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

• Published in: STAR protocols 2022-06, Vol.3 (2), p.101244-101244, Article 101244
• Main Authors: de Prisco, Nicola, Chemiakine, Alexei, Lee, Winston, Botta, Salvatore, Gennarino, Vincenzo A.
• Format: Article
• Language: English
• Subjects: Animals; Brain; Cell Biology; Cell culture; Cell isolation; Cell-based Assays; Genetics; Hippocampus; Mice; Model Organisms; Neurons; Neuroscience; Protocol
• ISSN: 2666-1667; 2666-1667
• DOI: 10.1016/j.xpro.2022.101244

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). [Display omitted] •Primary mouse hippocampal neuron isolation and immunohistochemistry•Transfecting hippocampal neurons with different gene variants•Sholl analysis of dendritic arborization•Nissl staining and morphological evaluation of mouse brain Publisher's note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. 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).