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M Srinivas

Publications and source records attributed to M Srinivas.

At least 55 records · Page 3Linked to original sources
PubMed2001

Growth-suppressive function of human connexin32 in a conditional immortalized mouse hepatocyte cell line.

Mouse hepatocytes immortalized with a temperature-sensitive allele of the SV40 large T-antigen (CHST8 cells) were found to lack the high expression of the gap junction proteins Cx26 and Cx32 that characterizes normal mouse hepatocytes, expressing instead Cx43 and Cx45 at minimal levels. In order to examine the growth suppressive function of Cx32 on hepatocytes, we transfected these CHST8 cells with human Cx32 complementary deoxyribonucleic acid and measured the growth rates at 33, 37, and 39 degrees C. Expression of human Cx32 and its messenger ribonucleic acid in the stable cell lines was confirmed by immunocytochemistry and by Western and Northern blots analyses. Dye transfer following lucifer yellow injection into the transfectants was extensive; Cx32 channels displayed unitary conductances of about 70 pS and were moderately voltage sensitive. When cultured at 33 and 39 degrees C, growth rates of both parental cells and transfectants were of the same level. When examined at 37 degrees C, growth rate of the transfectant, which highly expressed Cx32 at the membranes, was significantly decreased compared to the parental cells. However, no changes in the expression of Cx32 protein in the transfectants were observed between 33 and 37 degrees C. These results suggest that Cx32 expression could inhibit hepatocyte growth in vitro using the conditional immortalized cells. Cx32 transfectants using a conditional immortalized mouse hepatocyte may be useful for examining the mechanisms of growth and differentiation in hepatocytes by gap junction expression.

Animals↗
PubMed2001

Identification and functional expression of HCx31.9, a novel gap junction gene.

By combining in silico and bench molecular biology methods we have identified a novel human gap junction gene that encodes a protein designated HCx31.9. We have determined its human chromosomal location and gene structure, and we have identified a putative mouse ortholog, mCx30.2. We have observed the presence of HCx31.9 in human cerebral cortex, liver, heart, spleen, lung, and kidney and the presence of mCx30.2 in mouse cerebral cortex, liver and lung. Moreover, preliminary data on the electrophysiological properties of HCx31.9 have been obtained by functional expression in paired Xenopus oocytes and in transfected N2A cells.

Amino Acid Sequence↗
PubMed2001

Blindness in the Indian state of Andhra Pradesh.

PURPOSE: To determine the current prevalence and causes of blindness in the Indian state of Andhra Pradesh to assess if blindness has decreased since the last survey of 1986-1989. METHODS: A population-based epidemiology study, using a stratified, random, cluster, systematic sampling strategy, was conducted in the state of Andhra Pradesh in India. Participants of all ages (n = 10,293), 87.3% of the 11,786 eligible, from 94 clusters in one urban and three rural areas representative of the population of Andhra Pradesh, underwent interview and a detailed dilated ocular evaluation by trained professionals. Blindness was defined as presenting distance visual acuity < 6/60 or central visual field < 20(o) in the better eye. RESULTS: Two hundred seventy-five participants were blind, a prevalence of 1.84% (95% confidence interval, 1.49%-2.19%) when adjusted for the age, sex, and urban-rural distribution of the population in 2000. The causes of this blindness were easily treatable in 60.3% (cataract, 44%; refractive error, 16.3%). Preventable corneal disease, glaucoma, complications of cataract surgery, and amblyopia caused another 19% of the blindness. Blindness was more likely with increasing age and decreasing socioeconomic status, and in female subjects and in rural areas. Among the 76 million population of Andhra Pradesh, 714,400 are estimated to have cataract-related blindness (615,600 cataract, 53,200 cataract surgery-related complications, 45,600 aphakia), and 228,000 refractive error-related blindness (159,600 myopia, 22,800 hyperopia, 45,600 refractive error-related amblyopia). If 95% of the cataract and refractive error blindness in Andhra Pradesh had been treated effectively, 3.4 and 7.4 million blind-person-years, respectively, could have been prevented. If 90% of the blindness due to preventable corneal disease and glaucoma had been prevented, another 2.7 million blind-person-years could have been prevented. CONCLUSIONS: The prevalence of blindness in this Indian state has increased from 1.5% in the late 1980s to 1.84% currently, as against the target of the National Program for Control of Blindness to reduce the prevalence to 0.3% by 2000. The number of people with cataract-related blindness has not reduced even with the eye care policy focus on cataract. Reduction of blindness in India will require strategies that are more effective than those that have been pursued so far.

Adolescent↗
PubMed2001

Febrile seizures.

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Child, Preschool↗
PubMed2001

Effect of metronidazole on spermatogenesis and FSH, LH and testosterone levels of pre-pubertal rats.

Metronidazole, a 5-nitroimidazole drug has been reported to decrease testicular weight, testicular and epididymal spermatid counts and causes abnormal sperm morphology with degeneration of seminiferous tubules with 6 weeks treatment of metronidazole (400 mg/kg, day). In contrast to DNA flow cytometry (FCM), the histological and gravimetric parameters do not allow a rapid, sensitive, objective and multiparameteric evaluation of reproductive toxicants on spermatogenesis. Moreover, the exact mechanisms for such an effect are not entirely clear. The present study was therefore undertaken to assess the effects of intraperitoneal (i.p.) administration of metronidazole 400 mg/kg daily for 30 days on testicular germ cell changes assessed by DNA (FCM) and hormone levels of testosterone, FSH and LH in pre-pubertal rats. A significant reduction in the haploid cell population in metronidazole treated groups as compared to saline treated controls was observed. The mean serum FSH, LH and testosterone value were also lowered in treated animals. Thus, the spermatotoxic effects of metronidazole were probably mediated by decrease in the circulating hormones responsible for spermatogenesis.

Animals↗
PubMed2000

Molecular and functional diversity of neural connexins in the retina.

Electrical synapses (gap junctions) in neuronal circuits have become a major focus in the study of network properties such as synchronization and oscillation (Galarreta and Hestrin, 1999; Gibson et al., 1999). Despite the recent progress made in unraveling the contribution of gap junctions to network behavior, little is known about the molecular composition of the junctional constituents. By cloning gap junction proteins [connexins (Cxs)] from zebrafish retina and through functional expression, we demonstrate that the retina possesses a high degree of connexin diversity, which may account for differential functional properties of electrical synapses. Three new Cxs, designated as zebrafish Cx27.5 (zfCx27.5), zfCx44.1, and zfCx55.5, and the carp ortholog of mammalian Cx43 were cloned. By in situ hybridization and in situ RT-PCR, we demonstrate that the four fish connexin mRNAs show differential localization in the retina. Transient functional expression in paired Xenopus oocytes and in the neuroblastoma N2A cell line indicate an extreme range of electrophysiological properties of these connexins in terms of voltage dependence and unitary conductance. For instance, the new zfCx44.1 exhibited high sensitivity to voltage-induced closure with currents decaying rapidly for transjunctional potentials >10 mV, whereas zfCx55.5 channels showed an opposite voltage dependence in response to voltage steps of either polarity. Moreover, although zfCx44.1 channels showed unitary conductance as high as any previously reported for junctional channels (nearly 300 pS), zfCx55. 5 and zfCx27.5 exhibited much lower unitary conductances (<60 pS).

Animals↗
PubMed2000

Gastric teratoma in children.

Gastric teratoma (GT) comprises less than 1% of all teratomas in children. Though GT in the presence of immature neuroepithelial elements is regarded as malignant, the prognosis is excellent after complete excision of the tumor. Because of its rarity the world literature lacks a large study. Clinical experience with ten cases of GT is presented and discussed. Only one patient was female; the mean age at presentation was 3.2 months. Two cases were immature grade III GT; one of these had infiltrated the left lobe of the liver and the transverse colon while the other had metastasized to the regional lymph nodes and omentum. All the patients underwent complete excision. There were no deaths, and after a mean follow-up period of 4.2 years, all the patients had no recurrence and were healthy. Both the mature and immature types of GT have an excellent prognosis after complete excision of the tumor. Even when the immature type infiltrates surrounding structures, complete excision offers recurrence-free survival without requiring chemo- or radiotherapy.

Biopsy↗
PubMed2000

Congenital anterior urethral diverticulum in children.

Congenital anterior urethral diverticulum (CAUD) is an uncommon condition in children usually presenting as a fluctuant ventral penile swelling. Retrospective data of nine patients with CAUD were analyzed. Patients presented with penile swelling (n = 7), recurrent urinary tract infection (UTI) (n = 5), and poor urinary stream (n = 2). One patient who had gross pyuria was treated with initial marsupialization of the diverticulum and later underwent a definitive surgical procedure. One had deranged renal function, grade IV vesicourethral reflux (VUR), and UTI and was treated with initial urinary diversion followed later by excision of the diverticulum and urethral reconstruction. Of the seven patients who underwent primary repair of the diverticulum, all except one had a normal urethrogram on follow-up. One patient developed a stricture of the urethra that was treated with dilatations and is presently asymptomatic. In the two patients who had bilateral VUR, one grade III and the other grade IV, preoperatively, the reflux subsided and did not require antireflux surgery. Only one patient is on low-dose urinary antibiotic prophylaxis and presently has grade II reflux. Primary excision and repair is the preferred mode of treatment for CAUD. CAUD producing obstructive uropathy and VUR can be managed conservatively. In the presence of gross pyuria, marsupialization of the diverticulum followed by definitive surgery at a later date is safe and recommended.

Diverticulum↗
PubMed2000

Properties of gap junction channels formed by Cx46 alone and in combination with Cx50.

Gap junctions formed of connexin46 (Cx46) and connexin50 (Cx50) in lens fiber cells are crucial for maintaining lens transparency. We determined the functional properties of homotypic Cx46, heterotypic Cx46/Cx50, and heteromeric Cx46/Cx50 channels in a communication-deficient neuroblastoma (N2A) cell line, using dual whole-cell recordings. N2A cultures were stably and/or transiently transfected with Cx46, Cx50, and green fluorescent protein (EGFP). The macroscopic voltage sensitivity of homotypic Cx46 conformed to the two-state model (Boltzmann parameters: G(min) = 0.11, V(0) = +/- 48.1 mV, gating charge = 2). Cx46 single channels showed a main-state conductance of 140 +/- 8 pS and multiple subconductance states ranging from < or =10 pS to 60 pS. Conservation of homotypic properties in heterotypic Cx46/Cx50 cell pairs allowed the determination of a positive relative gating polarity for the dominant gating mechanisms in Cx46 and Cx50. Observed gating properties were consistent with a second gating mechanism in Cx46 connexons. Moreover, rectification was observed in heterotypic cell pairs. Some cell pairs in cultures simultaneously transfected with Cx46 and Cx50 exhibited junctional properties not observed in other preparations, suggesting the formation of heteromeric channels. We conclude that different combinations of Cx46 and Cx50 within gap junction channels lead to unique biophysical properties.

Animals↗
PubMed2000

Angle-closure glaucoma in an urban population in southern India. The Andhra Pradesh eye disease study.

OBJECTIVE: To assess the prevalence and features of angle-closure glaucoma (ACG) in an urban population in southern India. DESIGN: A population-based, cross-sectional study. PARTICIPANTS: A total of 2522 (85.4% of those eligible) persons of all ages, including 1399 persons 30 years of age or older, from 24 clusters representative of the population of Hyderabad city. TESTING: The participants underwent an interview and detailed eye examination that included logarithm of minimum angle of resolution visual acuity, refraction, slit-lamp biomicroscopy, applanation tonometry, and gonioscopy; pupil dilatation and stereoscopic fundus evaluation was performed if the risk of angle-closure as a result of dilatation was not believed to be imminent. Humphrey threshold 24-2 visual fields (Humphrey Instruments Inc., San Leandro, CA) were performed when indicated by standardized criteria for disc damage or if intraocular pressure (IOP) was 22 mmHg or more. MAIN OUTCOME MEASURES: An occludable angle was defined as pigmented posterior trabecular meshwork not visible by gonioscopy in three quarters or more of the angle circumference. Manifest primary angle-closure glaucoma (PACG) was defined as IOP of 22 mmHg or more or glaucomatous optic disc damage with visual field loss in the presence of an occludable angle. An IOP of 22 mmHg or more or glaucomatous optic disc damage in the presence of an occludable angle secondary to an obvious cause was defined as secondary ACG. RESULTS: Manifest PACG and occludable angles without ACG were present in 12 and 24 participants, respectively, with age- and gender-adjusted prevalence (95% confidence interval [CI]) of 0.71% (0.34%-1.31%) and 1.41% (0.73%-2.09%) in participants 30 years of age or older, and 1.08% (0.36%-1.80%) and 2.21% (1.15%-3.27%) in participants 40 years of age or older, respectively. With multivariate analysis, the prevalence of these two conditions considered together increased significantly with age (P < 0.001); although not statistically significant, these were more common in females (odds ratio 1.70; 95% CI, 0.82-3.54) and in those belonging to lower socioeconomic strata as compared with middle and upper strata (odds ratio, 1.82; 95% CI, 0.88-3.74). The odds of manifest PACG were higher in the presence of hyperopia of more than 2 diopters ([D]; odds ratio, 3.69; 95% CI, 0.89-15.2). Only four of 12 participants (33.3%) with manifest PACG had been previously diagnosed, and one of 12 (8.3%) had peripheral iridotomy performed previously. Manifest PACG had caused blindness in one or both eyes in five of these 12 participants (41.7%); best-corrected distance visual acuity less than 20/400 in one or both eyes in four patients, and acuity less than 20/200 in one eye in another patient. Most (83.3%) of those with manifest PACG could be classified as having chronic form of the disease. We may have underestimated manifest PACG because visual fields were per- formed only on those with clinical suspicion of optic disc damage. Secondary ACG was present in two participants. CONCLUSIONS: The prevalence of PACG in this urban population in southern India is close to that reported recently in a Mongolian population. A large proportion of the PACG in this population was undiagnosed and untreated. Because visual loss resulting from PACG is potentially preventable if peripheral iridotomy or iridectomy is performed in the early stage, strategies for early detection of PACG could reduce the high risk of blindness resulting from PACG seen in this urban population in India.

Adolescent↗
PubMed2000

Open-angle glaucoma in an urban population in southern India: the Andhra Pradesh eye disease study.

OBJECTIVE: To assess the prevalence and features of open-angle glaucoma in an urban population in southern India. DESIGN: A population-based cross-sectional study. PARTICIPANTS: A total of 2522 persons (85.4% of those eligible) of all ages, including 1399 persons 30 years of age or older, from 24 clusters representative of the population of Hyderabad city. TESTING: The participants underwent an interview and detailed eye examination that included logarithm of minimum angle of resolution visual acuity, refraction, slit-lamp biomicroscopy, applanation tonometry, gonioscopy, dilatation, cataract grading, and stereoscopic fundus evaluation. Automated Humphrey threshold 24-2 visual fields (Humphrey Instruments Inc., San Leandro, CA) and optic disc photography were performed when indicated by standardized criteria for disc damage or if intraocular pressure (IOP) was 22 mmHg or more. MAIN OUTCOME MEASURES: Definite primary open-angle glaucoma (POAG) was defined as obvious glaucomatous optic disc damage and visual field loss in the presence of an open-angle, and suspected POAG was defined as suspected glaucomatous optic disc damage without definite visual field loss. Ocular hypertension (OHT) was defined as IOP of 22 mmHg or more without glaucomatous optic disc damage or visual field loss in the presence of an open-angle. Glaucomatous optic disc damage or IOP of 22 mmHg or more secondary to an obvious cause and with an open-angle was defined as secondary open-angle glaucoma. RESULTS: Definite POAG, suspected POAG, and OHT were present in 27, 14, and 7 participants, respectively, with age- and gender-adjusted prevalence (95% confidence interval) of 1.62% (0.77%-2.48%), 0.79% (0.39%-1.41%), and 0.32% (0.10%-0.78%) in those 30 years of age or older, and 2.56% (1.22%-3.91%), 1.11% (0.43%-1.78%), and 0.42% (0.11%-1.12%) in those 40 years of age or older, respectively. The prevalence of POAG increased significantly with age using multivariate analysis (P < 0.001). Only two of 27 participants (7.4%) with definite POAG had been previously diagnosed and treated, and 66.7% of the previously undiagnosed had IOP less than 22 mmHg. Fourteen of 27 participants (51.9%) with definite POAG had severe glaucomatous damage based on optic disc and visual field criteria, of which five participants (18.5%) had at least one blind eye as a result of POAG (all with best-corrected distance visual acuity less than 20/400 or central visual field less than 10 degrees); the other 13 participants (48.1%) had moderate glaucomatous damage. Because visual fields and optic disc photography were not performed on all participants, the prevalence of POAG may have been underestimated. Secondary open-angle glaucoma was present in one participant as a result of angle recession. CONCLUSIONS: The prevalence of open-angle glaucoma in this urban population in southern India is at least as much as that reported recently from white populations in developed countries. However, the vast majority of persons with glaucoma were undiagnosed in this population, and a large proportion of those having definite POAG already had severe glaucomatous damage.

Adolescent↗
PubMed2000

Temporal expression of neuronal connexins during hippocampal ontogeny.

Communication through gap junction channels provides a major signaling mechanism during early brain histogenesis, a developmental time during which neural progenitor cells are inexcitable and do not express ligand-gated channel responses to the major CNS neurotransmitters. Expression of different gap junction types during neurogenesis may therefore define intercellular pathways for transmission of developmentally relevant molecules. To better understand the molecular mechanism(s) by which growth and differentiation of neurons are modulated by gap junction channels, we have been examining the developmental effects of a specific set of cytokines on differentiation and gap junction expression in a conditionally immortalized mouse embryonic hippocampal neuronal progenitor cell line (MK31). When multipotent MK31 cells are in an uncommitted state, they uniformly express the neuroepithelial intermediate filament class VI marker, nestin, are strongly coupled by gap junctions composed of connexin43 (Cx43) and express connexin45 (Cx45) at the mRNA level. As these cells undergo neuronal lineage commitment and exit from cell cycle, they begin to express the early neurofilament marker, NF66, and coupling strength and expression of Cx43 begin to decline with concurrent expression of other connexin proteins, including Cx26, Cx33, Cx36, Cx40 and Cx45. Terminal neuronal differentiation is heralded by the expression of more advanced neurofilament proteins, increased morphologic maturation, the elaboration of inward currents and action potentials that possess mature physiological properties, and changing profiles of expression of connexin subtypes, including upregulation of Cx36 expression. These important developmental transitions are regulated by a complex network of cell cycle checkpoints. To begin to examine the precise roles of gap junction proteins in traversing these developmental checkpoints and in thus regulating neurogenesis, we have focused on individual members of two classes of genes involved in these seminal events: ID (inhibitor of differentiation)-1 and GAS (growth arrest-specific gene)5. When MK31 cells were maintained in an uncommitted state, levels of ID-1 mRNA were high and GAS5 transcripts were essentially undetectable. Application of cytokines that promote neuronal lineage commitment and cell cycle exit resulted in down-regulation of ID-1 and upregulation of GAS5 transcripts, whereas additional cytokine paradigms that promoted terminal neuronal differentiation resulted in the delayed down-regulation of GAS5 mRNA. Stable MK31 transfectants were generated for ID-1 and GAS5. In basal conditions, cellular proliferation was enhanced in the ID-1 transfectants and inhibited in the GAS5 transfectants when compared with control MK31 cells. When cytokine-mediated neurogenesis was examined in these transfected cell lines, constitutive expression of ID-1 inhibited and constitutive expression of GAS5 enhanced initial and terminal stages of neuronal differentiation, with evidence that terminal neuronal maturation in both transfectant lines was associated with decreased cellular viability, possibly due to the presence of conflicting cell cycle-associated developmental signals. These experimental reagents will prove to be valuable experimental tools to help define the functional interrelationships between changing profiles of connexin protein expression and cell cycle regulation during neuronal ontogeny in the mammalian brain. The present review summarizes the current state of research involving the temporal expression of such connexin types in differentiating hippocampal neurons and speculates on the possible role of these intercellular channels in the development and plasticity of the nervous system. In addition, we describe the functional properties and expression pattern of the newly discovered neuronal-specific gap junctional protein, Cx36, in the developing mouse fetal hippocampus and in the rat retina and brain.

Animals↗
PubMed2000

Ocular trauma in an urban population in southern India: the Andhra Pradesh Eye Disease Study.

PURPOSE: To assess the cumulative prevalence of ocular trauma and presence of vision loss due to ocular trauma in an urban population in southern India. METHODS: As part of the population-based Andhra Pradesh Eye Disease Study, 2522 people of all ages from 24 clusters representative of the population of Hyderabad city in southern India, underwent a detailed interview and standardized dilated ocular evaluation. An eye was considered to be blind due to trauma if best corrected distance visual acuity was worse than 6/60 due to trauma. RESULTS: One hundred and thirteen subjects gave a history of ocular trauma and another two had evidence of ocular trauma by examination, a combined age sex-adjusted rate of 3.97% (95% CI 2.52-5.42%). Blindness in one eye due to trauma was present in 17 subjects, and in both eyes in one subject, a combined age-sex-adjusted prevalence of 0.60% (95% CI 0.23-1.04%). Visual acuity in all the blind eyes except one was worse than 3/60. With multiple logistic regression, the odds of blindness in at least one eye due to trauma were highest for current age range of 30-39 years (odds ratio 6.33, 95% CI 1.69-23.77 compared with a current age of less than 30 years), were significantly higher for lower socioeconomic status (3.74, 95% CI 1.18 -11.84), and were higher for males (2.48, 95% CI 0.91-6.82) though this did not reach statistical significance. Trauma resulting in blindness had occurred by the age of 15 years in 55% of subjects, and before the age of 40 years in 92.1% of subjects; this had occurred most commonly while playing (53.6% of the cases). With multiple logistic regression, the odds ratios for any ocular trauma were significantly higher for males (2.10, 95% CI 1.40-3.15), and for labourers than for other occupations (2.50, 95% CI 1.62-3.86). CONCLUSIONS: Ocular trauma affects one in 25 people in this urban population in India, and one in 167 people in this population are estimated to be blind in at least one eye due to trauma. The majority of the trauma resulting in blindness occurs during childhood and young adulthood, and slightly more than half occurs while playing. Targeting mothers and children of lower socioeconomic strata in eye health awareness strategies to reduce blindness due to trauma needs to be considered in urban India.

Adolescent↗
PubMed2000

Unilateral visual impairment in an urban population in southern India.

This study assessed the prevalence and causes of unilateral visual impairment in the urban population of Hyderabad city as part of the Andhra Pradesh Eye Disease Study. Stratified, random, cluster, systematic sampling was used to select 2,954 subjects from 24 clusters representative of the population of Hyderabad. Eligible subjects underwent detailed eye examination including logMAR visual acuity, refraction, slitlamp biomicroscopy, applanation tonometry, gonioscopy, dilatation, cataract grading, and stereoscopic evaluation of fundus. Automated threshold visual fields and slitlamp and fundus photography were done when indicated by standardised criteria. Unilateral visual impairment was defined as presenting distance visual acuity < 6/18 in the worse eye and > or = 6/12 in the better eye, which was further divided into unilateral blindness (visual acuity < 6/60 in the worse eye) and unilateral moderate visual impairment (visual acuity < 6/18-6/60 in the worse eye). A total of 2,522 subjects (85.4% of eligible), including 1,399 > or = 30 years old, participated in the study. In addition to the 1% blindness and 7.2% moderate visual impairment (based on bilateral visual impairment criteria) reported earlier in this sample, 139 subjects had unilateral visual impairment, an age-gender-adjusted prevalence of 3.8% (95% confidence interval 2.7-4.9%). The major causes of this visual impairment 39.9% were refractive error (42.9%), cataract (14.4%), corneal disease (11.5%), and retinal disease (11.2%). Of this unilateral visual impairment was blindness. The major causes of unilateral blindness were corneal disease (23.2%), cataract (22.5%), retinal disease (18%), and optic atrophy (12.9%). On the other hand, the predominant cause of unilateral moderate visual impairment was refractive error (67%) followed by cataract (9%). Of the total unilateral visual impairment, 34.3% was present in those < 30 years old and 36.2% in those 30-49 years old. Unilateral visual impairment afflicts approximately 1 in 25 persons in this urban population. A large proportion of this unilateral visual impairment is present in younger age groups. The causes of unilateral visual impairment, like those of bilateral visual impairment in this population, are varied, suggesting therefore, that in addition to the current focus of eye care in India predominantly on cataract, other causes of visual impairment need to be addressed as well.

Adolescent↗
PubMed1999

Functional properties of channels formed by the neuronal gap junction protein connexin36.

The expression and functional properties of connexin36 (Cx36) were examined in two communication-deficient cell lines (N2A-neuroblastoma and PC-12 cells) transfected with Cx36 and in hippocampal neurons that express the connexin endogenously. Transfected cells expressed the expected 2.9 kb Cx36 transcript and Cx36 immunoreactivity, whereas nontransfected cells were devoid of Cx36. The relationship between steady-state junctional conductance (g(j)) and transjunctional voltage was well described by a two-state Boltzmann equation. The half-inactivation voltage (V(0)), the ratio of minimal to maximal g(j) (g(min)/g(max)), and the equivalent gating charge were +/- 75 mV, 0.55, and 1.75, respectively, indicating that Cx36 exhibits very low voltage sensitivity. Conductance of single Cx36 channels measured with patch pipettes containing 130 mM CsCl was 10-15 pS (n = 15 cell pairs); despite this low unitary conductance, Cx36 channels were permeable to the dye Lucifer yellow. Hippocampal neurons expressed Cx36 both in vivo and in culture. The electrophysiological properties of channels in cultured hippocampal neurons were similar to those of the channels expressed by the transfected cell lines, and the neuronal channels were similarly permeable to Lucifer yellow. The unique combination of weak voltage sensitivity, small unitary conductance, and permeation by anions as large as second messenger molecules endows Cx36 gap junction channels with properties well suited for mediating flexible electrical and biochemical interactions between neurons.

Animals↗
PubMed1999

Voltage dependence of macroscopic and unitary currents of gap junction channels formed by mouse connexin50 expressed in rat neuroblastoma cells.

1. The macroscopic and single channel gating characteristics of connexin (Cx) 50 gap junction channels between pairs of N2A neuroblastoma cells transfected with mouse Cx50 DNA were investigated using the dual whole-cell voltage clamp technique. 2. The macroscopic junctional current (Ij) of Cx50-transfected cells decayed exponentially with time in response to transjunctional voltage (Vj) steps (time constant (tau) of approximately 4 s at a Vj of 30-40 mV and 100-200 ms at a Vj of 80-100 mV). The steady-state junctional conductance (gj) was well described by a two-state Boltzmann equation. The half-inactivation voltage (V0), the ratio of minimal to maximal gj (gmin/gmax) and the equivalent gating charge were +/- 37 mV, 0.21 and 4, respectively. 3. The conductance of single Cx50 channels measured using patch pipettes containing 130 mM CsCl was 220 +/- 13.1 pS (12 cell pairs). A prominent residual or subconductance state corresponding to 43 +/- 4. 2 pS (10 cell pairs) was also observed at large Vj s. 4. The relationship between channel open probability (Po) and Vj was well described by a Boltzmann relationship with parameters similar to those obtained for macroscopic gj (V0 = 34 mV, gating charge = 4.25, maximum P= 0.98). The ensemble average of single channel currents at Vj = 50 mV declined in a monoexponential manner (tau = 905 ms), a value similar to the decline of the macroscopic Ij of Cx50 channels at the same voltage. 5. Ion substitution experiments indicated that Cx50 channels have a lower permeability to anions than to cations (transjunctional conductance of KCl vs. potassium glutamate (gammaj, KCl/gammaj,KGlut), 1.2; 6 cell pairs). 6. The results have important implications for understanding the role of connexins in tissues where Cx50 is a major gap junction component, including the lens.

Animals↗