Abstract
BackgroundPGF2α is essential for the induction of the corpus luteum regression which in turn reduces progesterone production. Early growth response (EGR) proteins are Cys2-His2-type zinc-finger transcription factor that are strongly linked to cellular proliferation, survival and apoptosis. Rapid elevation of EGR1 was observed after luteolytic dose of PGF2α. EGR1 is involved in the transactivation of many genes, including TGFβ1, which plays an important role during luteal regression.MethodsThe current study was conducted in buffalo luteal cells with the aim to better understand the role of EGR1 in transactivation of TGFβ1 during PGF2α induced luteal regression. Luteal cells from mid stage corpus luteum of buffalo were cultured and treated with different doses of PGF2α for different time durations. Relative expression of mRNAs encoding for enzymes within the progesterone biosynthetic pathway (3βHSD, CYP11A1 and StAR); Caspase 3; AKT were analyzed to confirm the occurrence of luteolytic event. To determine if EGR1 is involved in the PGF2α induced luteal regression via induction of TGFβ1 expression, we knocked out the EGR1 gene by using CRISPR/Cas9.ResultThe present experiment determined whether EGR1 protein expression in luteal cells was responsive to PGF2α treatment. Quantification of EGR1 and TGFβ1 mRNA showed significant up regulation in luteal cells of buffalo at 12 h post PGF2α induction. In order to validate the role of PGF2α on stimulating the expression of TGFβ1 by an EGR1 dependent mechanism we knocked out EGR1. The EGR1 ablated luteal cells were stimulated with PGF2α and it was observed that EGR1 KO did not modulate the PGF2α induced expression of TGFβ1. In PGF2α treated EGR1 KO luteal cell, the mRNA expression of Caspase 3 was significantly increased compared to PGF2α treated wild type luteal cells maintained for 12 h. We also studied the influence of EGR1 on steroidogenesis. The EGR1 KO luteal cells with PGF2α treatment showed no substantial difference either in the progesterone concentration or in StAR mRNA expression with PGF2α-treated wild type luteal cells.ConclusionThese results suggest that EGR1 signaling is not the only factor which plays a role in the regulation of PGF2α induced TGFβ1 signaling for luteolysis.
Highlights
PGF2α is essential for the induction of the corpus luteum regression which in turn reduces progesterone production
These results suggest that EGR1 signaling is not the only factor which plays a role in the regulation of PGF2α induced transforming growth factor β1 (TGFβ1) signaling for luteolysis
Effects of Prostaglandin F2α on luteal cell steroidogenesis in buffalo To understand the effect of PGF2α in corpus luteum, luteal cells derived from the mid stage Corpus Luteum (CL) of buffalo were treated with PGF2α
Summary
PGF2α is essential for the induction of the corpus luteum regression which in turn reduces progesterone production. The mature corpus luteum is comprised of two steroidogenic cell populations, the large and small luteal cells, which are derived from ruptured follicular granulosa and thecal cell, respectively [1]. These large and small luteal cells have ProstaglandinF2α (PGF2α) receptors, which upon binding by PGF2α, can induce luteal regression [2]. PGF2α binds to a G-protein coupled receptor on the target cell and activates PKC signaling pathway [7]. ERK translocates to the nucleus where the binding of SRF/ Elk-1 is promoted which subsequently regulates EGR1 [9]
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