Animal Behaviour and Ecology Laboratory, School of Environmental and Rural Science, Faculty of Science, Agriculture, Business and Law, University of New England, Armidale, NSW, Australia.
Brain Behaviour Research Group, School of Science and Technology, Faculty of Science, Agriculture, Business and Law, University of New England, Armidale, NSW, Australia.
J Neurosci Methods. 2021 Mar 15;352:109081. doi: 10.1016/j.jneumeth.2021.109081. Epub 2021 Jan 20.
Recent anthropomorphic disturbances are occurring at an increasing rate leading to organisms facing a variety of challenges. This change is testing the information processing capacity (IPC) of all animals. Brain function is widely accepted to be influenced by a variety of factors, including relative size, number of neurons and neuronal densities. Therefore, in order to understand what drives an animals IPC, a methodological approach to analyze these factors must be established.
Here we created a protocol that allowed for high-throughput, non-biased quantification of neuronal density and size across six regions of the brain. We used the Isotropic Fractionator method in combination with flow cytometry to identify neuronal and non-neuronal cells in the brains of adult rats.
The results obtained were comparable to those identified using stereological counting methods.
By employing this new method, the number of nuclei in a specific brain region can be compared between replicate animals within an experiment. By calibrating the forward scatter channel of the flow cytometer with size standard beads, neuronal and non-neuronal nuclear sizes can be estimated simultaneously with nuclei enumeration. These techniques for nuclear counting and size estimation are technically and biologically reproducible.
Use of flow cytometry provides a methodological approach that allows for consistency in research, so that information on brain morphology, and subsequent function, will become comparable across taxa.
最近，拟人化干扰的发生频率呈上升趋势，导致生物面临着各种挑战。这种变化正在考验所有动物的信息处理能力（IPC）。人们普遍认为，大脑功能受到多种因素的影响，包括相对大小、神经元数量和神经元密度。因此，为了了解是什么驱动动物的 IPC，必须建立一种分析这些因素的方法。
在这里，我们创建了一种方案，可以在大脑的六个区域内进行高通量、无偏的神经元密度和大小的定量分析。我们使用各向同性分馏器方法结合流式细胞术来识别成年大鼠大脑中的神经元和非神经元细胞。
使用该新方法，可以在实验内的重复动物之间比较特定脑区的核数。通过用标准尺寸珠子校准流式细胞仪的前向散射通道，可以在核计数的同时估计神经元和非神经元核的大小。这些核计数和大小估计技术在技术和生物学上具有可重复性。
流式细胞术的使用提供了一种方法学方法，使研究具有一致性，从而使脑形态学及其后续功能的信息在分类群之间变得可比。
