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Phospholipase C inhibits apoptosis of porcine primary granulosa cells cultured in vitro.

ABSTRACT: Phospholipase C (PLC) can participate in cell proliferation, differentiation and aging. However, whether it has a function in apoptosis in porcine primary granulosa cells is largely uncertain. The objective of this study was to examine the effects of PLC on apoptosis of porcine primary granulosa cells cultured in vitro. The mRNA expression of BAK, BAX and CASP3, were upregulated in the cells treated with U73122 (the PLC inhibitor). The abundance of BCL2 mRNA, was upregulated, while BAX and CASP3 mRNA expression was decreased after treatment with m-3M3FBS (the PLC activator). Both the early and late apoptosis rate were maximized with 0.5??M?U73122 for 4?h. The rate of early apoptosis was the highest at 4?h and the rate of late apoptosis was the highest at 12?h in the m-3M3FBS group. The protein abundance of PLC?1, protein kinase C ? (PKC?), calmodulin-dependent protein kinaseII ? (CAMKII?) and calcineurinA (CalnA) were decreased by U73122, and CAMKII? protein abundance was increased by m-3M3FBS. The mRNA expression of several downstream genes (CDC42, NFATc1, and NF?B) was upregulated by PLC. Our results demonstrated that apoptosis can be inhibited by altering PLC signaling in porcine primary granulosa cells cultured in vitro, and several calcium-sensitive targets and several downstream genes might take part in the processes.

SUBMITTER: Chen H

PROVIDER: S-EPMC6761717 | biostudies-literature | 2019 Sep

REPOSITORIES: biostudies-literature

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Phospholipase C inhibits apoptosis of porcine primary granulosa cells cultured in vitro.

Chen Huali H Yang Youfu Y Wang Youlin Y Li Yuan Y He Yamei Y Duan Jiaxin J Xu Dejun D Pei Yifei Y Cheng Jianyong J Yang Li L Hua Rongmao R Li Xiaoya X Wang Jie J Jiang Xiaohan X He Huanshan H Wu Lin L Liu Dingbang D Li Qingwang Q

Journal of ovarian research 20190925 1

Phospholipase C (PLC) can participate in cell proliferation, differentiation and aging. However, whether it has a function in apoptosis in porcine primary granulosa cells is largely uncertain. The objective of this study was to examine the effects of PLC on apoptosis of porcine primary granulosa cells cultured in vitro. The mRNA expression of BAK, BAX and CASP3, were upregulated in the cells treated with U73122 (the PLC inhibitor). The abundance of BCL2 mRNA, was upregulated, while BAX and CASP3 ...[more]

PMID: 31554511

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Project description:The physiological processes that drive the development of ovarian follicle, as well as the process of oogenesis, are quite well known. Granulosa cells are major players in this occurrence, being the somatic element of the female gamete development. They participate directly in the processes of oogenesis, building the cumulus-oocyte complex surrounding the ovum. In addition to that, they have a further impact on the reproductive processes, being a place of steroid sex hormone synthesis and secretion. It is known that the follicle development creates a major need for angiogenesis and blood vessel development in the ovary. In this study, we use novel molecular approaches to analyze markers of these processes in porcine granulosa cultured primarily in vitro. The cells were recovered from mature sus scrofa specimen after slaughter. They were then subjected to enzymatic digestion and culture primarily for a short term. The RNA was extracted from cultures in specific time periods (0h, 24h, 48h, 96h, and 144h) and analyzed using expression microarrays. The genes that exhibited fold change bigger than |2|, and adjusted p-value lower than 0.05, were considered differentially expressed. From these, we have chosen the members of "angiogenesis," "blood vessel development," "blood vessel morphogenesis," "cardiovascular system development," and "vasculature development" for further selection. CCL2, FGFR2, SFRP2, PDPN, DCN, CAV1, CHI3L1, ITGB3, FN1, and LOX which are upregulated, as well as CXCL10, NEBL, IHH, TGFBR3, SCUBE1, IGF1, EDNRA, RHOB, PPARD, and SLITRK5 genes whose expression is downregulated through the time of culture, were chosen as the potential markers, as their expression varied the most during the time of culture. The fold changes were further validated with RT-qPCR. The genes were described, with special attention to their possible function in GCs during culture. The results broaden the general knowledge about GC's in vitro molecular processes and might serve as a point of reference for further in vivo and clinical studies.

Dataset Information

Phospholipase C inhibits apoptosis of porcine primary granulosa cells cultured in vitro.

Publications

Phospholipase C inhibits apoptosis of porcine primary granulosa cells cultured in vitro.

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