We study cholinergic neurons and astrocytes of the dorsal striatum that are important components of the nigrostriatal circuit. We have shown previously that these cell types lose primary cilia in 4 mouse models of LRRK2 Parkinson’s disease (Dhekne et al., 2019; Khan et al., 2021). Here we present a method to identify the same brain region in human postmortem tissue to evaluate ciliary status. The striatum is identified morphologically and also by immunofluorescence microscopy using anti-DARPP-32 antibodies (Ouimet & Greengard, 1990; Arlotta et al., 2008). Cholinergic cell bodies stain brightly with anti-choline acetyltransferase (ChAT) antibodies and these cells are also identifiable using antibodies that recognize the choline transporter, CHT. Auto-fluorescence is decreased using Sudan Black B.
We are grateful to our colleagues at the Banner Institute at the University of Arizona (Drs. Geidy E. Serrano and Ileana Lorenzini) for providing tissue and guidance (Beach et al., 2015).
Arlotta P, Molyneaux BJ, Jabaudon D, Yoshida Y, Macklis JD. Ctip2 controls the differentiation of medium spiny neurons and the establishment of the cellular architecture of the striatum. J Neurosci. 2008 Jan 16;28(3):622-32.
doi: 10.1523/JNEUROSCI.2986-07.2008.
Beach, T. G., Adler, C. H., Sue, L. I., Serrano, G., Shill, H. A., Walker, D. G., Lue, L., Roher, A. E., Dugger, B. N., Maarouf, C., Birdsill, A. C., Intorcia, A., Saxon-Labelle, M., Pullen, J., Scroggins, A., Filon, J., Scott, S., Hoffman, B., Garcia, A., Caviness, J. N., … Sabbagh, M. N. (2015). Arizona Study of Aging and Neurodegenerative Disorders and Brain and Body Donation Program. Neuropathology : official journal of the Japanese Society of Neuropathology, 35(4), 354–389. https://doi.org/10.1111/neup.12189
Dhekne, H. S., Yanatori, I., Gomez, R. C., Tonelli, F., Diez, F., Schüle, B., Steger, M., Alessi, D. R., & Pfeffer, S. R. (2018). A pathway for Parkinson's Disease LRRK2 kinase to block primary cilia and Sonic hedgehog signaling in the brain. eLife, 7, e40202. https://doi.org/10.7554/eLife.40202
Khan, S. S., Sobu, Y., Dhekne, H. S., Tonelli, F., Berndsen, K., Alessi, D. R., & Pfeffer, S. R. (2021). Pathogenic LRRK2 control of primary cilia and Hedgehog signaling in neurons and astrocytes of mouse brain. eLife, 10, e67900. https://doi.org/10.7554/eLife.67900
Ouimet CC, Greengard P. Distribution of DARPP-32 in the basal ganglia: an electron microscopic study. J Neurocytol. 1990 Feb;19(1):39-52.