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溶血磷脂对大鼠 PC12 细胞胞吐作用的差异影响。

Differential effects of lysophospholipids on exocytosis in rat PC12 cells.

机构信息

Department of Oral and Maxillofacial Surgery, National University of Singapore, Singapore, Singapore.

出版信息

J Neural Transm (Vienna). 2010 Mar;117(3):301-8. doi: 10.1007/s00702-009-0355-1.

Abstract

Secretory phospholipase A2 (sPLA2) activity is present in the CNS and the sPLA2-IIA isoform has been shown to induce exocytosis in cultured hippocampal neurons. However, little is known about possible contributions of various lysophospholipid species to exocytosis in neuroendocrine cells. This study was therefore carried out to examine the effects of several lysophospholipid species on exocytosis on rat pheochromocytoma-12 (PC12) cells. An increase in vesicle fusion, indicating exocytosis, was observed in PC12 cells after external infusion of lysophosphatidylinositol (LPI), but not lysophosphatidylcholine or lysophosphatidylserine by total internal reflection microscopy. Similarly, external infusion of LPI induced significant increases in capacitance, or number of spikes detected at amperometry, indicating exocytosis. Depletion of cholesterol by pre-incubation of cells with methyl beta cyclodextrin and depletion of Ca2+ by thapsigargin and incubation in zero external Ca2+ resulted in attenuation of LPI induced exocytosis, indicating that exocytosis was dependent on the integrity of lipid rafts and intracellular Ca2+. Moreover, LPI induced a rise in intracellular Ca2+ suggesting that this could be the trigger for exocytosis. It is postulated that LPI may be an active participant in sPLA2-mediated exocytosis in the CNS.

摘要

分泌型磷脂酶 A2(sPLA2)活性存在于中枢神经系统中,并且已经证明 sPLA2-IIA 同工型可诱导培养的海马神经元中的胞吐作用。然而,对于各种溶血磷脂种类对神经内分泌细胞中胞吐作用的可能贡献知之甚少。因此,本研究旨在研究几种溶血磷脂种类对大鼠嗜铬细胞瘤-12(PC12)细胞胞吐作用的影响。通过全内反射显微镜观察到,溶血磷脂酰肌醇(LPI)的外输注可增加囊泡融合,表明发生了胞吐作用,但溶血磷脂酰胆碱或溶血磷脂酰丝氨酸则没有。同样,LPI 的外输注诱导电容的显著增加,或在安培法中检测到的尖峰数量增加,表明发生了胞吐作用。用甲基-β-环糊精预先孵育细胞以耗竭胆固醇,用 thapsigargin 耗竭 Ca2+并在无外 Ca2+中孵育可减弱 LPI 诱导的胞吐作用,表明胞吐作用依赖于脂筏和细胞内 Ca2+的完整性。此外,LPI 诱导细胞内 Ca2+升高,表明这可能是胞吐作用的触发因素。据推测,LPI 可能是 sPLA2 介导的中枢神经系统胞吐作用的活跃参与者。

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