Exclusive labeling of direct and indirect pathway neurons in the mouse neostriatum by an adeno-associated virus vector with Cre/lox system
| Autor: | Fumino Fujiyama, Kenta Yamauchi, Takahiro Furuta, Hiroyuki Hioki, Shinichiro Okamoto, Jaerin Sohn, Yoko Ishida, Megumu Takahashi, Masato Koike |
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| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
viruses
Genetic Vectors Cre recombinase Gene Expression Biology medicine.disease_cause Indirect pathway of movement General Biochemistry Genetics and Molecular Biology Virus Green fluorescent protein Mice Transduction Genetic Neural Pathways Protocol medicine Animals Axon lcsh:Science (General) Adeno-associated virus Neurons Microscopy Integrases General Immunology and Microbiology Immunoperoxidase General Neuroscience Dependovirus Cell biology Neostriatum medicine.anatomical_structure Cre-Lox recombination Neuroscience lcsh:Q1-390 |
| Zdroj: | STAR Protocols, Vol 2, Iss 1, Pp 100230-(2021) STAR Protocols |
| ISSN: | 2666-1667 |
| Popis: | Summary We developed an adeno-associated virus (AAV) vector-based technique to label mouse neostriatal neurons comprising direct and indirect pathways with different fluorescent proteins and analyze their axonal projections. The AAV vector expresses GFP or RFP in the presence or absence of Cre recombinase and should be useful for labeling two cell populations exclusively dependent on its expression. Here, we describe the AAV vector design, stereotaxic injection of the AAV vector, and a highly sensitive immunoperoxidase method for axon visualization. For complete details on the use and execution of this protocol, please refer to Okamoto et al. (2020). Graphical Abstract Highlights • Exclusive labeling of two groups of neurons with different fluorescent proteins • Detailed protocols from stereotaxic virus injection to immunostaining methods • A signal enhancement method via peroxidase activity, BT-GO reaction • Significant improvement in a signal-to-noise ratio by the cost-effective reaction We developed an adeno-associated virus (AAV) vector-based technique to label mouse neostriatal neurons comprising direct and indirect pathways with different fluorescent proteins and analyze their axonal projections. The AAV vector expresses GFP or RFP in the presence or absence of Cre recombinase and should be useful for labeling two cell populations exclusively dependent on its expression. Here, we describe the AAV vector design, stereotaxic injection of the AAV vector, and a highly sensitive immunoperoxidase method for axon visualization. |
| Databáze: | OpenAIRE |
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