Sperm motility alterations occur in two stages; motility initiation during epididymal transit and transition to hyperactivated motility within the female reproductive tract. We have demonstrated that ejaculated primate sperm contain the serine/threonine PP-1, its endogenous regulators and that treatment of human sperm with the cell-permeable protein phosphatase inhibitor calyculin-a (CL-A) stimulates motility. Our objectives were to determine macaque caput and caudal epididymal sperm PP activity, caput and caudal epididymal sperm motility following CL-A treatment, and PP activity of ejaculated sperm treated with dibutyryl cAMP (dbcAMP) and caffeine which stimulates hyperactivated motility. Protein phosphatase activity was determined using 32P-labeled phosphorylase-` in the absence of divalent cations. Motility analysis was conducted using a computer-assisted motility analysis of system following sperm treatment with 0, 10, 20, 50 or 100 nM CL-A for 10 min. Macaque caput epididymal sperm sonicates contained 4-6-fold more PP activity in comparison to caudal epididymal sperm sonicates. Both caput and caudal epididymal sperm sonicates contained heat-stable PP inhibitor activity as well as Mg-ATP dependent glycogen synthase kinase-3 activity. Treatment of caput epididymal sperm with CL-A increased the percent of motile sperm in a dose-dependent manner from 9% (0 nM CL-A) to 30% (100 nM CL-A) whereas CL-A did not influence the percent of motile caudal epididymal sperm. Treatment of in vitro-capacitated macaque sperm with dbcAMP and caffeine for 1 hr resulted in a significant decrease (P<0.05) in protein phosphatase activity in comparison to nontreated in vitro-capacitated sperm (1.3 q 0.32 and 2.04 q 0.19 x 10-2 nM/min/106 sperm, respectively). These results implicate PP in regulating motility and begin to address mechanisms which control motility initiation of epididymal sperm and the transition to hyperactivated motility.

Agency
National Institute of Health (NIH)
Institute
National Center for Research Resources (NCRR)
Type
Primate Research Center Grants (P51)
Project #
5P51RR000163-37
Application #
5219814
Study Section
Project Start
Project End
Budget Start
Budget End
Support Year
37
Fiscal Year
1996
Total Cost
Indirect Cost
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