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Cre-assisted fine-mapping of neural circuits using orthogonal split inteins
Haojiang Luan1, Alexander Kuzin2, Ward F Odenwald2
1Laboratory of Molecular Biology, National Institute of Mental Health, NIH, Bethesda, United States.
Elife
|April 15, 2020
Summary
This study introduces a novel iterative method for precise neuronal targeting in Drosophila, refining brain circuit mapping. The technique enhances resolution by combining developmental lineage isolation with split Cre recombinase activation.
Area of Science:
- Neuroscience
- Genetics
- Developmental Biology
Background:
- Current genetic neuronal targeting methods lack the resolution for detailed brain circuit mapping.
- Improved techniques are essential for advancing our understanding of neural networks.
Purpose of the Study:
- To develop a novel, iterative method for refined neuronal targeting.
- To enhance the precision of brain circuit mapping in Drosophila.
Main Methods:
- Isolating neurons by developmental lineage within a targeted group (Gal4 pattern).
- Intersectionally limiting lineages by selecting for dual neuroblast enhancer activity.
- Utilizing split Cre recombinase fragments fused to split inteins for targeted activation.
Main Results:
- Demonstrated a two-step refinement process for neuronal targeting.
- Successfully activated Cre recombinase in specific neuroblasts, enabling Gal4 activity in derived cells.
- Facilitated enhanced neural circuit mapping in Drosophila.
Conclusions:
- The developed method offers improved resolution for neuronal targeting.
- This iterative approach provides a powerful tool for detailed neural circuit analysis.
- The system is applicable to complex brain circuit investigations in Drosophila.
