Electrically evoked calcium waves and membrane oscillations reveal astrocytic excitability in the neurostimulation range

Electrically evoked calcium waves and membrane oscillations reveal astrocytic excitability in the neurostimulation range # Authors Miguel Aroso (1), Sara C Silva (1), Domingos L Castro (1), Paulo Aguiar (1,2,*)(1) Neuroengineering and Computational Neuroscience Lab (NCN), i3S - Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Porto, Portugal(2) Faculdade de Medicina da Universidade do Porto, Porto, Portugal* Correspondence to: [e-mail] # Preprint https://doi.org/10.1101/2024.02.28.582466Currently (Apr 2026) under review in Journal of Neural Engineering # Overview This dataset contains recordings of calcium imaging and electrophysiology of rat primary astrocyte cultures, as described in https://doi.org/10.1101/2024.02.28.582466 # Folder Structure & Description The CalciumImaging folder contains representative TIFF/video files and associated metadata. All images were acquired at a sampling rate of 100 Hz. For all stimulated conditions, the electrical stimulus was applied 2 seconds after the start of the recording. This folder is organized into three subfolders: Evoked_Vs_Baseline: Comparative recordings of spontaneous vs. triggered activity- Basel.tif: Baseline recording (no stimulus).- Stim_800mV.tif: Recording with an 800 mV stimulusCa_Vs_NoCa: Experiments assessing the role of extracellular calcium.- Ca.tif: Stimulated recording (800 mV) in standard HBSS (with calcium).- No_Ca.tif: Stimulated recording (800 mV) in nominally calcium-free HBSS.400_800mV: Recordings testing stimulus-amplitude dependency.- The stimulation amplitude is indicated in the filename (e.g., 400mV.tif corresponds to a 400 mV stimulus). The Electrophysiology folder contains raw voltage recordings from astrocytes cultured on microelectrode arrays (MEAs). These recordings characterize the membrane voltage oscillations in response to electrical stimulation under controlled conditions and in the presence of pharmacological inhibitors. For all stimulated conditions, the initial stimulus was applied 1 minute after the start of the recording. This section contains subfolders labelled with abbreviated names of the chemical inhibitors used This folder contains the following subfolders: 400_800mV: Recordings testing stimulus-amplitude dependency.- TC_D1.h5: Recording with stimuli increasing amplitude. Stimuli were applied at 1-minute intervals, with each amplitude repeated three times. "D1" indicates the stimulation electrode. 800mV_3Stim: Recording assessing the reliability of astrocytic activation across repeated stimuli at 800 mV.- TS_A3.h5: Three 800 mV stimuli applied to electrode “A3”, interspaced by 1 minute, starting at the 1-minute mark. Ca_Vs_NoCa: Electrophysiological responses recorded in standard HBSS vs. nominally calcium-free HBSS.- Ca_A3.h5: Stimulated recording in standard HBSS- Ca_A3_NoCa.h5: Stimulated recording in nominally calcium-free HBSS. Cil (Cilnidipine): N-type and L-type VGCC blocker.- DC_A15.h5: control with response to stimulation at 800 mV.- DC_A15_Cil.h5: response to stimulation at 800 mV after Cilnidipine exposure. MA (Meclofenamic Acid): Inhibitor of gap junction communication.- DM_C16.h5: control with response to stimulation at 800 mV.- DM_C16_MA.h5: response to stimulation at 800 mV after Meclofenamic Acid exposure. Sur (Suramin): Purinergic receptor antagonist used to block ATP-mediated signaling.- DS_P16.h5: control with response to stimulation at 800 mV.- DS_P16_Sur.h5: response to stimulation at 800 mV after Suramin exposure. Ver (Verapamil): L-type VGCC blocker.- DV_O15.h5: control with response to stimulation at 800 mV.- DV_O15_Ver.h5: response to stimulation at 800 mV after Verapamil exposure.- Xest-C (Xestospongin C): Blocker of IP3 receptors.- DX_C16.h5: control with response to stimulation at 800 mV.- DX_C16_XC.h5: response to stimulation at 800 mV after Xestospongin C exposure.