Histological Phenotyping of the Brain

P.I. and Contact:   Dan Goldowitz, Ph.D.

Lab:   Developmental Neurogenetics

Location:    Department of Anatomy and Neurobiology, University of Tennessee, Health Science Center, Memphis TN 38163

Description:  Our group has extensive experience in the histological analysis of neurological mutant mice. We use histochemical, immunocytochemical, and in situ hybridization histochemical techniques to ascertain the developmental status and structural integrity of the nervous system. Our particular focus has been on the cerebellum, retina, and hippocampus. The PI is a member of the Tennessee Mouse Genome Consortium (http://tnmouse.org/) which is undertaking a large scale ENU-mutagenesis of the mouse and is PI of a NIH-U01 proposal to assess the neurological function of potential mutants produced from this program. The PI also works closely with Dr. Robert W. Williams for the phenotyping of the mouse eye and registering phenotypic data on his website. We have established a super team of experimental neuroanatomists in the Department of Anatomy and Neurobiology at the University of Tennessee to analyze the histological material. The group consists of the PI, Andrea Elberger, Kristin Hamre, Rich Smeyne (main affiliation: St. Jude Dept of Developmetnal Neurobiology), Dennis Steindler, Bob Waters, and Rob Williams.

Example:   See selected references below. Web-accessible images of histological material are under construction, see the brain stain library.

Screening Protocols and Techniques:    Four mice (two male and two female) are typically examined for each strain or potential mutant. Two mice are cervically dislocated and processed for fresh-frozen cryostat sectioning and two brains are immersion-fixed in Bouinās solution for paraffin sectioning. For screening purposes we have established a set of eight histological procedures to assay the cellular, nerve fiber, activity, neurotransmitter, and proliferative/degenerative states of the CNS. For the analysis of cytoarchitectonics we use Nissl stains and anti-NeuN immunocytochemistry. Enzyme histochemistry for acetylcholinesterase and, cytochrome oxidase provide a view of cholinergic neurotransmitter systems and mitochondrial activity. Nerve tracts are visualized with osmium tetroxide and astroglia with anti-glial fibrillary acidic protein immunocytochemistry. Cell production and death are highlighted with anti-BrdU immunohistochemistry and Fluoro-Jade histochemistry. New stains are being examined for high-throughput capability and ease of performance to demonstrate other aspects of nervous system structure and function. A particular focus for our screens will be on histological analysis of the aged nervous system.

Collaborative Interests:  We welcome all queries about the histological analysis of brain structure and development. Contact Dan Goldowitz at (901) 448–7019.

Selected References

1. Wille, W., Cremer, H, Barthels, D., and Goldowitz, D. (1992) Hybridization analyses of nervous system gene expression. In: Genetic Analysis of Brain and Behavior: Focus on the Mouse. D. Goldowitz, R. Wimer and D. Wahlsten, eds. Amsterdam, Elsevier, pp. 291–315.
2. Goldowitz, D., Moran, T.H., and Wetts, R. (1992) Mouse chimeras in the study of genetic and structural determinants of behavior. In: Genetic Analysis of Brain and Behavior: Focus on the Mouse. D. Goldowitz, R. Wimer and D. Wahlsten, eds. Amsterdam, Elsevier, pp. 271–290.
3. Rice, D.S., Williams, R.W., Harris, B., Bailey, P.W., Davisson, M.T., Goldowitz, D. (1995) Mapping the Bst locus on mouse chromosome 16: A candidate for human optic atropy 1 (OPA1). Mammalian Genome 6:546–548.
4. Williams, R.W., Strom, R.C., Rice, D.S., and Goldowitz, D. (1996) Genetic and environmental control of variation in retinal ganglion cell number in mice. J. Neuroscience 16:7193–7205.
5. Surmeier, D.J., Mermelstein, P. and Goldowitz, D. (1996) The weaver mutation of GIRK2 results in a loss of inwardly rectifying K+ current in cerebellar granule cells. Proc. Natl. Acad. Sci. USA 93:11191–11195.
6. Goldowitz, D., Rice, D.S., Williams, R.W. (1996) "Clonal architecture of the mouse retina" In: Neural Development and Plasticity, R.R. Mize and R.S. Erzurumlu, eds. Progress in Brain Research 108:3–15, Amsterdam, Elsevier.
7. Rice, D.S., Tang, Q., Williams, R.W., Harris, B., Davisson, M.T., Goldowitz, D. (1997) Retinal ganglion cell loss and misdirected axon growth associated with fissure defects in Bst/+ mutant mice. Invest. Ophth. Vis. Sci.
8. Chen, S.-C., Ehrhard, P., Goldowitz, D. and Smeyne, R.J. (1997) Developmental expression of the GIRK family of inward rectifying potassium channels: implication for annormalities inthe weaver mutant mouse. Brain Res. 778:251–264.
9. Hamre, K.M. and Goldowitz, D. (1997) Meander tail acts intrinsic to granule cell precursors to disrupt cerebellar development: Analysis of meander tail chimeric mice. Development 124:4201–4212.
10. Sheldon, M., Rice, D.S., D'Arcangelo, G., Yoneshima, H., Nakajima, K., Mikoshiba, K., Howell, B.W., Cooper, J.A., Goldowitz, D. and Curran, T. (1997) Scrambler and yotari disrupt the disabled gene and produce a reeler-like phenotype in mice. Nature 389:730–733.
11. Goldowitz, D., Cushing, R.C., Laywell, E., D'Arcangelo, G., Sheldon, M., Sweet, H.O., Davisson, M., Steindler, D., and T. Curran. (1997) Cerebellar disorganization characteristic of reeler in scrambler mutant mice despite presence of reelin. J Neurosci. 17:8767–8777.
12. Mullen, R.J., Hamre, K.M. and Goldowitz, D. (1997) "Cerebellar mutant mice and chimeras revisited" In: Perspectives on Developmental Neurobiology: The Cerebellum, C.A. Mason and C. Sotelo, eds. vol. 5 no. 1, Amsterdam:Gordon, pp.43–55.
13. Goldowitz, D. and Hamre, K.M. (1998) The cells and molecules that make a cerebellum. Trends in Neurosci. 21:375–382.
14. Rice, D.S., Sheldon, M., D'Arcangelo, G., Nakajima, K., Goldowitz, D., and T. Curran. (1998) Disabled-1 acts downstream of reelin in a signaling pathway that controls laminar organization in the mammalian brain. Development 125:3719–3729.
15. Tang, Q., Rice, D.S., and D. Goldowitz (1999) Disrupted retinal development in the embryonic belly spot and tail mutant mouse. Developmental Biology 207:239–255.
16. Jensen, P., Surmeier, D.J., and D. Goldowitz (1999) Rescue of cerebellar granule cells from death in weaver NR1 double knockout mutants. J. Neurosci. 19:7991–7998.