Gender specific click and tone burst evoked ABR datasets from mice lacking the Cav3.2 T-type voltage-gated calcium channel
BMC Research Notes volume 12, Article number: 157 (2019)
Voltage-gated Ca2+ channels (VGCCs) are of central relevance in regulating Ca2+ influx into living cells. The low-voltage activated (LVA) Cav3 T-type Ca2+ channels are widely distributed throughout the brain including the peripheral auditory system and ascending auditory tract. Their exact role in auditory information processing is still not fully understood. Within the LVA subgroup, Cav3.2 T-type Ca2+ channels seem to be of special importance as qPCR revealed a steady increase in Cav3.2 transcript levels over age, e.g. in the cochlea and spiral ganglion neurons (SGN). Furthermore, pharmacological studies suggested an association between Cav3.2 expression and both age-related and noise-induced hearing loss. Given the potential functional relevance of Cav3.2 VGGCs in sensorineural hearing loss, we recorded gender specific auditory evoked brainstem responses (ABRs) upon both click and tone burst presentation. Here we present auditory brainstem response (ABR) data from Cav3.2+/+, Cav3.2+/− and Cav3.2−/− mice from both genders which are of value for researchers who want to evaluate how Cav3.2 loss affects basic auditory parameters, e.g. click and tone burst based hearing thresholds, amplitude growth function and peak latencies.
Information presented here includes ABR data from age-matched female and male Cav3.2+/+, Cav3.2+/− and Cav3.2−/− mice and technical aspects of the auditory recording protocol. Data were recorded using a commercially available ABR setup from Tucker Davis Technologies Inc. (TDT). Raw data files (arf.-file format) were exported as txt.-files with free access for analysis.
Voltage-gated Ca2+ channels are key players in regulating cellular Ca2+ homeostasis. Only few Ca2+ channels have been functionally related to auditory information processing including Cav1.3 L-type Ca2+ channels, the ablation of which results in congenital deafness. Recent animal studies suggested that Cav3.2 T-type Ca2+ channels might play a role in age- and noise-induced hearing loss as ablation of the channel seemed to protect from sensorineural hearing loss. On the other hand, Cav3.2 Ca2+ channels were reported to exhibit increased expression with age in the inner ear/spiral ganglion neurons pointing to an important functional role in the auditory system. Interestingly, no auditory profiling of Cav3.2+/− and Cav3.2−/− mice had been performed to unravel the physiological involvement of Cav3.2 in the peripheral and ascending auditor tract. To do so, we carried out click and tone burst evoked auditory brainstem response (ABR) recordings from Cav3.2+/+, Cav3.2+/− and Cav3.2−/− mice from both genders. Monaural recording results in all three lines were analyzed for threshold alterations and differences in peak amplitudes and peak latencies and submitted elsewhere. Raw ABR data were exported as txt.-files to provide free access and to enable researchers to carry out their own ABR data analysis, including further investigation of binaural recordings or application of additional manual and/or automatic analytical tools (Table 1).
Cav3.2 transgenic mice  from Mutant Mouse Resource and Research Centers (MMRRC: 009979-MU; strain name: B6.129-Cacna1htm1Kcam/Mmmh) were maintained in the C57Bl/6J background. For subsequent ABR recordings, Cav3.2+/+ controls, heterozygous Cav3.2+/− and homozygous null mutant Cav3.2−/− mice (55 animals in total) were used from both age-matched genders with the following characteristics: Males: Cav3.2+/+: n = 11 (♂), weight 32.82 ± 0.58 g; Cav3.2+/−: n = 7 (♂), weight 33.11 ± 0.81 g; Cav3.2−/−: n = 9 (♂), weight 29.09 ± 0.75 g. Females: Cav3.2+/+: n = 12 (♀), weight 24.09 ± 0.41 g; Cav3.2+/−: n = 8 (♀), weight 23.50 ± 0.41 g; Cav3.2−/−: n = 8 (♀), weight 22.10 ± 0.43 g.
ABR recording procedure
For recording of monaural bioelectrical auditory potentials, subdermal stainless steel electrodes were inserted at the vertex, axial the pinnae [(+) electrode] and ventrolateral of the right pinna [(−) electrode]. The ground electrode was positioned at the hip of the animal. To verify proper electrode positioning/conductivity, impedance measurements of all electrodes (< 5 kΩ) were carried out prior to each recording [Lundt A, Seidel, Robin, Soos J, Henseler C, Müller R, Bakki M, Arshaad IM, Ehninger D, Hescheler J, Sachinidis A, Broich K, Wormuth C, Papazoglou A, Weiergräber M. Cav3.2 T-type calcium channels are physiologically mandatory for the auditory system despite their devastating role in sensorineural hearing loss. Neuroscience, unpublished].
All ABR recordings were performed under free field conditions using a single loudspeaker (MF1 Multi-Function Speaker, TDT, USA) which was positioned 10 cm opposite to the rostrum of the animals.
The SigGenRZ software (TDT) was used to program stimulus protocols for click and tone bursts. The bioelectrical ABR signals recorded from the subdermal electrodes were transferred to a head stage (RA4LI, TDT) and forwarded to the preamplifier (RA4PA, TDT) with 20-fold amplification.
ABR data acquisition was carried out at a sampling rate of 24.4 kHz and signals were bandpass filtered (high pass 300 Hz, low pass 5 kHz) using a 6-pole Butterworth filter. The individual ABR data acquisition time was 25 ms starting with a 5 ms baseline period prior to the individual acoustic stimulus onset (pre ABR baseline) and exceeding the 10 ms ABR section by another 10 ms baseline (post ABR baseline) [Lundt et al., unpublished].
Two types of acoustic stimuli were applied for ABR recordings using the SigGenRZ software (TDT) and applied via the TDT BioSigRZ platform. The first stimulus entity was a click of 100 µs duration, with alternating polarity (switching between condensation and rarefaction).
The second stimulus entity was a 4.5 ms tone burst (transient sinusoidal plus) of alternating polarity with Hann envelope rise and fall times of 1.5 ms duration. The frequency range covers 1–42 kHz in 6 kHz steps. All acoustic stimuli were applied 300 times at a rate of 20 Hz for averaging.
Sound pressure levels (SPL) were increased in 5 dB steps for clicks and 10 dB steps for tone bursts, starting from 0 dB up to 90 dB (increasing SPL mode). Sound pressure levels for tone bursts within the range of 1–42 kHz were calibrated each day prior to recording [Lundt et al., unpublished].
ABR data presented here were performed under standard free field conditions. Data were recorded from age-matched animals of ~ 20 weeks. We did not record from animals of different age.
auditory brainstem response
sound pressure level
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Lundt A, Seidel, Robin, Soos J, Henseler C, Müller R, Bakki M, Arshaad IM, Ehninger D, Hescheler J, Sachinidis A, Broich K, Wormuth C, Papazoglou A, Weiergräber M. Datasets from gender specific auditory brainstem responses (ABRs) from Cav3.2+/+, Cav3.2+/− and Cav3.2−/− mice following click and tone burst stimulation. Mendeley Data. http://dx.doi.org/10.17632/dms4hsd75s.1.
AL, data acquisition; CH, animal housing and genotyping; CW, data management; JS, data acquisition; RS, data management; RM, data management; IMA, data management; KB, drafting manuscript; JH, drafting manuscript; AS, drafting manuscript; DE, drafting manuscript; AP, data acquisition, drafting manuscript; MW, project management, project planning, data acquisition, writing manuscript. All authors read and approved the final manuscript.
The authors are grateful to Dr. Christina Kolb (German Center for Neurodegenerative Diseases, DZNE) and Dr. Robert Stark (DZNE) for assistance in animal breeding and animal health care.
The authors declare that they have no competing interests.
Availability of data materials
The data described in this Data note can be freely and openly accessed on Mendeley Data (http://dx.doi.org/10.17632/dms4hsd75s.1). Please see Table 1 and reference list for details and links to the data.
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Ethics approval and consent to participate
All animal procedures were performed according to the Guidelines of the German Council on Animal Care and all protocols were approved by the Local Institutional and National Committee on Animal Care (Landesamt für Natur, Umwelt und Verbraucherschutz, LANUV, Germany). The authors further certify that all animal experimentation was carried out in accordance with the European Communities Council Directive of November 24, 1986 (86/609/EEC). Specific effort was made to minimize the number of animals used and their suffering.
This work was financially supported by the Federal Institute for Drugs and Medical Devices (Bundesinstitut für Arzneimittel und Medizinprodukte (BfArM), Bonn, Germany). BfArM provided financial and experimental resources to carry out the study.
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Lundt, A., Henseler, C., Wormuth, C. et al. Gender specific click and tone burst evoked ABR datasets from mice lacking the Cav3.2 T-type voltage-gated calcium channel. BMC Res Notes 12, 157 (2019). https://doi.org/10.1186/s13104-019-4169-4