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Abstract

Neuroscience

Isolation and Culture of Primary Neurons and Glia from Adult Rat Urinary Bladder

Published: May 23rd, 2020

DOI:

10.3791/61177

1School of Pharmaceutical Sciences, Guangzhou University of Chinese Medicine, 2Dongguan & Guangzhou University of Chinese Medicine Cooperative Academy of Mathematical Engineering for Chinese Medicine, Guangzhou University of Chinese Medicine, 3School of Basic Medical Sciences, Guangzhou University of Chinese Medicine

Abstract

The lower urinary tract has two main functions, namely, periodic urine storage and micturition; these functions are mediated through central and peripheral neuroregulation. Although extensive research on the lower urinary tract nervous system has been conducted, most studies have focused on primary culture. This protocol introduces a method for the isolation and culture of bladder neurons and glia from Sprague–Dawley rats. In this method, the neurons and glia were incubated in a 37 °C, 5% CO2 incubator for 5–7 days. As a result, they grew into mature shapes suitable for related subsequent immunofluorescence experiments. Cells were morphologically observed using an optical microscope. Neurons, synaptic vesicles, and glia were identified by β-III-tubulin and MAP-2, Synapsin-1, and GFAP staining, respectively. Meanwhile, immunocytochemistry was performed on several neurotransmitter-related proteins, such as choline acetyltransferase, DYNLL2, and SLC17A9.

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Keywords Primary Neurons

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