et al. (2005) and Duan et al.(2008), both used two-dimensional electrophoresis
together with tandem mass spectrometry, and managed to identify 7 unique aqueous humor
proteins with high high significance, and in both studies interfering high abundance proteins
like albumin which limited the analysis.
Even today, there are only a small number of research done on the proteomics of human
aqueous humor. Earlier studies have had limitations of differend kinds and the human aqueous
humor proteome remained poorly defined besides all efforts. Limitations such as inadequate
animal models, interfering high abundance proteins and limited proteomic technologies
hindered the researchers. (Richardson et al., 2009) To make more investigations into human
aqueous humor functions, the human aqueous humor proteome was analyzed using advanced

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proteomic techniques. (Murthy et al., 2015) The maximum number of proteins identified from
previous studies was 676 (Chowdhury et al.,2010). Chodwury at al. used nano-LC-ESIMS/MS and identified proteins with catalytic, enzymatic, and structural properties. Using
antibody-based protein arrays, 328 cytokines, chemokines, and receptors were identified.
More than 80% of these proteins have been identified for the first time. (Chowdhury et
al.,2010)
Other studies usng mostly LC-MS/MS, but also MALDI-TOF/TOF MS method were used to
identify different amounts of protein, 135 (Anshu et al.), 198 (Bennett et al), 323 (Pollreisz et
al), 501 (Taube et al.) etc.
In this specific study, LC-MS/MS was used to identify the largest amount of protein in human
aqueous humor until now. Murthy et al. (2015) identified 763 proteins, including 386 proteins
identified for the first time in human aqueous humor. The study is confirming the annotated
translational start sites of 47 proteins from those 763, based on their N-terminal peptides.
They also provide evidence of signal peptide cleavage sites in 33 proteins. (Murthy et
al.,2015)
The advantage of this study is the number of patients and methods used to analyse the
samples. This is one of the rare studies working with a number of 250 patients. All of them
were undergoing cataract surgery to take the aqueous humor samples, which were further
processed by fractionating and subjected to mass spectometry. In Figure 3 is a description of
the procedure used by Murthy et al. to extract proteins from samples, going from depletion of
abundant proteins by MARS-14, over SDS-PAGE, strong cation exchange chromatography
and bRPLC, until mass spectrometry by the LTQ-Orbitrap Velos mass spectrometer. (Murthy
et al., 2015)

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Figure 3. Workflow employed in the study (Picture taken from: Murthy et al., 2015)

3.1 Classification of proteins found in Human Aqueous Humor

Earlier studies identified proteins in the human aqueous humor which had catalytic,
enzymatic, and structural functions. Figure 4 shows the classification of proteins identified in
the study done by Chowdhury et al. (2010). Shown are 355 proteins with known function,
other proteins in that study had unknown function. (Chowdhury et al., 2010) Our specific
study did a detailed classification of identified proteins according to domain information and
subcellular localization (Figure 5). (Murthy et al., 2015)

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Figure 4. Classification of identified

proteins according to their function (Picture
taken from: Chowdhury et al., 2010)

Figure 5. Classification of proteins based on domain information and subcellular localization

(Picture taken from: Murthy et al., 2015)

3.2 Proteins Previously Identified in Human Aqueous Humor

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As mentioned before, earlier studies identified different proteins in human aqueous

humor. Most of them were growth factors, cytokines, or receptors. Some of the proteins
identified in previous studies are shown in Table 1. (Chowdhury et al., 2010)

Table 1. Proteins identified in previous studies (Picture taken from: Chowdhury et al., 2010)

Besides Chadrwory et al. (2010), it is important to mention the finding of other research
groups (Richardson et al., 2009) as well as results from our specific study (Murthy et al.,
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2015). In the Supplementary Table 1 you will find the complete list of proteins identified in
the study done by Murthy et al. (2015).
Some of the most interesting proteins previously identified, are the vitamin D-binding protein
(GC), transforming growth factor beta 2 (TGFB2), components of the complement pathway
(C3, C4B, C5, C6, C7), osteopontin (SPP1), platelet derived growth factor D (PDGFD),
vasorin (VASN) and pigment epitelium derived factor (SERPINF1).
3.2.1

Vitamin D-binding protein (GC) and Pigment epithelium derived factor

(SERPINF1)

Anti-angiogenic proteins are key molecules in the human aqueous humor, because
there are some tissues that have to remain avascular in order to maintain the function they
have. One of them is the cornea which has a function of a clear window for the eye. Besides
that, it is known that the human aqueous humor suppresses angiogenesis, but the main
proteins which are responsible for this function are incompletely understood. Earlier studies
identified different proteins with high confidence, and some of them for the first time in
human aqueous humor, and they have been proven to be potent inhibitors of angiogenesis.
Until now this proteins have been confirmed in the human aqueous humor and mentioned in
this specific study. These include GC and Pigment epithelium-derived factor (SERPINF1).
GC is modified by membrane-bound -galactosidase and sialidase of activated B- and Tlymphocytes, and the molecule we get from it is known as GC-maf (macrophages activating
factor). This molecule is the key anti-angiogenic protein. SERPINF1 has been shown to
suppress endothelial cell migration in the presence of angiogenic proteins. The expression of
SERPINF1 in the human aqueous humor has been brought in connection with lens opacity as
well as antioxidant capacity. (Yoshida et al., 2007) Its levels are inversely correlated with age,

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and lack of SERPINF1 has been proposed to play a role in the development of cataracts.
(Richardson et al., 2009)
The presence of angiogenic inducers such as angiogenin, and inhibitors such as vitamin D
binding protein, show the presence of an balance within human aqueous humor between
proangiogenic and antiangiogenic molecules. The balance between those two, angiogenic and
antiangiogenic proteins, is important in the pathogenesis of different front chamber diseases.
(Chowdhury et al.,2010)
Vitamin D-binding protein, was also previously mentioned to have a role in the human
aqueous humor and is a multi-functional plasma protein known to transport vitamin D and
bind fatty acids. It also has a role in immunological processes. (Richardson et al., 2009)
3.2.2

Transforming growth factor beta 2 (TGFB2)

TGFB2 is a protein previously identified in human aqueous humor as a central

mediator regulating trabecular meshwork (TM) cell function. The TM cells secrete TGFB2
and also express TGFB2 receptors. Studies have shown that TGFB2 has an important role in
glaucoma, it also increases expression of other genes, PAI-1, TSP-1, tissue transglutaminase,
hyaluronan synthase etc. (Civan, 2008)
3.2.3

Components of the complement pathway

Although many of the proteins in the human aqueous humor come from blood,

cDNAs encoding plasma proteins are also present in the ciliary body. Some of these are
complement components of the complement pathway. Studies confirmed their presence in
human aqueous humor and suggests that the ciliary body could be one of the tissues in the
front chamber that can produce and secrete plasma proteins into human aqueous humor.
(Chowdhury et al., 2010)
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Murthy et al. (2015) suggest that the complement factors have an antimicrobial role in the eye
and have an still unclear function in the immune response of the human aqueous humor and
the phenomenon called anterior chamber associated immune deviation (ACAID).
3.2.4

Osteopontin and Vasorin

Osteopontin, a member of the matricellular protein family, originally identified in

hAH by the nano-LC-ESI-MS/MS studies had exhibited levels near 70 ng/mL. (Chowdhury et
al.,

2010)

Osteopontin

is

phosphorylated

acidic

arginine-glycine-aspartate-

(RGD-)containing glycoprotein that exists both as an immobilized ECM component and as a

soluble, multifunctional, proinflammatory cytokine that plays important roles in promoting
inflammation, tissue remodeling, fibrosis, and angiogenesis. (El-Asrar et al., 2012)
The roles of several other proteins identified here have already been purported in the AH. For
example, though the exact role of Glutathione in the AH is unclear, it is known to protect cells
and other proteins from oxidative damage and most likely serves a similar function in the AH.
(Richardson et al., 2009)
3.3 Proteins identified in human aqueous humor for the first time
Murthy et al. (2015) identified 386 unique proteins in the human aqueous humor and
put a partial table of those in their study (see Table 2).
Table 2. Partial list of uniquely identified proteins (Picture taken from: Murthy)
The most interesting proteins, with known function in the aqueous humor, are the first five
from the list: Sorbitol dehydrogenase (SORD), Beaded filament structural protein 1 (Filensin)
and 2 (Phakinin), platelet derived growth factor alpha and beta (PDGFA and PDGFB).
3.3.1

Sorbitol dehydrogenase (SORD)

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Sorbitol dehydrogenase (SORD) is the most importan enzyme in the metabolism of

glucose in the human lens. The lens derives its energy requirements by 70% from the (Murthy
et al., 2015) anaerobic glycolisis pathway, while the rest is derived from the aerobic Kreb's
cycle. Sorbitol dehydrogenase is a cytosolic enzyme converting the sugar alcohol form of
glucose (sorbitol), into fructose. In disease conditions SORD is accumulated and can cause
sugar-related cataract. (Murphy et al., 2015)
3.3.2

Filensin, Phakinin and Platelet derived growth factors

BSFP1 and BSFP2 are proteins unique to differentiated lens fiber cells, and their
function has been shown in the differentiation of epithelial lens cells to fiber lens cells.
Platelet derived growth factors are receptor tyrosine kinases, appearing in alpha and beta
form. Both of them have important functions in cell proliferation, migration, organogenesis,
while PDGFB is more important in cardiovascular system and angiogenesis. Theor
overexpression or reduction lead to different disorders such as glioblastomas, diabetic
retinopathy or vitreoretinopathy. (Murthy et al., 2015)
3.4 Enzymes identified in human aqueous humor

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The study made by Murthy et al. (2015), reports 22 enzymes in the human aqueous
humor. Those enzymes are involved in glycolysis, gluconeogenesis and pentose phosphate
pathway. In Figure 6.we see a pathway representation of proteins in the anaerobic energy
metabolism, and enzymes (highlighted in blue) that were identified in this study. A complete
list of enzymes is given in Table 3. (Murthy et al., 2015)

Figure 6. Pathway representation of proteins in the anaerobic energy metabolism (Picture

taken from: Murthy et al., 2015)

Enzymes identified in Human Aqueous Humor

Enzyme symbol
PGM1
PFKL
G6PD

Enzyme name
Phosphoglucomutase-1
6-phosphofructokinase, liver type
Glucose-6-phosphate dehydrogenase
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PGLS
PGD

6-Phosphogluconolactonase
Phosphogluconate dehydrogenase

TKT
TALDO1
GPI

Transketolase
Transaldolase 1
Glucose-6-phosphate isomerase

LDHA
Lactate dehydrogenase alpha
LDHB
Lactate dehydrogenase beta
BPGM
Bisphosphoglycerate mutase
TPI1
Triosephosphate isomerase 1
ALDOA
Aldolase A
ALDOC
Aldolase C
PKM2
ENO1
ENO2
PGAM1

Pyruvate kinase M2 (muscle)

Enolase 1
Enolase 2
Phosphoglycerate mutase 1

PGK1
Phosphoglycerate kinase 1
AKR1A1
Aldo-keto reductase family 1, member A1
ALDH3A1

ALDH9A1

Aldehyde dehydrogenase 3 family, member

A1
Aldehyde dehydrogenase 9 family, member A1

Table 3. List of 22 enzymes identified in Human Aqueous Humor ( Table made by Author)
Some of those enzymes identified in aqueous humor are catalyzing irreversible reactions in
the glycolysis pathway. Those are PFKL which catalyzes D fructose-6-phosphate to D
fructose-1, 6- bisphosphate conversion, and PKM2 which catalyzes transfer of a phosphoryl
group from phosphoenolpyruvate to adenosine diphosphate (ADP) to generate a molecule of
ATP and pyruvate at the end of the pathway. (Murthy et al., 2015)

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4. DISCUSSION
Studies on the human aqueous humor and it's proteomics have been done by several
research groups (Richardson et al, 2015; Chowdhury et al.,2010).
In 1998, Rohde et al. (1998) tried to identify lower abundance human aqueous humor
proteins by using membrane preconcentration together with capillary electrophoresis and
mass spectrometry (mPC-CE-MS). In 2005, Funding et al. (2005) used two-dimensional
electrophoresis (2-DE) to analyze human aqueous humor proteins and image analysis to find
differentially expressed human aqueous humor proteins in patients with acute corneal
rejection. By using nanoLC-MS/MS and MALDI-TOF MS they identified six proteins with
high significance. In 2007, Stastna et al. (2007) used an combination of 1-DE, 2-DE, and
reversed phase liquid chromatography (RPLC) to separate proteins and MALDITOF MS and
LC-MS/MS to identify human aqueous humor proteins.
Identifying of the protein composition of human aqueous humor showed molecules involved
in keeping a homeostatic environment for front chamber tissues. Changes in protein or ionic
amounts in the human aqueous humor can have significant risks for the cellular function and
cellmatrix communication. However, only approximately 676 proteins have been identified
in this time in human aqueous humor. (Chowdhury et al., 2010)
The present study was conducted to analyze the proteome of human aqueous humor. Using
an approach that included different proteomic techniques and several comparisons, Murthy et
al. (2015) have identified 763 proteins in human aqueous humor. From this number, 386 have
been uniquely identified, while other have been previously reported.
Aqueous humor keeps a normal homeostatic environment and is crucial to the proper
functioning of front chamber tissues. Because of that, it is not surprising that a big number of
the proteins identified by LC-MS/MS have catalytic and enzymatic functions. (Chowdhury et

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al., 2010). There is also a high amount of vasculogenic proteins and immunomodulators
which were identified in this study. This suggests the role of those proteins in balancing
between angiogenic and anti-angiogenic factors. The proteomic profiling of normal aqueous
humor in this study and an in-depth analysis using LC-MS/MS can serve as a template for
ongoing studies.

5. CONCLUSION
The exact number of human aqueous humor proteins is still unknown, and it may be
possible that there are tens if not hundreds of proteins in the human aqueous humor and many
of these could be under the nowdays detection limits. Muthy et al. (2015) managed to prove
the existance of 763 proteins in the human aqueous humor, which is the highest number of
proteins proven. This study provided a deep insight into the proteomics of the human aqueous
humor through applying various prefractionation techniques and mass spectrometry to
identify proteins and enzymes, and extensively characterize the human aqueous humor
proteome.
Although the study was thorough, 763 proteins are probably only a fraction of the whole
protein profile in the human aqueous humor. Nevertheless, the study provides a
comprehensive number of the human aqueous proteins. Still, there are some lacks in this
study in presenting the results, such as missing separate tables of previously identified and
uniquely identified proteins, or an explanation why they mentioned just some of the proteins
and considered them interesting and other not. The list of the 763 proteins provided by
Murthy et al. (2015) may serve as a reference to look for differences in protein expression in
many pathologic conditions of the fornt chamber and the eye with the possibility of
identifying novel biomarkers for diseases and novel therapeutic treatments.

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6. References
1. Abu El-Asrar, A., Imtiaz Nawaz, M., Kangave, D., Siddiquei, M., & Geboes, K.
(2012). Osteopontin and Other Regulators of Angiogenesis and Fibrogenesis in the
Vitreous from Patients with Proliferative Vitreoretinal Disorders. Mediators Of
Inflammation, 2012, 1-8. http://dx.doi.org/10.1155/2012/493043
2. Anshu Arundhati, Marianne O. Price, Matthew R. Richardson, Zaneer M. Segu,
Xianyin Lai, Mervin C. Yoder and Francis W. Price, Jr. (2011) Alterations in the
aqueous humor proteome in patients with a glaucoma shunt device. Molecular Vision,
17(2), 18911900.
3. Bennett KL, et al. 2011. Proteomic analysis of human cataract aqueous humour:
comparison of one-dimensional gel LCMS with two-dimensional LCMS of unlabelled
and iTRAQ(R)-labelled specimens. J. Proteomics 74:151166
4. Brubaker RF. (1991). Flow of aqueous humor in humans [The Friedenwald Lecture].
Invest Ophthalmol Vis Sci 32, 3145 3166.
5. Chowdhury, U., Madden, B., Charlesworth, M., & Fautsch, M. (2010). Proteome
Analysis of Human Aqueous Humor. Investigative Opthalmology & Visual Science,
51(10), 4921. http://dx.doi.org/10.1167/iovs.10-5531
6. Civan, Mortimer. (2008). The eye's aqueous humor. Current Topics in Membranes.
62(2): 449-453
7. Cole DF. 1974. Comparative aspects of the intraocular fluids. In: Davson H, Graham
LT Jr, editors. eds. The Eye. New York: Academic Press; 71121
8. De Berardinis, E., Tieri, O., Polzella, A., & Iuglio, N. (1965). The chemical
composition of the human aqueous humour in normal and pathological conditions.
Experimental Eye Research, 4(3), 179-186. http://dx.doi.org/10.1016/s00144835(65)80030-6
9. Duan X, Lu Q, Xue P, Zhang H, Dong Z, Yang F, Wang N. Proteomic analysis of
aqueous humor from patients with myopia. (2008). Mol Vis; 14:370-7
10. Freddo TF, Bartels SP, Barsotti MF, and Kamm RD. (1990). The source of proteins in
the aqueous humor of the normal rabbit. Invest Ophthalmol Vis Sci 31, 125137.
11. Funding M Vorum H Honore B Nexo E Ehlers N . (2005). Proteomic analysis of
aqueous humour from patients with acute corneal rejection. Acta Ophthalmol
Scand.;83:3139.

21 | P a g e

12. Healthline.com,. (2015). Eye Vitreous & Aqueous Humor Diagram & Function | Body
Maps. Retrieved 7 January 2016, from http://www.healthline.com/human-bodymaps/eye-vitreous-and-aqueous-humor
13. Johnson Mark, Erickson Kristine. (2010). Mechanisms and Routes of Aquous Humor
Drainage. Aqueous Humor and the Dynamics of its Flow. CHAPTER 191, 2577-2595
14. Lamb D.Trevor. (2011). Evolution of the Eye. Scientific American, 1(4), 351516
15. McBride Dyan. (2010). The Human Eye and Vision. Modern Miracle Medical
machines. Kansas State University Physics Education Research.
16. MBBS DOCTORS,. (2016). MBBS DOCTORS: Glaucoma- What is Glaucoma.
Retrieved 7 January 2016, from http://medicforyou.blogspot.ba/2015/09/what-isglaucoma-glaucome-glucoma.html
17. McCaa S. Connie. (1982). The Eye and Visual Nervous System: Anatomy, Physiology
and Toxicology. Environmental Health Perspectives. Vol 4, 1-8
18. Murthy Krishna R., Rajagopalan Pavithra, Pinto Sneha M., Advani Jayshree, Murthy
Praveen R., Goel Renu, Subbannayya Yashwanth, Balakrishnan Lavanya, Dash
Mahashweta, Anil Abhijith K., Manda Srikanth S., Nirujogi Raja Sekhar, Kelkar
Dhanashree S., Sathe Gajanan J., Dey Gourav, Chatterjee Aditi, Gowda Harsha,
Chakravarti Shukti, Shankar Subramanian, Sahasrabuddhe Nandini A., Nair Bipin,
Somani Babu Lal, Prasad T. S. Keshava, and Pandey Akhilesh. (2015). Proteomics of
Human Aqueous Humor. OMICS: A Journal Of Integrative Biology, 19(5), 283-93.
http://dx.doi.org/10.1089/omi.2015.0029.
19. Pollreisz Andreas, Marion Funk, Florian P Breitwieser, Katja Parapatics, Stefan Sacu,
Michael Georgopoulos, Roman Dunavoelgyi, Gerhard J Zlabinger, Jacques Colinge,
Keiryn L Bennett, Ursula Schmidt-Erfurth 2013.Quantitative proteomics of aqueous
and vitreous fluid from patients with idiopathic epiretinal membranes. Experimental
Eye Research, vol. 108, pp. 4858
20. Richardson MR, Price MO, Price FW, Pardo JC, Grandin JC, You J, Wang M, Yoder
MC. (2009). Proteomic analysis of human aqueous humor using multidimensional
protein identification technology. Mol Vis.;15:27402750
21. Rohde E, Tomlinson AJ, Johnson DH, Naylor S. Comparison of protein mixtures in
aqueous humor by membrane preconcentration - capillary electrophoresis - mass
spectrometry. (1998). Electrophoresis; 19:2361-70.
22. Stastna M Behrens A Noguera G Herretes S McDonnell P Van Eyk JE . (2007).
Proteomics of the aqueous humor in healthy New Zealand rabbits.
Proteomics.;7:43584375.
23. Taube, Amelie Botling, Wetterhall Magnus, Artemenko Konstantin, Andersson Marit,
Bergquist Jonas . (2013). Proteins in aqueous humor from cataract patients with and
without pseudoexfoliation syndrome. European journal of mass spectrometry, 18(6):
531-541

22 | P a g e

24. Yoshida Y, Yamagishi S, Matsui T, Nakamura K, Imaizumi T, Yoshimura K,

Yamakawa R. (2007). Positive correlation of pigment epithelium-derived factor and
total antioxidant capacity in aqueous humour of patients with uveitis and proliferative
diabetic retinopathy. Br J Ophthalmol; 91:1133-4
25. Wikipedia,. (2016). Aqueous humour. Retrieved 7 January 2016, from
https://en.wikipedia.org/wiki/Aqueous_humour#/media/File:Schematic_diagram_of_t
he_human_eye_en.svg

7. Supplemetary Material
Supplementary material available in printed version.

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