Eguchi Kohgaku, Velicky Philipp, Hollergschwandtner Elena, Itakura Makoto, Fukazawa Yugo, Danzl Johann Georg, Shigemoto Ryuichi
Institute of Science and Technology Austria (IST Austria), Klosterneuburg, Austria.
Department of Biochemistry, Kitasato University School of Medicine, Sagamihara-shi, Japan.
Front Cell Neurosci. 2020 Mar 19;14:63. doi: 10.3389/fncel.2020.00063. eCollection 2020.
Acute brain slice preparation is a powerful experimental model for investigating the characteristics of synaptic function in the brain. Although brain tissue is usually cut at ice-cold temperature (CT) to facilitate slicing and avoid neuronal damage, exposure to CT causes molecular and architectural changes of synapses. To address these issues, we investigated ultrastructural and electrophysiological features of synapses in mouse acute cerebellar slices prepared at ice-cold and physiological temperature (PT). In the slices prepared at CT, we found significant spine loss and reconstruction, synaptic vesicle rearrangement and decrease in synaptic proteins, all of which were not detected in slices prepared at PT. Consistent with these structural findings, slices prepared at PT showed higher release probability. Furthermore, preparation at PT allows electrophysiological recording immediately after slicing resulting in higher detectability of long-term depression (LTD) after motor learning compared with that at CT. These results indicate substantial advantages of the slice preparation at PT for investigating synaptic functions in different physiological conditions.
急性脑片制备是研究大脑突触功能特征的一种强大实验模型。尽管脑组织通常在冰冷温度（CT）下切割以利于切片并避免神经元损伤，但暴露于CT会导致突触的分子和结构变化。为了解决这些问题，我们研究了在冰冷和生理温度（PT）下制备的小鼠急性小脑切片中突触的超微结构和电生理特征。在CT下制备的切片中，我们发现显著的树突棘丢失和重建、突触小泡重排以及突触蛋白减少，而在PT下制备的切片中均未检测到这些情况。与这些结构发现一致，PT下制备的切片显示出更高的释放概率。此外，在PT下制备允许切片后立即进行电生理记录，与CT相比，运动学习后长期抑郁（LTD）的可检测性更高。这些结果表明在PT下制备脑片对于研究不同生理条件下的突触功能具有显著优势。
