Overexpression of Gjb4 impairs cell proliferation and insulin secretion in primary islet cells.,Molecular Metabolism

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Overexpression of Gjb4 impairs cell proliferation and insulin secretion in primary islet cells.
Molecular Metabolism ( IF 8.1 ) Pub Date : 2020-06-18 , DOI: 10.1016/j.molmet.2020.101042
Anneke Gässler ₁ , Charline Quiclet ₂ , Oliver Kluth ₂ , Pascal Gottmann ₂ , Kristin Schwerbel ₂ , Anett Helms ₂ , Mandy Stadion ₂ , Ilka Wilhelmi ₂ , Wenke Jonas ₂ , Meriem Ouni ₂ , Frank Mayer ₃ , Joachim Spranger ₄ , Annette Schürmann ₅ , Heike Vogel ₆

Affiliation

Department of Experimental Diabetology, German Institute of Human Nutrition Potsdam-Rehbruecke, Arthur-Scheunert-Allee 114-116, D-14558, Nuthetal, Germany; German Center for Diabetes Research (DZD), Ingolstädter Landstraße 1, D-85764, München-Neuherberg, Germany; Department of Endocrinology and Metabolism, Charité-Universitätsmedizin Berlin, Charitéplatz 1, D-10117, Berlin, Germany.
Department of Experimental Diabetology, German Institute of Human Nutrition Potsdam-Rehbruecke, Arthur-Scheunert-Allee 114-116, D-14558, Nuthetal, Germany; German Center for Diabetes Research (DZD), Ingolstädter Landstraße 1, D-85764, München-Neuherberg, Germany.
University Outpatient Clinic, Centre of Sports Medicine, University of Potsdam, Am Neuen Palais 10, D-14469, Potsdam, Germany.
Department of Endocrinology and Metabolism, Charité-Universitätsmedizin Berlin, Charitéplatz 1, D-10117, Berlin, Germany.
Department of Experimental Diabetology, German Institute of Human Nutrition Potsdam-Rehbruecke, Arthur-Scheunert-Allee 114-116, D-14558, Nuthetal, Germany; German Center for Diabetes Research (DZD), Ingolstädter Landstraße 1, D-85764, München-Neuherberg, Germany; Institute of Nutritional Sciences, University of Potsdam, D-14558, Nuthetal, Germany.
Department of Experimental Diabetology, German Institute of Human Nutrition Potsdam-Rehbruecke, Arthur-Scheunert-Allee 114-116, D-14558, Nuthetal, Germany; German Center for Diabetes Research (DZD), Ingolstädter Landstraße 1, D-85764, München-Neuherberg, Germany; Molecular and Clinical Life Science of Metabolic Diseases, University of Potsdam, Arthur-Scheunert-Allee 114-116, D-14558, Nuthetal, Germany.

Objective

Altered gene expression contributes to the development of type 2 diabetes (T2D); thus, the analysis of differentially expressed genes between diabetes-susceptible and diabetes-resistant mouse models is an important tool for the determination of candidate genes that participate in the pathology. Based on RNA-seq and array data comparing pancreatic gene expression of diabetes-prone New Zealand Obese (NZO) mice and diabetes-resistant B6.V-ob/ob (B6-ob/ob) mice, the gap junction protein beta 4 (Gjb4) was identified as a putative novel T2D candidate gene.

Methods

Gjb4 was overexpressed in primary islet cells derived from C57BL/6 (B6) mice and INS-1 cells via adenoviral-mediated infection. The proliferation rate of cells was assessed by BrdU incorporation, and insulin secretion was measured under low (2.8 mM) and high (20 mM) glucose concentration. INS-1 cell apoptosis rate was determined by Western blotting assessing cleaved caspase 3 levels.

Results

Overexpression of Gjb4 in primary islet cells significantly inhibited the proliferation by 47%, reduced insulin secretion of primary islets (46%) and INS-1 cells (51%), and enhanced the rate of apoptosis by 63% in INS-1 cells. Moreover, an altered expression of the miR-341-3p contributes to the Gjb4 expression difference between diabetes-prone and diabetes-resistant mice.

Conclusions

The gap junction protein Gjb4 is highly expressed in islets of diabetes-prone NZO mice and may play a role in the development of T2D by altering islet cell function, inducing apoptosis and inhibiting proliferation.

中文翻译：

Gjb4 的过表达会损害原代胰岛细胞的细胞增殖和胰岛素分泌。

客观的

改变的基因表达有助于 2 型糖尿病 (T2D) 的发展；因此，对糖尿病易感和糖尿病抗性小鼠模型之间差异表达基因的分析是确定参与病理的候选基因的重要工具。基于比较易患糖尿病的新西兰肥胖 (NZO) 小鼠和抗糖尿病 B6.V- ob/ob (B6- ob/ob ) 小鼠的胰腺基因表达的 RNA-seq 和阵列数据，间隙连接蛋白 4 ( Gjb4 ) 被鉴定为推定的新型 T2D 候选基因。

方法

Gjb4在源自 C57BL/6 (B6) 小鼠的原代胰岛细胞和 INS-1 细胞中通过腺病毒介导的感染过度表达。通过 BrdU 掺入评估细胞的增殖率，并在低 (2.8 mM) 和高 (20 mM) 葡萄糖浓度下测量胰岛素分泌。INS-1 细胞凋亡率通过蛋白质印迹评估裂解的半胱天冬酶 3 水平来确定。