Evaluation of growth, cell proliferation, and cell death in bovine corpora lutea throughout the estrous cycle
1994
Zheng, J. | Fricke, P.M. | Reynolds, P. | Redmer, D.A.
To evaluate the kinetics of luteal growth, bovine CL were obtained from four stages (stage 1 Days 1-4; stage 11 Days 5-10; stage III, Days 11-17: stage IV, Days 18-21) of the estrous cycle and luteal fresh weight as well as DNA protein and progesterone contents was determined. To evaluate the relative rate of cell proliferation proliferating cell nuclear antigen (PCNA: a specific marker for cell proliferation) was immunolocalized in paraffin-embedded luteal tissue sections. To evaluate the relative rate of cell death nucleosomal fragmentation of DNA (a specific marker for apoptosis) was detected by agarose gel electrophoresis and also by histochemical localization in paraffin-embedded luteal tissue sections. Luteal fresh weight and DNA protein and progesterone contents increased (p < 0.01) from stage I to stage II were similar between stages II and III and then decreased (p < 0.01) from stage III to stage IV . The ratio of protein to DNA (an index of average cell size) was similar for stages I, II, and III and then decreased (p < 0.01) at stage IV. For stage I (corpora hemorrhagica) most proliferating (PCNA-positive) cells were located in or around the core of the tissue infoldings (presumably thecal-derived areas) whereas relatively few proliferating cells were located at the periphery of the tissue infoldings (presumably granulosa derived areas). For stages II, III, and IV the majority of proliferating cells appeared to be small cells (i.e. small parenchymal cells fibroblasts and endothelial cells). The labeling index (LI; percentage of cells that were PCNA-positive) was greatest at stage I (20.3 +/- 1.1%); it then decreased (p < 0.01) by stage II and was similar at stages II, III, and IV (3.4 +/- 1.1%). Apoptosis as determined by evaluation of nucleosomal DNA fragmentation by agarose gel electrophoresis and in situ localization was detectable only at stage IV. These data demonstrate that luteal growth from stage I to stage II resulted from cell proliferation as shown by a high LI at stage I accompanied by increased luteal DNA content but no change in average cell size and by similar protein:DNA ratios. Luteal regression from stage III to stage IV was primarily associated with cell deletion and decreased cell size as shown by a decrease in luteal DNA content and the appearance of apoptosis along with a decrease in the luteal protein:DNA ratio. In addition analysis of the estimated hnetics of cell turnover revealed that the number of proliferating luteal cells remained high until stage III and then decreased at stage IV in association with luteal regression.
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