A Novel Rat Model with Obesity-Associated Retinal

Degeneration
1. Geereddy Bhanuprakash Reddy * , 1 , 2 , 3 ,
2. Vidyullatha Vasireddy 2 , 3 , 4 ,
3. Md Nawajes A. Mandal 2 , 5 ,
4. Mrudula Tiruvalluru 1 ,
5. Xiaofei F. Wang 6 ,
6. Monica M. Jablonski 6 ,
7. Giridharan Nappanveettil 1 and
8. Radha Ayyagari * , 2 , 4

+ Author Affiliations

1. From the National Institute of Nutrition, Hyderabad, India;

1

2. Department of Ophthalmology and Visual Sciences, University of Michigan, Ann Arbor,

2

Michigan; and
3. 6Department of Ophthalmology, Hamilton Eye Center, University of Tennessee, Knoxville,
Tennessee.
4. 4Present affiliation: Department of Ophthalmology, University of California at San Diego, La
Jolla, California; and
5. 5Department of Ophthalmology, Dean A. McGee Eye Institute, University of Oklahoma Health
Sciences Center, Oklahoma City, Oklahoma.
 
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Abstract
PURPOSE. A strong association between retinal degeneration and obesity has been shown in humans.
However, the molecular basis of increased risk for retinal degeneration in obesity is unknown. Thus,
an animal model with obesity and retinal degeneration would greatly aid the understanding of
obesity-associated retinal degeneration. The retinal abnormalities in a novel rat model (WNIN-Ob)
with spontaneously developed obesity are described.

METHODS. Histologic and immunohistochemical examination were performed on retinal sections of 2-

to 12-month-old WNIN-Ob rats, and findings were compared with those of lean littermate controls.
RNA from retinas of 12-month-old WNIN-Ob and lean littermate rats was used for microarray and
qRT-PCR analysis.

RESULTS. The WNIN-Ob rats developed severe obesity, with an onset at approximately 35 days.
Evaluation of retinal morphology in 2- to 12-month-old WNIN-Ob and age-matched lean littermate
controls revealed progressive retinal degeneration, with an onset between 4 to 6 months of age.
Immunohistochemical analysis with anti–rhodopsin, anti–cone opsin, and PSD-95 antibodies further
confirmed retinal degeneration, particularly rod cell loss and thinner outer plexiform layer, in the
obese rat retina. Gene expression by microarray analysis and qRT-PCR established activation of
stress response, tissue remodeling, impaired phototransduction, and photoreceptor degeneration in
WNIN-Ob rat retina.

CONCLUSIONS. WNIN-Ob rats develop increased stress in retinal tissue and progressive retinal
degeneration after the onset of severe obesity. The WNIN-Ob rat is the first rat model to develop
retinal degeneration after the onset of obesity. This novel rat model may be a valuable tool for
investigating retinal degeneration associated with obesity in humans.

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Introduction
The problems of excess weight and obesity have been recognized globally both in developed and in
developing countries.1 2 3 4 5
Excess weight is the second leading modifiable risk factor for death in
the United States, 3 6
where 64% of adults age 20 years and older are overweight and 30% are obese.
In the past 20 years, the rates of obesity have tripled in developing countries. 6 7
Eye problems are
probably the latest addition to the list of complications associated with obesity. 1 8
The ocular
complications of obesity include diabetic retinopathy, high intraocular pressure, cataracts, macular
degeneration, floppy lid syndrome, and exophthalmos. 1 8
Although these complications can have
serious consequences, the effects of overweight and obesity on the eye are not well known to
patients or to health care providers.

There is evidence of the increased risk for retinal degeneration and diabetic retinopathy in obese
persons. Obese men are more than twice as likely to have dry macular degeneration than men with a
normal basal metabolic index.9 An association between higher basal metabolic index and early
macular degeneration has been reported.10 Similarly, abdominal obesity in patients with early or
intermediate stages of macular degeneration increases the risk for progression to advanced macular
degeneration.11 Reduction in the macular pigments, lutein, and zeaxanthin in obese persons further
supports the association between obesity and retinal degeneration. 12 Patients with Bardet-Biedl
syndrome (BBS) develop retinal dystrophy, obesity, polydactyly, renal malformation, and learning
disabilities.13 14
In several populations, obesity is one of the basic components of metabolic
syndrome, which is implicated in microvascular changes in the retina. 15

Retinal degeneration, including age-related macular degeneration (AMD) and diabetic retinopathy
(DR), is a major cause of irreversible blindness in the developed and the developing world. 16 17
The
alarming increase in the prevalence of obesity further exacerbates the concern about retinal
degeneration. In the past 20 years, considerable progress has been made in our understanding of
the molecular basis of retinal dystrophies. However, there is no experimental evidence to establish
the mechanism by which obesity increases the risk for retinal dystrophies. Although retinal
abnormalities are found in tubby mouse and mouse models of BBS, 18 19 20
obesity develops along with
or much later than retinal changes. Therefore, an animal model that develops retinal degeneration
after the onset of obesity, similar to obesity-associated retinopathy in patients, would immensely aid
our basic understanding of the relationship between retinal degeneration and obesity.

Wistar is the oldest strain of rats used in biomedical research. The National Institute of Nutrition
(NIN) in India has an inbred stock of Wistar rats dating back to 1920 that is christened WNIN (Wistar
maintained at NIN).21 A spontaneously developed obese rat was isolated from WNIN rats, and a
colony of WNIN-Obese (WNIN-Ob) rats was generated by selective breeding. These rats (Fig. 1(i)) are
maintained at the National Center for Laboratory Animal Sciences at NIN. 21 22 23
The inheritance
pattern and the biochemical and phenotypic characteristics of obesity in WNIN-Ob rats have been
characterized in detail. 21 22 23
Starting at 35 to 40 days of age, rats of the WNIN-Ob phenotype are
different from their lean littermates in terms of body weight. Their body weight increased
progressively until the age of 12 months (Fig. 1 (ii)). By this age they weigh as much as 1.2 kg, in
contrast to their lean littermates that weigh 500 to 600 g. WNIN-Ob rats demonstrate low fertility,
and their average lifespan is 20 to 24 months. The colony is maintained by mating carriers (+/−),
which, on crossing, gives three genotypes: lean (+/+), carrier (+/−), and WNIN-Ob (−/−)—in a
classical Mendelian ratio of 1:2:1, respectively. 21 22 23
The present study describes retinal
degeneration in this spontaneous obese rodent model.

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FIGURE 1.

(i) WNIN-Ob rat in comparison with its littermate lean control at the age of 12 months. (A) WNIN-Ob
rat. (B) Lean control. (ii) Relative increase in body weights of WNIN-Ob rat compared with lean
control and Wistar controls. Values represent mean ± SD of 12 animals in each group.

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Materials and Methods
Reagents and Antibodies

The details of antibodies used for this study are as follows: rabbit-anti–opsin, red/green (1:250
dilution; Chemicon, Temecula CA), mouse monoclonal anti–rhodopsin (1:200 dilution; Chemicon),
mouse postsynaptic density protein-95 (PSD-95; 1:250 dilution; Sigma-Aldrich, St. Louis, MO), anti–
rabbit Alexa Fluor-555 (1:2500 dilution; Invitrogen-Molecular Probes, Carlsbad, CA), and anti–rabbit
Alexa Fluor-488 (1:1000 dilution; Invitrogen-Molecular Probes).

Animals and Tissue Collection

All procedures involving rats were performed in accordance with the ARVO Statement for the Use of
Animals in Ophthalmic and Vision Research and were approved by the Institutional Animal Ethics
Committee at the National Institute of Nutrition. Animals were kept in a 12-hour light/12-hour dark
cycle with ambient light intensity and temperature at the National Center for Laboratory Animal
Science, National Institute of Nutrition. Two- to 12-month-old WNIN-Ob rats and their lean
littermates were fasted overnight before they were euthanatized at the end of the dark cycle. All
studies described here were carried out by evaluating at least three animals.

Histology and Immunohistochemistry

Eyeballs were collected from 2- to 12-month-old animals and fixed in 4% paraformaldehyde in

phosphate-buffer (pH 7.2), followed by embedding and sectioning for plastic and cryosections using
standard protocols, as described previously. 24 25 26 27
Three eyes were used for each age group. Plastic
sections were stained with hematoxylin and eosin. Immunohistochemistry was performed on
cryosections using respective primary antibody, as described previously. 24 25 26 27
The slides were
mounted in antifade reagent containing DAPI (ProlongGold; Invitrogen) and were observed under an
epifluorescence microscope (E800; Nikon, Tokyo, Japan). Images were captured using appropriate
filters. Higher magnification images were captured with a confocal microscope (Zeiss, Oberkochen,
Germany).

RNA Preparation

Retinas dissected from the eyes of WNIN-Ob rats and age-matched littermate controls were collected
in reagent (Trizol; Invitrogen) for RNA isolation. RNA was isolated according to the manufacturer’s
instructions.

Global Gene Expression by Microarray Analysis

Five micrograms of total retinal RNA isolated from two pooled retinas was used to synthesize
double-stranded cDNA with the use of first- and second-strand cDNA synthesis kits (Affymetrix,
Santa Clara, CA) according to the manufacturer’s guidelines. Purification of ds-cDNA, labeling,
hybridization, washing, and staining were carried out according to the Affymetrix protocol. The rat
gene expression chip (230.20; Affymetrix) was used for hybridization. Three independent samples
for each WNIN-Ob and control group were analyzed.

Gene chips were scanned (Microarray Suite 5.0 [Affymetrix] operated by GCOS software version 2.0),
and expression data were normalized with the use of robust multiarray average. 28 Data were based
on three gene chips per each control and WNIN-Ob groups. Ratios of average signal intensity (log 2-
transformed data) were then calculated for the probe sets (WNIN-Ob relative to lean control) and
were converted to fold change. 28 The contrast between experimental and control was extracted
selecting those genes with an adjusted P of 0.05 and a threefold difference.

Quantitative Real-Time PCR

To remove any trace of genomic DNA, RNA was treated with RNase-free DNase (RQ1; Promega,
Madison, WI) according to the manufacturer’s protocol. Equal amounts of RNA (7 μg) were reverse
transcribed for each sample (control and WNIN-Ob) using oligo-dT (12–18) primer to first-stand
cDNA with reverse transcriptase (SS II; Invitrogen) according to the manufacturer’s
recommendations. Quantitative real-time PCR was then performed on cDNA templates using gene
specific primers, as described previously. 24 27
Specificity of primers was confirmed by melt curve
analysis and by gel electrophoresis. Primer sequences are available from the authors on request. PCR
data were analyzed to estimate the relative expression of different genes based on the difference in
threshold cycle (Ct) between control and WNIN-Ob groups after normalization to various
housekeeping genes—hypoxanthine phosphoribosyltransferase ( Hgprt), β-actin, succinate
dehydrogenase (SdhA), and ribosomal protein L19 (Rpl19)—as described.24 25 26 27
Mean (±SD) values
of percentage expression over control (lean) calculated from at least three independent samples are
presented.

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Results
Morphologic Evaluation

We have evaluated the retinal morphology of 2- to 12-month-old WNIN-Ob rats and compared our
findings with those of lean littermate controls. No significant changes were observed in the
morphology of 2-month-old WNIN-Ob animals compared with controls (Figs. 2(i) 2A 2B) , whereas
the retinas of 4- to 12-month-old WNIN-Ob rats had significant alterations. At 4 months, the
thickness of the outer nuclear layer (ONL) in the central retina of WNIN-Ob rats was reduced to
approximately 7 to 8 nuclei compared with 10 to 12 nuclei in lean littermate controls (Figs. 2(i) 2C
2D) . The retinal changes found in WNIN-Ob rat at 6 months were not much different from the
changes seen at 4 months age (data not shown). By 9 to 12 months, ONL thickness was reduced to 4
to 6 rows of nuclei in WNIN-Ob retinas compared with 10 to 12 nuclei in age-matched lean
littermate controls (Figs. 2(i) 2E 2F) . Evaluation of retinal cell loss in the central and peripheral
regions of 2- to 12-month-old WNIN-Ob and control rat retina demonstrated progressive
degeneration of the photoreceptor cells in the central retina. In peripheral retina, a concomitant
decrease occurred in the number of photoreceptor nuclei (Figs. 2(ii) 2(iii)) .

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FIGURE 2.

Representative histology of control rat retina (A, C, E) and WNIN-Ob rat retina (B, D, F) at various
ages: (A) 2-month-old control; (B) 2-month-old WNIN-Ob; (C) 4-month-old control; (D) 4-month-
old WNIN-Ob; (E) 12-month-old control; (F) 12-month-old WNIN-Ob. Histograms summarizing the
progression of retinal degeneration in central (ii) and peripheral (iii) retina of WNIN-Ob rats
compared with lean controls. Values represent mean ± SD of at least seven independent
observations. OS, outer segments; IS, inner segments; INL, inner nuclear layer; IPL, inner plexiform
layer; GC, ganglion cell layer.

Further histologic evaluation of retinas at age 9 to 12 months demonstrated abnormalities in the

inner retinal layers. The gross structure of the inner retina did not demonstrate significant
abnormalities at ages 4 to 6 months, but the thickness of the outer plexiform layer (OPL) and inner
plexiform layer appeared to decrease in the 9- to 12-month-old WNIN-Ob rat retina compared with
controls (Figs. 2(i) 2E 2F) . Concomitant with the decrease in the thickness of the OPL, a few
photoreceptor nuclei appeared to migrate into this layer, suggesting disruption of its structure. By
morphologic evaluation, the onset of retinal degeneration in the WNIN-Ob rat appeared to occur
between 4 and 6 months age, and variation was observed in the severity of degeneration at a given
age.

In addition to analyzing WNIN-Ob rats and lean littermate controls, we evaluated the retinal
histology of 3- to 12-month-old WNIN parent strain rats. These rats did not show significant
changes in retinal morphology (data not shown), suggesting that the retinal degeneration observed
in the WNIN-Ob rat is neither sporadic nor a nonspecific observation in this strain of rats.

Immunohistochemistry

Immunohistochemical analysis of WNIN-Ob and lean littermate control rat retinas was carried out
using rod and cone photoreceptor specific marker antibodies and anti–PSD-95 antibodies.
Immunostaining with rhodopsin antibodies showed reduction in the brightness of the rhodopsin-
specific signal in the WNIN-Ob rat retina compared with that of lean controls at 9 months (data not
shown) and 12 months (Figs. 3A 3B 3C 3D) . Immunostaining with cone opsin (MW) antibodies
indicated no significant alteration in the number of cones in the WNIN-Ob rat retina (Figs. 3E 3F 3G
3H) ; staining the retinal sections with shorter wavelength cone opsin (SW) gave similar results (data
not shown). The number of cones in the retinas of WNIN-Ob and lean control rats was found to be
similar at ages 9 to 12 months (Fig. 3(ii)) .
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FIGURE 3.

Immunohistochemical evaluation of rod and cone photoreceptor markers in WNIN-Ob rat compared
with controls at 12 months. (i) Retinal sections of (A, C, E, G) control and (B, D, F, H) WNIN-Ob rats
were labeled with (A–D) rhodopsin and (E–H) cone opsin. Nuclei are labeled with DAPI. Scale bar, 50
μm. (ii) Histogram demonstrates the number of cones in WNIN-Ob and control rat retinas. Values
represent mean ± SD of three independent observations (P = 0.2).

To further evaluate the decrease observed histologically in the thickness of OPL, we labeled the
retinal sections from WNIN-Ob and control rats with postsynaptic density protein-95 (PSD-95) and
compared the immunofluorescence (Fig. 4) . In the 12-month-old WNIN-Ob rat retina, the
immunofluorescence of PSD-95 was weaker (Figs. 4C 4D) than the PSD-95 signal observed in age-
matched lean littermates (Figs. 4A 4B) . This indicated a loss of synaptic terminals and further
supported the observed reduction in the thickness of OPL in WNIN-Ob rat retina.

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FIGURE 4.

Immunofluorescence images of 12-month-old control (A, B) and WNIN-Ob rat (C, D) retinal sections
labeled with PSD-95. Labeling of PSD-95 is prominent and intense in OPL layer of controls, whereas
in the WNIN-Ob rat the fluorescence signal of PSD-95 is less intense.

Differential Expression of Genes in WNIN-Ob Retina

Having observed retinal degeneration in the WNIN-Ob rat, microarray analysis was performed on the
WNIN-Ob rat retina at age 12 months to assess the global gene expression profile of the WNIN-Ob
rat retina under degenerative conditions. The rat gene expression chip (230.2.0; Affymetrix) used in
this experiment contains 30,000 probes representing 26,000 genes. Based on absent and present
calls, approximately 60% of the probe sets were reported as present. We have calculated the
expression values (log2 transformed) for each gene using robust multiarray average. Principal
component analysis confirmed that biological replicate samples were grouped together. After gene
filtering, 13,570 genes were identified by removing any that did not appear to be differentially
expressed in any of the three independent samples in each group. We then extracted the contrast of
experimental versus control, selecting those genes with an adjusted P of 0.05 and a threefold
difference. This resulted in 423 genes, of which 369 showed lower levels of expression, whereas the
expression of 54 genes was found to be significantly higher than in controls. Most downregulated
genes were involved in phototransduction pathways or related to retinal structural proteins, carrier
proteins, or transcription factors (Supplementary Table S1; both Supplementary Tables are online at
http://www.iovs.org/cgi/content/full/50/7/3456/DC1). Most upregulated genes were related to
apoptotic stress response and inflammatory mechanisms (Supplementary Table S2).

Quantitative real-time PCR analysis was carried out to validate the observations by microarray
analysis and to verify the changes observed by histologic and immunohistochemical examination by
quantifying the expression of selected retinal genes in 12-month-old lean and WNIN-Ob rats.
Expression of rod-specific genes such as rhodopsin ( Rho), rod transducin (Gnat1), rod arrestin (Sag),
peripherin (Rds), retinal outer segment membrane protein (Rom1), and cGMP-dependent
phosphodiesterase 6B (Pde6B) was found to be significantly lower in WNIN-Ob rat retina than in lean
controls (Fig. 5) . Levels of expression of the retinal transcription factor, cone-rod homeobox gene
(Crx), and elongation of very long chain fatty acids 4 ( Elovl4) genes expressed in rods and cones also
decreased significantly in 12-month-old WNIN-Ob rats (Fig. 6) . Obesity-associated genes tubby and
Bbs7 also showed a lower level of expression in the retinas of WNIN-Ob rats than in those of
littermate controls (Fig. 6) , but the decrease was not significant.

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FIGURE 5.

Quantitative expression of rod-specific genes in 12-month-old WNIN-Ob rats and their lean
littermate controls. Expression values were determined by qRT-PCR and presented on an arbitrary
scale (y-axis) after normalization with rat Hgprt. Data represent the mean (±SD) on an arbitrary scale
and were calculated from at least three independent observations.

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FIGURE 6.

Expression of Crx, Elovl4, and obesity-related genes in the retinas of 12-month-old WNIN-Ob rats
and their lean littermate controls. Expression values were determined by qRT-PCR and presented on
an arbitrary scale (y-axis) after normalization with rat Hgprt. Data represent the mean (±SD) on an
arbitrary scale and were calculated from at least three independent observations.

Interestingly, expression of cone cell marker genes, SW cone opsin ( Opn1SW), MW cone opsin
(Opn1MW), cone arrestin (Arr3), and cone transducin ( Gnat2) was unaltered or significantly increased
(Fig. 7) . Expression of stress response genes, glial fibrillary acidic protein ( Gfap), annexin-1,
ceruloplasmin (Cp), hemeoxygenase-1 (Ho-1), and vascular endothelial growth factor (Vegf) were
significantly higher in 12-month-old WNIN-Ob rat retinas than in those of lean controls (Fig. 8) .
These results indicate the preservation of cone cells, degeneration of rod photoreceptors, activation
of stress response, and remodeling of tissue pathways in the WNIN-Ob rat retina at 12 months.
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FIGURE 7.

Quantitative expression of cone-specific genes in 12-month-old WNIN-Ob rats and their lean
littermate controls. Expression values were determined by qRT-PCR and presented on an arbitrary
scale (y-axis) after normalization with rat Hgprt. Data represent the mean (±SD) on an arbitrary scale
and were calculated from at least three independent observations.

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FIGURE 8.

Quantitative expression of selected stress-related genes in retinas of 12-month-old WNIN-Ob rats

and their lean littermate controls. Expression values were determined by qRT-PCR and presented on
arbitrary scale (y-axis) after normalization with rat Hgprt. Data represent the mean (±SD) on an
arbitrary scale and were calculated from at least three independent observations.

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Discussion
Retinal degeneration, including age-related macular degeneration (AMD) and diabetic retinopathy
(DR), is a major cause of blindness in the developed and the developing world. The molecular events
that contribute to retinal degeneration are highly complex because of the involvement of different
retinal cell types. Mutations in approximately 150 genes have been implicated in retinal
degeneration (RetNet, www.sph.uth.tmc.edu/Retnet/home.htm). Despite extensive knowledge about
the disease-causing genes,29 30 31 32
the molecular basis underlying many forms of retinal
degeneration are not understood. The interplay between multiple genetic and environmental factors
could add to the complexity for the pathogenesis of retinal dystrophies. A strong association found
between retinal degeneration and obesity further adds to this complexity. The alarming increase in
the prevalence of obesity in Southeast Asian countries 6 7 33 34
further exacerbates concern about
retinal degeneration globally. Therefore, identifying a suitable animal model will aid in investigating
the underlying mechanism for the association between retinal degeneration, obesity, and metabolic
syndrome.

Evaluation of the morphology, photoreceptor-specific protein, and gene expression profile of the
WNIN-Ob rat retina indicated significant and progressive rod photoreceptor degeneration with
preservation of cone cells. The preservation of cone photoreceptors is remarkable in WNIN-Ob rats
even at 12 months, when ONL thickness is reduced to 50% to 60% of controls. Survival of cones
beyond the age of 12 months in these rats is yet to be investigated. In WNIN-Ob rats, preservation of
the number of cone cells is intriguing. In several types of retinal degeneration, in patients and in
animal models, rod cell death precedes cone loss. 35 36
It is likely that the molecular defect underlying
retinal degeneration in these rats is associated with a biological pathway critical for rod cell survival.
Cone viability is believed to depend on diffusible or contact-mediated survival factors that are
synthesized by rods.37 38 39
Therefore, this rat model with unaltered cone cell number, despite
significant rod cell loss, may aid in understanding possible means to preserve cones in the absence
of rods.

Retinal abnormalities were observed in a few rodent models with obesity. The tubby mice develop
neurosensory abnormalities and obesity, but neurosensory abnormalities develop much earlier than
the onset of obesity. The obesity of tubby mice is relatively mild, has late onset, and progresses
slowly.19 20
Mouse models for Bardet-Biedl syndrome develop obesity and retinal abnormalities in
addition to a spectrum of other phenotypic features. 40 41
Among other obese rodent models, ocular
complications, particularly pronounced retinal changes, were found in diabetic obese (fa/fa) rats
when they were fed a sucrose diet for 68 weeks. 42 Mice lacking functional tubby-like protein-1
(TULP-1) develop retinal degeneration but not obesity. 43 The phenotype observed in WNIN-Ob rats is
unique because of the early age of onset and the severity of obesity and retinal degeneration.

The pattern of gene expression in the WNIN-Ob rat retina supports photoreceptor degeneration.
Downregulated genes appear to be involved predominantly in transcriptional regulation ( Crx, neural
fold homeobox, high mobility group box, zinc finger proteins) and visual transduction ( Gnb, Gnat,
Cng, Pde6B, Rho, phosphatases, retinol binding protein). In addition, rod cell structural proteins
(Rho, Rom, peripherin) were also observed to be significantly decreased. A number of these
downregulated genes are also implicated in various forms of inherited retinal degeneration (RetNet,
www.sph.uth.tmc.edu/Retnet/home.htm). Lower levels of expression of rod cell specific genes may
reflect the loss of photoreceptors rather than specific downregulation of these genes.

The upregulated genes are mostly related to apoptotic, stress response, tissue remodeling pathways
(annexin-1, cathepsin, glutathione S-transferase, Cp, Gfap) indicating activation of the stress
response in the WNIN-Ob rat retina compared with the lean littermate control rat retina. The
expression pattern of some genes observed in the WNIN-Ob rat retina is similar to the gene
expression changes observed in rd1 mice and injured rat retina. 44 45
Levels of expression of obesity-
related genes, tubby-like, and Bbs7 were found to be lower on microarray and qRT-PCR analysis,
indicating a possible direct role for these obesity genes in maintaining normal retinal physiology. 41 43

46
Unlike many studies involving microarray analysis, we found that only a limited number of genes
are differentially expressed in the Ob-rat retina compared with lean littermate controls. Most of the
genes showing altered expression are likely to have a significant role in retinal physiology. The
expression profile of genes observed in the WINN-Ob rat retina is similar to the findings reported in
Bbs4 knockout mice.41 These observations suggest a direct association between obesity and retinal
degeneration in these rats.

Further studies are needed to determine the causal relationship between obesity and retinal
degeneration in the WNIN-Ob rat and to determine whether obesity, either in itself or through its
metabolic consequences, alters retinal structure and function in these rats. No reports relate
metabolic changes (similar to those seen in this model, such as hyperphagia, hypertriglyceridemia,
hypercholesterolemia) to retinal changes, particularly photoreceptor degeneration, in a temporal
manner. Nevertheless, studies are under way in our laboratories wherein we are inducing these
metabolic changes in the parent strain of rat, which are not obese, by manipulating the diet. The
effect of these metabolic changes without overt obesity might help us to address their relationship
with retinal degeneration. This animal model may aid in investigating the association between
obesity and retinal degeneration and the possibility of modifying obesity-associated risk by altering
diet. The WNIN-Ob rat may thus be explored in greater detail for its usefulness as a model to study
obesity-associated retinal degeneration. Evaluation of retinal vasculature and lipofuscin content in
WNIN-Ob rats may provide valuable information. Given the impact of retinal dystrophies on
blindness and the increased risk for retinal dystrophy caused by obesity, it is highly desirable that
we understand the molecular basis of obesity-associated retinal degeneration. In this context, the
observations made in this study provided such an opportunity.

Although obesity may be a risk factor for many ocular conditions, including retinal degeneration, the
present literature is inadequate in establishing any convincing association. Nevertheless, the reduced
levels of macular pigments in obese persons 12 47
support the association of increased risk for retinal
degeneration in this population. Reductions in macular lutein and zeaxanthin may be caused by
decreased dietary intake in obese persons or by competition between retina and adipose tissue for
the uptake of lutein and zeaxanthin. 12 47
However, whether weight loss, by physical activities and
dietary manipulations, reduces the risk for eye diseases remains unresolved. Because of the potential
public health impact of obesity, there is a greater need to understand its ocular effects. The WNIN-
Ob rat may aid investigators to address these issues.

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Acknowledgments
The authors thank Boindla Sesikeran (National Institute of Nutrition) for assistance in examining the
histology slides, Krishnakumar Sarada and Pasupuleti Lopamudra (National Institute of Nutrition) for
assistance in preparing histologic specimens, Motha Satyavani (National Institute of Nutrition) for
providing animal maintenance and technical assistance, Austra Liepa (University of Michigan) for
assistance in manuscript preparation, and Mitchell Gillet (University of Michigan) for histologic tissue
preparation.

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Footnotes
  ↵3 These authors contributed equally to the work presented here and should therefore be
regarded as equivalent authors.

 Supported in part by the Indian Council of Medical Research and Department of

Biotechnology, New Delhi; National Institutes of Health Grants EY13198 (RA) and EY015208
(MMJ); National Eye Institute Core Grants EY013080 (University of Tennessee Health Science
Center), P30EY007003 (Department of Ophthalmology, University of Michigan), and
P30EY07060 (Department of Vision Research, University of Michigan); an unrestricted grant
from Research to Prevent Blindness, Inc. (Department of Ophthalmology, University of
Tennessee Health Science Center); Foundation Fighting Blindness (RA); and Research to
Prevent Blindness Inc. (RA, GBR). GBR is the recipient of an International Fellowship from the
Indian Council of Medical Research and an Overseas Associateship (Niche Areas) from the
Department of Biotechnology, New Delhi.

 Submitted for publication June 26, 2008; revised November 24, 2008, and January 26, 2009;
accepted May 15, 2009.

 Disclosure: G.B. Reddy, None; V. Vasireddy, None; M.N.A. Mandal, None; M. Tiruvalluru,
None; X.F. Wang, None; M.M. Jablonski, None; G. Nappanveettil, None; R. Ayyagari, None

 The publication costs of this article were defrayed in part by page charge payment. This
article must therefore be marked “ advertisement” in accordance with 18 U.S.C. §1734 solely to
indicate this fact.

 ↵* Each of the following is a corresponding author: Radha Ayyagari, Department of

Ophthalmology, #206, Jacob’s Retina Center (Shiley Eye Center), University of California at San
Diego, C9415 Campus Point Drive, La Jolla, CA 92093; [email protected]. Geereddy
Bhanuprakash Reddy, National Institute of Nutrition, Hyderabad 500007, India;
[email protected].

 Copyright 2009 The Association for Research in Vision and Ophthalmology, Inc.

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