AI Article Synopsis
- The lower urinary tract is responsible for urine storage and release, controlled by the nervous system.
- Research on bladder neurons and glial cells has typically focused on primary cultures, but this new protocol details how to isolate and culture these cells from Sprague-Dawley rats.
- The cultured cells were grown at 37 °C for 5-7 days, allowing for their identification and study through various staining techniques and immunocytochemistry for neurotransmitter-related proteins.
Article 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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| http://dx.doi.org/10.3791/61177 | DOI Listing |
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