cholesterol free bridge membrane. NP-40was expected to extract
mainly the fragmented membrane and cytosol. The testicular
tissue was fixed with 4% paraformaldehyde in 0.1M
phosphate buffer (PB) at 4 ◦C for 6 h. The tissue was embedded
in OCT compound (Miles), and frozen sections were cut
using a cryostat (Histostat, AO). The sections were stained
with 1M TRITC-phalloidin (Sigma, P-1951) for 30 min in
the dark and were washed with phosphate buffered saline
(PBS) and were then mounted in GEL/MOUNT (Biomeda,
Co.). Specimens were observed under a FLUOVIEW laser
scanning confocal microscope (Olympus) with X100 objective
(NA 1.40).