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Reliable Identification of Living Dopaminergic Neurons in Midbrain Cultures Using RNA Sequencing and TH-promoter-driven eGFP Expression

Reliable Identification of Living Dopaminergic Neurons in Midbrain Cultures Using RNA Sequencing and TH-promoter-driven eGFP Expression

Published on: February 10, 2017

Distinct transcriptomes define rostral and caudal serotonin neurons.

Christi J Wylie¹, Timothy J Hendricks, Bing Zhang

¹Department of Neurosciences, Case Western Reserve University, Cleveland, Ohio 44106, USA.

The Journal of Neuroscience : the Official Journal of the Society for Neuroscience

|January 15, 2010

Summary

Researchers identified distinct molecular subtypes of developing serotonin (5HT) neurons using gene expression profiling. These subtypes, defined by homeodomain gene expression, offer new insights into brain development and neurological disorders.

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Published on: September 23, 2015

Area of Science:

Neuroscience
Molecular Biology
Genetics

Background:

The molecular underpinnings of developing serotonin (5HT) neurons are largely unknown, hindering understanding of their functional diversity and roles in disease.
Elucidating this molecular architecture is crucial for addressing serotonergic dysfunction in various neuropathologies.

Purpose of the Study:

To characterize the molecular profiles of developing embryonic 5HT neurons.
To identify novel markers and genetic networks defining 5HT neuron subtypes.
To investigate the heterogeneity of rostral and caudal 5HT neuron populations.

Main Methods:

Purification of postmitotic embryonic 5HT neurons via flow cytometry.
Whole-genome microarray expression profiling to analyze gene expression patterns.
Verification studies using homeodomain and engrailed gene expression analysis.

Main Results:

Identification of hundreds of uniquely enriched genes in 5HT neurons, serving as potential new markers.
Discovery of differential gene enrichment in rostral and caudal 5HT neuron subtypes, including homeodomain, axon guidance, and imprinted genes.
Defined rostral (Hmx+) and caudal (Hox+) 5HT neuron subtypes, with further rostral subtypes (Hmx+En+, Hmx+En-) identified.

Conclusions:

Developing 5HT neurons exhibit significant molecular and pathway heterogeneity, particularly between rostral and caudal populations.
A homeodomain code (Hmx and Hox genes) distinguishes these major 5HT neuron subtypes.
The findings provide a valuable resource for understanding 5HT neuron development, function, and associated neurodevelopmental disorders.