Figure 9.

The effect of E2 on GVBD of normal and antisense oligonucleotide-injected zebrafish oocytes. A, Expression of GPR30 mRNA in zebrafish ovarian tissue. B, Effects of 20 nm E2 on spontaneous (CTL) and DHP-induced (10 nm) GVBD. *, P < 0.05 compared with respective control not treated with E2. C, Comparison of the effects of microinjection of zebrafish oocytes with either antisense morpholino oligonucleotide to zebrafish GPR30 or its 5-mispair control oligonucleotide during the hCG priming stage on spontaneous GVBD (hCG-primed only, Veh), DHP-induced (10 nm) GVBD, or DHP-induced GVBD in the presence of 100 nm E2. *, P < 0.05 compared with respective 5-mispair CTL group; +, P < 0.05 compared with DHP-treated groups. A total of 20–30 oocytes were counted in each treatment group for each experiment. D, Relative intensity of GPR30-immunoreactive bands on a Western blot of cell membranes prepared from zebrafish oocytes microinjected with antisense and 5-mispair oligonucleotides. Integrin loading control was used for the membrane samples and detected with integrin antibody. *, P < 0.05 compared with 5-mispair oligonucleotide control. Microinjection experiments were repeated three times with oocytes from different donors and similar results were obtained in each assay.