Effects of membrane potential and tension on prestin, the outer hair cell lateral membrane motor protein

Joseph Santos-Sacchi, Weixing Shen, Jing Zheng, Peter Dallos

Research output: Contribution to journalArticle

64 Citations (Scopus)

Abstract

1. Under whole-cell voltage clamp, the effects of initial voltage conditions and membrane tension on gating charge and voltage-dependent capacitance were studied in human embryonic kidney cells (TSA201 cell line) transiently transfected with the gene encoding the gerbil protein prestin. Conformational changes in this membrane-bound protein probably provide the molecular basis of the outer hair cell (OHC) voltage-driven mechanical activity, which spans the audio spectrum. 2. Boltzmann characteristics of the charge movement in transfected cells were similar to those reported for OHCs (Q max = 0.99 ± 0.16 pC, z = 0.88 ± 0.02; n = 5, means ± S.E.M.). Unlike that of the adult OHC, the voltage at peak capacitance (V pkcm) was very negative (-74.7 ± 3.8 mV). Linear capacitance in transfected cells was 43.7 ± 13.8 pF and membrane resistance was 458 ± 123 MΩ. 3. Voltage steps from the holding potential preceding the measurement of capacitance-voltage functions caused a time- and voltage-dependent shift in V pkcm. For a prepulse to -150 mV, from a holding potential of 0 mV, V pkcm shifted 6.4 mV, and was fitted by a single exponential time constant of 45 ms. A higher resolution analysis of this time course was made by measuring the change in capacitance during a fixed voltage step and indicated a double exponential shift (τ 0 = 51.6 ms, τ 1 = 8.5 s) similar to that of the native gerbil OHC. 4. Membrane tension, delivered by increasing pipette pressure, caused a positive shift in V pkcm. A maximal shift of 7.5 mV was obtained with 2 kPa of pressure. The effect was reversible. 5. Our results show that the sensitivity of prestin to initial voltage and membrane tension, though present, is less than that observed in adult OHCs. It remains possible that some other interacting molecular species within the lateral plasma membrane of the native OHC amplifies the effect of tension and prior voltage on prestin's activity.

Original languageEnglish (US)
Pages (from-to)661-666
Number of pages6
JournalJournal of Physiology
Volume531
Issue number3
DOIs
StatePublished - Mar 15 2001

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Outer Auditory Hair Cells
Membrane Potentials
Membrane Proteins
Cell Membrane
Membranes
Gerbillinae
Pressure
Kidney
Cell Line
Genes
Proteins

ASJC Scopus subject areas

  • Physiology

Cite this

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title = "Effects of membrane potential and tension on prestin, the outer hair cell lateral membrane motor protein",
abstract = "1. Under whole-cell voltage clamp, the effects of initial voltage conditions and membrane tension on gating charge and voltage-dependent capacitance were studied in human embryonic kidney cells (TSA201 cell line) transiently transfected with the gene encoding the gerbil protein prestin. Conformational changes in this membrane-bound protein probably provide the molecular basis of the outer hair cell (OHC) voltage-driven mechanical activity, which spans the audio spectrum. 2. Boltzmann characteristics of the charge movement in transfected cells were similar to those reported for OHCs (Q max = 0.99 ± 0.16 pC, z = 0.88 ± 0.02; n = 5, means ± S.E.M.). Unlike that of the adult OHC, the voltage at peak capacitance (V pkcm) was very negative (-74.7 ± 3.8 mV). Linear capacitance in transfected cells was 43.7 ± 13.8 pF and membrane resistance was 458 ± 123 MΩ. 3. Voltage steps from the holding potential preceding the measurement of capacitance-voltage functions caused a time- and voltage-dependent shift in V pkcm. For a prepulse to -150 mV, from a holding potential of 0 mV, V pkcm shifted 6.4 mV, and was fitted by a single exponential time constant of 45 ms. A higher resolution analysis of this time course was made by measuring the change in capacitance during a fixed voltage step and indicated a double exponential shift (τ 0 = 51.6 ms, τ 1 = 8.5 s) similar to that of the native gerbil OHC. 4. Membrane tension, delivered by increasing pipette pressure, caused a positive shift in V pkcm. A maximal shift of 7.5 mV was obtained with 2 kPa of pressure. The effect was reversible. 5. Our results show that the sensitivity of prestin to initial voltage and membrane tension, though present, is less than that observed in adult OHCs. It remains possible that some other interacting molecular species within the lateral plasma membrane of the native OHC amplifies the effect of tension and prior voltage on prestin's activity.",
author = "Joseph Santos-Sacchi and Weixing Shen and Jing Zheng and Peter Dallos",
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Effects of membrane potential and tension on prestin, the outer hair cell lateral membrane motor protein. / Santos-Sacchi, Joseph; Shen, Weixing; Zheng, Jing; Dallos, Peter.

In: Journal of Physiology, Vol. 531, No. 3, 15.03.2001, p. 661-666.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Effects of membrane potential and tension on prestin, the outer hair cell lateral membrane motor protein

AU - Santos-Sacchi, Joseph

AU - Shen, Weixing

AU - Zheng, Jing

AU - Dallos, Peter

PY - 2001/3/15

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N2 - 1. Under whole-cell voltage clamp, the effects of initial voltage conditions and membrane tension on gating charge and voltage-dependent capacitance were studied in human embryonic kidney cells (TSA201 cell line) transiently transfected with the gene encoding the gerbil protein prestin. Conformational changes in this membrane-bound protein probably provide the molecular basis of the outer hair cell (OHC) voltage-driven mechanical activity, which spans the audio spectrum. 2. Boltzmann characteristics of the charge movement in transfected cells were similar to those reported for OHCs (Q max = 0.99 ± 0.16 pC, z = 0.88 ± 0.02; n = 5, means ± S.E.M.). Unlike that of the adult OHC, the voltage at peak capacitance (V pkcm) was very negative (-74.7 ± 3.8 mV). Linear capacitance in transfected cells was 43.7 ± 13.8 pF and membrane resistance was 458 ± 123 MΩ. 3. Voltage steps from the holding potential preceding the measurement of capacitance-voltage functions caused a time- and voltage-dependent shift in V pkcm. For a prepulse to -150 mV, from a holding potential of 0 mV, V pkcm shifted 6.4 mV, and was fitted by a single exponential time constant of 45 ms. A higher resolution analysis of this time course was made by measuring the change in capacitance during a fixed voltage step and indicated a double exponential shift (τ 0 = 51.6 ms, τ 1 = 8.5 s) similar to that of the native gerbil OHC. 4. Membrane tension, delivered by increasing pipette pressure, caused a positive shift in V pkcm. A maximal shift of 7.5 mV was obtained with 2 kPa of pressure. The effect was reversible. 5. Our results show that the sensitivity of prestin to initial voltage and membrane tension, though present, is less than that observed in adult OHCs. It remains possible that some other interacting molecular species within the lateral plasma membrane of the native OHC amplifies the effect of tension and prior voltage on prestin's activity.

AB - 1. Under whole-cell voltage clamp, the effects of initial voltage conditions and membrane tension on gating charge and voltage-dependent capacitance were studied in human embryonic kidney cells (TSA201 cell line) transiently transfected with the gene encoding the gerbil protein prestin. Conformational changes in this membrane-bound protein probably provide the molecular basis of the outer hair cell (OHC) voltage-driven mechanical activity, which spans the audio spectrum. 2. Boltzmann characteristics of the charge movement in transfected cells were similar to those reported for OHCs (Q max = 0.99 ± 0.16 pC, z = 0.88 ± 0.02; n = 5, means ± S.E.M.). Unlike that of the adult OHC, the voltage at peak capacitance (V pkcm) was very negative (-74.7 ± 3.8 mV). Linear capacitance in transfected cells was 43.7 ± 13.8 pF and membrane resistance was 458 ± 123 MΩ. 3. Voltage steps from the holding potential preceding the measurement of capacitance-voltage functions caused a time- and voltage-dependent shift in V pkcm. For a prepulse to -150 mV, from a holding potential of 0 mV, V pkcm shifted 6.4 mV, and was fitted by a single exponential time constant of 45 ms. A higher resolution analysis of this time course was made by measuring the change in capacitance during a fixed voltage step and indicated a double exponential shift (τ 0 = 51.6 ms, τ 1 = 8.5 s) similar to that of the native gerbil OHC. 4. Membrane tension, delivered by increasing pipette pressure, caused a positive shift in V pkcm. A maximal shift of 7.5 mV was obtained with 2 kPa of pressure. The effect was reversible. 5. Our results show that the sensitivity of prestin to initial voltage and membrane tension, though present, is less than that observed in adult OHCs. It remains possible that some other interacting molecular species within the lateral plasma membrane of the native OHC amplifies the effect of tension and prior voltage on prestin's activity.

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