title: Mechanisms of intracellular calcium homeostasis in developing and mature bovine corpora lutea.
abstract: Although calcium (Ca(2+)) is accepted as an intracellular mediator of prostaglandin F2 alpha (PGF2alpha) actions on luteal cells, studies defining mechanisms of Ca(2+) homeostasis in bovine corpora lutea (CL) are lacking. The increase in intracellular Ca(2+) concentration ([Ca(2+)]i) induced by PGF2alpha in steroidogenic cells from mature CL is greater than in those isolated from developing CL. Our hypothesis is that differences in signal transduction associated with developing and mature CL contribute to the increased efficacy of PGF2alpha to induce a Ca(2+) signal capable of inducing regression in mature CL. To test this hypothesis, major genes participating in Ca(2+) homeostasis in the bovine CL were identified, and expression of mRNA, protein, or activity, in the case of phospholipase Cbeta (PLCbeta), in developing and mature bovine CL was compared. In addition, we examined the contribution of external and internal Ca(2+) to the PGF2alpha stimulated rise in [Ca(2+)]i in LLCs isolated from developing and mature bovine CL. Three differences were identified in mechanisms of calcium homeostasis between developing and mature CL, which could account for the lesser increase in [Ca(2+)]i in response to PGF2alpha in developing than in mature CL. First, there were lower concentrations of inositol 1,4,5-trisphosphate (IP3) after similar PGF2alpha challenge, indicating reduced phospholipase C beta (PLCbeta) activity, in developing than mature CL. Second, there was an increased expression of sorcin (SRI) in developing than in mature CL. This cytoplasmic Ca(2+) binding protein modulates the endoplasmic reticulum (ER) Ca(2+) release channel, ryanodine receptor (RyR), to be in the closed configuration. Third, there was greater expression of ATP2A2 or SERCA, which causes calcium reuptake into the ER, in developing than in mature CL. Developmental differences in expression detected in whole CL were confirmed by Western blots using protein samples from steroidogenic cells isolated from developing and mature CL. Localization of these genes in steroidogenic luteal cells was confirmed by immunohistochemistry. Therefore, it is concluded that the cellular mechanisms that allow PGF2alpha to induce a calcium signal of greater magnitude in mature than in developing CL involve 1) greater PLCbeta activity with enhanced generation of IP3, 2) an enhanced Ca(2+) release from the ER via unrestrained RYR2 due to a decrease in SRI expression, and 3) a reduction in calcium reuptake to the ER due to lower expression of ATP2A2. Accordingly, the increase in [Ca(2+)]i induced by PGF2alpha in mature large steroidogenic cells had less dependency from extracellular calcium than in those isolated from immature CL.
authors: Division of Animal and Nutritional Sciences, Davis College of Agriculture, Natural Resources and Design, West Virginia University, Morgantown, West Virginia.
authors: Marietta F
authors: MF
authors: Wright
authors: Elizabeth
authors: E
authors: Bowdridge
authors: Erica L
authors: EL
authors: McDermott
authors: Samuel
authors: S
authors: Richardson
authors: James
authors: J
authors: Scheidler
authors: Qaisar
authors: Q
authors: Syed
authors: Taylor
authors: T
authors: Bush
authors: E Keith
authors: EK
authors: Inskeep
authors: Jorge A
authors: JA
authors: Flores
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002118:Calcium / Q000378:metabolism / Q000502:physiology*; D020013:Calcium Signaling / Q000235:genetics / Q000502:physiology*; D002417:Cattle; D002462:Cell Membrane / Q000235:genetics; D003338:Corpus Luteum / Q000254:growth & development* / Q000502:physiology*; D018076:DNA, Complementary / Q000096:biosynthesis / Q000235:genetics; D015237:Dinoprost / Q000502:physiology; D004721:Endoplasmic Reticulum / Q000235:genetics; D005260:Female; D006706:Homeostasis / Q000235:genetics / Q000502:physiology; D007150:Immunohistochemistry; D007295:Inositol Phosphates / Q000378:metabolism; D054799:Phospholipase C beta / Q000378:metabolism; D011247:Pregnancy; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D043705:Receptor, Adenosine A2A / Q000378:metabolism; D019837:Ryanodine Receptor Calcium Release Channel / Q000378:metabolism; D013256:Steroids / Q000096:biosynthesis
chemical_list: D018076:DNA, Complementary; D007295:Inositol Phosphates; D012333:RNA, Messenger; D043705:Receptor, Adenosine A2A; D019837:Ryanodine Receptor Calcium Release Channel; D013256:Steroids; D015237:Dinoprost; D054799:Phospholipase C beta; D002118:Calcium
keywords: calcium homeostasis; corpus luteum; luteal physiology; luteal sensitivity to PGF2α; ovary