Viral-mediated Labeling and Transplantation of Medial Ganglionic Eminence (MGE) Cells for In Vivo Studies.
GABAergic cortical interneurons, derived from the embryonic medial and caudal ganglionic eminences (MGE and CGE), are functionally and morphologically diverse. Inroads have been made in understanding the roles of distinct cortical interneuron subgroups, however, there are still many mechanisms to be worked out that may contribute to the development and maturation of different types of GABAergic cells. Moreover, altered GABAergic signaling may contribute to phenotypes of autism, schizophrenia and epilepsy. Specific Cre-driver lines have begun to parcel out the functions of unique interneuron subgroups. Despite the advances in mouse models, it is often difficult to efficiently study GABAergic cortical interneuron progenitors with molecular approaches in vivo. One important technique used to study the cell autonomous programming of these cells is transplantation of MGE cells into host cortices. These transplanted cells migrate extensively, differentiate, and functionally integrate. In addition, MGE cells can be efficiently transduced with lentivirus immediately prior to transplantation, allowing for a multitude of molecular approaches. Here we detail a protocol to efficiently transduce MGE cells before transplantation for in vivo analysis, using available Cre-driver lines and Cre-dependent expression vectors. This approach is advantageous because it combines precise genetic manipulation with the ability of these cells to disperse after transplantation, permitting greater cell-type specific resolution in vivo.
Department of Psychiatry, University of California San Francisco; daniel.vogt@ucsf.edu.
Daniel
D
Vogt
Department of Psychiatry, University of California San Francisco.
Pei-Rung
PR
Wu
Department of Neurological Surgery, University of California San Francisco.
Shawn F
SF
Sorrells
Department of Neurological Surgery, University of California San Francisco.
Christine
C
Arnold
Department of Neurological Surgery, University of California San Francisco.
Arturo
A
Alvarez-Buylla
Department of Psychiatry, University of California San Francisco.
John L R
JL
Rubenstein
D000818:Animals; D017690:Cell Transplantation / Q000379:methods*; D005260:Female; D059330:GABAergic Neurons / Q000166:cytology / Q000502:physiology / Q000637:transplantation* / Q000821:virology; D057809:HEK293 Cells; D006801:Humans; D007395:Interneurons / Q000166:cytology / Q000502:physiology* / Q000637:transplantation / Q000821:virology*; D016086:Lentivirus / Q000235:genetics; D008473:Median Eminence / Q000166:cytology / Q000502:physiology* / Q000637:transplantation / Q000821:virology*; D051379:Mice; D058953:Neural Stem Cells / Q000166:cytology; D011247:Pregnancy; D015398:Signal Transduction; D014161:Transduction, Genetic