The potassium channel subunit Kv1.8 (Kcna10) is essential for the distinctive outwardly rectifying conductances of type I and II vestibular hair cells
Data files
Jan 12, 2024 version files 1.96 MB
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F1_sourcedata.xlsx
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F2_sourcedata.xlsx
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F3_sourcedata.xlsx
380.56 KB
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F4_sourcedata.xlsx
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F6_sourcedata.xlsx
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F7_sourcedata.xlsx
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README.md
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SF1_sourcedata.xlsx
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SF2_sourcedata.xlsx
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SF3_sourcedata.xlsx
55.59 KB
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SF4_sourcedata.xlsx
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SF5_sourcedata.xlsx
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SF6_sourcedata.xlsx
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Abstract
In amniotes, head motions and tilt are detected by two types of vestibular hair cells (HCs) with strikingly different morphology and physiology. Mature type I HCs express a large and very unusual potassium conductance, gK,L, which activates negative to resting potential, confers very negative resting potentials and low input resistances, and enhances an unusual non-quantal transmission from type I cells onto their calyceal afferent terminals. Following clues pointing to KV1.8 (KCNA10) in the Shaker K channel family as a candidate gK,L subunit, we compared whole-cell voltage-dependent currents from utricular hair cells of KV1.8-null mice and littermate controls. We found that KV1.8 is necessary not just for gK,L but also for fast-inactivating and delayed rectifier currents in type II HCs, which activate positive to resting potential. The distinct properties of the three KV1.8-dependent conductances may reflect different mixing with other KV1 subunits, such as KV1.4 (KCNA4). In KV1.8-null HCs of both types, residual outwardly rectifying conductances include KV7 (KCNQ) channels.
Current clamp records show that in both HC types, KV1.8-dependent conductances increase the speed and damping of voltage responses. Features that speed up vestibular receptor potentials and non-quantal afferent transmission may have helped stabilize locomotion as tetrapods moved from water to land.
README: The potassium channel subunit KV1.8 (Kcna10) is essential for the distinctive outwardly rectifying conductances of type I and II vestibular hair cells
This README file was generated on 2023-10-06 by Hannah R. Martin. It can be read with Markdown syntax.\
These datasets contain processed (not raw) data used in the analyses of patch clamp electrophysiology in semi-intact utricles from mice.\
The study investigates the role of Kv1.8 conductances in vestibular hair cells.
Description of the data and file structure
Datasets are in excel or csv format. Datasets have headers, and additional information is provided in this document.
GENERAL INFORMATION
Title of Dataset: The potassium channel subunit KV1.8 (Kcna10) is essential for the distinctive outwardly rectifying conductances of type I and II vestibular hair cells
Author Information
A. Principal Investigator Contact Information
Name: Ruth Anne Eatock
Institution: The University of Chicago
Address: Chicago, IL USA
Email: eatock@uchicago.eduB. First Author Contact Information
Name: Hannah R. Martin
Institution: The University of Chicago
Address: Chicago, IL USA
Institutional Email: hrmartin@uchicago.edu
Permanent Email: hannah.m2357@gmail.comDate of data collection: 2017-2023
Information about funding sources that supported the collection of the data: National Institutes of Health (NIDCD) grant R01DC012347 (to R.A.E.), National Science Foundation GRFP (to H.R.M.)
SHARING/ACCESS INFORMATION
Licenses/restrictions placed on the data: CC BY 1.0
Links to publications that cite or use the data:
https://doi.org/10.1101/2023.11.21.563853Links to other publicly accessibly locations of the data: None
Links/relationships to ancillary data sets: None
Was data derived from another source? No\
A. If yes, list source(s): NARecommended citation for this dataset:
[Dryad repository citation]Code/Software: NA
Blank cells: Any blank cells in the excel/csv files indicates a lack of data for that variable and observation.
DATA & FILE OVERVIEW
Each file corresponds to a figure in the manuscript, and contains sufficient data to recreate all statistical tests. Each .xlsx file contains sheets that are explicitly labelled to refer to a panel within that figure. Each sheet has unambiguous header information describing the parameter (details on analysis methods are found in the Methods section of the manuscript), condition (genotype, age, zone, drug), and the unique CellID of every cell used in the analysis.
- Voltage-Clamp Data F1_sourcedata.xlsx F3_sourcedata.xlsx F6_sourcedata.xlsx SF1_sourcedata.xlsx SF2_sourcedata.xlsx SF3_sourcedata.xlsx SF4_sourcedata.xlsx SF5_sourcedata.xlsx SF6_sourcedata.xlsx
- Current-Clamp Data F2_sourcedata.xlsx F4_sourcedata.xlsx
- Pharmacology Data F7_sourcedata.xlsx
Relationship between files: Some data is repeated in different files if it is used in multiple figures. This is easily seen if when the unique CellID for each data entry is found in multiple files. Below are the files that share some data between them:
- F1_sourcedata, SF1_sourcedata: SF1_sourcedata shows data from a wider age range than F1_sourcedata
- F3_sourcedata, SF2_sourcedata, SF23_sourcedata: SF2_sourcedata contains the individual data points that are represented by mean and SEM in F3_sourcedata; SF3_sourcedata shows data from a wider age range than SF2_sourcedata
Additional related data collected that was not included in the current data package: Immunohistochemistry data was included because it was not used quantitatively.
Are there multiple versions of the dataset? No
A. If yes, name of file(s) that was updated: N/A
i. Why was the file updated? N/A
ii. When was the file updated? N/A
DATA-SPECIFIC INFORMATION
Additional information for some sheets within files is provided below.
Voltage-Clamp Data
Filename: F1_sourcedata.xlsx
Sheetname: F1C
Number of Variables: 12
- V_het: mV, step voltage of the representative Kv1.8+/- type I HC
- G_het: nS, tail conductance of the representative Kv1.8+/- type I HC
- Gnorm_het: normalized tail conductance of the representative Kv1.8+/- type I HC
- V_ko: mV, step voltage of the representative Kv1.8-/- type I HC
- G_ko: nS, tail conductance of the representative Kv1.8-/- type I HC
- Gnorm_ko: normalized tail conductance of the representative Kv1.8-/- type I HC
- V_het_fit: mV, voltages of Boltzmann fit of representative Kv1.8+/- type I HC
- G_het_fit: nS, fit conductance values of the representative Kv1.8+/- type I HC
- Gnorm_het_fit: normalized fit conductance of the representative Kv1.8+/- type I HC
- V_ko_fit: mV, voltages of Boltzmann fit of representative Kv1.8-/- type I HC 11 G_ko_fit: nS, fit conductance values of the representative Kv1.8-/- type I HC
- Gnorm_ko_fit: normalized fit conductance of the representative Kv1.8-/- type I HC
Sheetname: F1D
Number of variables: 16
- CellID: Unique cell identifier
- Genotype: Kcna10-TM45
- Age: Postnatal Days
- Weight: Grams. Blank means the weight was not recorded.
- Sex: Male or Female. Blank means the sex was not determined.
- Zone: Zone location within the utricle
- Vhalf: in mV, the midpoint of activation of the tail G-V curve
- S: in mV, the slope factor of the tail G-V curve
- gmin: in nS, the minimum of the tail G-V curve
- gmax: in nS, the maximum of the tail G-V curve
- Cm: in pF, the capacitance of the cell measured in HEKA
- Gdensity: in nS/pF, (gmax - gmin)/Cm
- Rs: in megaOhms, the series resistance
- Rs_comp: in percent, the percent compensation of series resistance
- VCfile1: the name of the best quality voltage-clamp file used for these parameters
- VCfile2: the name of the second-best quality voltage-clamp file
Filename: F3_sourcedata.xlsx
Sheetname: F3A
Number of variables: 15
Variable List:
- Time: ms
- I_WT_LES: current normalized by max
- I_WT_LES_fit: current fit to Equation 4, normalized to max
- I_WT_MES: current normalized by max
- I_WT_MES_fit: current fit to Equation 4, normalized to max
- I_WT_Striola: current normalized by max
- I_WT_Striola_fit: current fit to Equation 4, normalized to max
- Genotype: KCNA10-TM45
- Zone: utricular location
- Tau_FastInactivation_Mean_ms: ms, mean fast time constant of inactivation
- Tau_FastInactivation_SEM: ms, standard error of the mean
- Tau_FastInactivation_N: number of cells
- Fraction_FastInactivation_Mean: mean relative amplitude of fast inactivation to total inactivation
- Fraction_FastInactivation_SEM: standard error of the mean
- Fraction_FastInactivation_N: number of cells
Sheetname: F3C
Number of Variables: 5
- Genotype: KCNA10-TM45
- Zone: utricular location
- PercentInactivation_Mean: %, inactivating current as a percent of total current
- PercentInactivation_SEM: standard error of the mean
- PercentInactivation_N: number of cells
Sheetname: F3D
Number of variables: 43
Variable List:
For panel Fig 3D1
- Time: ms
- I_WT_LES: nA, representative current response from Kv1.8+/+ LES type II HC
- I_WT_LES_fit: nA, fit curve
- I_HET_LES: nA, representative current response from Kv1.8+/- LES type II HC
- I_HET_LES_fit: nA, fit curve
- I_KO_LES: nA, representative current response from Kv1.8-/- LES type II HC
- I_KO_LES_fit: nA, fit curve
- VpkMean_WT: mV, mean of step voltages of Kv1.8+/+ LES type II HCs
- GpkMean_WT: nS, mean of peak conductances of Kv1.8+/+ LES type II HCs
- GpkSEM_WT: nS, SEM of peak conductances of Kv1.8+/+ LES type II HCs
- Npk_WT: number of Kv1.8+/+ LES type II HCs
- GpkMeanNorm_WT: peak conductances of Kv1.8+/+ LES type II HCs, normalized by Gmax
- GpkSEMNorm_WT: SEM of peak conductances of Kv1.8+/+ LES type II HCs, normalized by Gmax
- VpkFit_WT: mV, voltage of fit curve to mean peak conductance
- GpkFit_WT: nS, conductance of fit curve to mean peak conductance
- GpkFitNorm_WT: conductance of fit curve to mean peak conductance, normalized by Gmax
- VssMean_WT: mV, mean of step voltages of Kv1.8+/+ LES type II HCs
- GssMean_WT: nS, mean of steady-state conductances of Kv1.8+/+ LES type II HCs
- GssSEM_WT: nS, SEM of steady-state conductances of Kv1.8+/+ LES type II HCs
- Nss_WT: number of Kv1.8+/+ LES type II HCs
- GssMeanNorm_WT: steady-state conductances of Kv1.8+/+ LES type II HCs, normalized by Gmax
- GssSEMNorm_WT: SEM of steady-state conductances of Kv1.8+/+ LES type II HCs, normalized by Gmax
- VssFit_WT: mV, voltage of fit curve to mean steady-state conductance
- GssFit_WT: nS, conductance of fit curve to mean steady-state conductance
- GssFitNorm_WT: conductance of fit curve to mean steady-state conductance, normalized by Gmax
- VpkMean_KO: mV, mean of step voltages of Kv1.8-/- LES type II HCs
- GpkMean_KO: nS, mean of peak conductances of Kv1.8+-/- LES type II HCs
- GpkSEM_KO: nS, SEM of peak conductances of Kv1.8-/- LES type II HCs
- Npk_KO: number of Kv1.8-/- LES type II HCs
- GpkMeanNorm_KO: peak conductances of Kv1.8-/- LES type II HCs, normalized by Gmax
- GpkSEMNorm_KO: SEM of peak conductances of Kv1.8-/- LES type II HCs, normalized by Gmax
- VpkFit_KO: mV, voltage of fit curve to mean peak conductance
- GpkFit_KO: nS, conductance of fit curve to mean peak conductance
- GpkFitNorm_KO: conductance of fit curve to mean peak conductance, normalized by Gmax
- VssMean_KO: mV, mean of step voltages of Kv1.8-/- LES type II HCs
- GssMean_KO: nS, mean of steady-state conductances of Kv1.8-/- LES type II HCs
- GssSEM_KO: nS, SEM of steady-state conductances of Kv1.8-/- LES type II HCs
- Nss_KO: number of Kv1.8-/- LES type II HCs
- GssMeanNorm_KO: steady-state conductances of Kv1.8-/- LES type II HCs, normalized by Gmax
- GssSEMNorm_KO: SEM of steady-state conductances of Kv1.8-/- LES type II HCs, normalized by Gmax
- VssFit_KO: mV, voltage of fit curve to mean steady-state conductance
- GssFit_KO: nS, conductance of fit curve to mean steady-state conductance
- GssFitNorm_KO: conductance of fit curve to mean steady-state conductance, normalized by Gmax
Filename: F6_sourcedata.xlsx
Sheetname: F6B
Number of Variables: 12
- Vpeak: mV, for x-axis of representative activation curve in h-infinity plot
- Gpeak: nS, for y-axis
- Gpeak_norm: Gpeak/Gmax
- Vpeakfit: mV, x-values for fit curves
- Gpeakfit: nS
- Gpeak_normfit
- Vinac: mV, for x-axis of representative inactivation curve in h-infinity plot
- Ginac: nS
- Ginac_norm: Ginac/Gmax
- Vinacfit: mV
- Ginacfit: nS
- Ginax_normfit
Filename: SF1_sourcedata.xlsx
Sheetname: SF1A
Number of Variables: 14
Variable List:
- CellID: Unique cell identifier
- Genotype: KCNA10-TM45 genotype
- Age: postnatal day
- Weight: grams
- Sex
- Zone: Utricular zone
- Vhalf: mV, tail Vhalf
- S: mV, tail slope factor
- gmin: nS, tail Gmin
- gmax: nS, tail Gmax
- Cm: pF, cell capacitance 12: Gdensity: nS/pF, (gmax - gmin)/Cm
- VCfile1: best voltage clamp file name 14: VCfile2: second best voltage clamp file name
Sheetname: SF1B
Number of Variables: 8
Variable List:
- CellID: unique cell identifier
- Genotype: KCNA10-TM45 genotype
- Age: postnatal day
- Zone: Utricular zone
- Vhalf: mV, tail Vhalf
- Gdensity: nS/pF, (gmax - gmin)/Cm
- Group: see Variable 8
- GroupName: informative name that describes which group the cells are in (age range, with or without gKL, without or without Kv1.8)
Filename: SF2_sourcedata.xlsx
Sheetname: SF2
Number of variables: 15
Variable List:
- CellID: Unique cell identifier
- Genotype: KCNA10-TM45 genotype
- Age: Postnatal Days
- Weight: Grams. Blank means the weight was not recorded.
- Sex: Male or Female. Blank means the sex was not determined.
- Zone: Zone location within the utricle
- Cm: in pF, the capacitance of the cell measured in HEKA
- PercentInactivation: %, percent inactivation at 30 mV
- tau_Activation: ms, from fit to Eq. 3 or Eq. 4
- tau_FastInactivation: ms, from fit to Eq. 4
- Inactivation_FastFraction: from fit to Eq. 4
- Rs: in megaOhms, the series resistance
- Rs_comp: in percent, the percent compensation of series resistance
- VCfile1: the name of the best quality voltage-clamp file used for these parameters
- VCfile2: the name of the second-best quality voltage-clamp file
Filename: SF3_sourcedata.xlsx
Sheetname: SF3
Number of variables: 21
Variable List:
- CellID: Unique cell identifier
- Genotype: KCNA10-TM45 genotype
- Age: Postnatal Days
- Weight: Grams. Blank means the weight was not recorded.
- Sex: Male or Female. Blank means the sex was not determined.
- Zone: Zone location within the utricle
- Cm: in pF, the capacitance of the cell measured in HEKA
- peak_Vhalf: mV, midpoint of activation of peak conductance voltage curve
- peak_S: mV, slope factor of peak conductance-voltage fit
- peak_Gmax: nS, gmax of peak conductnace-voltage fit
- peakGdensity: nS/pF, gmax/Cm
- ss_Vhalf: mV
- ss_S: mV
- ss_Gmax: nS
- ss_Gdensity: nS/pF
- inac_Gdensity: nS/pF
- PercentInactivation: %
- Rs: megaOhm, total series resistance
- Comp: %, percent compensatin of series resistance
- VCfile1: the name of the best quality voltage-clamp file used for these parameters
- VCfile2: the name of the second-best quality voltage-clamp file
Filename: SF4_sourcedata.xlsx
Sheetname: SF4A
Number of variables: 3
- IA30WT: %, percent inactivation in Kv1.8+/+ type II HCs >P12
- IA30HET: %, percent inactivation in Kv1.8+/- type II HCs >P12
- IA30KO: %, percent inactivation in Kv1.8-/- type II HCs >P12
Sheetname: SF4B
Number of variables: 20
- Time: ms
- I_fit: nA, fit curve of double exponential decay fit to current at 30 mV
- CellID: unique cell identifier
- Genotype: KCNA10-TM45
- Age: Postnatal day
- Weight: g
- Sex
- Zone: Utricular location
- Cm: pF, cell capacitance
- peakVhalf: mV, midpoint of activation of peak conductance voltage curve
- peakS: mV, slope factor of peak conductance-voltage fit
- peakGdensity: nS/pF, gmax/Cm
- ssVhalf: mV
- ssS: mV
- ssGDensity: nS/pF
- inacGdensity: nS/pF
- PercentInactivation: %
- tau_activatiom: ms
- tau_inactivationFast: mS
- fastFraction: amplitude of fast inactivation relative to total inactivation
Filename: SF5_sourcedata.xlsx
Sheetname: SF5A
Number of variables: 2
- Time: ms
- I_fit: nA, exponential decay fit of representative XE991-sensitive current in Kv1.8-/- type I HC from the striola
Sheetname: SF5B
Number of variables: 4
- Vtail: mV
- Gtail: nS, representative XE991-sensitive tail conductance in Kv1.8-/- type I HC from the striola
- Vtail_fit: mV
- Gtail_fit: nS, from double Boltzmann fit
SHeetname: SF5C
Number of variables: 2
- KO_ES_TypeI_tailVhalf: tail Vhalf values of Kv1.8-/- type I HCs (>P12) from the extrastriola
- KO_Striola_TypeI_tailVhalf: tail Vhalf values (including single and double Boltzmann fits) of Kv1.8-/- type I HCs (>P12) from the striola
Filename: SF6_sourcedata.xlsx
Sheetname: SF6A
Number of variables: 12
Variable List:
- Time: ms
- WH_TypeI_ES: nA, representative Kv1.8+/+,+/- type I HC from the extrastriola
- WH_TypeI_Striola: nA, representative Kv1.8+/+,+/- type I HC from the striola
- KO_TypeI_ES: nA, representative Kv1.8-/- type I HC from the extrastriola
- KO_TypeI_Striola1: nA, first representative Kv1.8-/- type I HC from the striola
- KO_TypeI_Striola2: nA, second representative Kv1.8-/- type I HC from the striola
- KO_TypeI_Striola3: nA, third representative Kv1.8-/- type I HC from the striola
- WH_TypeII_ES: nA, representative Kv1.8+/+,+/- type II HC from the extrastriola
- WH_TypeII_Striola: nA, representative Kv1.8+/+,+/- type II HC from the striola
- KO_TypeII_ES: nA, representative Kv1.8-/- type II HC from the extrastriola
- KO_TypeII_Striola: nA, representative Kv1.8-/- type II HC from the striola
- V: mV, voltage waveform for representative currents
Sheetname: SF6B
CellID: Unique cell identifier
HCType: Type I or II
Genotype: KCNA10-TM45 genotype
Age: Postnatal Day
Zone: utricular
Cm: pF, cell size
IHandIKirDensity: -1*pA/pF
Current-Clamp Data
Filename: F2_sourcedata.xlsx
Sheetname: F2A
Number of variables: 12
Variable List:
- CellID: Unique cell identifier
- Genotype: KCNA10-TM45 genotype
- Age: Postnatal Days
- Weight: Grams. Blank means the weight was not recorded.
- Sex: Male or Female. Blank means the sex was not determined.
- Zone: Zone location within the utricle
- Cm: pF, the capacitance of the cell measured in HEKA
- Vm: mV, the resting membrane potential at 0 pA current injection
- Rm: megaOhms, the input resistance is the ratio of steady-state voltage to injected current
- cTauRC: ms, the membrane time constant is the product of Cm and Rm
- CCfile1: best quality current-clamp file used for these parameters
- CCfile2: second-best quality current-clamp file
Sheetname: F2B3
Number of variables: 4
Variable List:
- Time: ms
- Vnorm_het: voltage response of representative Kv1.8+/- type I HC normalized by peak response
- Vnorm_KO: voltage response of representative Kv1.8-/- type I HC normalized by peak response
- I_inj: step of injected current, normalized by current amplitude
Filename: F4_sourcedata.xlsx
Sheetname: F4AE
Number of variables: 12
Variable List:
- CellID: Unique cell identifier
- Genotype: KCNA10-TM45 genotype
- Age: Postnatal Days
- Zone: Zone location within the utricle
- Cm: pF, the capacitance of the cell measured in HEKA
- Vrest: mV, the resting membrane potential at 0 pA current injection 7: I_inj: pA, the small step used to fit with an exponential growth equation 8: deltaV: mV, a fit parameter, the amplitude of the voltage response
- tau: ms, time constant of fit to exponential growth function
- V0: mV, baseline of fit
- R2: R-squared value of fit
- FitEndTime: user-selected time to stop the fit
- Rinput: GigaOhms, deltaV divided by I_inj
- peakTime: ms, delay of the peak of the depolarizing transient upon 170 pA current injection
- peakHeight: mV, absolute height of the depolarizing transient upon 170 pA current injection
Sheetname: F4C2
Number of variables: 8
Variable List:
- CellID: Unique cell identifier
- Zone: Location within the utricle
- Genotype: KCNA10-TM45 genotype
- Age: Postnatal Days
- I_Inj: pA, magnitude of current injection
- f: Hz, frequency of resonance from fit to Equation 5
- Q: Quality factor from Equation 6
- R2: R-squared value of fit to Equation 5
Sheetname: F4D
Number of variables: 4
- Time: ms
- Vnorm_HET: voltage response in Kv1.8+/- type II HC, normalized by peak
- Vnorm_KO: voltage response in Kv1.8-/- type II HC, normalized by peak
- Inorm_inj: current step waveform, normalized by step amplitude
Pharmacology with XE991
Filename: F7_sourcedata
Sheetname: F7B
Number of variables: 10\
Variable List:
- CellID: Unique cell id
- Zone: Utricular zone
- Age: Postnatal day
- Genotype: KCNA10-TM45
- Weight: in grams, the weight of the mouse
- Sex: Male or Female
- HCType: Type I or II HCCm: in pF, cell capacitance
- XE991_sensitive_Vhalf: in mV, the midpoint voltage of activation of the difference current tail G-V curve (current in 0 uM XE991 - current in 10 uM XE991)
- XE991_sensitive_gDensity: in nS/pF, the (gmax - gmin)/Cm of the difference current tail G-V curve
- PercentBlock: in percent, percent that the gDensity was of the total before drug
Sheetname: F7B_GV
Number of variables: 9
- V_KOTypeI: mV, step voltage of Kv1.8-/- type I HC
- Gfit_mean_KOTypeI: nS, mean Boltzmann fits to XE991-sensitive tail conductances of Kv1.8-/- type I HCs
- Gfit_sem_KOTypeI: nS, sem of Boltzmann fits to XE991-sensitive tail conductances of Kv1.8-/- type I HCs
- V_KOTypeII: mV, step voltage of Kv1.8-/- type II HC
- Gfit_mean_KOTypeII: nS, mean Boltzmann fits to XE991-sensitive tail conductances of Kv1.8-/- type II HCs
- Gfit_sem_KOTypeII: nS, sem of Boltzmann fits to XE991-sensitive tail conductances of Kv1.8-/- type II HCs
- V_WTTypeII: mV, step voltage of Kv1.8+/+ type II HC
- Gfit_mean_WTTypeII: nS, mean Boltzmann fits to XE991-sensitive tail conductances of Kv1.8+/+ type II HCs
- Gfit_sem_WTTypeII: nS, sem of Boltzmann fits to XE991-sensitive tail conductances of Kv1.8+/+ type II HCs
Methods
We compared whole-cell voltage-activated K+ currents in type I and type II hair cells from homozygous knockout (KV1.8–/–) animals and their wildtype (KV1.8+/+) or heterozygote (KV1.8+/–) littermates. We immunolocalized KV1.8 subunits in the utricular epithelium and pharmacologically characterized the residual K+ currents of KV1.8–/– animals. We performed current clamp experiments to assess the impact of KV1.8-dependent currents on passive membrane properties.