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Enhancement of mouse sperm motility by trophinin-binding peptide.

ABSTRACT: BACKGROUND: Trophinin is an intrinsic membrane protein that forms a complex in the cytoplasm with bystin and tastin, linking it microtubule-associated motor dynein (ATPase) in some cell types. Previously, we found that human sperm tails contain trophinin, bystin and tastin proteins, and that trophinin-binding GWRQ (glycine, tryptophan, arginine, glutamine) peptide enhanced motility of human sperm. METHODS: Immunohistochemistry was employed to determine trophinin protein in mouse spermatozoa from wild type mouse, by using spermatozoa from trophinin null mutant mice as a negative control. Multivalent 8-branched GWRQ (glycine, tryptophan, arginine, glutamine) peptide or GWRQ-MAPS, was chemically synthesized, purified by HPLC and its structure was confirmed by MALDI-TOF mass spectrometry. Effect of GWRQ-MAPS on mouse spermatozoa from wild type and trophinin null mutant was assessed by a computer-assisted semen analyzer (CASA). RESULTS: Anti-trophinin antibody stained the principal (central) piece of the tail of wild type mouse sperm, whereas the antibody showed no staining on trophinin null sperm. Phage particles displaying GWRQ bound to the principal piece of sperm tail from wild type but not trophinin null mice. GWRQ-MAPS enhanced motility of spermatozoa from wild type but not trophinin null mice. CASA showed that GWRQ-MAPS enhanced both progressive motility and rapid motility in wild type mouse sperm. CONCLUSIONS: Present study established the expression of trophinin in the mouse sperm tail and trophinin-dependent effect of GWRQ-MAPS on sperm motility. GWRQ causes a significant increase in sperm motility.

SUBMITTER: Park SK

PROVIDER: S-EPMC3551822 | biostudies-other | 2012

REPOSITORIES: biostudies-other

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Enhancement of mouse sperm motility by trophinin-binding peptide.

Park Seong Kyu SK Yoon Jiwon J Wang Ling L Shibata Toshiaki K TK Motamedchaboki Khatereh K Shim Kyung Jun KJ Chang Mun Seog MS Lee Seung Ho SH Tamura Naoaki N Hatakeyama Shingo S Nadano Daita D Sugihara Kazuhiro K Fukuda Michiko N MN

Reproductive biology and endocrinology : RB&E 20121129

<h4>Background</h4>Trophinin is an intrinsic membrane protein that forms a complex in the cytoplasm with bystin and tastin, linking it microtubule-associated motor dynein (ATPase) in some cell types. Previously, we found that human sperm tails contain trophinin, bystin and tastin proteins, and that trophinin-binding GWRQ (glycine, tryptophan, arginine, glutamine) peptide enhanced motility of human sperm.<h4>Methods</h4>Immunohistochemistry was employed to determine trophinin protein in mouse spe ...[more]

PMID: 23194061

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Project description:Background and purposeNumerous studies have shown that N-arachidonoylethanolamine (AEA) can inhibit sperm motility and function but the ability of cannabinoids to inhibit sperm motility is not well understood. We investigated the effects of WIN 55,212-2, a CB(1) cannabinoid receptor agonist, and ?(9) -tetrahydracannabinol (?(9) -THC) on the ATP levels and motility of murine sperm in vitro. In addition, the effects of acute administration of ?(9) -THC on male fecundity were determined.Experimental approachEffects of ?(9) -THC on basal sperm kinematics were determined using computer-assisted sperm analysis (CASA). Stop-motion imaging was performed to measure sperm beat frequency. The effect of ?(9) -THC on sperm ATP was determined using a luciferase assay. Male fertility was determined by evaluating the size of litters sired by ?(9) -THC-treated males. KEY RESULTS Pretreatment of sperm for 15 min with 1 µM ?(9) -THC reduced their basal motility and attenuated the ability of bicarbonate to stimulate flagellar beat frequency. Treatment with 5 µM WIN 55,212-2 or 10 µM ?(9) -THC for 30 min reduced sperm ATP levels. In sperm lacking CB(1) receptors this inhibitory effect of WIN 55,212-2 on ATP was attenuated whereas that of ?(9) -THC persisted. Administration of 50 mg·kg(-1) ?(9) -THC to male mice just before mating caused a 20% decrease in embryonic litter size.Conclusions and implications?(9) -THC inhibits both basal and bicarbonate-stimulated sperm motility in vitro and reduces male fertility in vivo. High concentrations of WIN 55,212-2 or ?(9) -THC inhibit ATP production in sperm; this effect of WIN 55,212-2 is CB(1) receptor-dependent whereas that of ?(9) -THC is not. LINKED ARTICLES This article is part of a themed section on Cannabinoids in Biology and Medicine. To view the other articles in this section visit http://dx.doi.org/10.1111/bph.2012.165.issue-8. To view Part I of Cannabinoids in Biology and Medicine visit http://dx.doi.org/10.1111/bph.2011.163.issue-7.

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Enhancement of mouse sperm motility by trophinin-binding peptide.

Publications

Enhancement of mouse sperm motility by trophinin-binding peptide.

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