PKCepsilon and an increase in intracellular calcium concentration are necessary for PGF2alpha to inhibit LH-stimulated progesterone secretion in cultured bovine steroidogenic luteal cells |
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Authors: | Madhusudan P Goravanahally Aritro Sen Emmet K Inskeep and Jorge A Flores |
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Institution: | (1) Department of Biology, West Virginia University, Morgantown, West Virginia, USA;(2) Department of Animal Sciences, Michigan State University, East Lansing, Michigan, USA;(3) Animal and Veterinary Sciences, West Virginia University, Morgantown, West Virginia, USA |
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Abstract: | The hypotheses that PKCepsilon is necessary for: 1) PGF2alpha to inhibit LH-stimulated progesterone (P4) secretion, and 2)
for the expression of key prostaglandin synthesizing/metabolizing enzymes were tested in bovine luteal cells in which PKCepsilon
expression had been ablated using a validated siRNA protocol. Steroidogenic cells from Day -6 bovine corpus luteum (CL) were
isolated and transfected to reduce PKCepsilon expression after 48, 72 and 96 h. A third tested hypothesis was that an increase
in intracellular calcium concentration (Ca(2+)]i) is the cellular mechanism through which PGF2alpha inhibits luteal progesterone.
The hypothesis was tested with two pharmacological agents. In the first test, the dose-dependent effects on raising the Ca(2+)]i
with the ionophore, A23187, on basal and LH-stimulated P4 secretion in cells collected from early (Day -4) and mid-cycle (Day
-10) bovine CL was examined. In the second test, the ability of PGF2alpha to inhibit LH-stimulated P4 secretion in Day-10
luteal cells was examined under conditions in which an elevation in Ca(2+)]i had been buffered by means of the intracellular
calcium chelator, Bapta-AM. |
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