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快速,生物化学标记细胞活动史 in vivo

Run Zhang^1,2, Maribel Anguiano^2,3, Isak K Aarrestad^2,3,4

¹Biomedical Engineering Graduate Group, University of California, Davis, Davis, CA, USA.

|August 5, 2024

概括

我们开发了CaST,这是一种新的方法,可以在体内生物化学标记具有高 (Ca2+) 活性的细胞. 这种酶催化方法能够非侵入性地实时监测自由移动的动物的细胞信号和行为.

科学领域:

神经科学是一个神经科学.
生物化学生物化学
细胞生物学细胞生物学

背景情况:

细胞内 (Ca2+) 对于细胞信号传递至关重要.
目前的Ca2+传感器通常需要侵入性植入物用于深层组织成像.
对于研究自由行为动物的细胞活性,需要使用非侵入性方法.

研究的目的:

设计一种非侵入性,酶催化方法,在体内标记具有高Ca2+活性的细胞.
开发一个快速和灵敏的通报器,用于总细胞内Ca2+信号传输.
实时将Ca2+活动与行为反应相关联.

主要方法:

开发了Ca2+激活分裂-化物 (CaST),一种在Ca2+激活后用生物素标记细胞的酶.
在几分钟内给予外源生物素用于激活细胞的酶标记.
测量CaST信号强度作为Ca2+度和标记时间的函数.
应用CaST标记前额叶皮层神经元在小鼠在psilocybin管理期间.

主要成果:

使用外源生物素分子,CaST在10分钟内快速标记Ca2+激活细胞.
CaST信号与Ca2+度和标签持续时间成比例,作为一个时间隔离的集成器.
与较慢的转录记者不同,CaST提供了即时的读数.
在前额叶皮层神经元中成功关联了CaST信号,与小鼠的psilocybin诱导的头部抽反应.

结论:

CaST提供了一种非侵入性,快速和敏感的方法来监测体内Ca2+活动.
这项技术可以实时将细胞活动与自由移动的受试者的行为相关联.
在神经科学研究,药物发现和理解复杂的生物过程中,CaST具有潜在的应用.

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