Characterization of Oligophrenin -1, a RhoGAP Lost in Patients Affected with Mental Retardation: Lentiviral Injection in Organotypic Brain Slice Cultures

Nadif Kasri, N., Govek, E. E., Van Aelst, L. (March 2008) Characterization of Oligophrenin -1, a RhoGAP Lost in Patients Affected with Mental Retardation: Lentiviral Injection in Organotypic Brain Slice Cultures. Methods Enzymol, 439. pp. 255-266.

URL: http://www.ncbi.nlm.nih.gov/pubmed/18374170
DOI: 10.1016/S0076-6879(07)00419-3

Abstract

Mutations in regulators and effectors of the Rho GTPases underlie various forms of mental retardation (MR). Among them, oligophrenin[hyphen (true graphic)]1 (OPHN1), which encodes a Rho[hyphen (true graphic)]GTPase activating protein, was one of the first Rho[hyphen (true graphic)]linked MR genes identified. Upon characterization of OPHN1 in hippocampal brain slices, we obtained evidence for the requirement of OPHN1 in dendritic spine morphogenesis and neuronal function of CA1 pyramidal neurons. Organotypic hippocampal brain slice cultures are commonly used as a model system to investigate the morphology and synaptic function of neurons, mainly because they allow for the long[hyphen (true graphic)]term examination of neurons in a preparation where the gross cellular architecture of the hippocampus is retained. In addition, maintenance of the trisynaptic circuitry in hippocampal slices enables the study of synaptic connections. Today, a multitude of gene transfer methods for postmitotic neurons in brain slices are available to easily manipulate and scrutinize the involvement of signaling molecules, such as Rho GTPases, in specific cellular processes in this system. This chapter covers techniques detailing the preparation and culturing of organotypic hippocampal brain slices, as well as the production and injection of lentivirus into brain slices.

Item Type: Paper
Subjects: diseases & disorders
Investigative techniques and equipment
organs, tissues, organelles, cell types and functions > tissues types and functions > hippocampus
organism description > virus > lentivirus
diseases & disorders > congenital hereditary genetic diseases > mental retardation
organs, tissues, organelles, cell types and functions
organs, tissues, organelles, cell types and functions > tissues types and functions
organism description > virus
CSHL Authors:
Communities: CSHL labs > Van Aelst lab
Depositing User: Matt Covey
Date: 26 March 2008
Date Deposited: 26 Feb 2013 21:46
Last Modified: 26 Feb 2013 21:46
PMCID: PMC3556824
Related URLs:
URI: http://repository.cshl.edu/id/eprint/27500

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