This document describes immunohistochemistry staining of mouse brain tissue to label various proteins. It includes details on primary and secondary antibodies used to stain for glial fibrillary acidic protein (GFAP) in astrocytes of the hippocampus and aquaporin 1 and 4 (AQP1, AQP4) in the choroid plexus and ventricles. References are provided on the role of aquaporins and the choroid plexus in cerebrospinal fluid production and homeostasis. Future work proposed includes RNA sequencing of choroid plexus from LXRβ knockout and wild type mice.

1. Mouse Brain
Immunohistochemistry (IHC)
Staining
Andrew Roman
Margaret Warner

2. Glial fibrillary acidic
protein (anti-GFAP)
Primary antibody:
#17, goat polyclonal
IgG, Santa Cruz
Biotechnology
1:500uL
Secondary antibody:
Biotinylated #11,
anti-goat, Invitrogen
1:200uL
V3
chpl
astrocytes
CA1
HIP
TH

3. Aquaporin 1 (AQP1)
Primary antibody:
rabbit, 1:500uL
Secondary antibody:
Anti-rabbit IgG, Zymed,
1:200uL
DGCA1
chpl
V3

4. chpl
V3
midbrain

5. CTX

6. Aquaporin 4 (AQP4)
Primary antibody:
rabbit, 1:500uL
Secondary antibody:
Anti-rabbit IgG, Zymed,
1:200uL
chpl
V3
RSP

7. Bibliography
Dai, Y.-B., Wu, W.-F., Huang, B., Miao, Y.-F., Nadarshina, S., Warner, M., & Gustafsson, J.-Å. (2016, June 21). Liver X receptors
regulate cerebrospinal fluid production. Retrieved from https://www.ncbi.nlm.nih.gov/pubmed/26324101
Lein, E.S. et al. (2007) Genome-wide atlas of gene expression in the adult mouse brain, Nature 445: 168-176.
doi:10.1038/nature05453
Lun, M. P., Johnson, M. B., Broadbelt, K. G., Watanabe, M., Kang, Y.-J., Chau, K. F., … Lehtinen, M. K. (2015, March 25).
Spatially heterogeneous choroid plexus transcriptomes encode positional identity and contribute to regional CSF production.
Retrieved from https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4389594/
Monnot, A. D., & Zheng, W. (2013, April 10). Culture of choroid plexus epithelial cells and in vitro model of blood-CSF barrier.
Retrieved from https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3982224/
Trillo-Contreras, J. L., Toledo-Aral, J. J., Echevarría, M., & Villadiego, J. (2019, February 24). AQP1 and AQP4 Contribution to
Cerebrospinal Fluid Homeostasis. Retrieved from https://www.mdpi.com/2073-4409/8/2/197

8. Future directions
• RNA-seq of choroid plexus from LXRβ
knockout and wild type mice