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Regular Article |
-Induced Luteal Regression in the Buffalo Cow (Bubalus bubalis): Involvementof Mitogen-Activated Protein Kinases1
a Department of Molecular Reproduction, Development and Genetics, and
b Primate Research Laboratory,Indian Institute of Science, Bangalore 560012, India
The present study was conducted to evaluate whether the corpus luteum (CL) of the water buffalo (Bubalus bubalis) cow undergoes luteal regression by the process of apoptosis and to examine the involvement of mitogen-activated protein (MAP) kinases during prostaglandin (PG) F2
-induced luteolysis. Sections of CL from late in the estrous cycle, i.e., during spontaneous luteolysis, stained for 4',6'-diamidino-2-phenylindole revealed increased numbers of condensed nuclei, indicating cell death by apoptosis, which was confirmed further by the occurrence of pronounced oligonucleosome formation. For morphological and biochemical characterization during PGF2-induced apoptosis, CL were collected at 0, 4, 12, and 18 h after injection of 750 µg of Tiaprost, a synthetic analogue of PGF2, to midestrous buffalo cows. Serum progesterone concentrations fell within 4 h and decreased (P < 0.05) maximally by 18 h. Concomitant decreases (P < 0.05) in the levels of steroidogenic acute regulatory mRNA and protein were observed in CL during 12–18 h, with the more profound effect on mRNA levels. Quantitative analysis of the genomic DNA showed a >5-fold increase (P < 0.05) in the low molecular weight DNA fragments by 18 h postinjection. Immunoblot analysis of CL tissue lysates showed increased (P < 0.05) levels of phospho-Jun N-terminal kinase (JNK) 1 (4- to 14-fold during 4–18 h) and phospho-p38 (2- to 4-fold at 18 h). Immunohistochemical evaluation of CL sections revealed an increased nuclear localization of phospho-JNK after treatment. These findings demonstrate that the CL of the buffalo cow undergoes cell death by the process of apoptosis both during spontaneous and PGF2-induced luteolysis and that MAP kinases are involved during PGF2-mediated apoptosis in the CL.
1 This work was supported financially by Council of Scientific and Industrial Research, New Delhi.
2 Correspondence: R. Medhamurthy, Indian Institute of Science, Malleswarum, KA, Bangalore 560012, India. FAX: 91 80 360 0999;rmm{at}mrdg.iisc.ernet.in
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