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Tipo do documento: Artigo
Título: Quantitative and morphological study of preantral follicles from prepubertal gilts
Autor: Alves, Benner Geraldo
Alves, Kele Amaral
Araújo, Valdevane Rocha
Beletti, Marcelo Emílio
Gambarini, Maria Lúcia
Jacomini, José Octavio
Abstract: 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 μm 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 μm thick. At each 1000 μm interval, 2 slides were removed per block, totaling 200 slides that were dyed with periodic acid Schiff (PAS) and hematoxylin. The total PAF calculated in situ for each follicular stage (primordial, primary and secondary) was estimated according with the preantral follicles number evaluated on histological sections. The number of isolated preantral follicles per ovary was (599,160 ± 74,089), presented positive correlation (P < 0.05) between ovarian weight. After isolation, 76.4% of preantral follicles were viable (P < 0.05), while 64.3% had granulosa cells around the oocyte (P < 0.05). The average number of PAF estimated in histological cuts was 131,938 ± 14,615 per ovary, oscillating between 67,599 and 291,898, out of which 117,968 (89.41%) primordial follicles, 3,448 (2.61%) primary follicles, and 10,522 (7.98%) secondary follicles. The verifi ed follicles diameter was primordial (33.1 μm), primary (47.6 μm) and secondary (79.1 μm). Discussion: The PAF quantifi cation estimated in situ per ovary was different from results obtained on mechanical isolation, showing that used techniques do not complement. However, both procedures highlighted the specie reproductive potencial. The greater percentage of oocytes was included in primordial follicles (89.4%; P < 0.05). Follicular diameter and the number of granulosa cells increased (P < 0.05) according to the development stage. These data are important to defi ne which interval of cut can be adopted before utilizing the mechanical isolation procedure with tissue chopper, avoiding sections damages to the follicular structure, which contributes to the smaller viability of isolated PAF. In summary, the results reported that it is possible to obtain a great number of viable PAF from of prepubertal gilts ovaries after mechanical isolation, providing material to supply the demand for studies relating to folliculogenesis and in vitro culture techniques.
Palavras-chave: Primordial follicles
País: Brasil
Unidade acadêmica: Escola de Veterinária e Zootecnia - EVZ (RG)
Citação: ALVES; Benner Geraldo; ALVES, Kele Amaral; ARAÚJO, Valdevane Rocha; BELETTI, Marcelo Emílio; GAMBARINI, Maria Lúcia; JACOMINI, José Octávio. Quantitative and morphological study of Preantral follicles from prepubertal gilts. Acta Scientiae Veterinariae, Porto Alegre, v. 40, n. 4, p. 1079, 2012.
Tipo de acesso: Acesso Aberto
Data de publicação: 2012
Aparece nas coleções:EVZ - Artigos publicados em periódicos

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