Characterization of the tail current of Channelrhodopsin-2 variants

IF 2.3 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Biochemistry and Biophysics Reports Pub Date : 2024-07-16 DOI:10.1016/j.bbrep.2024.101787
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Abstract

Our study focused on specific ChR2 variants, particularly those with the Step function Opsins (SFO) mutation at the D156-C128 gate. These are widely used in optogenetics due to their heightened sensitivity to light and bi-stable prolonged activation. However, in some ChR2 variants, specifically D156 mutants, a tail current occurs when continuous light exposure is stopped. We specifically examined the D156H-T159S ChR2 variant, which demonstrated a tail current that was somewhat responsive to light and voltage, with a single-channel current of around 9fA, similar to wt-ChR2 as determined by stationary noise analysis. To further investigate, we used nonstationary noise analysis in cell-attached patching mode, which revealed that the tail current's single-channel current falls within the same range as the peak current, albeit with mild contamination from adaptation and desensitization. This finding strongly supports the notion that a portion of the ChR2 molecules open or re-open at the end of illumination, leading to further membrane depolarization.

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通道发光素-2 变体的尾流特征
我们的研究侧重于特定的 ChR2 变体,特别是那些在 D156-C128 门处发生阶跃功能眼动素(SFO)突变的变体。这些变体因其对光的高度敏感性和双稳态长时间激活而被广泛用于光遗传学。然而,在某些 ChR2 变体,特别是 D156 突变体中,当停止连续光照射时会出现尾电流。我们特别研究了 D156H-T159S ChR2 变体,它表现出对光照和电压有一定反应的尾电流,单通道电流约为 9fA,与通过静态噪声分析确定的 wt-ChR2 类似。为了进一步研究,我们在细胞连接的修补模式下使用了非稳态噪声分析,结果发现尾电流的单通道电流与峰值电流在同一范围内,尽管有轻微的适应和脱敏污染。这一发现有力地支持了这样一种观点,即部分 ChR2 分子在光照结束时打开或重新打开,导致膜进一步去极化。
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来源期刊
Biochemistry and Biophysics Reports
Biochemistry and Biophysics Reports Biochemistry, Genetics and Molecular Biology-Biophysics
CiteScore
4.60
自引率
0.00%
发文量
191
审稿时长
59 days
期刊介绍: Open access, online only, peer-reviewed international journal in the Life Sciences, established in 2014 Biochemistry and Biophysics Reports (BB Reports) publishes original research in all aspects of Biochemistry, Biophysics and related areas like Molecular and Cell Biology. BB Reports welcomes solid though more preliminary, descriptive and small scale results if they have the potential to stimulate and/or contribute to future research, leading to new insights or hypothesis. Primary criteria for acceptance is that the work is original, scientifically and technically sound and provides valuable knowledge to life sciences research. We strongly believe all results deserve to be published and documented for the advancement of science. BB Reports specifically appreciates receiving reports on: Negative results, Replication studies, Reanalysis of previous datasets.
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