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Characterization of rabbit follicular oocytes and their ability to mature in vitro

Characterization of rabbit follicular oocytes and their ability to mature in vitro

Archivos de Zootecnia 45(169): 25-35

The present study was undertaken to characterize the cellular morphology, meiotic configuration, and the ability to mature in vitro of rabbit follicular oocytes in relation to the method of recovery. Oocytes (n = 433) were recovered from the ovaries of unstimulated does using either aspiration or follicle slicing and subsequent rupture. Oocytes were classified according to the characteristics of the cumulus cells and ooplasm into one of five groups: (A) surrounded by layers of cumulus cells and intact cytoplasm; (B) naked and with intact cytoplasm; (C) with degeneration in the cumulus cells; (D) with degeneration in the cytoplasm, and (E) with degeneration in both, cytoplasm and cumulus-calls. In experiment 1, oocytes were morphologically classified into five groups, and fixed and stained with acetic-orcein to asses their nuclear status. In experiment 2, oocytes were recovered (n = 96 and n = 105, for aspiration and slicing, respectively), classified as above, and they were then matured in vitro in Brackett medium for 12 h. At the end of culture oocytes were assessed for maturation by staining with acetic-orcein. When aspiration was used, 105 oocytes were recovered from 146 follicles, whereas 127 oocytes were recovered by rupturing 139 follicles. The proportions of the five types of recovered oocytes were similar in the two methods of oocyte collection. Only oocytes without morphological degeneration (A and B types) showed high values of normal germinal vesicle configuration and nuclear maturation. The maturation rates using aspiration were 83.3 p.100 (type A), 52.3 p.100 (type B), 7.6p.100 (type C), 5.8p.100 (type D), and Op.100 (type E), while using follicular slicing were: 87.8 p.100 (type A), 47.3 p.100 (type B), 5.2 p.100 (type C), and 0 p.100 (types D and E). In conclusion, (i) follicular aspiration and follicle slicing offered a similar quality in the population of the obtained oocytes. Follicle rupture can yield more number of oocytes than aspiration, but follicle aspiration has the advantage of higher speed operation; (ii), oocytes without morphological degeneration presented higher maturation rates (87.8p.100, p lt 0.01) than degenerate oocytes, and (iii) the extremes in investment and ooplasm appearance were the only obvious indicators of an oocyte's potential to mature in vitro.

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Accession: 002775896

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