Vazetdinova Alina, Valiullina-Rakhmatullina Fliza, Rozov Andrei, Evstifeev Alexander, Khazipov Roustem, Nasretdinov Azat
Laboratory of Neurobiology, Kazan Federal University, Kazan, Russia.
Institut für Physiologie und Pathophysiologie, Heidelberg, Germany.
Front Mol Neurosci. 2022 Aug 11;15:979479. doi: 10.3389/fnmol.2022.979479. eCollection 2022.
Cell-attached current-clamp (CA/CC) recordings have been proposed to measure resting membrane potential and synaptic/agonist responses in neurons without disrupting the cell membrane, thus avoiding the intracellular dialysis that occurs in conventional whole-cell recordings (WC). However, the accuracy of CA/CC recordings in neurons has not been directly assessed. Here, we used concomitant CA and WC current clamp recordings from cortical neurons in brain slices. Resting membrane potential values and slow voltage shifts showed variability and were typically attenuated during CA/CC recordings by ~10-20% relative to WC values. Fast signals were slowed down and their amplitude was greatly reduced: synaptic potentials by nearly 2-fold, and action potentials by nearly 10-fold in CA/CC mode compared to WC. The polarity of GABAergic postsynaptic responses in CA/CC mode matched the responses in WC, and depolarising GABAergic potentials were predominantly observed during CA/CC recordings of intact neonatal CA3 hippocampal pyramidal neurons. Similarly, CA/CC recordings reliably detected neuronal depolarization and excitation during network-induced giant depolarizing potentials in the neonatal CA3 hippocampus, and revealed variable changes, from depolarization to hyperpolarization, in CA1 pyramidal cells during sharp wave ripples in the adult hippocampus. Thus, CA/CC recordings are suitable for assessing membrane potential but signal distortion, probably caused by leakage via the seal contact and RC filtering should be considered.
细胞贴附式电流钳(CA/CC)记录法被提议用于测量神经元的静息膜电位以及突触/激动剂反应,而不会破坏细胞膜,从而避免了传统全细胞记录(WC)中发生的细胞内透析。然而,CA/CC记录法在神经元中的准确性尚未得到直接评估。在这里,我们使用了脑片皮层神经元的同步CA和WC电流钳记录。静息膜电位值和缓慢的电压变化显示出变异性,并且在CA/CC记录过程中相对于WC值通常衰减约10-20%。快速信号变慢,其幅度大大降低:与WC相比,CA/CC模式下的突触电位降低近2倍,动作电位降低近10倍。CA/CC模式下GABA能突触后反应的极性与WC中的反应相匹配,并且在完整新生CA3海马锥体神经元的CA/CC记录过程中主要观察到去极化GABA能电位。同样,CA/CC记录在新生CA3海马体网络诱导的巨大去极化电位期间可靠地检测到神经元去极化和兴奋,并揭示了成年海马体尖波涟漪期间CA1锥体细胞从去极化到超极化的可变变化。因此,CA/CC记录适用于评估膜电位,但应考虑可能由密封接触处的泄漏和RC滤波引起的信号失真。