A Novel GJA8 Mutation (p.V44A) Causing Autosomal Dominant Congenital Cataract
A Novel GJA8 Mutation (p.V44A) Causing Autosomal Dominant Congenital Cataract
Yanan Zhu 0 1 4
Hao Yu 2 4
Wei Wang 0 1 4
Xiaohua Gong 3 4
Ke Yao 0 1 4
0 Eye center, 2nd Affiliated Hospital of Medical College, Zhejiang University , Hangzhou , China,
1 Key Laboratory of Ophthalmology of Zhejiang Province , Wenzhou , China,
2 Department of Neurobiology, Key Laboratory of Neurobiology of Zhejiang Province, Zhejiang University School of Medicine , Hangzhou , China,
3 School of Optometry and Vision Science Program, University of California , Berkeley, California , United States of America
4 Editor: Roy A. Quinlan, University of Durham , United Kingdom
Purpose: To examine the mechanism by which a novel connexin 50 (Cx50) mutation, Cx50 V44A, in a Chinese family causes suture-sparing autosomal dominant congenital nuclear cataracts. Methods: Family history and clinical data were recorded and direct gene sequencing was used to identify the disease-causing mutation. The Cx50 gene was cloned from a human lens cDNA library. Connexin protein distributions were assessed by fluorescence microscopy. Hemichannel functions were analyzed by dye uptake assay. Formation of functional channels was assessed by dye transfer experiments. Results: Direct sequencing of the candidate GJA8 gene revealed a novel c.131T.C transition in exon 2, which cosegregated with the disease in the family and resulted in the substitution of a valine residue with alanine at codon 44 (p. V44A) in the extracellular loop 1 of the Cx50 protein. Both Cx50 and Cx50V44A formed functional gap junctions, as shown by the neurobiotin transfer assay. However, unlike wild-type Cx50, Cx50V44A was unable to form open hemichannels in dye uptake experiments. Conclusion: This work identified a unique congenital cataract in the Chinese
Congenital cataracts are a worldwide cause of childhood blindness, accounting for
more than 1 million cases of pediatric blindness in Asia and approximately 10% of
the worldwide cases of childhood blindness . Genetic mutations may be the
most common cause of this condition, particularly in cases of bilateral cataracts.
Thirty-seven gene mutations are currently known to be associated with congenital
or childhood cataracts, approximately one-quarter of which are located in the
connexins [2, 3].
Both connexin 46 (Cx46) and connexin 50 (Cx50), which are encoded by the
GJA3 and GJA8 genes, respectively, have been reported to be associated with
autosomal dominant congenital cataracts . Several strains of mice or rats that
develop cataracts also harbor mutations of the GJA3 (Cx46) or GJA8 (Cx50) genes
[5, 6]. Studies have shown that these transport membrane proteins are important
for the correct embryological development of the lens.
Gap junctions, which are channels that connect the cytoplasm of neighboring
cells, are well documented to play an important role in intercellular
communication in the lens, which is avascular. These junctions are formed by two
hemichannels (connexons), which consist of six connexin molecules. Prior to
assembly into gap junctional channels, hemichannels transiently reside in the
plasma membrane and can open in response to various triggers . Recent studies
have proposed a potential role of hemichannels in autocrine/paracrine signaling
that is unrelated to their role as precursors in the formation of gap junction
In this study, we identified a novel heterozygous mutation, the c.131T.C
(p.V44A) mutation, in exon2 of GJA8 in a Chinese family with suture-sparing
nuclear cataracts. In addition, we examined the mechanism of cataract formation
in patients carrying the Cx50 V44A mutation. Functional study showed that this
mutation caused abnormal alterations in hemichannel function, which may
represent a possible mechanism for the formation of cataracts.
The study protocol was in accord with the ethical guidelines of the Declaration of
Helsinki and was approved by the Ethics Committee of Zhejiang University.
Appropriate written informed consent was obtained from all of the adult
participants; consent was obtained from adult guardians on behalf of the pediatric
Clinical Evaluation and Examination
A four-generation Chinese pedigree diagnosed with autosomal dominant
congenital cataracts was recruited at the Second Affiliated Hospital of Zhejiang
University (Hangzhou, China). Thirty-four individuals (eight affected and 26
unaffected) from the family participated in the study. The affected status was
determined by a history of cataract surgery or by ophthalmic examinations.
Genomic DNA Preparation
We collected blood specimens (5 mL) of all the participants into EDTA tubes. The
peripheral blood leukocytes were then used to extract genomic DNA.
We performed the mutation screening using the functional candidate gene
analysis approach. Gene-specific PCR primers that flanked each exon and
intronexon junction were designed for genes that were related to nuclear cataract, such
as CRYAA, CRYAB, CRYBA3/A1, CRYBB1, CRYBB2, CRYGC, CRYGD, GJA3,
GJA8 and MIP. Details of the strategy used for PCR and sequencing are described
in our previous study . The results were compared to the NCBI GenBank
sequences. Nucleotide numbering reflected the cDNA numbering, with +1
corresponding to the A of the ATG translation initiation codon in the reference
sequence. The initiation codon was assigned as codon 1.
Sequencing and Cloning of Wild-type and Mutant Connexins
The GJA8 cDNA sequence was cloned from a human lens cDNA library (provided
by Graeme Wistow from the NIH as a gift) by PCR using PrimeSTAR HS DNA
polymerase (Takara Ltd., Dalian, China) and the following oligo nucleotide
(59-GCATTACTCGAGATGGGCGACTGGAGTTTCCTGGG-39) and antisense-primer
(59-CCATTGAATTCGTACGGTTAGATCGTCTGACCTGGCTCG-39). After digestion with XhoI and EcoRI (NEB,
Ipswich, MA, USA), the PCR product was cloned into the expression vector
pEGFP-N1 (Invitrogen, Carlsbad, CA, USA). The resultant construct, Cx50WILD,
was confirmed by DNA sequencing.
A Quik-Change Site-Directed Mutagenesis Kit (Stratagene, LaJolla, CA, USA)
was used to construct the mutant Cx50V44A plasmid using the following
59-GATTGCTCATCCCCCCACGCGAACTCTGCGGCCGTG39. The Cx50 mutants were sequenced to verify the target mutation.
Cell Culture and Transfections
Human epithelial carcinoma cells (HeLa, ATCC) were grown in DMEM (Gibco,
Carlsbad, CA, USA) with 10% FBS. Transfection was performed on HeLa cells
using the transfection reagent Lipofectamine 2000 (Invitrogen), according to the
manufacturers instructions. HeLa cell monoclones that stably expressed human
wild-type Cx50, mutant Cx50V44A or the EGFP control were isolated based on
their resistance to geneticin (G418) (1 mg/mL). To maintain stability of
Identification the Connexin43 Status of Transfected HeLa Cells
Cx50WILD-transfected, Cx50V44A-transfected, EGFP-transfected HeLa cells and
human retinal epithelial cells (ARPE-19, ATCC) were seeded in 100-mm dishes
for 24 h. ARPE-19, which has been reported to express Cx43 , served as a
control. The cells were collected in ice-cold PBS and were then centrifuged at
1000 rpm at 4C for 5 min. After removing the supernatant, total protein was
extracted from the cells with a lysis buffer (Sangon Biotech, Shanghai, China).
After incubation on ice for 30 min, the extracts were centrifuged at 14,000 rpm at
4C for 15 min. Then, the supernatant of each sample was transferred to a fresh
tube and boiled with a protein-loading buffer (Sangon Biotech). The proteins
were separated by 10% SDS-polyacrylamide gel electrophoresis, transferred to
polyvinylidene difluoride membranes and blotted with anti-Cx43 rabbit
polyclonal antibody (1:200 dilution, Santa Cruz) and anti-GAPDH rabbit
monoclonal antibody (1:5000 dilution, Cell Signaling Technology, USA). After
incubation in fluorescent secondary antibodies (1:5000 dilution, Cell Signaling),
the blots were analyzed using the Bio-Rad ChemiDoc MP imaging system. The
target protein expression levels were normalized relative to GAPDH expression.
Identification of Cx50 Status of Transfected Cells
The expression levels of Cx50WILD and Cx50V44A in stably transfected HeLa
cells were verified by Western blot, and the EGFP-transfected cells served as
controls. The target protein was blotted with anti-Cx50 rabbit polyclonal antibody
(1:200 dilution, Santa Cruz) and with fluorescent secondary antibodies, and its
expression level was normalized relative to GAPDH expression.
Stably transfected cells that expressed the GFP-tagged connexins were plated on
four-well chamber slides (Lab Tek; Nalge Nunc International, Naperville, IL,
USA). Fluorescence images were obtained when the cultured cells reached 80% to
90% confluence. Cx50 protein distribution and gap junction formation were
monitored based on tagged GFP signaling in these cells using a confocal laser
scanning microscope (640, Zeiss LSM 510, Zurich, Switzerland).
Dye Uptake Experiments
For hemichannel function analysis, three groups of cells (Cx50WILD, Cx50V44A,
and control) were placed in a 35-mm collagen-coated glass-bottom culture dish
(50% confluence). The cells were washed twice with Ca2+-free HBSS (Gibco). Dye
uptake assays were performed by incubating each group of cells in Ca2+-free HBSS
(Gibco) and in HBSS containing 1.2 mmol/L Ca2+ or a specific hemichannel
blocker, flufenamic acid (FFA, 300 mmol/L, Sigma, Saint Louis, MO, USA) .
All of the solutions were supplemented with 0.1% propidium iodide (PI, Sigma)
or 0.1% 49,6-diamidino-2-phenylindole (DAPI, Sigma). After 30 min, the
solutions were removed; the cells were then washed with PBS, twice, and finally
incubated in normal culture solutions for counting. Excluding the dead cells that
either lost their normal shape or became unattached, the total numbers of live
cells and dye-loading live cells were counted using fluorescence microscopy
(640). Thirty fields were applied each time, and the dye uptake experiments were
repeated three times. The results are shown as percentages of loading cells and are
expressed as the means the standard deviations (SDs). Statistical analysis was
performed using Students t-test.
In situ Dye Transfer Experiments
The cells were cultured with normal culture medium on glass coverslips until
80%90% confluence was reached. Then, the glass coverslips were transferred to a
bath solution containing 140 mM NaCl, 1 mM MgCl2, 5.4 mM KCl, 1.2 mM
CaCl2, and 10 mM HEPES (pH 7.2). The cells were impaled with a micropipette
filled with a patch pipette solution and 4% neurobiotin dye (Mr5322.8, charge
5+1; Vector Laboratories, Burlingame, CA, USA) or 5% lucifer yellow dye
(Mr5457, charge 522; Invitrogen). The patch pipette solution contained
140 mM KCl, 1 mM MgCl2, 5 mM NaCl, and 10 mMHEPES (pH 7.4). The dye
solution was microinjected for 3 min with a Picospritzer (model PLI-188; Nikon,
After neurobiotin dye injection, the cells were fixed, permeabilized, and stained
with Cy3-streptavidin conjugate (Sigma) for tracer detection. However, after
lucifer yellow dye injection, the cells required only fixation. Both types of cells
were then photographed with fluorescence microscopy. The adjacent cells stained
by the dye were counted to determine the extent of the intercellular tracer transfer.
Three independent stable-cell clones were all used to perform the dye transfer
assay. The number of coupled cells was expressed as the means SDs. Statistical
analysis was performed with Students t-test.
We identified a four-generation Chinese family with autosomal dominant
congenital cataracts. The majority of the patients realized their visual impairment
before the age of 10, particularly during the daytime. The visual acuity decreased
gradually, and cataract surgery was eventually performed at approximately 40
years of age (Fig. 1A). The affected individuals all presented with symmetrical,
homogeneous nuclear opacification in both eyes. The anterior and posterior
Ysutures were not involved (Fig. 1B, C). The patients had best-corrected visual
acuity (BCVA) values that ranged from finger counting to 0.2 prior to surgery,
Fig. 1. Family data collection, cataract phenotype, and mutation sequencing of Cx50. A: Pedigree of the proband. The black symbols indicate the
affected individuals. The arrow indicates the proband. All the living members shown in this figure participated in this project. B: Diffuse illumination of the left
eye of the proband (the arrows show transparent anterior suture). C: Slit lamp photograph of the lens of the proband showing the nuclear opacities; anterior
and posterior sutures are not involved (arrowheads). D: Partial DNA sequence of GJA8 from one normal individual and one affected individual. The black
box shows a heterozygous mutation (c.131T.C) in exon 2. E: Conservation analysis of p.V44A of the Cx50 gene.
and they achieved BCVA values ranging from 0.5 to 0.8 after surgery. In their
family, the unaffected members had good BCVA values ranging from 0.6 to 1.0.
There was no family history of other ocular or systemic abnormalities.
Fig. 2. Subcellular localization of Cx50WILD and Cx50V44A in stably transfected HeLa cells. Both Cx50WILD (A) and Cx50V44A (B) were observed at
appositional membranes and perinuclear cytoplasmic locations (the arrows show the gap junction plaques). Scale bar: A-B 20 mm. C: Endogenous
expression of Cx43 in transfected HeLa cells. The transfected cells did not express Cx43 protein, but the ARPE-19 cells (RPE) did. (P,0.001, marked with
***) D: The stable transfected cells expressed similar protein levels for Cx50WILD or Cx50V44A.
Using gene sequencing, we identified a novel heterozygous c.131T.C mutation in
the GJA8 gene (RefSeq NM_005267.4). This variation occurred in all of the
affected individuals (Fig. 1D), but it was not observed in the unaffected family
members or in the 100 unrelated healthy individuals who served as controls. This
mutation resulted in a substitution of a valine residue for alanine at codon 44 of
connexin 50 (p.V44A), which is located in the first extracellular loop of this
protein . Multiple sequence alignment indicates that the mutation is located
within a highly conserved region (Fig. 1E).
HeLa cells from ATCC have been reported to express Cx43 in early generations,
but in our experiment, the transfected HeLa cells that were used did not express
the Cx43 protein (Fig. 2C). Therefore, these transfected HeLa cells were good
models because of their connexin deficiency . GFP-tagged versions of
Cx50WILD or Cx50V44A were expressed in HeLa cells, and similar levels of the
expression of wild-type protein or mutant protein of approximately 98 kDa were
detected by Western blot, whereas no expression of Cx50 was detected in the
control cells (Fig. 2D). Cellular distributions of the GFP-tagged Cx50WILD and
Cx50V44A proteins were determined based on their GFP fluorescence. As
expected, the wild-type Cx50 proteins formed gap junctional plaques on
appositional membranes and were present in the cytoplasmic and perinuclear
regions (Fig. 2A). The Cx50V44A mutants also formed typical gap junctions and
were present in the cytoplasmic and perinuclear regions, similar to wild-type Cx50
(Fig. 2B). The results indicate that the V44A mutation did not affect Cx50 protein
trafficking and gap junction formation.
We then examined hemichannel function. Dye (such as DAPI or PI) uptake
assays revealed that wild-type Cx50 hemichannels opened in Ca2+-free HBSS
medium after 30 min of treatment and closed after incubation in HBSS
containing 1.2 mmol/L Ca2+ or 300 mmol/L FFA for the same period of time. As
expected, the DAPI uptake assay confirmed that almost all of the Cx50WILD cells
(97.53.63%) contained high-intensity blue fluorescence in their nuclei, whereas
a low percentage of DAPI-stained cells was observed in the Cx50V44A cells
(3.725.91%) and the control cells (1.101.48%). After incubation in HBSS
containing 1.2 mmol/L Ca2+ for the same duration, very few Cx50WILD cells
(3.003.07%) contained fluorescent dye, which was similar to the Cx50V44A
cells (1.343.04%) and the control cells (0.170.52%) (Figs. 3 and 4). The PI
uptake assay revealed similar results for the Cx50, Cx50V44A and control cells.
Fig. 3. DAPI dye uptake in HeLa cells stably transfected with Cx50WILD and Cx50V44A. DAPI was
abosorbed by most of the Cx50WILD-transfected cells, but not by the Cx50V44A- and pEGFP-N1-transfected
cells (Mock), after hemichannel opening. DAPI loading in Ca2+-free HBSS was blocked by 1.2 mM Ca2+ and
300 mM FFA. The three pictures in the last line show the Cx50 protein, DAPI-stained nuclei and merged look
of the wild-type cells in Ca2+-free HBSS. Scale bar: 20 mm.
Most of the Cx50WILD cells(73.578.21%) showed high-intensity red
fluorescence in their nuclei, particularly in the nucleoli, and low-intensity red
fluorescence in the cytoplasm, whereas a very low percentage of the Cx50V44A
(2.595.33%) and control cells (0.670.96%) were positive for PI staining. After
incubation in HBSS containing 1.2 mmol/L Ca2+ for the same amount of time,
very few of the Cx50WILD cells (1.462.86%), Cx50V44A cells (2.443.50%),
and control cells (0.841.59%) absorbed PI (Figs. 4 and 5). Hemichannel
function was assured using FFA. Dye uptake was completely blocked by
300 mmol/L FFA. In the FFA-treated cells, only 0.321.39% of the Cx50WILD
cells, 0.672.41% of the Cx50V44A cells and 0.320.74% of the control cells
absorbed the PI dye, while 0.411.34% of Cx50WILD cells, 0.560.95% of the
Cx50V44A cells and 0.410.80% of the control cells absorbed the DAPI dye.
Fig. 4. PI dye uptake in HeLa cells stably transfected with Cx50WILD and Cx50V44A. The PI dye uptake assay revealed similar results as the DAPI dye
uptake assay. The three pictures in the last line show the Cx50 protein, PI-stained nuclei (nucleoli)/cytoplasm, and merged look of the wild-type cells in
Ca2+free HBSS. Scale bar: 20 mm.
(Figs. 3 and 5) Thus, these results suggest that the Cx50V44A mutant proteins
were unable to form calcium-sensitive hemichannels, such as those formed by
We further tested the ability of Cx50V44A to form functional gap junction
channels by using dye microinjection of gap junction tracers in individual cells.
We quantified the intercellular gap junction communication by counting the
number of tracer-containing neighboring cells. Neurobiotin transfer was detected
in the Cx50WILD cells (26.751.89coupled cells; the number of microinjected
cells (n)520). Surprisingly, the neurobiotin transfer data were similar in the
Cx50V44A cells (26.333.98 coupled cells; n520). The control cells showed
almost no transfer of neurobiotin (0.150.37 coupled cells, n520) (Fig. 6A, C).
As expected, both the Cx50 and Cx50V44A cells did not transfer Lucifer yellow
dye (Fig. 6B, C). These results indicated that the V44A mutation did not affect the
intercellular dye transfer of the Cx50 gap junction channels.
In this study, we described the suture-sparing nuclear cataract phenotype in a
Chinese population and identified a novel TRC mutation in the second exon of
the GJA8 gene that was associated with this phenotype. Interestingly, this special
cataract phenotype was also previously reported to be related to the CRYBA3/A1
gene , indicating that similar types of cataracts could be caused by different
Our data indicated that the p.V44Amutation in the GJA8 gene was the causative
mutation in this ADCC family, rather than a rare polymorphism in the normal
population. First, the mutation cosegregated with the disease phenotype in the
Fig. 5. Statistical analysis of DAPI (A) and PI (B) loading in different solutions. Data are presented as
meanSDs. There was a significant difference in the percentage of dye-stained cells between the Cx50WILD
and Cx50V44A groups in the Ca2+-free environment (P,0.001, marked with ***); there was also a significant
difference between the Ca2+-free Cx50WILD and the 1.2 mM Ca2+/300 mM FFA Cx50WILD group (P,0.001,
marked with ***).
family and it was absent in the 100 unaffected, unrelated individuals who were
screened as the control. Second, the valine residue at this site was highly conserved
because it was present in the sequences of all of the Cx50 orthologs that were
deposited in the database. Third, the mutant Cx50 displayed a dysfunctional
The GJA8 gene, which encodes Cx50, has been heavily investigated in
congenital cataracts. To date, 22 Cx50 mutations have been identified in human
congenital cataract pedigrees [4, 15, 16]. Cx50, like other connexins, includes an
amino terminus (NT), a carboxyl terminus (CT), and four transmembrane
domains (TMs) that are linked by a single cytoplasmic loop (CL) and by two
extracellular domains (E) . The V44A mutation was located in the
extracellular loop 1 near the TM1/E1 border. The importance of this region was
Fig. 6. Dye transfer assay in HeLa cells stably transfected with Cx50WILD and Cx50V44A. A: Cx50WILD- and Cx50V44A-transfected cells displayed
extensive transfer of neurobiotin, whereas the pEGFP-N1-transfected cells did not allow transfer of this tracer. The white stars show the injected cells. (B)
Neither cell line showed significant intercellular transfer of Lucifer yellow. The orange stars show the injected cells. Scale bar: A-B 20 mm. (C) The number of
three groups of cells absorbing neurobiotin (NB) and lucifer yellow (LY) from the injected cell. Data are presented as meanSDs; n520; the neurobiotin
transfer assay showed significant differences in the percentage of coupled cells between the control group and the Cx50WILD/Cx50V44A groups (P,0.001,
marked with ***).
further emphasized by its identification at position 4251 of Cx50 (V44E, W45S,
G46V, D47A, D47Y, D47N, D47H, E48K, S50P) [5, 16, 1824]. Among the Cx50
human mutations previously described, Cx50E48K and Cx50D47N showed a
decreased ability to form functional channels [23, 24], while Cx50G46V formed
enhanced functional channels . Recently, Rubinos reported that Cx50V44E,
which was at the same location as the mutation described in the present study,
failed to form functional homotypic or heterotypic gap junctions and strongly
inhibited WT connexin function .
We examined the effects of the novel cataract-associated mutation Cx50V44A.
Our results showed that Cx50V44Amutant proteins were able to traffic and
oligomerize into functional gap junctions in the plasma membrane in transfected
HeLa cells. However, the Cx50V44A mutants failed to form appropriate
functional hemichannels. The electrophysiological data collected in vitro
demonstrated that hemichannels opened in reduced external calcium [26, 27] and
closed in a solution containing 1.2 mmol/L Ca2+ . Here, we showed that the
opened Cx50V44A hemichannel could not absorb either DAPI dye
(MW5350 Da, charge, +2) or PI dye (MW5668 Da, charge, +2), whereas
Cx50WILD could. These results suggested that, in a Ca2+-free environment, the
pores of the mutant hemichannels could be closed or be smaller in diameter than
the size of the DAPI dye.
The dysfunctional hemichannel caused by Cx50V44A plays an important role in
cataract formation. Although the concentration of calcium in the human aqueous
humor is ,1.3 mM , calcium homeostasis in the central mature fibers of the
lens appears to be more complex. The interfiber calcium concentration in the lens
remains unknown. Gao reported that the intracellular calcium concentration in
mouse lens varied from 700 nM in the center to 300 nM at the surface . In the
physiological environment, perhaps, extracellular calcium concentrations in
human lenses are somewhere between ,1.3 mM calcium in the aqueous humor
and intracellular calcium concentration (300700 nM) in the lens fiber cells.
Thus, wild-type hemichannels are probably regulated by the dynamic ranges of
extracellular calcium in the lens. However, the Cx50V44A mutated hemichannels
are unable to be gated by extracellular calcium levels in the lens. Future studies
will be needed to determine the extracellular calcium concentrations in different
parts (superficial region, deep cortex and nucleus) of the lens.
The valine at position 44 is known to form part of a hydrophobic core that
participates in intraprotomer interactions, which are believed to be important in
the stabilization of the connexin structure . Rubinos suggested that
substitution of the valine with a charged residue (glutamicacid) could interfere
with connexin oligomerization, resulting in a loss of function . However,
unlike Cx50V44E, the ability of our mutation to transfer neurobiotin indicated
that the Cx50V44A mutation succeeded in achieving functional gap junctions
. Moreover, our mutation caused a suture-sparing nuclear cataract, which was
much milder than the total cataracts that are caused by Cx50V44E. Alanine, which
is an uncharged residue like valine, could not influence connexin oligomerization
or twist the orientation of docking, so the mutant connexin remained capable of
forming gap junctions.
Hemichannels were once believed to function only when two such channels
formed an intercellular channel (the gap junction) between closely apposed cells,
containing an aqueous pore exclusive of the extracellular space [32, 33]. However,
not all hemichannels are destined to become components of a gap junction
channel, and research has suggested that hemichannels play an important role in
cellular processes, such as volume regulation , the influx/efflux of
metabolically relevant solutes such as ATP , and cell death [36, 37].
Furthermore, a recent review also suggested that hemichannels are a significant
source of autocrine and paracrine messengers . Dysfunctional connexin
hemichannels have been reported to be related to cataracts . Cx50G46V forms
normal gap junctions, but it increased hemichannel activity . Cx46G2N
reduces hemichannel permeability, but it still forms gap junction plaques .
Moreover, Cx46G143R increases hemichannel function and causes apoptosis,
even when the transfected cells are cultured at low density with minimal gap
junctions . Because the V44A mutations cause a loss and/or a reduction in
hemichannel activity, they could also cause a metabolic imbalance within the lens,
such as a disruption of the lens circulation current that could trigger the
development of cataracts.
In summary, we identified a novel heterozygous c.131T.C (p.V44A) mutation
in GJA8 in a family of Chinese origin with congenital cataracts. Our study
indicated that residue V44 of Cx50 might be vital in hemichannel formation;
however, this mutation did not impair the function of the gap junction channels.
Abnormal activity of the hemichannels could disrupt cellular homeostasis,
subsequently causing cataracts.
Conceived and designed the experiments: YZ KY. Performed the experiments: YZ
HY WW. Analyzed the data: YZ XG KY. Contributed reagents/materials/analysis
tools: KY. Wrote the paper: YZ XG KY.
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