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BOR - Papers in Press, published online ahead of print December 15, 2004.
Biol Reprod 2004, 10.1095/biolreprod.104.037432
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BIOLOGY OF REPRODUCTION 72, 976–984 (2005)
DOI: 10.1095/biolreprod.104.037432
© 2005 by the Society for the Study of Reproduction, Inc.

Effects of Selective Protein Kinase C Isozymes in Prostaglandin2{alpha}-Induced Ca2+ Signaling and Luteinizing Hormone-Induced Progesterone Accumulation in the Mid-Phase Bovine Corpus Luteum1

Aritro Sen3, Ekta Choudhary3, E. Keith Inskeep4, and Jorge A. Flores2,3

Department of Biology,3 Eberly College of Arts and Sciences; Division of Animal and Veterinary Sciences,4 College of Agriculture, Forestry and Consumer Sciences, West Virginia University, Morgantown, West Virginia 26506

A single-cell approach for measuring the concentration of cytoplasmic calcium ions ([Ca2+]i) and a protein kinase C-epsilon (PKC{epsilon})-specific inhibitor were used to investigate the developmental role of PKC{epsilon} in the prostaglandin F2{alpha}(PGF2{alpha})-induced rise in [Ca2+]i and the induced decline in progesterone accumulation in cultures of cells isolated from the bovine corpus luteum. PGF2{alpha} increased [Ca2+]i in Day 4 large luteal cells (LLCs), but the response was significantly lower than in Day 10 LLCs (4.3 ± 0.6, n = 116 vs. 21.3 ± 2.3, n = 110). Similarly, the fold increase in the PGF2{alpha}-induced rise in [Ca2+]i in Day 4 small luteal cells (SLCs) was lower than in Day 10 SLCs (1.6 ± 0.2, n = 198 vs. 2.7 ± 0.1, n = 95). A PKC{epsilon} inhibitor reduced the PGF2{alpha}-elicited calcium responses in both Day 10 LLCs and SLCs to 3.5 ± 0.3 (n = 217) and 1.3 ± 0.1 (n = 205), respectively. PGF2{alpha} inhibited LH-stimulated progesterone (P4) accumulation only in the incubation medium of Day 10 luteal cells. Both conventional and PKC{epsilon}-specific inhibitors reversed the ability of PGF2{alpha} to decrease LH-stimulated P4 accumulation, and the PKC{epsilon} inhibitor was more effective at this than the conventional PKC inhibitor. In conclusion, the evidence indicates that PKC{epsilon}, an isozyme expressed in corpora lutea with acquired PGF2{alpha} luteolytic capacity, has a regulatory role in the PGF2{alpha}-induced Ca2+ signaling in luteal steroidogenic cells, and that this in turn may have consequences (at least in part) on the ability of PGF2{alpha} to inhibit LH-stimulated P4 synthesis at this developmental stage.

1 Supported in part by a U.S. Department of Agriculture CREES award 2002-35203-12230 to E.K.I. and J.A.F.

2 Correspondence: Jorge A. Flores, Department of Biology, Eberly College of Arts and Sciences, West Virginia University, P.O. Box 6057, Morgantown, WV 26506-6057. FAX: 304 293 6363; jflores{at}wvu.edu






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Copyright © 2005 by the Society for the Study of Reproduction.