Protocol to assess the effect of disease-driving variants on mouse brain morphology and primary hippocampal neurons

Nicola de Prisco; Alexei Chemiakine; Winston Lee; Salvatore Botta; Vincenzo A Gennarino

doi:10.1016/j.xpro.2022.101244

Protocol to assess the effect of disease-driving variants on mouse brain morphology and primary hippocampal neurons

STAR Protoc. 2022 Mar 15;3(2):101244. doi: 10.1016/j.xpro.2022.101244. eCollection 2022 Jun 17.

Authors

Nicola de Prisco¹, Alexei Chemiakine¹, Winston Lee^{1

2}, Salvatore Botta^{1

3}, Vincenzo A Gennarino^{1

4

5

6}

Affiliations

¹ Department of Genetics and Development, Columbia University Irving Medical Center, New York, NY 10032, USA.
² Department of Ophthalmology, Columbia University Irving Medical Center, New York, NY 10032, USA.
³ Department of Translational Medical Science, University of Campania Luigi Vanvitelli, 81100 Caserta, Italy.
⁴ Departments of Pediatrics and Neurology, Columbia University Irving Medical Center, New York, NY 10032, USA.
⁵ Columbia Stem Cell Initiative, Columbia University Irving Medical Center, New York, NY 10032, USA.
⁶ Initiative for Columbia Ataxia and Tremor, Columbia University Irving Medical Center, New York, NY 10032, USA.

Abstract

Genetic variants that affect neurological function will often produce changes visible at the level of gross morphology, either of the whole brain or of specific neuronal types. Here we describe how to perfuse and dissect the brain in preparation for Nissl staining. Then we outline steps for culturing mouse primary hippocampal neurons to evaluate dendritic arborization (Sholl analysis). For complete details on the use and execution of this protocol, please refer to Gennarino et al. (2018).

Keywords: Cell Biology; Cell culture; Cell isolation; Cell-based Assays; Genetics; Model Organisms; Neuroscience.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Animals
Brain
Hippocampus*
Mice
Neurons*

Grants and funding

R01 NS109858/NS/NINDS NIH HHS/United States