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Measuring Intracellular Ca2+ Changes in Human Sperm using Four Techniques: Conventional Fluorometry, Stopped Flow Fluorometry, Flow Cytometry and Single Cell Imaging

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Flow cytometry analysis reveals a decrease in intracellular sodium during sperm capacitation.

Jessica Escoffier¹, Dario Krapf, Felipe Navarrete

¹Department of Veterinary and Animal Science, Integrated Sciences Building, University of Massachusetts, Amherst, MA, USA.

Journal of Cell Science

|February 4, 2012

Summary

This summary is machine-generated.

Sperm capacitation, essential for fertilization, involves decreased intracellular sodium ([Na+](i)) via epithelial sodium channels (ENaC) inhibition. This process is regulated by protein kinase A (PKA) and cystic fibrosis transmembrane conductance regulator (CFTR) activation.

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Published on: November 28, 2018

Area of Science:

Reproductive biology
Cell physiology
Ion channel function

Background:

Mammalian sperm undergo capacitation in the female reproductive tract to acquire fertilizing ability.
Capacitation involves physiological changes including altered intracellular pH, calcium, protein phosphorylation, and plasma membrane potential hyperpolarization.
Epithelial sodium channels (ENaC) are present in sperm and are reportedly blocked during capacitation.

Purpose of the Study:

To investigate changes in intracellular sodium concentration ([Na+](i)) during sperm capacitation.
To determine the role of ENaC and its regulation by CFTR and PKA in capacitation-associated membrane potential changes.

Main Methods:

Flow cytometry was used to analyze intracellular Na+ concentration ([Na+](i)) in individual sperm cells.
Experiments utilized sperm with fluorescently tagged acrosomes to assess the integrity of sperm with altered [Na+](i).
Pharmacological agents including amiloride (ENaC inhibitor), genistein (CFTR activator), and PKA inhibitors were employed.

Main Results:

Capacitated sperm exhibited lower intracellular sodium concentrations ([Na+](i)).
This reduction in [Na+](i) was observed only in sperm with intact acrosomes.
In non-capacitated sperm, ENaC inhibition and CFTR activation decreased [Na+](i), indicating crosstalk.
PKA inhibition prevented the decrease in [Na+](i) in capacitated sperm.

Conclusions:

Capacitation-associated hyperpolarization involves a decrease in intracellular sodium ([Na+](i)).
This [Na+](i) decrease is mediated by the inhibition of epithelial sodium channels (ENaC).
The process is regulated by protein kinase A (PKA) through the activation of cystic fibrosis transmembrane conductance regulator (CFTR) channels.