Journal of Neuroscience Methods ( IF 3 ) Pub Date : 2021-09-03 , DOI: 10.1016/j.jneumeth.2021.109351 Sara Spadini 1 , Gabriella Racchetti 2 , Alice Adiletta 3 , Jacopo Lamanna 1 , Andrea Stefano Moro 4 , Mattia Ferro 4 , Vincenzo Zimarino 2 , Antonio Malgaroli 1
Background
Mitochondria and their dynamics fuel most cellular processes both in physiological and pathological conditions. In the central nervous system, mitochondria sustain synaptic transmission and plasticity via multiple mechanisms which include their redistribution and/or expansion to higher energy demanding sites, sustaining activity changes and promoting morphological circuit adaptations.
New Method
To be able to evaluate changes in mitochondrial number and protein phenotype, we propose a novel methodological approach where the simultaneous analysis of both mitochondrial DNA and protein content is performed on each individual microsample, avoiding non-homogeneous loss of material.
Results
We validated this method on neuronal-like cells and tissue samples and obtained estimates for the mitochondrial/genomic DNA ratio as well as for the abundance of protein counterparts. When the mitochondrial content per cell was evaluated in different brain areas, our results matched the known regional variation in aerobic-anaerobic metabolism. When long-term potentiation (LTP) was induced on hippocampal neurons, we detected increases in the abundance of mitochondria that correlated with the degree of synaptic enhancement.
Conclusions
Our approach can be effectively used to study the mitochondrial content andits changes in different brain cells and tissues.
中文翻译:
一种估计神经元细胞和脑组织线粒体含量的新型综合方法
背景
线粒体及其动力学在生理和病理条件下为大多数细胞过程提供燃料。在中枢神经系统中,线粒体通过多种机制维持突触传递和可塑性,包括将它们重新分配和/或扩展到更高能量需求的部位、维持活动变化和促进形态回路适应。
新方法
为了能够评估线粒体数量和蛋白质表型的变化,我们提出了一种新的方法学方法,其中对每个单独的微量样品同时分析线粒体 DNA 和蛋白质含量,避免材料的非均质损失。
结果
我们在神经元样细胞和组织样本上验证了这种方法,并获得了线粒体/基因组 DNA 比率以及蛋白质对应物丰度的估计值。当在不同的大脑区域评估每个细胞的线粒体含量时,我们的结果与已知的有氧-无氧代谢区域变化相匹配。当在海马神经元上诱导长时程增强 (LTP) 时,我们检测到与突触增强程度相关的线粒体丰度的增加。
结论
我们的方法可以有效地用于研究不同脑细胞和组织中线粒体的含量及其变化。