Quantitative and morphological study of preantral follicles from prepubertal gilts
Geraldo Alves, BennerAmaral Alves, KeleRocha Araújo, ValdevaneEmílio Beletti, MarceloLúcia Gambarini, MariaOctavio Jacomini, José
Background: The development of animal reproductive biotechniques is intended of raising reproduction effi ciency. The mammal ovary contains thousands of follicles, of which approximately 99.9% are eliminated by means of the atresia, apoptosis and cellular necrosis process. In other to reduce this follicular loss, the methods for isolation and characterization of preantral follicles have been studied as a premise to culture systems of these structures. The purpose of this study was quantify and evaluate the quality of preantral ovarian follicles from prepubertal gilts after mechanical isolation procedure. Furthermore, it aims to analyze the preantral follicular histological morphology.Materials, Methods & Results: Ovaries (n = 20) from prepubertal gilts were divided in two halves and used each one for isolation and histological processes. The tissue chopper previously regulated for the performance of seriated cuts at 200 m intervals was used for mechanical isolation. The marker bisbenzimidine Hoechst 33342 was added to the follicular pool for evidence the presence of granulosa cells around oocyte and the viability of the isolated PAF was evaluated by using propidium iodide. For the histological evaluation, the ovarian halves were fi xed, dehydrated and diaphanized, and after enclosed in paraffi n, each of them was divided into 5 blocks and sectioned in series 6 m thick. At
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