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J Cell Sci. 2019 Jul 26;132(14). pii: jcs230565. doi: 10.1242/jcs.230565.

MRP4-mediated cAMP efflux is essential for mouse spermatozoa capacitation.

Author information

1
Centro de Estudios Farmacológicos y Botánicos (CEFYBO) (UBA-CONICET), Facultad de Medicina, Universidad de Buenos Aires, Buenos Aires C1121ABG, Argentina.
2
Instituto de Biología y Medicina Experimental (IBYME), Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Buenos Aires C1428ADN, Argentina.
3
Instituto de Investigaciones Farmacológicas (ININFA) (UBA-CONICET), Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires, Buenos Aires C1113AAD, Argentina.
4
Department of Veterinary and Animal Sciences, University of Massachusetts, Amherst, MA 01003, United States.
5
Centro de Estudios Farmacológicos y Botánicos (CEFYBO) (UBA-CONICET), Facultad de Medicina, Universidad de Buenos Aires, Buenos Aires C1121ABG, Argentina perezms@fmed.uba.ar.

Abstract

Mammalian spermatozoa must undergo biochemical and structural changes to acquire the capacity for fertilization, in a process known as capacitation. Activation of PKA enzymes is essential for capacitation, and thus cAMP levels are tightly regulated during this process. Previously, we demonstrated that during capacitation, bovine spermatozoa extrude cAMP through multidrug resistance-associated protein 4 (MRP4, also known as ABCC4), which regulates intracellular levels of the nucleotide and provides cAMP to the extracellular space. Here, we report the presence of functional MRP4 in murine spermatozoa, since its pharmacological inhibition with MK571 decreased levels of extracellular cAMP. This also produced a sudden increase in PKA activity, with decreased tyrosine phosphorylation at the end of capacitation. Blockade of MRP4 inhibited induction of acrosome reaction, hyperactivation and in vitro fertilization. Moreover, MRP4 inhibition generated an increase in Ca2+ levels mediated by PKA, and depletion of Ca2+ salts from the medium prevented the loss of motility and phosphotyrosine inhibition produced by MK571. These results were supported using spermatozoa from CatSper Ca2+ channel knockout mice. Taken together, these results suggest that cAMP efflux via MRP4 plays an essential role in mouse sperm capacitation.This article has an associated First Person interview with the first author of the paper.

KEYWORDS:

ABCC4; Ca2+; MRP4; Mouse; PKA activity; Sperm capacitation; cAMP efflux

PMID:
31253671
DOI:
10.1242/jcs.230565

Conflict of interest statement

Competing interestsThe authors declare no competing or financial interests.

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