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PMID: 25180269 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

Evidence for the involvement of proline-rich tyrosine kinase 2 in tyrosine phosphorylation downstream of protein kinase A activation during human sperm capacitation.

Molecular human reproduction ·Vol. 20 ·No. 11 ·2014-11-00 ·Pages 1054-66

Battistone MA, Alvau A, Salicioni AM, Visconti PE, Da Ros VG, Cuasnicú PS

Abstract

Sperm capacitation involves an increase in intracellular Ca(2+) concentration as well as in protein kinase A (PKA)-dependent protein tyrosine (Tyr) phosphorylation. Interestingly, in humans, a decrease in extracellular Ca(2+) concentration ([Ca(2+)]e) during capacitation induces an increase in Tyr phosphorylation indicating the complexity of Ca(2+) signaling during this process. In view of this, in the present study we further investigated the Ca(2+)-mediated signaling pathways implicated in Tyr phosphorylation during human sperm capacitation. Results revealed that sperm incubation in a medium without added Ca(2+) (⊖ Ca(2+)) increased Tyr phosphorylation but did not modify PKA-mediated phosphorylation. Moreover, inhibition of either PKA or Src family kinase signaling cascades in ⊖ Ca(2+) down-regulated both PKA substrate and Tyr phosphorylations, indicating that the [Ca(2+)]e effects on Tyr phosphorylation depend on PKA targets. Inhibition of calmodulin or Ser/Thr protein phosphatase 2B also increased Tyr phosphorylation without affecting PKA-mediated phosphorylation, supporting the potential role of these Ca(2+) downstream effectors in the increase in Tyr phosphorylation observed in ⊖ Ca(2+). Experiments aimed to identify the kinase responsible for these observations revealed the presence of proline-rich tyrosine kinase 2 (PYK2), a focal adhesion kinase (FAK) family member, in human sperm, and the use of PF431396, an FAK inhibitor, supported the involvement of PYK2 in Tyr phosphorylation downstream of PKA activation. Results also showed that PYK2 was activated in ⊖ Ca(2+) as well as during capacitation and that PF431396 affected capacitated sperm motility, acrosome reaction and ability to penetrate both mouse cumulus matrix and zona-free hamster eggs. Together, our observations support PYK2 as an intermediary component of Ca(2+) signaling between PKA-mediated and Tyr phosphorylations that is required for achieving functional human sperm capacitation.

Keywords
calcium capacitation human sperm tyrosine phosphorylation
MeSH Terms
Calcium Signaling Cyclic AMP-Dependent Protein Kinases/metabolism Enzyme Activation Focal Adhesion Kinase 2/metabolism,physiology Humans Phosphorylation Sperm Capacitation/physiology Tyrosine/metabolism
Chemicals
Tyrosine Focal Adhesion Kinase 2 Cyclic AMP-Dependent Protein Kinases
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Battistone M A
Instituto de Biología y Medicina Experimental (IByME-CONICET), Ciudad Autónoma de Buenos Aires C1428ADN, Argentina.
Alvau A
Department of Veterinary and Animal Science, University of Massachusetts-Amherst, Amherst, MA 01003, USA.
Salicioni A M
Department of Veterinary and Animal Science, University of Massachusetts-Amherst, Amherst, MA 01003, USA.
Visconti P E
Department of Veterinary and Animal Science, University of Massachusetts-Amherst, Amherst, MA 01003, USA.
Da Ros V G
Instituto de Biología y Medicina Experimental (IByME-CONICET), Ciudad Autónoma de Buenos Aires C1428ADN, Argentina.
Cuasnicú P S
Instituto de Biología y Medicina Experimental (IByME-CONICET), Ciudad Autónoma de Buenos Aires C1428ADN, Argentina [email protected].
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Article Info
Journal
Molecular human reproduction
Abbr.
Mol Hum Reprod
ISSN
1460-2407
DOI
10.1093/molehr/gau073
Email
[email protected]
Published
2014-11-00
Epub
2014-00-01
Pages
1054-66
Language
English
Region
England
NLM ID
9513710
PMCID
PMC4209883
Subset
IM
Grants
NICHD NIH HHS · R01 HD038082 · United States
NICHD NIH HHS · R01 HD38082 · United States
NICHD NIH HHS · HD44044 · United States
External Links
PubMed source DOI: 10.1093/molehr/gau073 PMC Full Text View journal details
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