Miduturu Srinivas

State University of New York College of Optometry, New York, New York, United States

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Publications (48)197.48 Total impact

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    ABSTRACT: Maintenance of adequate levels of glutathione (GSH) in the lens nucleus is critical for protection of lens proteins from the effects of oxidative stress and for lens transparency. How GSH is transported to the nucleus is unknown. We show that GSH diffuses to the nucleus from the outer cortex, where a high concentration of the anti-oxidant is established by synthesis or uptake, via the network of gap junctions. Using electrophysiological measurements, we found that channels formed by Cx46 and Cx50, the two connexin isoforms expressed in the lens, were moderately cation-selective (PNa/PCl ∼5 for Cx46 and ∼3 for Cx50). Single channel permeation of the larger GSH anion was low but detectable (PNa/PGSH ∼12 for Cx46 and ∼8 for Cx50), whereas permeation of divalent anion glutathione disulfide (GSSG) was undetectable. Measurement of GSH levels in the lenses from connexin knock-out (KO) mice indicated Cx46, and not Cx50, is necessary for transport of GSH to the core. Levels of GSH in the nucleus were markedly reduced in Cx46 KO, whereas they were unaffected by Cx50 KO. We also show that GSH delivery to the nucleus is not dependent on the lens microcirculation, which is believed to be responsible for extracellular transport of other nutrients to membrane transporters in the core. These results indicate that glutathione diffuses from cortical fiber cells to the nucleus via gap junction channels formed by Cx46. We present a model of GSH diffusion from outer cells to inner fiber cells through gap junctions.
    Journal of Biological Chemistry 10/2014; 289(47). DOI:10.1074/jbc.M114.597898 · 4.57 Impact Factor
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    ABSTRACT: Keratitis-ichthyosis-deafness (KID) syndrome is an ectodermal dysplasia caused by dominant mutations of connexin26 (Cx26). Loss of Cx26 function causes non-syndromic sensorineural deafness, without consequence in the epidermis. Functional analyses have revealed that a majority of KID-causing mutations confer a novel expansion of hemichannel activity, mediated by connexin channels in a non-junctional configuration. Inappropriate Cx26 hemichannel opening is hypothesized to compromise keratinocyte integrity and epidermal homeostasis. Pharmacological modulators of Cx26 are needed to assess the pathomechanistic involvement of hemichannels in the development of hyperkeratosis in KID syndrome. We have used electrophysiological assays to evaluate small molecule analogs of quinine for suppressive effects on aberrant hemichannel currents elicited by KID mutations. Here, we show that mefloquine inhibits several mutant hemichannel forms implicated in KID syndrome when expressed in Xenopus laevis oocytes (IC50≈16 μM), using an extracellular divalent cation, zinc (Zn(++)), as a non-specific positive control for comparison (IC50≈3 μM). Furthermore, we used freshly isolated transgenic keratinocytes to show that micromolar concentrations of mefloquine attenuated increased macroscopic membrane currents in primary mouse keratinocytes expressing human Cx26-G45E, a mutation causing a lethal form of KID syndrome.Journal of Investigative Dermatology accepted article preview online, 17 September 2014. doi:10.1038/jid.2014.408.
    Journal of Investigative Dermatology 09/2014; 135(4). DOI:10.1038/jid.2014.408 · 7.22 Impact Factor
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    Miduturu Srinivas
    Journal of Ophthalmic & Vision Research 07/2014; 9(1):148-9.
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    ABSTRACT: Excessive opening of undocked Cx26 hemichannels in the plasma membrane is associated with disease pathogenesis in keratitis-ichthyosis-deafness (KID) syndrome. Thus far, excessive opening of KID mutant hemichannels has been attributed, almost solely, to aberrant inhibition by extracellular Ca2+. This study presents two new possible contributing factors, pH and Zn2+. Plasma pH levels and micromolar concentrations of Zn2+ inhibit WT Cx26 hemichannels. However, A40V KID mutant hemichannels show substantially reduced inhibition by these factors. Using excised patches, acidification was shown to be effective from either side of the membrane, suggesting a protonation site accessible to H+ flux through the pore. Sensitivity to pH was not dependent on extracellular aminosulfonate pH buffers. Single channel recordings showed that acidification did not affect unitary conductance or block the hemichannel but rather promoted gating to the closed state with transitions characteristic of the intrinsic loop gating mechanism. Examination of two nearby KID mutants in the E1 domain, G45E and D50N, showed no changes in modulation by pH or Zn2+. N-bromo-succinimide, but not thiol-specific reagents, attenuated both pH and Zn2+ responses. Individually mutating each of the five His residues in WT Cx26 did not reveal a key His residue that conferred sensitivity to pH or Zn2+. From these data and the crystal structure of Cx26 that suggests that Ala-40 contributes to an intrasubunit hydrophobic core, the principal effect of the A40V mutation is probably a perturbation in structure that affects loop gating, thereby affecting multiple factors that act to close Cx26 hemichannels via this gating mechanism.
    Journal of Biological Chemistry 06/2014; 289(31). DOI:10.1074/jbc.M114.578757 · 4.57 Impact Factor
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    ABSTRACT: Mutations in Cx50 cause dominant cataracts in both humans and mice. The exact mechanisms by which mutations cause these variable phenotypes are poorly understood. We examined the functional properties of gap junctions made by three Cx50 mutations, V44E, D47N and V79L, expressed in mammalian cell lines. V44E trafficked to the plasma membrane properly and formed gap junctional plaques. However, the mutant did not form functional gap junctions when expressed alone, or with wildtype (WT) Cx46 and Cx50, indicating that V44E is a dominant negative inhibitor of WT connexin function. In contrast, D47N subunits did not localize to junctional plaques or form functional homotypic gap junctions; however, mixed expression of D47N and WT subunits of either Cx50 or Cx46 resulted in functional intercellular channels, with high levels of coupling. Single channel studies indicated that D47N formed heteromeric channels with WT Cx46 with unique properties. Unlike either V44E or D47N, V79L formed functional homotypic intercellular channels. However, the mutation caused an alteration in voltage gating and a reduction in the open probability, resulting in much lower levels of conductance in cells expressing V79L alone, or together with WT connexin subunits. Thus, each mutation produced distinct changes in the properties of junctional coupling. V44E failed to form intercellular channels in any configuration, D47N formed only heteromeric channels with WT connexins and V79L formed homotypic and heteromeric channels with altered properties. These results suggest that unique interactions between mutant and wild-type lens connexins might underlie the development of various cataract phenotypes in humans.
    AJP Cell Physiology 09/2013; 306(3). DOI:10.1152/ajpcell.00098.2013 · 3.78 Impact Factor
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    ABSTRACT: Mutations in the GJB2 gene, which encodes Cx26, are the most common cause of sensorineural deafness. In syndromic cases, such as keratitis-ichthyosis-deafness (KID) syndrome, in which deafness is accompanied by corneal inflammation and hyperkeratotic skin, aberrant hemichannel function has emerged as the leading contributing factor. We found that D50N, the most frequent mutation associated with KID syndrome, produces multiple aberrant hemichannel properties, including loss of inhibition by extracellular Ca(2+), decreased unitary conductance, increased open hemichannel current rectification and voltage-shifted activation. We demonstrate that D50 is a pore-lining residue and that negative charge at this position strongly influences open hemichannel properties. Examination of two putative intersubunit interactions involving D50 suggested by the Cx26 crystal structure, K61-D50 and Q48-D50, showed no evidence of a K61-D50 interaction in hemichannels. However, our data suggest that Q48 and D50 interact and disruption of this interaction shifts hemichannel activation positive along the voltage axis. Additional shifts in activation by extracellular Ca(2+) remained in the absence of a D50-Q48 interaction but required an Asp or Glu at position 50, suggesting a separate electrostatic mechanism that critically involves this position. In gap junction (GJ) channels, D50 substitutions produced loss of function, whereas K61 substitutions functioned as GJ channels but not as hemichannels. These data demonstrate that D50 exerts effects on Cx26 hemichannel and GJ channel function as a result of its dual role as a pore residue and a component of an intersubunit complex in the extracellular region of the hemichannel. Differences in the effects of substitutions in GJ channels and hemichannels suggest that perturbations in structure occur upon hemichannel docking that significantly impact function. Collectively, these data provide insight into Cx26 structure-function and the underlying bases for the phenotypes associated with KID syndrome patients carrying the D50N mutation.
    The Journal of General Physiology 07/2013; 142(1):3-22. DOI:10.1085/jgp.201310962 · 4.79 Impact Factor
  • Vytas K Verselis · Miduturu Srinivas
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    ABSTRACT: Gap junction channels and hemichannels formed by the connexin family of proteins play important roles in many aspects of tissue homeostasis in the brain and in other organs. In addition, connexin channels have been proposed as pharmacological targets in the treatment of a number of human disorders. In this review, we describe the connexin-subtype selectivity and specificity of pharmacological agents that are commonly used to modulate connexin channels. We also highlight recent progress made towards identifying new agents for connexin channels that act in a selective and specific manner. Finally, we discuss developing insights into possible mechanisms by which these agents modulate connexin channel function.
    Neuropharmacology 04/2013; 75. DOI:10.1016/j.neuropharm.2013.03.020 · 5.11 Impact Factor
  • Jack Kronengold · Miduturu Srinivas · Vytas K Verselis
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    ABSTRACT: Connexins form channels with large aqueous pores that mediate fluxes of inorganic ions and biological signaling molecules. Studies aimed at identifying the connexin pore now include a crystal structure that provides details of putative pore-lining residues that need to be verified using independent biophysical approaches. Here we extended our initial cysteine-scanning studies of the TM1/E1 region of Cx46 hemichannels to include TM2 and TM3 transmembrane segments. No evidence of reactivity was observed in either TM2 or TM3 probed with small or large thiol-modifying reagents. Several identified pore residues in E1 of Cx46 have been verified in different Cx isoforms. Use of variety of thiol reagents indicates that the connexin hemichannel pore is large and flexible enough, at least in the extracellular part of the pore funnel, to accommodate uncommonly large side chains. We also find that that gating characteristics are largely determined by the same domains that constitute the pore. These data indicate that biophysical and structural studies are converging towards a view that the N-terminal half of the Cx protein contains the principal components of the pore and gating elements, with NT, TM1 and E1 forming the pore funnel.
    Journal of Membrane Biology 07/2012; 245(8):453-63. DOI:10.1007/s00232-012-9457-z · 2.46 Impact Factor
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    ABSTRACT: The paucity of specific pharmacological agents has been a major impediment for delineating the roles of gap junction (GJ) channels formed by connexin proteins in physiology and pathophysiology. Here, we used the selective optimization of side activities (SOSA) approach, which has led to the design of high affinity inhibitors of other ion channels, to identify a specific inhibitor for channels formed by Cx50, a connexin subtype that is primarily expressed in the lens. We initially screened a library of common ion channel modulating pharmacophores for their inhibitory effects on Cx50 GJ channels, and identified four new classes of compounds. The triarlymethane (TRAM) clotrimazole was the most potent Cx50 inhibitor and we therefore used it as a template to explore the structure activity relationship (SAR) of the TRAMs for Cx50 inhibition. We describe the design of T122 (N-[(2-methoxyphenyl)diphenylmethyl]-1,3-thiazol-2-amine) and T136 (N-[(2-iodophenyl)diphenylmethyl]-1,3-thiazol-2-amine), which inhibit Cx50 with IC(50)s of 1.2 and 2.4 μM. Both compounds exhibit at least 10-fold selectivity over other connexins as well as major neuronal and cardiac voltage-gated K(+) and Na(+) channels. The SAR studies also indicated that the TRAM pharmacophore required for connexin inhibition is significantly different from the pharmacophore required for blocking the calcium-activated KCa3.1 channel. Both T122 and T136 selectively inhibited Cx50 GJ channels in lens epithelial cells, suggesting that they could be used to further explore the role of Cx50 in the lens. In addition, our results indicate that a similar approach may be used to find specific inhibitors of other connexin subtypes.
    Frontiers in Pharmacology 06/2012; 3:106. DOI:10.3389/fphar.2012.00106 · 3.80 Impact Factor
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    Clio Rubinos · Helmuth A Sánchez · Vytas K Verselis · Miduturu Srinivas
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    ABSTRACT: The anti-malarial drug quinine and its quaternary derivative N-benzylquininium (BQ(+)) have been shown to inhibit gap junction (GJ) channels with specificity for Cx50 over its closely related homologue Cx46. Here, we examined the mechanism of BQ(+) action using undocked Cx46 and Cx50 hemichannels, which are more amenable to analyses at the single-channel level. We found that BQ(+) (300 µM-1 mM) robustly inhibited Cx50, but not Cx46, hemichannel currents, indicating that the Cx selectivity of BQ(+) is preserved in both hemichannel and GJ channel configurations. BQ(+) reduced Cx50 hemichannel open probability (P(o)) without appreciably altering unitary conductance of the fully open state and was effective when added from either extracellular or cytoplasmic sides. The reductions in P(o) were dependent on BQ(+) concentration with a Hill coefficient of 1.8, suggesting binding of at least two BQ(+) molecules. Inhibition by BQ(+) was voltage dependent, promoted by hyperpolarization from the extracellular side and conversely by depolarization from the cytoplasmic side. These results are consistent with binding of BQ(+) in the pore. Substitution of the N-terminal (NT) domain of Cx46 into Cx50 significantly impaired inhibition by BQ(+). The NT domain contributes to the formation of the wide cytoplasmic vestibule of the pore and, thus, may contribute to the binding of BQ(+). Single-channel analyses showed that BQ(+) induced transitions that did not resemble pore block, but rather transitions indistinguishable from the intrinsic gating events ascribed to loop gating, one of two mechanisms that gate Cx channels. Moreover, BQ(+) decreased mean open time and increased mean closed time, indicating that inhibition consists of an increase in hemichannel closing rate as well as a stabilization of the closed state. Collectively, these data suggest a mechanism of action for BQ(+) that involves modulation loop gating rather than channel block as a result of binding in the NT domain.
    The Journal of General Physiology 01/2012; 139(1):69-82. DOI:10.1085/jgp.201110678 · 4.79 Impact Factor
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    ABSTRACT: An estimated 10% to 15% of sudden infant death syndrome (SIDS) cases may stem from channelopathy-mediated lethal arrhythmias. Loss of the GJA1-encoded gap junction channel protein connexin43 is known to underlie formation of lethal arrhythmias. GJA1 mutations have been associated with cardiac diseases, including atrial fibrillation. Therefore, GJA1 is a plausible candidate gene for premature sudden death. GJA1 open reading frame mutational analysis was performed with polymerase chain reaction, denaturing high-performance liquid chromatography, and direct DNA sequencing on DNA from 292 SIDS cases. Immunofluorescence and dual whole-cell patch-clamp studies were performed to determine the functionality of mutant gap junctions. Immunostaining for gap junction proteins was performed on SIDS-associated paraffin-embedded cardiac tissue. Two rare, novel missense mutations, E42K and S272P, were detected in 2 of 292 SIDS cases, a 2-month-old white boy and a 3-month-old white girl, respectively. Analysis of the E42K victim's parental DNA demonstrated a de novo mutation. Both mutations involved highly conserved residues and were absent in >1000 ethnically matched reference alleles. Immunofluorescence demonstrated no trafficking abnormalities for either mutation, and S272P demonstrated wild-type junctional conductance. However, junctional conductance measurements for the E42K mutation demonstrated a loss of function not rescued by wild type. Moreover, the E42K victim's cardiac tissue demonstrated a mosaic immunostaining pattern for connexin43 protein. This study provides the first molecular and functional evidence implicating a GJA1 mutation as a novel pathogenic substrate for SIDS. E42K-connexin43 demonstrated a trafficking-independent reduction in junctional coupling in vitro and a mosaic pattern of mutational DNA distribution in deceased cardiac tissue, suggesting a novel mechanism of connexin43-associated sudden death.
    Circulation 12/2011; 125(3):474-81. DOI:10.1161/CIRCULATIONAHA.111.057224 · 14.43 Impact Factor
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    Comprehensive Physiology, 12/2010; , ISBN: 9780470650714
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    ABSTRACT: Mutations in GJB2, which encodes Cx26, are one of the most common causes of inherited deafness in humans. More than 100 mutations have been identified scattered throughout the Cx26 protein, most of which cause nonsyndromic sensorineural deafness. In a subset of mutations, deafness is accompanied by hyperkeratotic skin disorders, which are typically severe and sometimes fatal. Many of these syndromic deafness mutations localize to the amino-terminal and first extracellular loop (E1) domains. Here, we examined two such mutations, A40V and G45E, which are positioned near the TM1/E1 boundary and are associated with keratitis ichthyosis deafness (KID) syndrome. Both of these mutants have been reported to form hemichannels that open aberrantly, leading to "leaky" cell membranes. Here, we quantified the Ca(2+) sensitivities and examined the biophysical properties of these mutants at macroscopic and single-channel levels. We find that A40V hemichannels show significantly impaired regulation by extracellular Ca(2+), increasing the likelihood of aberrant hemichannel opening as previously suggested. However, G45E hemichannels show only modest impairment in regulation by Ca(2+) and instead exhibit a substantial increase in permeability to Ca(2+). Using cysteine substitution and examination of accessibility to thiol-modifying reagents, we demonstrate that G45, but not A40, is a pore-lining residue. Both mutants function as cell-cell channels. The data suggest that G45E and A40V are hemichannel gain-of-function mutants that produce similar phenotypes, but by different underlying mechanisms. A40V produces leaky hemichannels, whereas G45E provides a route for excessive entry of Ca(2+). These aberrant properties, alone or in combination, can severely compromise cell integrity and lead to increased cell death.
    The Journal of General Physiology 07/2010; 136(1):47-62. DOI:10.1085/jgp.201010433 · 4.79 Impact Factor
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    Silke B. Bodendiek · Clio Rubinos · Miduturu Srinivas · Heike Wulff
    Biophysical Journal 01/2010; 98(3). DOI:10.1016/j.bpj.2009.12.531 · 3.97 Impact Factor
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    ABSTRACT: Both connexins and signal transduction pathways have been independently shown to play critical roles in lens homeostasis, but little is known about potential cooperation between these two intercellular communication systems. To investigate whether growth factor signaling and gap junctional communication interact during the development of lens homeostasis, we examined the effect of mitogen-activated protein kinase (MAPK) signaling on coupling mediated by specific lens connexins by using a combination of in vitro and in vivo assays. Activation of MAPK signaling pathways significantly increased coupling provided by Cx50, but not Cx46, in paired Xenopus laevis oocytes in vitro, as well as between freshly isolated lens cells in vivo. Constitutively active MAPK signaling caused macrophthalmia, cataract, glucose accumulation, vacuole formation in differentiating fibers, and lens rupture in vivo. The specific removal or replacement of Cx50, but not Cx46, ameliorated all five pathological conditions in transgenic mice. These results indicate that MAPK signaling specifically modulates coupling mediated by Cx50 and that gap junctional communication and signal transduction pathways may interact in osmotic regulation during postnatal fiber development.
    Molecular biology of the cell 04/2009; 20(10):2582-92. DOI:10.1091/mbc.E08-12-1257 · 4.47 Impact Factor
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    ABSTRACT: Unapposed connexin hemichannels exhibit robust closure in response to membrane hyperpolarization and extracellular calcium. This form of gating, termed "loop gating," is largely responsible for regulating hemichannel opening, thereby preventing cell damage through excessive flux of ions and metabolites. The molecular components and structural rearrangements underlying loop gating remain unknown. Here, using cysteine mutagenesis in Cx50, we demonstrate that residues at the TM1/E1 border undergo movement during loop gating. Replacement of Phe(43) in Cx50 with a cysteine resulted in small or no appreciable membrane currents. Bath application of dithiothreitol or TPEN (N,N,N',N'-tetrakis(2-pyridylmethyl) ethylenediamine), reagents that exhibit strong transition metal chelating activity, led to robust currents indicating that the F43C substitution impaired hemichannel function, producing "lock-up" in a closed or poorly functional state due to formation of metal bridges. In support, Cd(2+) at submicromolar concentrations (50-100 nm) enhanced lock-up of F43C hemichannels. Moreover, lock-up occurred under conditions that favored closure, indicating that the sulfhydryl groups come close enough to each other or to other residues to coordinate metal ions with high affinity. In addition to F43C, metal binding was also found for G46C, and to a lesser extent, D51C substitutions, positions found to be pore-lining in the open state using the substituted-cysteine accessibility method, but not for A40C and A41C substitutions, which were not found to reside in the open pore. These results indicate that metal ions access the cysteine side chains through the open pore and that closure of the loop gate involves movement of the TM1/E1 region that results in local narrowing of the large aqueous connexin pore.
    Journal of Biological Chemistry 02/2009; 284(7):4484-93. DOI:10.1074/jbc.M807430200 · 4.57 Impact Factor
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    Biophysical Journal 02/2009; 96(3). DOI:10.1016/j.bpj.2008.12.1417 · 3.97 Impact Factor
  • Miduturu Srinivas
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    ABSTRACT: Connexin channels play a wide variety of roles in different cell types and tissues. Genetic and molecular approaches have proven useful for understanding the roles of these proteins in tissue function. Identification and characterization of specific and high-affinity inhibitors of these channels would greatly assist investigation of their physiological function; however, progress in this area has been slow. Nevertheless, recent studies have identified a number of small molecules and peptides that inhibit connexin channels. Although the specificity of these new drugs for connexin channels remains problematic, several of these reagents inhibit channels in an isoform-specific manner and do so with reasonable potency. These reagents are likely to be useful for acute studies that investigate the physiological roles of different connexin channels. In addition, some agents appear to bind within the permeability pathway and may be useful in structure-function studies of the pore. KeywordsBlockers-Carbenoxolone-Polyamines-Fenamates-2-Aminoethoxydiphenyl borate-Octanol-Oleamide-Halothane-Cyclodextrin-Glycyrrhetinic acid-Quinine-Mefloquine-Cx26-Cx32-Cx35-Cx36-Cx37-Cx40-Cx43-Cx46-Cx50
    Connexins, 12/2008: pages 207-224;
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    Vytas K Verselis · Miduturu Srinivas
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    ABSTRACT: Connexin hemichannels are robustly regulated by voltage and divalent cations. The basis of voltage-dependent gating, however, has been questioned with reports that it is not intrinsic to hemichannels, but rather is derived from divalent cations acting as gating particles that block the pore in a voltage-dependent manner. Previously, we showed that connexin hemichannels possess two types of voltage-dependent gating, termed V(j) and loop gating, that in Cx46 operate at opposite voltage polarities, positive and negative, respectively. Using recordings of single Cx46 hemichannels, we found both forms of gating persist in solutions containing no added Mg(2+) and EGTA to chelate Ca(2+). Although loop gating persists, it is significantly modulated by changing levels of extracellular divalent cations. When extracellular divalent cation concentrations are low, large hyperpolarizing voltages, exceeding -100 mV, could still drive Cx46 hemichannels toward closure. However, gating is characterized by continuous flickering of the unitary current interrupted by occasional, brief sojourns to a quiet closed state. Addition of extracellular divalent cations, in this case Mg(2+), results in long-lived residence in a quiet closed state, suggesting that hyperpolarization drives the hemichannel to close, perhaps by initiating movements in the extracellular loops, and that divalent cations stabilize the fully closed conformation. Using excised patches, we found that divalent cations are only effective from the extracellular side, indicative that the binding site is not cytoplasmic or in the pore, but rather extracellular. V(j) gating remains essentially unaffected by changing levels of extracellular divalent cations. Thus, we demonstrate that both forms of voltage dependence are intrinsic gating mechanisms in Cx46 hemichannels and that the action of external divalent cations is to selectively modulate loop gating.
    The Journal of General Physiology 10/2008; 132(3):315-27. DOI:10.1085/jgp.200810029 · 4.79 Impact Factor
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    Thomas W White · Yang Gao · Leping Li · Caterina Sellitto · Miduturu Srinivas
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    ABSTRACT: Gap junctions between epithelial cells are essential for normal lens growth. In mice, knockout of Cx50 or targeted replacement of Cx50 with Cx46 (knockin) caused smaller lenses because of decreased epithelial cell proliferation. However, it remains unclear whether Cx50 functionally contributes to lens epithelial coupling during maximal proliferation on postnatal day 2 (P2) and P3. To determine which connexins functionally contribute to epithelial cell coupling and proliferation, junctional coupling from epithelial cells of wild-type and knockin mice was examined. Epithelial cells were isolated from wild-type or knockin mice at different developmental ages. Junctional currents were measured by dual whole cell voltage clamp. Cell proliferation was assayed by BrdU incorporation. Connexins were immunolocalized using specific antibodies. Junctional currents between lens epithelial cells exhibited a developmentally regulated sensitivity to quinine, a drug that blocks Cx50 gap junctions, but not Cx43 or Cx46. Single-channel currents had a unitary conductance of 210 pS, typical of Cx50. Immunocytochemical staining showed Cx43 and Cx50 were abundantly expressed in wild-type cells, and Cx46 replaced Cx50 in knockin cells. A correlation between functional activity of Cx50 and maximal proliferation was also found. In epithelial cells from P3 wild-type mice, there was a high density of BrdU-labeled nuclei in both the central epithelium and the equatorial epithelium, and 60% or more of total coupling was provided by Cx50. In older cells, proliferation was greatly reduced, and the contribution of Cx50 to total coupling was progressively reduced (45% or less on P12; 25% or less on P28). Coupling between epithelial cells of Cx46 knockin mice was similar in magnitude to that of wild-type mice but had pharmacologic and biophysical characteristics of Cx46. This functional replacement of Cx50 with Cx46 was correlated with 71% and 13% reductions in BrdU-labeled cells in the P3 central epithelium and equatorial epithelium, respectively. These results reconcile previous genetic studies showing that Cx50 influences epithelial cell proliferation, with numerous studies suggesting that Cx43 was the principal epithelial cell connexin. They further show that the contribution of Cx50 is highest during peak postnatal proliferation but progressively declines with age thereafter.
    Investigative Ophthalmology &amp Visual Science 01/2008; 48(12):5630-7. DOI:10.1167/iovs.06-1540 · 3.40 Impact Factor

Publication Stats

2k Citations
197.48 Total Impact Points


  • 2005–2014
    • State University of New York College of Optometry
      New York, New York, United States
  • 2006
    • State University of New York
      New York City, New York, United States
  • 1999–2006
    • Albert Einstein College of Medicine
      • "Dominick P. Purpura" Department of Neuroscience
      New York City, NY, United States
  • 2001
    • Sapporo Medical University
      • School of Medicine
      Sapporo, Hokkaidō, Japan
  • 2000
    • Ruhr-Universität Bochum
      Bochum, North Rhine-Westphalia, Germany
    • Harvard Medical School
      • Department of Neurobiology
      Boston, Massachusetts, United States