Suppression of the expression of a pancreatic beta-cell form of the kinesin heavy chain by antisense oligonucleotides inhibits insulin secretion from primary cultures of mouse beta-cells
Suppression of the expression of a pancreatic beta-cell form of the kinesin heavy chain by antisense oligonucleotides inhibits insulin secretion from primary cultures of mouse beta-cells
复制标题
DOI：
10.1210/en.138.5.1979
复制
发表时间：
1997-05-01
期刊：
影响因子：
4.8
通讯作者：
Balczon, R
中科院分区：
文献类型：
--
作者：
Meng, YX;Wilson, GW;Balczon, R
颗粒/囊泡运输被认为是由微管为基础的力产生的腺苷三磷酸酶，如驱动蛋白。驱动蛋白是一种在脑和其他神经组织中已被充分研究的运动分子。虽然囊泡运输对于胰腺β细胞分泌活动是重要的，但驱动蛋白在β细胞功能中的作用尚未研究。据推测，驱动蛋白作为转运蛋白发挥作用，其与β细胞中的微管和含胰岛素的颗粒相关联，并在分泌刺激时将分泌颗粒从细胞质深处（胰岛素在细胞质中合成和加工）转运到β细胞的表面。为了验证这一假设，克隆并测序了小鼠β细胞驱动蛋白重链互补DNA。然后通过抗小鼠β细胞驱动蛋白重链反义寡核苷酸处理选择性地抑制小鼠β细胞的原代培养物中的驱动蛋白表达。胰岛素分泌的分析确定来自处理的细胞的胰岛素分泌的基础水平降低了50%。此外，葡萄糖刺激的胰岛素释放从处理的P-细胞减少了近70%后，抑制驱动蛋白表达的反义治疗。这项研究的结果提供了第一个直接证据，驱动蛋白，微管为基础的运动蛋白，在胰岛素分泌中发挥重要作用。
Granular/vesicular transport is thought to be supported by microtubule-based force-generating adenosine triphosphatases such as kinesin. Kinesin is a motor molecule that has been well studied in brain and other neuronal tissues. Although vesicular transport is important for pancreatic beta-cell secretory activities, the role of kinesin in beta-cell function has not been investigated. It is hypothesized that kinesin functions as a translocator that associates with both microtubules and insulin-containing granules in beta-cells and transports the secretory granules from deep within the cytoplasm, where insulin is synthesized and processed, to the surface of beta-cells upon secretory stimulation. To test this hypothesis, a mouse beta-cell kinesin heavy chain complementary DNA was cloned and sequenced. Kinesin expression in primary cultures of mouse beta-cells then was selectively suppressed by antimouse beta-cell kinesin heavy chain antisense oligonucleotide treatment. Analysis of insulin secretion determined that the basal level of insulin secretion from the treated cells was decreased by 50%. Furthermore, glucose-stimulated insulin release from treated P-cells was reduced by almost 70% after suppression of kinesin expression by antisense treatment. The findings from this study provide the first direct evidence that kinesin, a microtubule-based motor protein, plays an important role in insulin secretion.