Hi everyone,
I have some questions regarding astrocytes morphology, primary culture, and MBP expression pattern. I am new in the field of neuroscience.
I am working on knockin mice of ALS model.
I dissected 8 months old mutant mice, and lumbar spinal cord showed increased astrocytes signals as compared to age matched WT mice. I crossed check with iba1 (Microglial activation marker) that supports the case.
At the same time, i did dissection of 9 months old (3 mice), in which we observed significant reduction of astrocytes in mutant mice mostly at peripheral region of spinal cord and +ve caspase signals as well. However the ventral region of lumbar spinal cord in 9 months mutant mice also reduced and it seems their morphology is compromised as well. It might be reactive astrocytes.
Now i wanted to do primary culture and to see the effect of this mutation on motor neurons and glial cells.
My question is, whether or not i can see the effect of this mutation in primary culture, if its phenotype comes around 9 months.
And can we see some radial glial cells in 9 months old mice if we stain with GFAP, as mostly we corelate GFAP with astrocytes in adult mice. Regarding MBP pattern, i am thinking that it should be at the peripheral region like it has the particular boundary between ventral and periphery. Or its pattern is at the extreme periphery appearing as a layer.
And finally if someone has a protocol for spinal cord primary culture.
It would be highly appreciated.
Thanks in advance.
Regards
shabbir
Dear colleague,
Yes, it does. Here are some literature regarding your question:
https://www.ncbi.nlm.nih.gov/pubmed/24796693
https://www.ncbi.nlm.nih.gov/pubmed/25287860
https://www.researchgate.net/publication/289287747_Morphology_of_reactive_astrocytes_in_culture
https://jmolecularpsychiatry.biomedcentral.com/articles/10.1186/2049-9256-1-18
http://m.pnas.org/content/103/46/17513.full
Sincerely,
Hércules Freitas
Dear Hercules,
Thanks for your comments and providing materials on astrocytes morphology.
Need a protocol for spinal cord primary culture.
It would be highly appreciated.
Thanks
Best Regards
Shabbir
Hello Shabbir,
Since you are looking for protocols for primary culture of spinal cord cells, you can read and follow :
article 1: Cao, D., Jing, P., Jiang, B. and Gao, Y. (2017). Primary Culture of Mouse Neurons from the Spinal Cord Dorsal Horn. Bio-protocol 7(1): e2098. DOI: 10.21769/BioProtoc.2098.
article 2: Kerstetter, A. E., & Miller, R. H. (2012). Isolation and Culture of Spinal Cord Astrocytes. Methods in Molecular Biology (Clifton, N.J.), 814, 93–104. http://doi.org/10.1007/978-1-61779-452-0_7
Best wishes!
LY
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