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Multiplex Detection of Gene Expression in the Intact Drosophila Brain Using Expansion-Assisted Iterative Fluorescence In Situ Hybridization
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Dynamic neurotransmitter specific transcription factor expression profiles during Drosophila development
Alicia Estacio-Gómez1, Amira Hassan1, Emma Walmsley1
1Department of Life Sciences, Imperial College London, Sir Ernst Chain Building, London SW7 2AZ, UK.
Biology Open
|June 5, 2020
Summary
Researchers identified 86 transcription factors involved in specifying neuron types, finding no simple code for neurotransmitter fate. Some factors regulate key genes like ChAT, but adult brains show non-overlapping expression, suggesting limited combinatorial control.
Area of Science:
- Neuroscience
- Developmental Biology
- Genetics
Background:
- Neuronal diversity arises during development through specification of cell morphology, receptor profiles, and neurotransmitter identities.
- Understanding the molecular mechanisms of neurotransmitter specification is crucial for deciphering nervous system development.
Purpose of the Study:
- To profile the transcription state of cholinergic, GABAergic, and glutamatergic neurons during development.
- To identify transcription factors involved in specifying neurotransmitter identity.
- To investigate the regulatory roles of specific transcription factors and their binding sites.
Main Methods:
- In vivo transcriptional profiling of specific neuron types at three developmental time points.
- Identification and analysis of differentially expressed transcription factors.
- Chromatin immunoprecipitation followed by sequencing (ChIP-seq) to profile transcription factor binding sites.
- Analysis of transcription factor expression patterns in the adult brain.
Main Results:
- Identified 86 differentially expressed transcription factors uniquely enriched or depleted in specific neurotransmitter neuron types.
- Some transcription factors exhibited consistent profiles across development, while others showed stage-specific enrichment or depletion.
- Acj6 and Ets65A were found to directly bind the ChAT locus and other neuronal differentiation genes.
- Cholinergic-enriched transcription factors were largely expressed in non-overlapping populations in the adult brain.
Conclusions:
- Neurotransmitter type specification is complex, with no simple transcription factor codes identified, similar to findings in Caenorhabditis elegans.
- While some transcription factors directly regulate key genes like ChAT, combinatorial regulation of neurotransmitter fate appears limited in the adult brain.
- The study provides insights into the dynamic regulation of transcription factors during neuronal development and neurotransmitter specification.
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