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• W1555358730 abstract "Sea urchin spermatozoa movements are controlled by intracellular pH that is in turn regulated by a Na+/H+ exchanger present in the flagellar membrane. The flagellar exchanger is insensitive to amildride but is inhibited by membrane depolarization. The voltage sensitivity is because of exchanger being turned off by depolarization. Data from both the isolated flagella and the plasma membrane vesicles derived from them support the idea that the exchanger is regulated by a voltage-sensitive gating mechanism. The physiological role of the voltage sensitivity of the exchanger appears to be related to the action of speract, a peptide. This peptide can modulate the flagellar Na+/H+ exchanger by hyperpolarizing the membrane potential. This is because of an increase in K+ permeability of the flagellar membrane through activation of K+ channels. Isolated flagella and flagellar membrane vesicles are single-compartment systems that allow unambiguous analysis and interpretation of ion transport measurements. These subcellular systems are of value for investigating the ionic mechanisms underlying sperm motility initiation. In this chapter, methods for measuring membrane potential and Na+ and H+ transport in these preparations are described. The methods detail—the media to be used, the process of fluorometric measurement of membrane potential, fluorometric measurement of internal pH, and measurement of Na+ uptake—is provided. The membrane potential of intact sperm, isolated flagella, and flagellar vesicles can be monitored with a lipophilic fluorescent probe, diS-C3-(5) (Molecular Probes). The presence of an internally negative membrane potential results in uptake of the cationic probe and quenching of its fluorescence. An alternative method for measuring membrane potential in intact sperm and isolated flagella is to determine the equilibrium distributions of lipophilic ions. The advantage of the fluorometric method is that it monitors the membrane potential continuously and is also a much more sensitive assay." @default.
• W1555358730 created "2016-06-24" @default.
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• W1555358730 date "1995-01-01" @default.
• W1555358730 modified "2023-09-27" @default.
• W1555358730 title "Chapter 56 Measurement of Membrane Potential and Na+ and H+ Transport in Isolated Sea Urchin Sperm Flagella and Their Membrane Vesicles" @default.
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• W1555358730 doi "https://doi.org/10.1016/s0091-679x(08)60837-7" @default.
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