Gómez-Gálvez, Y and Gates, MA (2020) Paclitaxel is effective for controlling astrocyte proliferation in vitro: Implications for generating ventral mesencephalic cultures enriched with dopamine neurons. Journal of Neuroscience Methods, 351. 109065 - ?. ISSN 0165-0270

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BACKGROUND: Primary embryonic ventral mesencephalic (VM) cultures are a high throughput tool for understanding and manipulating dopamine neurons, to study the mechanisms that trigger their degeneration during Parkinson's disease (PD), and to test new drugs aimed at treating the disease. Unfortunately, primary cell cultures are often quickly overwhelmed by dividing astrocytes which both obscure neuronal cells and distort the cellular composition that exists in vivo. NEW METHOD: To develop a new in vitro system whereby astrocyte division can be readily controlled while maintaining neuronal integrity, VM cultures were treated with different doses (1.75, 3.5, 7, 14 nM) of the anti-mitotic drug paclitaxel for up to seven days in vitro. The study subsequently sought to determine the importance of astrocytes in dopamine neuron survival when challenged with an exposure to the toxin 6-hydroxydopamine (6-OHDA). RESULTS: Optical density (O.D.) measures of GFAP expression and counts of β-III tubulin and tyrosine hydroxylase positive neurons reveals that a low dose of 3.5 nM of paclitaxel significantly reduced the density of GFAP + astrocytes in primary VM cultures, while maintaining the viability of neurons and dopamine neurons. Interestingly, a reduction of GFAP + astrocytes within primary VM cultures did not reveal any statistically significant differences in the number of dopamine neurons surviving treatment with 6-OHDA. CONCLUSIONS: These findings detail a quick and simple method for stabilising astrocyte numbers in primary VM cultures, without affecting the viability of dopamine neurons, and suggest that astrocytes may not enhance the survival of dopamine neurons when challenged with the 6-OHDA toxin.

Item Type: Article
Additional Information: The final version of this accepted manuscript and all relevant information can be found online at; https://www.sciencedirect.com/science/article/pii/S016502702030488X?via=ihub
Uncontrolled Keywords: 6-OHDA; Astrocyte; Dopamine; Paclitaxel; Parkinson’s disease; Primary cell culture.
Subjects: R Medicine > R Medicine (General) > R735 Medical education. Medical schools. Research
R Medicine > RA Public aspects of medicine > RA0421 Public health. Hygiene. Preventive Medicine
R Medicine > RC Internal medicine > RC346 Neurology. Diseases of the nervous system, including speech disorders
Divisions: Faculty of Medicine and Health Sciences > School of Pharmacy and Bioengineering
Related URLs:
Depositing User: Symplectic
Date Deposited: 05 Feb 2021 08:58
Last Modified: 30 Jun 2022 01:30
URI: https://eprints.keele.ac.uk/id/eprint/9130

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