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Department of Animal Sciences
The Robert H. Smith Faculty
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The Hebrew University of Jerusalem.

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miR-210 and GPD1L regulate EDN2 in primary and immortalized human granulosa-lutein cells

Citation:

Shrestha, K. ; Onasanya, A. E. ; Eisenberg, I. ; Wigoda, N. ; Yagel, S. ; Yalu, R. ; Meidan, R. ; Imbar, T. miR-210 and GPD1L regulate EDN2 in primary and immortalized human granulosa-lutein cells. Reproduction 2018, 155, 197-205.

Date Published:

2018 02

Abstract:

Endothelin-2 (EDN2), expressed at a narrow window during the periovulatory period, critically affects ovulation and corpus luteum (CL) formation. LH (acting mainly via cAMP) and hypoxia are implicated in CL formation; therefore, we aimed to elucidate how these signals regulate using human primary (hGLCs) and immortalized (SVOG) granulosa-lutein cells. The hypoxiamiR, microRNA-210 (miR-210) was identified as a new essential player in expression. Hypoxia (either mimetic compound-CoCl, or low O) elevated hypoxia-inducible factor 1A (HIF1A), miR-210 and Hypoxia-induced miR-210 was suppressed in HIF1A-silenced SVOG cells, suggesting that miR-210 is HIF1A dependent. Elevated miR-210 levels in hypoxia or by miR-210 overexpression, increased Conversely, miR-210 inhibition reduced levels, even in the presence of CoCl, indicating the importance of miR-210 in the hypoxic induction of A molecule that destabilizes HIF1A protein, glycerol-3-phosphate dehydrogenase 1-like gene-, was established as a miR-210 target in both cell types. It was decreased by miR-210-mimic and was increased by miR-inhibitor. Furthermore, reducing by endogenously elevated miR-210 (in hypoxia), miR-210-mimic or by siRNA resulted in elevated HIF1A protein and levels, implying a vital role for in the hypoxic induction of Under normoxic conditions, forskolin (adenylyl cyclase activator) triggered changes typical of hypoxia. It elevated , and miR-210 while inhibiting Furthermore, HIF1A silencing greatly reduced forskolin's ability to elevate and miR-210. This study highlights the novel regulatory roles of miR-210 and its gene target, GPD1L, in hypoxia and cAMP-induced by human granulosa-lutein cells.