The calcium‐sensing receptor regulates protein tyrosine phosphorylation through PDK1 in boar spermatozoa

Regulation of protein tyrosine phosphorylation is required for sperm capacitation and oocyte fertilization. The objective of the present work was to study the role of the calcium‐sensing receptor (CaSR) on protein tyrosine phosphorylation in boar spermatozoa under capacitating conditions. To do this...

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Bibliographic Details
Published in:Molecular reproduction and development 2019-07, Vol.86 (7), p.751-761
Main Authors: Macías‐García, Beatriz, García‐Marín, Luis J., Bragado, María J., González‐Fernández, Lauro
Format: Article
Language:English
Subjects:
Animals
boar
Calcium
Calcium-sensing receptors
calcium‐sensing receptor
Capacitation
Cyclic AMP-Dependent Protein Kinases - antagonists & inhibitors
Cyclic AMP-Dependent Protein Kinases - metabolism
Fertilization
Indazoles - pharmacology
Inhibition
Isoquinolines - pharmacology
Kinases
Male
Naphthalenes - pharmacology
Phenylmethylsulfonyl Fluoride - pharmacology
Phosphorylation
Phosphorylation - drug effects
Protein kinase A
Protein Kinase Inhibitors - pharmacology
protein kinases
protein tyrosine phosphorylation
Proteins
Pyrimidines - pharmacology
Pyruvate Dehydrogenase (Acetyl-Transferring) Kinase - antagonists & inhibitors
Pyruvate Dehydrogenase (Acetyl-Transferring) Kinase - metabolism
Receptors, Calcium-Sensing - antagonists & inhibitors
Receptors, Calcium-Sensing - metabolism
Reproductive system
Serine
Signal transduction
Signal Transduction - drug effects
Signal Transduction - physiology
Sperm
Sperm Capacitation - physiology
Sperm Motility - drug effects
spermatozoa
Spermatozoa - metabolism
Sulfonamides - pharmacology
Sus scrofa - metabolism
Threonine
Tyrosine
Tyrosine - metabolism
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Summary:Regulation of protein tyrosine phosphorylation is required for sperm capacitation and oocyte fertilization. The objective of the present work was to study the role of the calcium‐sensing receptor (CaSR) on protein tyrosine phosphorylation in boar spermatozoa under capacitating conditions. To do this, boar spermatozoa were incubated in Tyrode's complete medium for 4 hr and the specific inhibitor of the CaSR, NPS2143, was used. Also, to study the possible mechanism(s) by which this receptor exerts its function, spermatozoa were incubated in the presence of specific inhibitors of the 3‐phosphoinositide dependent protein kinase 1 (PDK1) and protein kinase A (PKA). Treatment with NPS2143, GSK2334470, an inhibitor of PDK1 and H‐89, an inhibitor of PKA separately induced an increase in tyrosine phosphorylation of 18 and 32 kDa proteins, a decrease in the serine/threonine phosphorylation of the PKA substrates together with a drop in sperm motility and viability. The present work proposes a new signalling pathway of the CaSR, mediated by PDK1 and PKA in boar spermatozoa under capacitating conditions. Our results show that the inhibition of the CaSR induces the inhibition of PDK1 that blocks PKA activity resulting in a rise in tyrosine phosphorylation of p18 and p32 proteins. This novel signalling pathway has not been described before and could be crucial to understand boar sperm capacitation within the female reproductive tract. Calcium‐sensing receptor (CaSR) regulates PY in boar sperm.
ISSN:1040-452X
1098-2795
DOI:10.1002/mrd.23160