dlx+ interneuron-specific transsynaptic tracing  (Disect Systems Ltd)

Journal: Frontiers in Neural Circuits

Article Title: Anatomical identification of a corticocortical top-down recipient inhibitory circuitry by enhancer-restricted transsynaptic tracing

doi: 10.3389/fncir.2023.1245097

Figure Lengend Snippet: Comparison of DISECT with the AAV1-mediated anterograde transsynaptic tracing method. (A) Schematic of the conventional AAV1-mediated method shown in a previous study . Injections of AAV1-Cre into the presynaptic region enable trans- synaptical spread of infection of the vector, allowing Cre-dependent transduction of genes at the postsynaptic neurons. However, the subtype-independent spread induces gene expression at excitatory neurons, which disturbs the post hoc analysis of subtype specificity of inhibitory neurons and decreases the interpretability of the results (see also Section “4. Discussion”). (B) Schematic of a complex use of the AAV1-mediated method shown in a previous study . Using transgenic animals (such as VGAT-Cre mice, which express Cre specifically in inhibitory neurons) allows us to distinguish excitatory projecting neurons from postsynaptic neurons, increasing interpretability. However, this method requires complex procedures, such as preparation of transgenic animals. (C) Schematic of DISECT developed in this study. By injecting a Cre-inducible Dlx enhancer-specific AAV vector into the postsynaptic region, transduction can be restricted only to the postsynaptic inhibitory neurons with simple procedures, ensuring simplified efficient tracing with high interpretability in wild-type animals.

Article Snippet: By combining this method with the conventional AAV1-mediated transsynaptic tracing, we developed “Dlx+ Interneuron-Specific Transsynaptic tracing” (abbreviated as DISECT), a method that allows more efficient transsynaptic tracing of postsynaptic inhibitory neurons ( ).

Techniques: Comparison, Infection, Plasmid Preparation, Transduction, Gene Expression, Transgenic Assay

Journal: Frontiers in Neural Circuits

Article Title: Anatomical identification of a corticocortical top-down recipient inhibitory circuitry by enhancer-restricted transsynaptic tracing

doi: 10.3389/fncir.2023.1245097

Figure Lengend Snippet: Comparison of DISECT with the AAV1-mediated anterograde transsynaptic tracing method. (A) Schematic of the conventional AAV1-mediated method shown in a previous study . Injections of AAV1-Cre into the presynaptic region enable trans- synaptical spread of infection of the vector, allowing Cre-dependent transduction of genes at the postsynaptic neurons. However, the subtype-independent spread induces gene expression at excitatory neurons, which disturbs the post hoc analysis of subtype specificity of inhibitory neurons and decreases the interpretability of the results (see also Section “4. Discussion”). (B) Schematic of a complex use of the AAV1-mediated method shown in a previous study . Using transgenic animals (such as VGAT-Cre mice, which express Cre specifically in inhibitory neurons) allows us to distinguish excitatory projecting neurons from postsynaptic neurons, increasing interpretability. However, this method requires complex procedures, such as preparation of transgenic animals. (C) Schematic of DISECT developed in this study. By injecting a Cre-inducible Dlx enhancer-specific AAV vector into the postsynaptic region, transduction can be restricted only to the postsynaptic inhibitory neurons with simple procedures, ensuring simplified efficient tracing with high interpretability in wild-type animals.

Article Snippet: We called this method “Dlx enhancer-restricted Interneuron-SpECific transsynaptic Tracing” (DISECT).

Techniques: Comparison, Infection, Plasmid Preparation, Transduction, Expressing, Transgenic Assay