GJA1 rs2071165 A > G Variant Increased Gastric Cancer Risk in Females of Northwest China: A Case-Control Study

Gastric cancer (GC) is one of the most common malignancies, and its incidence rates vary widely between men and women. Previous studies have suggested that connexin 43 (Cx43, encoded by gap junction protein alpha 1 (GJA1)) and secretory carrier membrane protein 1 (SCAMP1) are key functional proteins in tumors. Herein, the association between GJA1 and SCAMP1 polymorphisms and GC susceptibility and prognosis was evaluated. A total of three single-nucleotide polymorphisms among 681GC patients and 756 controls were tested using the Agena MassARRAY RS1000 system, including GJA1 rs2071165, SCAMP1 rs4530741, and SCAMP1 rs6874309. The strength of the association with GC risk was assessed by the odds ratios (ORs) and 95% confidence intervals (CIs) generated from the logistic regression model. Kaplan–Meier curve, long-rank tests, and a multivariate Cox proportional hazard model were used for prognosis analysis. The expression of GJA1 was assessed by immunohistochemistry. The GJA1 rs2071165 AA/AG genotype significantly increased the risk of GC in the female Chinese population (OR = 1.55, 95% CI = 1.03–2.32, p = 0.034 ). Furthermore, the risk effect of GJA1 rs2071165 was more evident in the subgroups of female patients with GC, stratified by age, clinical stage, tumor size, and recurrence/metastasis. However, no obvious differences in Cx43 expression in GC tissues were observed between males and females. Furthermore, no significant association between SCAMP1 rs4530741 and rs6874309 polymorphisms and GC risk or prognosis was observed. In conclusion, this study suggests for the first time that the GJA1 rs2071165 polymorphism is associated with increased GC risk in females, revealing a potential new clinical marker for assessing GC risk in females.

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GJA1 rs2071165 A>G Variant Increased Gastric Cancer Risk in Females of Northwest China: A Case-control Study

10.21203/rs.3.rs-115856/v1 ◽

2020 ◽

Author(s):

lijuan yuan ◽

Gang Wei ◽

Ping Yang ◽

Jianguo Lu ◽

Shujia Peng ◽

...

Keyword(s):

Gastric Cancer ◽

Cancer Risk ◽

Clinical Stage ◽

Northwest China ◽

Incidence Rates ◽

Cx43 Expression ◽

Cox Proportional Hazard ◽

Kaplan Meier ◽

Risk Effect ◽

First Time

Abstract Background: Gastric cancer (GC) is one of the most common malignancies and its incidence rates vary widely between men and women. Previous studies have suggested that Cx43 and SCAMP1 are key functional proteins in tumors. Herein, the association between GJA1 and SCAMP1 polymorphisms and GC susceptibility and prognosis was evaluated.Methods: A total of three SNPs among 681 GC patients and 756 controls were tested using the Agena Mass ARRAY RS1000 system including GJA1 rs2071165, SCAMP1 rs4530741, and rs6874309. The strength of the association with cancer risk was assessed by the odds ratios and 95% confidence intervals generated from the logistic regression model. Kaplan-Meier Curve, long-rank tests, and a multivariate Cox proportional hazard model were used for prognosis analysis. The expression of GJA1 was assessed by immunohistochemistry.Results: The GJA1 rs2071165 AA/AG genotype significantly increased the risk of GC in the female Chinese population (odds ratio [OR] =1.55, 95% confidence interval, 95%CI] = 1.03–2.32, p = 0.034). Furthermore, the risk effect of rs2071165 was more evident in the subgroups of female patients with GC, stratified by age, clinical stage, tumor size, and recurrence/metastasis. However, no obvious differences in Cx43 expression in GC tissues were observed between males and females. Furthermore, no significant association between rs4530741 and rs6874309 polymorphisms in SCAMP1 and GC risk or prognosis was observed. Conclusion: In conclusion, this study suggests that GJA1 rs2071165 polymorphisms are associated with increased GC risk in females for the first time, which showed a potential new clinical marker for assessing GC risk in females.

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Phosphorylation of connexin 43 induced by traumatic brain injury promotes exosome release

Journal of Neurophysiology ◽

10.1152/jn.00654.2017 ◽

2018 ◽

Vol 119 (1) ◽

pp. 305-311 ◽

Cited By ~ 12

Author(s):

Wei Chen ◽

Yijun Guo ◽

Wenjin Yang ◽

Lei Chen ◽

Dabin Ren ◽

...

Keyword(s):

Traumatic Brain Injury ◽

Brain Injury ◽

Gap Junction ◽

Connexin 43 ◽

Hippocampal Slices ◽

Long Term Potentiation ◽

Cx43 Expression ◽

Term Potentiation ◽

Junction Protein

Traumatic brain injury (TBI) caused by the external force leads to the neuronal dysfunction and even death. TBI has been reported to significantly increase the phosphorylation of glial gap junction protein connexin 43 (Cx43), which in turn propagates damages into surrounding brain tissues. However, the neuroprotective and anti-apoptosis effects of glia-derived exosomes have also been implicated in recent studies. Therefore, we detected whether TBI-induced phosphorylation of Cx43 would promote exosome release in rat brain. To generate TBI model, adult male Sprague-Dawley rats were subjected to lateral fluid percussion injury. Phosphorylated Cx43 protein levels and exosome activities were quantified using Western blot analysis following TBI. Long-term potentiation (LTP) was also tested in rat hippocampal slices. TBI significantly increased the phosphorylated Cx43 and exosome markers expression in rat ipsilateral hippocampus, but not cortex. Blocking the activity of Cx43 or ERK, but not JNK, significantly suppressed TBI-induced exosome release in hippocampus. Furthermore, TBI significantly inhibited the induction of LTP in hippocampal slices, which could be partially but significantly restored by pretreatment with exosomes. The results imply that TBI-activated Cx43 could mediate a nociceptive effect by propagating the brain damages, as well as a neuroprotective effect by promoting exosome release. NEW & NOTEWORTHY We have demonstrated in rat traumatic brain injury (TBI) models that both phosphorylated connexin 43 (p-Cx43) expression and exosome release were elevated in the hippocampus following TBI. The promoted exosome release depends on the phosphorylation of Cx43 and requires ERK signaling activation. Exosome treatment could partially restore the attenuated long-term potentiation. Our results provide new insight for future therapeutic direction on the functional recovery of TBI by promoting p-Cx43-dependent exosome release but limiting the gap junction-mediated bystander effect.

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A Novel Ferroptosis-Related Long Non-coding RNA Prognostic Risk Model for Gastric Cancer

10.21203/rs.3.rs-718468/v1 ◽

2021 ◽

Author(s):

Shenglan Huang ◽

Dan Li ◽

Lingling Zhuang ◽

Liying Sun ◽

Jianbing Wu

Keyword(s):

Gastric Cancer ◽

Risk Score ◽

Roc Curve ◽

Cox Regression ◽

Risk Group ◽

Clinical Stage ◽

Curve Analysis ◽

Clinicopathological Features ◽

Roc Curve Analysis ◽

Kaplan Meier

Abstract Background：Gastric cancer (GC) is one of the most common malignant tumors with a poor prognosis. Ferroptosis is a novel and distinct type of non-apoptotic cell death that is closely associated with metabolism, redox biology, and tumor prognosis. Recently, ferroptosis-related long non-coding RNAs (lncRNAs) have received increasing attention in predicting cancer prognosis. Thus, we aimed to construct an ferroptosis-related lncRNAs signature for predicting the prognosis of patients with gastric cancer.Methods：We built an ferroptosis-related lncRNA risk signature by using Cox regression based on TCGA database. Kaplan-Meier survival analysis was conducted to compare the overall survival (OS) in different risk groups. Cox regression was performed to explore whether the signature could be used as an independent factor. A nomogram was built involving the risk score and clinicopathological features. Furthermore, we explored the biological functions and immune states in two groups.Results：Eight ferroptosis-related lncRNAs were obtained for constructing the prognosis model in gastric cancer. Kaplan–Meier curve analysis revealed that patients in the high-risk group had worse survival than those in the low-risk group. The survival outcome was also appropriate for subgroup analysis, including age, sex, grade, and clinical stage. Multivariate Cox regression analysis and receiver operating characteristic (ROC) curve analysis demonstrated that the risk score was an independent prognostic factor and superior to traditional clinicopathological features in predicting GC prognosis. Next, we established a nomogram according to clinical parameters (age, sex, grade, and clinical stage) and risk score. All the verified results, including ROC curve analysis, calibration curve, and decision curve analysis, demonstrated that the nomogram could accurately predict the survival of patients with gastric cancer. Gene set enrichment analysis revealed that these lncRNAs were mainly involved in cell adhesion, cancer pathways, and immune function regulation.Conclusion: We established a novel ferroptosis-related prognostic risk signature including eight lncRNAs and constructed a nomogram to predict the prognosis of gastric cancer patients, which may improve prognostic predictive accuracy and guide individualized treatment for patients with GC.

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Abstract 253: HSPB7 is Required for Maintaining the Intercalated Disc Structure to Prevent Cardiac Conduction System Failure in Mouse

Circulation Research ◽

10.1161/res.121.suppl_1.253 ◽

2017 ◽

Vol 121 (suppl_1) ◽

Author(s):

Wern-Chir Liao ◽

Liang-Yi Juo ◽

Yen-Hui Chen ◽

Yu-Ting Yan

Keyword(s):

Heart Failure ◽

Gap Junction ◽

Connexin 43 ◽

Adherens Junction ◽

Small Heat Shock Protein ◽

Cardiac Conduction ◽

Nucleotide Polymorphisms ◽

Intercalated Disc ◽

Junction Protein ◽

And Function

HSPB7 is belonged to small heat-shock protein (HSPB) family and considered to function as a co-chaperone, which prevents protein aggregation and maintains protein structure. Single-nucleotide polymorphisms of HSPB7 associated with sporadic cardiomyopathy and heart failure have been identified in human patients. Additionally, HSPB7 is constitutively expressed in heart and rapidly increased in blood plasma after myocardial infarction, suggesting a functional role in the heart. In this study, we found that HSPB7 is highly colocalized with N-cadherin during the assembly and maturation of intercalated disc, suggesting that HSPB7 may involve in organizing and maintaining the cardiac cytoarchitecture. To elucidate the physiological function of HSPB7 in the adult heart, we generated a cardiac-specific inducible HSPB7 knockout mouse. Ablation of HSPB7 in the cardiomyocyte rapidly leads to heart failure, abnormal conduction properties and sudden arrhythmias death. Loss of HSPB7 did not cause significant changes in the organization of contractile proteins in sarcomeres, whereas severe abnormality in the intercalated disc was detected. The expression of connexin 43, a gap-junction protein located at the intercalated disc, was downregulated in HSPB7 knockout cardiomyocytes. Mislocalizations of desmoplakin (desmosomal proteins), and N-cadherin (adherens junction proteins) were also observed in the HSPB7 CKO hearts. Furthermore, filamin C, the interaction protein of HSPB7, was mislocalized and aggregated in HSPB7 mutant cardiomyocytes. The expressivity of the phenotype in the HSPB7 CKO mice is similar to human arrhythmogenic cardiomyopathy patients. Conclusively, we provide the first study characterizing HSPB7 as an intercalated disc protein. Our findings demonstrate that HSPB7 plays an essential role to maintain the structure and function of gap-junction complexes and intercalated disc and has vital implications for human heart disease.

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Abstract TP97: Improved Long-Term Outcome after Transient Cerebral Ischemia in Aquaporin-4 Knockout Mice

Stroke ◽

10.1161/str.44.suppl_1.atp97 ◽

2013 ◽

Vol 44 (suppl_1) ◽

Author(s):

Andrew M Fukuda ◽

Lorenz Hirt ◽

Kamalakar Ambadipudi ◽

Alan S Verkman ◽

Devin K Binder ◽

...

Keyword(s):

Connexin 43 ◽

Aquaporin 4 ◽

Lesion Volume ◽

Post Injury ◽

Edema Formation ◽

Cx43 Expression ◽

Long Term Outcome ◽

Junction Protein ◽

Network Properties

Introduction: Aquaporin-4 (AQP4) is an abundant aquaporin in brain that has been hypothesized to play a central role in edema formation. Furthermore, AQP4 downregulation has been correlated with connexin 43 (Cx43) downregulation, a gap junction protein important for ion and water transport through astrocyte networks. AQP4 deletion in mice was shown to be beneficial at 24 hours after permanent occlusion of the middle cerebral artery (MCAO). However, the long-term consequences of AQP4 deletion on behavior, edema, and tissue properties after stroke have not been studied. Objective: Following transient MCAO, behavior, lesion volumes, blood brain barrier (BBB) integrity, neuroinflammation, and astrocyte network properties with Cx43 expressions were assessed for 14 days in wild type (WT) and AQP4 -/- mice. Methods: Transient (30-min) filament MCAO was performed on AQP4 -/- (n=5) and WT (n=8) mice on a CD1 genetic background. Behavioral outcomes were assessed by rotarod, beam balance, and foot fault tests from 1 to 14 days (d) post injury. Temporal magnetic resonance imaging (MRI) was undertaken to assess lesion volume. Mice were transcardially perfused at 15 d and the brains were extracted and frozen to perform immunohistochemical analysis for IgG, AQP4, Cx43, and glial fibrillary acidic protein (GFAP). Results: MRI showed significantly smaller lesion volume in AQP4 -/- at 1, 3, 7, and 14d. AQP4 -/- mice also had improved motor function recovery with 54% and 75% decreased number of foot-faults than WT mice respectively at 1 and 3 d; and 60% increased time spent on the rotarod compared to WT over the 14 days after stroke onset. AQP4 -/- mice had reduced IgG extravasation and Cx43 expression but increased GFAP staining compared to WT at 14d, suggesting, respectively, less BBB disruption and modified astrocyte network properties combined with altered neuroinflammation fates. Conclusion: AQP4 deletion resulted in improved long-term functional recovery associated with decreased lesion volumes and improved BBB integrity. Although more work must be done, the beneficial outcomes in AQP4 -/- may partly be due to decreased Cx43 as well, resulting in not only decreased edema formation through AQP4, but also in decreased spread of edema through the astrocyte network.

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Survival rates of patients with cancer of the lip, mouth and pharynx: a cohort study of 10 years

Revista Brasileira de Epidemiologia ◽

10.1590/1809-4503201400030009 ◽

2014 ◽

Vol 17 (3) ◽

pp. 680-691 ◽

Cited By ~ 5

Author(s):

Ione Jayce Ceola Schneider ◽

Mayara Eloisa Flores ◽

Daniela Alba Nickel ◽

Luiz Gustavo Teixeira Martins ◽

Jefferson Traebert

Keyword(s):

Cohort Study ◽

Clinical Stage ◽

Descriptive Analysis ◽

Survival Rates ◽

Incidence Rates ◽

Advanced Clinical Stage ◽

Death Risk ◽

Patients With Cancer ◽

Kaplan Meier ◽

Using Data

Introduction: Cancer of the lip, mouth and pharynx is a serious health problem. High incidence rates are found worldwide. In Brazil, the Southern and Southeastern regions have the highest incidences in the country. Objective: To describe 5 and 10-year survival rates in patients with cancer of the lip, mouth and pharynx at a referral center in Florianopolis, Santa Catarina, Brazil. Methods: Retrospective cohort study using data from patients diagnosed between January 1st and December 31st, 2001, with follow-up until December 31st, 2011. Descriptive analysis was performed and survival was estimated by Kaplan-Meier method. Cox semi-parametric model was used to estimate death risk. Results: Survival rates at 5 and 10 years were 33.3 and 26.9%, respectively. Advanced clinical stage in the diagnosis increased death risk by 2.88 and 2.51, respectively. Sex, ethnicity, level of education, previous diagnosis and treatment, as well as age, did not show significant association. Conclusion: Survival rate at 5 years was 33.3% and, at 10 years, was 26.9%. Advanced stage was an independent risk factor for death due to cancer of the lip, mouth and pharynx in both periods analyzed.

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Prognostic values of E2F mRNA expression in human gastric cancer

Bioscience Reports ◽

10.1042/bsr20181264 ◽

2018 ◽

Vol 38 (6) ◽

Cited By ~ 12

Author(s):

Theasha Manicum ◽

Fubiao Ni ◽

Yiming Ye ◽

Xuhui Fan ◽

Bi-Cheng Chen

Keyword(s):

Gastric Cancer ◽

Transcription Factors ◽

Mrna Expression ◽

Prognostic Significance ◽

Family Members ◽

Incidence Rates ◽

Human Gastric Cancer ◽

Kaplan Meier ◽

E2f Transcription Factors ◽

Transcription Activators

Gastric cancer (GC) is the second most frequent cause of cancer-related mortality in the world, with Eastern Asia having the highest incidence rates. E2F is a family of transcription factor proteins that has a variety of functions, which include control of cell cycle, cell differentiation, DNA damage response and cell death. E2F transcription factors are divided into two subfamilies: transcription activators (E2F transcription factors 1 (E2F1), 2 (E2F2) and 3a (E2F3a)) and repressors (E2F3b, E2F transcription factors 4 (E2F4), 5 (E2F5), 6 (E2F6), 7 (E2F7) and 8 (E2F8)). Studies have demonstrated that E2F had prognostic significance in a number of cancers. However, the entirety of the prognostic roles of E2F mRNA expression in GC has not yet been apparently determined. In the present study, the prognostic value of individual family members of E2F mRNA expression for overall survival (OS) was evaluated by using online Kaplan–Meier Plotter (KM Plotter) database. Our result demonstrated that high expressions of three family members of E2F (E2F1, E2F3, E2F4) mRNA were significantly associated with unfavourable OS in all GC patients. However, increased expressions of E2F2, E2F5, E2F6 and E2F7 were significantly associated with favourable OS, especially for higher clinical stages in GC patients. These results provided a better insight into the prognostic functions of E2F mRNA genes in GC. Although the results should be further verified in clinical trials, our findings may be a favourable prognostic predictor for the development of newer therapeutic drugs in the treatment of GC.

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Amyloid-β regulates gap junction protein connexin 43 trafficking in cultured primary astrocytes

Journal of Biological Chemistry ◽

10.1074/jbc.ra120.013705 ◽

2020 ◽

Vol 295 (44) ◽

pp. 15097-15111

Author(s):

Mahua Maulik ◽

Lakshmy Vasan ◽

Abhishek Bose ◽

Saikat Dutta Chowdhury ◽

Neelanjana Sengupta ◽

...

Keyword(s):

Gap Junctions ◽

Gap Junction ◽

Connexin 43 ◽

Cx43 Expression ◽

Chemical Chaperone ◽

Junction Protein ◽

Gap Junction Coupling ◽

Gap Junction Protein ◽

Junction Coupling ◽

And Function

Altered expression and function of astroglial gap junction protein connexin 43 (Cx43) has increasingly been associated to neurotoxicity in Alzheimer disease (AD). Although earlier studies have examined the effect of increased β-amyloid (Aβ) on Cx43 expression and function leading to neuronal damage, underlying mechanisms by which Aβ modulates Cx43 in astrocytes remain elusive. Here, using mouse primary astrocyte cultures, we have examined the cellular processes by which Aβ can alter Cx43 gap junctions. We show that Aβ25-35 impairs functional gap junction coupling yet increases hemichannel activity. Interestingly, Aβ25-35 increased the intracellular pool of Cx43 with a parallel decrease in gap junction assembly at the surface. Intracellular Cx43 was found to be partly retained in the endoplasmic reticulum-associated cell compartments. However, forward trafficking of the newly synthesized Cx43 that already reached the Golgi was not affected in Aβ25-35-exposed astrocytes. Supporting this, treatment with 4-phenylbutyrate, a well-known chemical chaperone that improves trafficking of several transmembrane proteins, restored Aβ-induced impaired gap junction coupling between astrocytes. We further show that interruption of Cx43 endocytosis in Aβ25-35-exposed astrocytes resulted in their retention at the cell surface in the form of functional gap junctions indicating that Aβ25-35 causes rapid internalization of Cx43 gap junctions. Additionally, in silico molecular docking suggests that Aβ can bind favorably to Cx43. Our study thus provides novel insights into the cellular mechanisms by which Aβ modulates Cx43 function in astrocytes, the basic understanding of which is vital for the development of alternative therapeutic strategy targeting connexin channels in AD.

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Chemotherapy Effectiveness and Prognosis of Gastric Cancer Influenced by PTPN11 Polymorphisms

Cellular Physiology and Biochemistry ◽

10.1159/000447856 ◽

2016 ◽

Vol 39 (4) ◽

pp. 1537-1552 ◽

Cited By ~ 3

Author(s):

Chuanjun Zhuo ◽

Mingjing Shao ◽

Ce Chen ◽

Chongguang Lin ◽

Deguo Jiang ◽

...

Keyword(s):

Gastric Cancer ◽

Fold Increase ◽

Axillary Lymph Node Metastasis ◽

Axillary Lymph ◽

Nucleotide Polymorphisms ◽

Positive Interaction ◽

Kaplan Meier ◽

Pcr Rflp ◽

Differentiation Degree ◽

H Pylori

Objective: Since gastric cancer (GC) cells exhibited higher grades of SHP-2 encoded by PTPN11 than normal cells, it would be intriguing to explore whether PTPN11 single nucleotide polymorphisms (SNPs) would influence chemotherapy effectiveness and GC prognosis among a Chinese population. Methods: Altogether 430 late-stage GC patients and 960 healthy controls matched with age and sex were incorporated. Three PTPN11 SNPs (i.e. rs7958372, rs12229892 and rs2301756) were genotyped by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Chemotherapies of cisplatin and 5-fluorouracil were performed for 4 cycles. Odds ratios (ORs) and 95% confidence intervals (CIs) were calculated using the logistic regression. Survival curves were plotted with Kaplan-Meier method and the COX proportional hazard model was used to analyze independent factors for GC prognosis. Results: For rs12229892, AA and GA genotypes would cause 1.60-fold increase of GC risk in comparison to homozygote GG (OR = 1.60; 95% CI = 1.23-2.07; P < 0.001). The A allele of rs2301756 was significantly associated with a decrease in the risk of GC when compared with G allele (OR = 0.81; 95% CI = 0.65-0.99; P = 0.043). Results from both 2-cycle and 4-cycle chemotherapy suggested that chemotherapy was significantly more effective for GA and AA genotypes of rs2301756 compared with homozygote GG (P < 0.001). Besides, the joint impact of rs12229892 (AA) and environmental factors (i.e. smoking, family history, intake of processed food and H .pylori infection) on GC risk was considered as positive interaction, while that of rs2301756 (AA) and the above parameters was deemed as negative interaction. Finally, differentiation degree, axillary lymph node metastasis, rs12229892 and rs2301756 appeared as independent risk factors for GC development (all P < 0.05). Conclusion: Since rs2301756 polymorphism of PTPN11 was associated with reduced risk of GC and better effects of chemotherapy on GC, it can be considered as a predictor of GC prognosis and the treatment target for GC.

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Involvement of connexin 43 in meiotic maturation of bovine oocytes

Reproduction ◽

10.1530/rep.0.1220619 ◽

2001 ◽

pp. 619-628 ◽

Cited By ~ 54

Author(s):

C Vozzi ◽

A Formenton ◽

A Chanson ◽

A Senn ◽

R Sahli ◽

...

Keyword(s):

Gap Junction ◽

Oocyte Maturation ◽

Connexin 43 ◽

Lucifer Yellow ◽

Cumulus Cells ◽

Lower Amount ◽

Cx43 Expression ◽

Gap Junction Channels ◽

Junction Protein ◽

Bovine Oocytes

In ovarian follicles, cumulus cells provide the oocyte with small molecules that permit growth and control maturation. These nutrients reach the germinal cell through gap junction channels, which are present between the cumulus cells and the oocyte, and between the cumulus cells. In this study the involvement of intercellular communication mediated by gap junction channels on oocyte maturation of in vitro cultured bovine cumulus-oocyte complexes (COCs) was investigated. The stages of oocyte maturation were determined by Hoechst 33342 staining, which showed that 90% of COCs placed in the maturation medium for 24 h progress to the metaphase II stage. Bovine COC gap junction communication was disrupted initially using n-alkanols, which inhibit any passage through gap junctions. In the presence of 1-heptanol (3 mmol l(-1)) or octanol (3.0 mmol l(-1) and 0.3 mmol l(-1)), only 29% of the COCs reached metaphase II. Removal of the uncoupling agent was associated with restoration of oocyte maturation, indicating that treatment with n-alkanols was neither cytotoxic nor irreversible. Concentrations of connexin 43 (Cx43), the major gap junction protein expressed in the COCs, were decreased specifically using a recombinant adenovirus expressing the antisense Cx43 cDNA (Ad-asCx43). The efficacy of adenoviral infection was > 95% in cumulus cells evaluated after infection with recombinant adenoviruses expressing the green fluorescence protein. RT-PCR performed on total RNA isolated from Ad-asCx43-infected COCs showed that the rat Cx43 cDNA was transcribed. Western blot analysis revealed a three-fold decrease in Cx43 expression in COCs expressing the antisense RNA for Cx43. Injection of cumulus cells with Lucifer yellow demonstrated further that the resulting lower amount of Cx43 in infected COCs is associated with a two-fold decrease in the extent of coupling between cumulus cells. In addition, oocyte maturation was decreased by 50% in the infected COC cultures. These results indicate that Cx43-mediated communication between cumulus cells plays a crucial role in maturation of bovine oocytes.

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