Human ovarian granulosa cells use clathrin-mediated endocytosis for LDL uptake: immunocytochemical and electron microscopic study.

IF 1.1 4区 医学 Q4 MICROSCOPY
Aynur Abdulova, Merjem Purelku, Hakan Sahin, Gamze Tanrıverdi
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引用次数: 1

Abstract

The steroidogenic activity of the granulosa cells is important for the reproductive cycle, and lipoproteins are involved in this process. The clathrin-mediated endocytosis pathway for LDL transport is considered to be the main one in eukaryotic cells. However, there are no studies that elucidate LDL internalization in human granulosa cells clarifying whether the clathrin-mediated endocytic pathway is functional in this process. The aim of this study is to investigate the role of clathrin and v-SNARE proteins in the formation of vesicles in human granulosa cells. In this study, the COV434 human granulosa cells were cultured and divided into four groups where in some of the groups Dil-conjugated LDL and Icarugamycin (ICA) a clathrin-mediated endocytosis inhibitor were added. From the collected mediums pregnenolone and progesterone levels were measured using ELISA. Oil red O staining was performed to show the intracellular lipids in the cells. Clathrin-coated vesicles believed to be responsible for carrying LDL, and v-SNARE proteins that direct the vesicles to their target molecules were also labeled and investigated by histological and ultrastructural methods. Our results show that human granulosa cells as well use the LDL cholesterol for steroid biosynthesis and they may prefer the clathrin-mediated endocytotic pathway to internalize it.

人卵巢颗粒细胞使用网格蛋白介导的内吞作用来摄取LDL:免疫细胞化学和电镜研究。
颗粒细胞的类固醇生成活性对生殖周期具有重要意义,而脂蛋白参与了这一过程。网格蛋白介导的内吞作用途径被认为是真核细胞中LDL运输的主要途径。然而,目前还没有研究阐明LDL在人颗粒细胞中的内化,阐明网格蛋白介导的内吞途径是否在这一过程中起作用。本研究的目的是探讨网格蛋白和v-SNARE蛋白在人颗粒细胞囊泡形成中的作用。在本研究中,我们培养了COV434人颗粒细胞,并将其分为四组,其中一些组添加了dl偶联LDL和icarugycin(一种网格蛋白介导的内吞抑制剂)。从收集的培养基中采用ELISA法测定孕烯醇酮和孕酮水平。油红O染色显示细胞内脂质。网格蛋白包被的囊泡被认为负责携带LDL和引导囊泡到达目标分子的v-SNARE蛋白,也被标记并通过组织学和超微结构方法进行了研究。我们的研究结果表明,人颗粒细胞也使用低密度脂蛋白胆固醇进行类固醇生物合成,它们可能更倾向于通过网格蛋白介导的内吞途径将其内化。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Ultrastructural Pathology
Ultrastructural Pathology 医学-病理学
CiteScore
2.00
自引率
10.00%
发文量
40
审稿时长
6-12 weeks
期刊介绍: Ultrastructural Pathology is the official journal of the Society for Ultrastructural Pathology. Published bimonthly, we are the only journal to be devoted entirely to diagnostic ultrastructural pathology. Ultrastructural Pathology is the ideal journal to publish high-quality research on the following topics: Advances in the uses of electron microscopic and immunohistochemical techniques Correlations of ultrastructural data with light microscopy, histochemistry, immunohistochemistry, biochemistry, cell and tissue culturing, and electron probe analysis Important new, investigative, clinical, and diagnostic EM methods.
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