LABORATORY SCIENCE

Unfolded protein response activation

in cataracts
Beatriz E. Torres-Bernal, MS, Luis Fernando Torres-Bernal, PhD, Rafael R. Guti!errez-Campos, PhD,
David D. Kershenobich Stalnikowitz, PhD, Luis Fernando Barba-Gallardo, PhD, Arturo A. Chayet, MD,
Javier Ventura-Ju!arez, PhD

PURPOSE: To analyze the expression of 78 kDa glucose-regulated protein (GRP78) and activating
transcription factor 6 (ATF6), 2 factors in the unfolded protein response (UPR), in age-related and
diabetes-associated cataract.
SETTING: Universidad Aut!
onoma de Aguascalientes, Aguascalientes, M!exico.
DESIGN: Experimental study.
METHODS: The qualitative and quantitative expression of GRP78 and ATF6 were measured in
surgical samples from 11 senile cataracts, 9 diabetic-associated cataracts, and 3 normal lenses.
Both proteins were detected by immunofluorescence and immunogold-conjugated antibodies.
Quantitative morphometry was used to analyze the differences in GRP78 and ATF6 between
samples. The Mann-Whitney test was used for statistical analysis.
RESULTS: Scanning electron microscopy showed the characteristic organization of fibers in normal
lenses with regular alignment and interdigitation between them. On the other hand, lenses from eyes
with senile or diabetic cataract showed the same pattern of misalignment and disorganization of the
fibers. Both proteins were detected through immunofluorescence in senile and diabetic cataracts,
but not in normal lenses. Immunogold-conjugated antibodies and transmission electron
microscopy showed that GRP78 and ATF6 grains were 30% higher and 35% higher,
respectively, in diabetic cataracts than in senile cataracts (P<.05).
CONCLUSIONS: These data show for the first time in humans that GRP78 and ATF6 are present in
lens fibers of senile cataracts and diabetic cataracts, establishing that the UPR may be important in
the process of cataractogenesis.
Financial Disclosure: No author has a financial or proprietary interest in any material or method
mentioned.
J Cataract Refract Surg 2014; 40:16971705 Q 2014 ASCRS and ESCRS

Cataract is defined as any opacification of the lens that
causes symptoms, including decreased visual acuity,
decreased color perception, decreased contrast sensi-
tivity, and glare disability, which eventually result in
blindness.1 According to the World Health Organi-
zation, 25 million people are affected with cataract,
which is also the leading cause of blindness world-
wide. Age-related cataracts are responsible for nearly
one half of all blindness worldwide.2,3 Cataract is
also a multifactorial disease that has been associated
with systemic diseases such as diabetes, environ-
mental factors, genetics, and other factors such as
smoking4; however, the physiopathology remains
unknown.5
Recently, researchers have sought to determine the
role of progressive oxidative damage, protein
aggregation, and glucose levels. The prevalence of
cataract is 4 to 5 times higher in diabetic patients
than in normal controls; high levels of glucose in the
aqueous humor produce glycosylation of crystalline
proteins, a process that results in the formation of
advanced glycation end products and superoxide rad-
icals. In diabetic lenses, there is an increase in the
activity of aldose reductase, with the subsequent
reduction of glucose to sorbitol. The increased levels
of sorbitol in lens epithelial cells (LECs) cause major
osmotic changes5 that may induce stress in the endo-
plasmic reticulum. This stress in the endoplasmic

Q 2014 ASCRS and ESCRS 0886-3350/$ - see front matter 1697

Published by Elsevier Inc. http://dx.doi.org/10.1016/j.jcrs.2014.02.038
1698 LABORATORY SCIENCE: MOLECULAR BIOLOGY OF CATARACTS
reticulum could start a response to malformed pro-
teins called the unfolded protein response (UPR).6
The UPR is essential to maintain cellular homeo-
stasis,7 and the basic components of this response
are the transducers transmembrane protein
kinaseendoribonuclease (IRE1), activating transcrip-
tion factor 6 (ATF6), and double-stranded RNA acti-
vated protein kinase kinase-like endoplasmic
reticulum (PERK), as well as the master regulator
BiP/GRP78. The first transducer is the IRE1, which
has 2 isoforms in mammals (IRE1-a and IRE1-b).
This protein has an N terminal domain of endoplasmic
reticulum stress sensor within the lumen of the organ-
elle and a C-terminal domain with endoribonuclease
activity in the cytosol. The second transducer, ATF6,
is a transcription factor with an N terminal b-ZIP
(basic leucine zipper) in the cytosol and a C terminal
domain stress censor in the endoplasmic reticulum.
In addition, PERK contains a domain stress sensor of
endoplasmic reticulum and a cytosolic domain that
phosphorylates eIF2a. The master controller in all
this response is the endoplasmic reticulum chaperone
protein, GRP78, which in nonstress conditions binds
and inactivates the 3 transducers, keeping them within
the endoplasmic reticulum.8 Several stimuli within a
cell, such as osmotic changes and oxidation, may
converge in cellular stress. The accumulation of mal-
formed proteins in endoplasmic reticulum formed by
several stimuli may activate the UPR.
The UPR response has 2 main steps. The first
begins when the accumulation of malformed proteins
in endoplasmic reticulum activate 78 kDa glucose-
regulated protein (GRP78), separating it from the
Submitted: October 18, 2013.
Final revision submitted: February 10, 2014.
Accepted: February 22, 2014.
From Inova Vision Quirurgica (B.E. Torres-Bernal, L.F. Torres-
Bernal), the Departamento de Qu!mica (Guti!errez-Campos) and
the Departamento de Morfolog!a (Ventura-Ju!arez), Centro de
Ciencias B!asicas, the Departamento de Optometr!a (Barba-Gallardo)
and Departamento de Cirurgia (L.F. Torres-Bernal), Centro de Cien-
cias de la Salud, Universidad Aut!onoma de Aguascalientes, Aguasca-
lientes, the Instituto Nacional de Ciencias M!edicas y de Nutrici! on
Salvador Zubir!an (Kershenobich Stalnikowitz), Mexico City, and
CODET (Chayet), Vision Institute, Tijuana, Baja California, Mexico.
Ruth I. Solis-Arias, Inova Vision Quirurgica, Aguascalientes,
Mexico, corrected the English translation of this paper.
Corresponding author: Javier Ventura-Ju!arez, PhD, Departamento
de Morfolog!a, Centro de Ciencias B!asicas, Universidad Aut!
onoma
de Aguascalientes, Avenida Universidad 940, Colonia Ciudad Uni-
versitaria, Edificio 202, Aguascalientes 20131, Mexico. E-mail:
jventur@correo.uaa.mx.
J CATARACT REFRACT SURG - VOL 40, OCTOBER 2014
UPR transducers. This results in the activation of the
XBP1 transcription factor, which acts as a potent
activator of the transcription of various genes in
UPR. These genes share within their promoter a
sequence consensus known as the endoplasmic reticu-
lum stress response element.911 All this leads to the
transcription of chaperones and proteins whose
function is essential to maintain the homeostasis of
endoplasmic reticulum. The aim of this study was to
analyze the expression of GRP78 and ATF6 in senile
cataracts and diabetic cataracts.
MATERIALS AND METHODS
This prospective observational comparative trial included
tissue samples of lenses obtained from healthy donors
postmortem as well as lenses from patients with senile or
diabetes-associated cataract who had extracapsular cataract
extraction (ECCE) at the Department of Anterior Segment,
Inova Vision Quirurgica. The study was approved by the
Institutional Bioethics Committee, Autonomous University
of Aguascalientes (CIB-UAA-02). Lenses were obtained
from diabetic patients and nondiabetic patients who met
inclusion and exclusion criteria. All patients provided writ-
ten consent after receiving a detailed explanation of the
procedure and the possible risks. The normal lenses were
obtained postmortem from healthy donors through the Eye
Bank of Aguascalientes and were used with the relatives'
permission. Table 1 shows the epidemiological data for the
surgical samples.
Tissue Processing
After ECCE, the cataract nucleus was divided into
quadrants; each one was placed immediately in parafor-
maldehyde 4.0% and glutaraldehyde 3.0% solution in
phosphate-buffered saline (PBS). Analysis was performed
using 2 techniques; that is, immunofluorescence in paraffin
sections (sensitivity in microns) and immunogold conjuga-
tion through inclusion of samples in acrylic resin (LR-White,
London Resin Products) (sensitivity in nanometers)12 to
localize and quantify the presence of GRP78 and ATF6 in
the samples.13
Immersion of Sample in Paraffin
Once the lens tissue was fixed, it was washed with water
for 30 minutes and processed in histoquinet. The first drying
step was performed with ascending alcohol concentrations,
with each step lasting approximately 1 hour. Next, the
xylene was clarified in 2 steps, each lasting 1 hour. Finally,
the tissue was blocked by embedding it in paraffin in 2 steps
of 2 hours each. The slides were prepared with a microtome.
They were 5 mm thick and mounted on slides pretreated with
3-aminopropyltriethoxysilane (Sigma-Aldrich Co.)
Paraffin Slides Immunofluorescence
The paraffin was removed from slides by placing them in
an oven at 56! C for approximately 2 hours and then in a
xylene concentrate in a series of ethyl alcohol concentrations
ranging from 70% to 100%. They were rinsed immediately in
 LABORATORY SCIENCE: MOLECULAR BIOLOGY OF CATARACTS
Table 1. Epidemiologic data.
Senile Diabetic Normal
Parameter Cataracts Cataracts Lenses
Cases (n) 11 9 3
Sex
Male 6 5 1
Female 5 4 2
Mean age (y) 78 62 33
Cataract type Nuclear Nuclear NA
Cataract grade 4 4 NA
Medication Captopril Metformin NA
glibenclamide
Diabetes NA 11 NA
duration (y)
NA Z not applicable
running water for 5 minutes. The slides were placed for 2
minutes in 2 L of sodium citrate buffer pH 6.0 (buffer condi-
tions) and then passed to the PBS 1X pH 7.4. For the perme-
ation of the membrane, the slides were placed in a solution of
methanol and hydrogen peroxide and were again washed
with PBS 1X. They were left for 30 minutes in PBS 0.2%
Triton X-100 and then incubated for 1 hour at room temper-
ature with fetal bovine serum 20.0% in PBSTriton X-100
0.2% to avoid intersections to diminish crossing among the
proteins. The slides were then placed in a humid chamber
with the first antibody to incubate overnight. The antibody
comprised rabbit polyclonal antibody anti-GRP78/BiP
(AB21685) and ATF6 (AB21685) (Abcam) in concentrations
of 1:400 and 1:600, respectively, which dissolves when
placed in PBSTriton X-100 0.2%bovine serum albumin
(BSA) 3.0%. The following day, the slides were washed
with PBSTriton X-100 and incubated with the secondary
antibody bound to diluted chromogen (Alexa fluor 488
goat secondary antibody labeled, Molecular Probes) for 1
hour at room temperature. They were washed with PBS,
and staining of the nuclei was performed for 10 minutes
with Hoechst dye (1:1000 in PBS 1X, 33258 Sigma Aldrich).
They were washed again with PBS and finally covered
with an aqueous histologic mounting medium (Glycergel,
Dako), with care taken not to leave bubbles.
Inclusion of Samples in Acrylic Resin
Once attached, the sample was washed with PBS and
post-fixed with osmium tetroxide 1.0% dissolved in calcium
chloride 1.0%. The samples were placed in this solution for
2 to 4 hours for cooling. After 2 hours, they were washed 3
times for 10 minutes with PBS. Next, the samples were dehy-
drated through a series of graded alcohols in increasing
concentrations of 60%, 70%, 80%, 90%, and 100% for 15 mi-
nutes each. Then, inclusion in the premixtures of various
proportions of alcohol resin was performed. The first step
was in the acrylic resin with a ratio of 1:1 ethyl alcohol for
2 hours at 4! C. Then, they were changed to an alcohol resin
with a ratio of 2:1 and left at 4! C overnight. The next morn-
ing, the resin was changed with absolute ethyl alcohol 3:1
and incubated for 4 hours at 4! C. In the last step, the change
was to pure resin, which was left overnight at 4! C. Final
inclusion was performed the next day and consisted of
J CATARACT REFRACT SURG - VOL 40, OCTOBER 2014
1699
changing the samples to Eppendorf tubes and leaving
them to polymerize for 24 to 48 hours at 56! C in an oven.
Immunohistochemistry with Antibodies
for Immunogold
To increase the sensitivity of the study, immunohisto-
chemistry with colloidal gold antibodies was performed.
Slides of the lens tissue with a thickness of 80 nm were ob-
tained with the ultramicrotome, placed on 200-mesh nickel
grids (Structure Probe, Inc.), and incubated with PBSBSA
1.0% for 45 minutes. Then, they were immediately placed
in the primary antibody ATF6 (AB37149) and GRP78
(AB21685) diluted 1:20 in PBS C BSA 1.0% and incubated
for 2 hours at 37! C. Next, they were washed 10 times with
PBS C BSA 1.0%. They were then incubated with the second
antibody marked with colloidal gold (EM goat anti-rabbit
this immunoglobulin G 10 nm conjugated, BBI Solutions)
that was diluted 1:50 for 1 hour at 37! C and for 30 minutes
at room temperature. After, they were washed with PBS
BSA 1.0% 10 times, fixed with glutaraldehyde 1.0% for 15
minutes, and washed with distilled water and filtered.
Contrasting Uranyl and Lead
The last step was to stain the sections with uranyl and lead
to improve the contrast of the samples. The sections were
placed on racks on a drop of 50 mL uranyl acetate at 5.0%
for 20 minutes. Then, they were washed thoroughly with
bi-distilled water and then placed for 1 minute in a drop of
citrate lead. They were immediately washed again in
distilled water.
Quantification Colloidal Gold Grains
To compare the GRP78 and ATF6 detection in the samples
(ie, normal lenses and cataract lenses of patients with senile
cataract and patients with diabetic-associated cataract), the
grains of colloidal gold on images obtained with TEM
were counted and statistical analysis was performed using
the Mann-Whitney test.
RESULTS
Ultrastructural Morphological Analysis of Lenses
Tissue scanning electron microscopy used to eval-
uate the ultrastructural changes in the lenses showed
that the capsules and nuclei of normal lenses from
young patients were undamaged, with a characteristic
hexagonal organization of fibers (Figure 1) and inter-
digitations in the form of cellular protrusions on the
lateral surface of the lens fibers (Figure 1, B). In contrast,
the lenses of patients with senile or diabetic cataract
showed similar damage with structural alterations,
such as misalignment and disorganization of fibers
and loss of interdigitations (Figure 2). The formation
of protein aggregates in the fiber was also seen
(Figure 2, B).
Presence of GRP78 in Lenses
Immunofluorescence techniques did not show GRP78
in normal lenses (Figure 3, a). Immunohistochemistry
1700 LABORATORY SCIENCE: MOLECULAR BIOLOGY OF CATARACTS

Figure 1. Scanning electron microscopy images of normal lens. A: Lens panoramic view showing characteristic organization and interdigitation
of normal lens fibers (arrow), junctions (*), and normal intracellular connections known as ball and socket (original magnification "2000).
B: Higher magnification showing lateral extension (PL) and interdigitations (ID) of fibers (original magnification "7500).

with colloidal gold antibodies found minimal GRP78 on
normal lenses (Figure 4, a) confirming the immuno-
fluorescence results. In senile cataract tissue, GRP78 in
lens cells was detected by immunofluorescence
(Figure 4, b). This finding was confirmed using transmis-
sion electron microscopy (TEM) with the immunogold
technique (Figure 4, b). Both techniques detected
GRP78 in the fibers of the diabetic cataract lenses. The
fluorescence for GRP78 was the most intense in diabetic
cataract lenses (Figure 3, c).
Presence of ATF6 in Lenses
In normal lenses, neither immunofluorescence nor
immunohistochemistry detected ATF6 (Figures 5, a,
and 6, a). All techniques showed ATF6 in senile cata-
ract tissue (Figures 5, b, and 6, b). The transducer
ATF6 was strongly detected in diabetic cataracts in a
pattern similar to that of GRP78. The presence of
ATF6 was more intense in diabetic lens tissues than
in senile cataract tissue and normal lens tissue
(Figure 5, b and c, and Figure 6, b and c).

Figure 2. Scanning electron microscopy images of senile cataract. A: Flat areas (*) corresponding to loss of organization of lens fibers and absence
of interdigitations between fibers (arrow), appearing compacted (C) (original magnification "2000). B: Cataract tissue with same findings at
lower magnification (original magnification "1500).

J CATARACT REFRACT SURG - VOL 40, OCTOBER 2014

 LABORATORY SCIENCE: MOLECULAR BIOLOGY OF CATARACTS 1701

Figure 3. Immunofluorescence for GRP78. a: Normal lens (NL) without detection. b: Senile cataract (SCL) with detection of GRP78. c: Diabetic
cataract (DCL) has the most intense GRP78 signal. The arrows represent positive detection.

Quantification Colloidal Gold Grains
Analysis of the grains of colloidal gold on images
obtained with TEM confirmed that the presence of
GRP78 and ATF6 was greater in diabetic cataract tis-
sue (Figure 7). The mean number of colloidal gold
per image in GRP78 was 162 grains in diabetic lenses
and 96 grains in senile lenses. Similarly, for ATF6
(Figure 7, B), the mean was 98 grains in diabetic lenses,
which was statistically significantly higher than the 76
grains in the senile lenses (P!.05).
DISCUSSION
Our first goal was to document the morphologic
changes in normal lenses and compare them with
those in senile cataractous lenses and metabolic cata-
ractous lenses at a macroscopic and ultrastructural
level. Morphologic analysis performed on postmortem
lenses of healthy donors confirmed that their structure
was intact and in perfect condition, coinciding with
findings reported by Taylor et al.14 and Jongebloed
et al.15 This also confirmed that our preservation
methods were correct. Macroscopic and ultrastruc-
tural comparison of the lenses detected obvious
morphologic differences, with cataractous lenses
showing a loss of fiber alignment and organization
as well as degeneration of the interdigitations of the
lens fibers. Freel et al.16 found significant macroscopic
differences between the size of the nucleus of normal
lenses and that of cataractous lenses, with the trans-
parent nucleus being larger than the cataractous lens.
The smaller size of the cataractous lens is the result
of water loss, which leads to cytoplasmic cellular
dehydration because the nuclei of the lens fibers are
shorter. Other authors17 suggest that the shortening
of fibers contributes to hardness of cataracts. We
confirmed these findings in our study because in the
ultrastructural images of diabetic cataractous lenses,
we observed fusion or compacting of the lens fibers.
Also, we did not observe organization and interdigita-
tion of the fibers seen in the normal lenses, for which
the images were similar to a syncytium. Similarly, we

Figure 4. Detection of GRP78 by 10 nm immunogold. a: Normal lens (NL) with a scant amount of colloidal gold in the cytoplasm only (arrows). b:
Senile cataract (SCL) with positive detection of GRP78 in multiple cytoplasm areas (arrows). c: Diabetic cataract (DCL) with abundant colloidal
gold marks accumulated in cytoplasm and interdigitations (ID) (arrows) (TEM images).

J CATARACT REFRACT SURG - VOL 40, OCTOBER 2014

1702 LABORATORY SCIENCE: MOLECULAR BIOLOGY OF CATARACTS

Figure 5. Immunofluorescence detection of ATF6. a: Normal lens (NL) with no fluorescence. b: Senile cataract (SCL) with weak positive detection
(arrow). c: Diabetic cataract (DCL) with intense ATF6 detection (arrow).

observed through macroscopic morphology that the to-
tal diameter of a whole cataractous lens was smaller
than the total diameter of the normal lens of healthy do-
nors. Previous morphologic studies observed an
increased rate of compaction of lens fibers of the nu-
cleus in diabetic patients.16,18 Others report specific ul-
trastructural changes seen on TEM, such as the
formation of vacuoles in the case of diabetic cataract
lenses.19 However, we were not able to confirm these
results. Moreover, we detected protein aggregates in fi-
bers of both senile cataracts and diabetic cataracts.
The absence of GRP78 and ATF6 in normal lenses
may indicate that the UPR is activated as a result of
endoplasmic reticulum stress within lens cells under
pathologic conditions. In normal lenses, cell homeosta-
sis is maintained through several systems, including
glutathione and other chaperones. In contrast, we de-
tected both GRP78 and ATF6 in senile cataracts and
diabetic cataracts. It is possible that at some point dur-
ing lens opacification, the metabolic and senile
pathogenic pathways converge. To our knowledge,
there are no published studies of the UPR in senile
cataract. However, we believe that the UPR may be
the result of the oxidative environment during the
pathogenesis of age-related cataracts. Firtina et al.4
found ATF6 expressed in lenses of transgenic mice ex-
pressing Col4A, an animal cataract model. They also
observed an expanded, rough endoplasmic reticulum
in lens cells, irregular and poor alignment of the fibers,
and abnormalities in the lateral membrane. Elanchez-
hian et al.20 induced endoplasmic reticulum stress in
LECs using homocysteine, generating reactive oxygen
species that resulted in further oxidation and death of
LECs as well as nuclear factor-like 2 (Nrf2) degrada-
tion. Then, downstream enzymes of Nrf2, catalase,
and glutathione reductase were significantly
decreased. They found that the Nrf2-dependent anti-
oxidant defense protection in LECs was diminished,
resulting in highly oxidized lenses and age-related
cataract.

Figure 6. Detection of ATF6 transducer by immunogold 10 nm. a: Normal lens (NL) with no detection and cell interdigitations preserved (arrows).
b: In senile cataracts (SCL), ATF6 is seen in several areas of cytoplasm and endoplasmic reticulum (ER). c: In diabetic cataract (DCL), abundant
colloidal gold marks (arrows) are seen in the cisternae of endoplasmic reticulum (ER) (TEM images).

J CATARACT REFRACT SURG - VOL 40, OCTOBER 2014

 LABORATORY SCIENCE: MOLECULAR BIOLOGY OF CATARACTS
Figure 7. Quantification of GRP78 and ATF6 immunogold grains in
cataracts.
In adults, cataract-associated diabetes are often
difficult to differentiate from cataracts in patients
without diabetes because adult-onset diabetic cata-
racts often contain nuclear sclerosis, which closely
resembles the typical senile cataracts in patients
without diabetes.21 Similar morphologic changes
have also been observed in comparisons of the inner
nuclear fiber cells from diabetic lenses and nondia-
betic lenses with nuclear sclerosis. However, the
epithelial cell densities were lower in cataractous
lenses from diabetic patients than in those from pa-
tients without diabetes.22
In agreement with our findings, Mulhern et al.23
detected GRP78 in the ocular tissues of galacto-
semic rats, an animal model of diabetic damage,
suggesting that the UPR might play an important
J CATARACT REFRACT SURG - VOL 40, OCTOBER 2014
1703
role in cataract formation in animal models. We
believe that higher detection of UPR proteins in
diabetic cataracts may be the result of accumu-
lated damage of the lens caused by oxidation and
by altered metabolic pathways, such as sorbitol-
related and aldose reductase alterations in the inter-
nal lens. In recent years, UPR has been closely
linked to diabetes mellitus.24 Evidence suggests it
could play an important role in lens cell damage
in diabetic cataract development due to osmotic
stress.23 This suggests that in patients with hyper-
glycemia, the polyol pathway is activated, with
subsequent activation of aldose reductase that
causes sorbitol accumulation in the LECs, placing
stress on the endoplasmic reticulum, which in
turn activates the UPR.6 Our findings suggest that
the UPR may play an important role not only in
age-related cataracts but also in diabetes-associated
cataracts. The UPR may be activated by the
accumulation of malformed proteins in the endo-
plasmic reticulum of lens fibers that is caused by
oxidative damage, sorbitol-related stress, or both.
This response may trigger different mechanisms to
relieve stress in the endoplasmic reticulum in lens
cells.25 Our study is the first to evaluate this in
lenses of human patients with cataract. Previously,
Engler et al.26 studied the UPR in Fuchs dystrophy.
They also found that the UPR might play an
important role in the unresolved pathogenic mecha-
nism of this dystrophy.
Our study has several limitations. First, obtaining
biologic samples of normal lenses has ethical implica-
tions. Therefore, we included only 3 normal lenses
from 3 postmortem donors. However, morphologic
studies using TEM27,28 previously used a similar num-
ber of samples to assess the ultrastructure of cataract.
Our sample size of cataracts was large enough to
obtain statistically significant results. Second, we
used 2 standardized techniques with different sensi-
tivities to confirm and validate our results; this method
has been used in other studies.29,30 The sensitivity of
the technique is limited by tissue processing. Inclusion
material may be a key element in this. An ideal inclu-
sion material preserves tissue integrity and allows
antigen retrieval. Paraffin techniques combined with
fluorochromes (immunofluorescence in paraffin
sections) have a sensitivity of resolution that is limited
to 4 mm slides; however, resins such as glycol methac-
rylate for TEM allow 60 nm slides. This is probably
why we detected minimal quantities of GRP78 and
ATF6 on normal lenses using immunogold but none
using immunofluorescence on the same samples.
This also validates our results and those in other
studies.12,13,31
1704 LABORATORY SCIENCE: MOLECULAR BIOLOGY OF CATARACTS
WHAT WAS KNOWN
# Osmotic and oxidative stress in rat LECs induce the UPR,
which might play an important role in pathogenesis of dia-
betic cataracts.
# In transgenic mice lenses, activation and expression of
IRE1 and ATF6 transducers are associated with cataract
formation and expansion of the rough endoplasmic
reticulum.
WHAT THIS STUDY ADDS
# The study detected GRP78 and ATF6 expression in human
cataracts, suggesting that the UPR may be involved in
pathogenesis of senile and diabetic cataracts.
# In humans, GRP78 and ATF6 were present in lens fibers of
senile and diabetic cataracts, establishing that the UPR
may be important in the process of cataractogenesis.
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First author:
Beatriz E. Torres-Bernal, MS
Inova Vision Quirurgica,
Aguascalientes, Mexico