Immunolocalization of rHamOVGP1-enhanced tyrosine-phosphorylated proteins in hamster sperm.

<p>Sperm, after being cultured in modified TALP medium alone (A, C, E) or supplemented with 20 μg/ml rHamOVGP1 (B, D, F) for 0 h (A, B), 2 h (C, D) and 4 h (E, F), were subjected to indirect immunofluorescence as described in Materials and Methods. Immunofluorescent staining of tyrosine phosphorylated proteins was observed in the mid-piece (MP) and principal piece (PP). (A1-F1) are corresponding bright field photomicrographs of (A2-F2) respectively. (G) shows intense immunostaining over the principle piece (PP) with a weaker staining intensity over the mid-piece (MP) after sperm were demembranated with 2% Triton X-100 for 10 min following incubation for 4 h in modified TALP medium supplemented with 20 μg/ml rHamOVGP1. Scale bars = 50 μm