Data from: GCaMP6f-expressing chromaffin cells in murine adrenal slices exhibit dynamic spontaneous calcium responses that do not require nerve input
Data files
Jan 30, 2026 version files 118.76 MB
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ATR_Female_Subject_1.xlsx
1.80 MB
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ATR_Female_Subject_2.xlsx
1.33 MB
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ATR_Female_Subject_3.xlsx
1.89 MB
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ATR_Male_Subject_1.xlsx
2.03 MB
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ATR_Male_Subject_2.xlsx
5.59 MB
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ATR_Male_Subject_3.xlsx
1.04 MB
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CPA_Female_Subject_1.xlsx
2.50 MB
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CPA_Female_Subject_2.xlsx
1.25 MB
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CPA_Female_Subject_3.xlsx
1.43 MB
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CPA_Male_Subject_1.xlsx
2.85 MB
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CPA_Male_Subject_2.xlsx
5.58 MB
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CPA_Male_Subject_3.xlsx
2.74 MB
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DMSO_Female_Subject_1.xlsx
1.95 MB
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DMSO_Female_Subject_2.xlsx
4.16 MB
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DMSO_Female_Subject_3.xlsx
3.04 MB
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DMSO_Male_Subject_1.xlsx
3 MB
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DMSO_Male_Subject_2.xlsx
3.88 MB
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DMSO_Male_Subject_3.xlsx
2.70 MB
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EGTA_Female_Subject_1.xlsx
1.25 MB
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EGTA_Female_Subject_2.xlsx
3.33 MB
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EGTA_Female_Subject_3.xlsx
3.54 MB
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EGTA_Male_Subject_1.xlsx
309.87 KB
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EGTA_Male_Subject_2.xlsx
421.84 KB
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EGTA_Male_Subject_3.xlsx
305.77 KB
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ETH_Female_Subject_1.xlsx
1.06 MB
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ETH_Female_Subject_2.xlsx
495.64 KB
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ETH_Female_Subject_3.xlsx
2.69 MB
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ETH_Male_Subject_1.xlsx
2.51 MB
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ETH_Male_Subject_2.xlsx
4.67 MB
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ETH_Male_Subject_3.xlsx
5.57 MB
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HEX_Female_Subject_1.xlsx
2.25 MB
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HEX_Female_Subject_2.xlsx
1.08 MB
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HEX_Female_Subject_3.xlsx
1.03 MB
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HEX_Male_Subject_1.xlsx
2.16 MB
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HEX_Male_Subject_2.xlsx
2.50 MB
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HEX_Male_Subject_3.xlsx
523.45 KB
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NIF_Female_Subject_1.xlsx
1.56 MB
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NIF_Female_Subject_2.xlsx
4.19 MB
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NIF_Female_Subject_3.xlsx
465.12 KB
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NIF_Male_Subject_1.xlsx
4.46 MB
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NIF_Male_Subject_2.xlsx
1.28 MB
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NIF_Male_Subject_3.xlsx
1.56 MB
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NO_DRUG_Female_Subject_1.xlsx
989.94 KB
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NO_DRUG_Female_Subject_2.xlsx
5.12 MB
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NO_DRUG_Female_Subject_3.xlsx
2.03 MB
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NO_DRUG_Male_Subject_1.xlsx
1.09 MB
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NO_DRUG_Male_Subject_2.xlsx
2.62 MB
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NO_DRUG_Male_Subject_3.xlsx
933.61 KB
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README.md
4.81 KB
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TTX_Female_Subject_1.xlsx
1.06 MB
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TTX_Female_Subject_2.xlsx
307.45 KB
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TTX_Female_Subject_3.xlsx
2.17 MB
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TTX_Male_Subject_1.xlsx
708.04 KB
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TTX_Male_Subject_2.xlsx
2.15 MB
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TTX_Male_Subject_3.xlsx
1.62 MB
Abstract
Chromaffin cells of the adrenal medulla exhibit spontaneous action potentials and intracellular Ca2+ responses that, when dissociated in vitro, result in catecholamine secretion. However, spontaneous activity in these cells is nearly abolished when splanchnic nerve input is blocked in vivo. To address this discrepancy, we examined spontaneous Ca2+ responses in chromaffin cells using an intermediate preparation: adrenal slices from transgenic mice expressing the genetically encoded Ca2+ indicator GCaMP6f selectively in these cells. Within this preparation, the three-dimensional structure and extracellular environment surrounding chromaffin cells are retained, as is input from distal splanchnic nerve terminals, which spontaneously release the neurotransmitter acetylcholine. Chromaffin cells within these slices displayed spontaneous Ca2+ responses with frequencies and amplitudes that varied greatly within and between individual cells. However, population averages remained stable, providing a tool to measure the cellular and molecular mechanisms underlying these responses. While overall average frequencies and amplitudes of spontaneous Ca2+ responses depended on the influx of extracellular Ca2+ through voltage-gated Ca2+ channels, they did not require Ca2+ release from intracellular stores, splanchnic nerve input, or activation of voltage-gated sodium channels. Together, these results suggest that spontaneous activity in chromaffin cells in adrenal slices is generated autonomously, similar to that in dissociated chromaffin cells. By contrast, spontaneous activity in chromaffin cells in intact animals likely represents a distinct form that depends on basal input from the intact splanchnic nerve. This study provides a foundation to further explore the diverse mechanisms mediating chromaffin cell activation.
Dataset DOI: 10.5061/dryad.zpc866tm1
Description of the data and file structure
We collected spontaneous calcium fluorescence transients from chromaffin cells within adrenal slices from mice expressing the calcium indicator GCaMP6f. To determine which mechanisms contributed to this spontaneous activity, we conducted several experiments in which we blocked various ion channels and receptors while recording calcium activity in six one-minute videos spaced five minutes apart. The first three videos were recorded prior to (pre-), and the last three videos after (post-), perfusing the blocker of choice. The drugs we used included EGTA to deplete extracellular calcium, cyclopiazonic acid (CPA) to deplete intracellular calcium stores, nifedipine (NIF) to selectively block L-type voltage-gated calcium channels, tetrodotoxin (TTX) to block voltage-gated sodium channels, hexamethonium (HEX) to block nicotinic acetylcholine receptors, and atropine (ATR) to block muscarinic acetylcholine receptors. We also included control experiments in which we demonstrated the lack of effect of ethanol (ETH) and DMSO, the solvents in which NIF and CPA were dissolved, respectively.
For each drug experiment, we have provided 6 Excel .xlsx data files, including 3 from male and 3 from female mouse subjects/slices, to which we added the drug indicated in the file title. In turn, each Excel .xlsx file contains 6 tabs, the data on each tab representing that from each of 6 videos that were acquired at 5 minute intervals. Tabs 1-3 contain fluorescence transients from chromaffin cells (1 cell per column) captured prior to, and tabs 4-6 contain fluorescence transients captured after, drug perfusion. The transients were obtained after background subtraction was performed in Image J. Only transients that met the criteria described in the Methods Section of the manuscript were considered active and included in analyses.
Files and variables
File: NO_DRUG
Description: spontaneous calcium transients from 3 male and 3 female subjects (6 excel files), 6 videos each (tabs), in the absence of drug
File: EGTA
Description: spontaneous calcium transients from 3 male and 3 female subjects (6 excel files), 6 videos each (tabs), before (tabs 1-3) and after (tabs 4-6) EGTA perfusion to deplete extracellular calcium
File: CPA
Description: spontaneous calcium transients from 3 male and 3 female subjects (6 excel files), 6 videos each (tabs), before (tabs 1-3) and after (tabs 4-6) CPA perfusion to deplete intracellular calcium stores from the ER
File: NIF
Description: spontaneous calcium transients from 3 male and 3 female subjects (6 excel files), 6 videos each (tabs), before (tabs 1-3) and after (tabs 4-6) nifedipine perfusion to block L-type voltage-gated calcium channels
File: TTX
Description: spontaneous calcium transients from 3 male and 3 female subjects (6 excel files), 6 videos each (tabs), before (tabs 1-3) and after (tabs 4-6) tetrodotoxin perfusion to block voltage-gated sodium channels
File: HEX
Description: spontaneous calcium transients from 3 male and 3 female subjects (6 excel files), 6 videos each (tabs), before (tabs 1-3) and after (tabs 4-6) hexamethonium perfusion to block nicotinic acetylcholine receptors
File: ATROPINE
Description: spontaneous calcium transients from 3 male and 3 female subjects (6 excel files), 6 videos each (tabs), before (tabs 1-3) and after (tabs 4-6) atropine perfusion to block muscarinic acetylcholine receptors
File: ETH
Description: spontaneous calcium transients from 3 male and 3 female subjects (6 excel files), 6 videos each (tabs), before (tabs 1-3) and after (tabs 4-6) ethanol perfusion to demonstrate the lack of effect of this solvent
File: DMSO
Description: spontaneous calcium transients from 3 male and 3 female subjects (6 excel files), 6 videos each (tabs), before (tabs 1-3) and after (tabs 4-6) DMSO perfusion to demonstrate the lack of effect of this solvent
[ATR-Atropine, CPA- Cyclopentyladenosine, DMSO- Dimethyl Sulfoxide, EGTA- Ethylene Glycol Tetraacetic Acid, ETH- Ethanol, HEX- Hexamethonium, NIF- Nifedipine, TTX- Tetrodotoxin, NoDrug- Control]
Code/software
Image J was used to obtain fluorescence transients from chromaffin cells recorded in videos of adrenal slices from GCaMP6f mice. Transients were transferred into Excel files, 1 column per chromaffin cell transient. Excel files were read into Matlab, which was used to identify calcium events (troughs and peaks) and to calculate frequencies and amplitudes.
