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First published online March 2, 2006
Journal of Experimental Biology 209, 1147-1156 (2006)
Published by The Company of Biologists 2006
doi: 10.1242/jeb.02094
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Hydration of rainbow trout oocyte during meiotic maturation and in vitro regulation by 17,20ß-dihydroxy-4-pregnen-3-one and cortisol

Sylvain Milla, Bernard Jalabert, Helene Rime, Patrick Prunet and Julien Bobe*

Institut National de la Recherche Agronomique, INRA-SCRIBE, IFR 140, Campus de Beaulieu, 35000 Rennes, France


Figure 1
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  		Fig. 1. Changes in wet mass (WM) and dry mass (DM) of oocytes obtained from rainbow trout females at different days before and after ovulation. Oocytes were defolliculated as previously described (Finet et al., 1988Go). Values are mean ± s.d. (N=14–36). Different letters indicate significant differences between days (P<0.05).

 

Figure 2
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  		Fig. 2. Oocyte water content (WC)/water content before maturation (WC0, PreGVBD) ratio of oocytes sampled at three ovarian stages: before oocyte maturation with visible germinal vesicle (PreGVBD), during oocyte maturation (MAT) and after ovulation (OV). Oocytes were defolliculated as previously described (Finet et al., 1988Go). Values are mean ± s.d. (N=4). Different letters indicate significant differences between stages (P<0.05).

 

Figure 3
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  		Fig. 3. Representative SDS–PAGE (12%) of yolk proteins from preGVBD oocytes and ovulated eggs of rainbow trout. Each lane had 0.5 mg of yolk proteins originating from three oocytes/eggs. The position of molecular size markers is indicated.

 

Figure 4
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  		Fig. 4. Oocyte wet mass (WM)/`initial control' wet mass (WM0) ratio after a 60-h in vitro incubation. Follicle-enclosed oocytes were incubated in the presence of different steroids: 11-deoxycorticosterone (DOC, 5 ng ml–1); 17,20ß-dihydroxy-4-pregnen-3-one (MIS, 40 ng ml–1); cortisol (12 ng ml–1); cortisol (12 ng ml–1) plus 17,20ß-dihydroxy-4-pregnen-3-one (40 ng ml–1). IC, initial control; ethanol vehicle was used as the negative control (NC). Values are mean ± s.d. (N=7). Different letters indicate significant differences between treatments (P<0.05). The percentage of germinal vesicle breakdown (GVBD) after a 60 h incubation is shown below for each group.

 

Figure 5
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  		Fig. 5. Oocyte wet mass (WM)/`initial control' (IC) wet mass (WM0) ratio after a 60 h in vitro incubation. Follicle-enclosed oocytes were incubated in the presence of different steroids: 17,20ß-dihydroxy-4-pregnen-3-one (MIS40; 40 ng ml–1); cortisol (5, 12 or 50 ng ml–1); 11-deoxycorticosterone (DOC, 5, 12 or 50 ng ml–1); cortisol (5, 12 or 50 ng ml–1)+MIS40; DOC (5, 12 or 50 ng ml–1)+MIS40. IC, initial control; ethanol vehicle was used as the negative control (NC). Values are means ± s.d. (N=5). Asterisks indicate significantly different from NC. Triangles indicate significantly different from MIS40 value (P<0.05). The percentage of germinal vesicle breakdown (GVBD) after 60-h incubation is shown below for each group.

 

Figure 6
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  		Fig. 6. Abundance of messenger RNA of (A) glucocorticoid receptor 1 (rtGR1), (B) glucocorticoid receptor 2 (rtGR2) and (C) 11ß-hydroxysteroid dehydrogenase (11ß-HSD) in the whole ovary of rainbow trout females sampled at three stages of ovarian development: late vitellogenesis (LV, 3–4 weeks before expected ovulation, N=6), prior to germinal vesicle breakdown (preGVBD, N=8) and during oocyte maturation (MAT, N=6). Values are mean ± s.e.m. Different letters indicate significant differences between stages (P<0.05).

 




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