Transcriptional regulation by nicotine in dopaminergic neurons.
Dopaminergic neurons in the substantia nigra pars compacta (SNc) degenerate in Parkinson's disease. These neurons robustly express several nicotinic acetylcholine receptor (nAChR) subtypes. Smoking appears to be neuroprotective for Parkinson's disease but the mechanism is unknown. To determine whether chronic nicotine-induced changes in gene expression contribute to the neuroprotective effects of smoking, we develop methods to measure the effect of prolonged nicotine exposure on the SNc neuronal transcriptome in an unbiased manner. Twenty neurons were collected using laser-capture microscopy and transcriptional changes were assessed using RNA deep sequencing. These results are the first whole-transcriptome analyses of chronic nicotine treatment in SNc neurons. Overall, 129 genes were significantly regulated: 67 upregulated, 62 downregulated. Nicotine-induced relief of endoplasmic reticulum (ER) stress has been postulated as a potential mechanism for the neuroprotective effects of smoking. Chronic nicotine did not significantly affect the expression of ER stress-related genes, nor of dopamine-related or nAChR genes, but it did modulate expression of 129 genes that could be relevant to the neuroprotective effects of smoking, including genes involved in (1) the ubiquitin-proteasome pathway, (2) cell cycle regulation, (3) chromatin modification, and (4) DNA binding and RNA regulation. We also report preliminary transcriptome data for single-cell dopaminergic and GABAergic neurons isolated from midbrain cultures. These novel techniques will facilitate advances in understanding the mechanisms taking place at the cellular level and may have applications elsewhere in the fields of neuroscience and molecular biology. The results give an emerging picture of the role of nicotine on the SNc and on dopaminergic neurons.
California Institute of Technology, 156-29 Caltech, Pasadena, CA 91125, USA. Electronic address: bhenley@caltech.edu.
Beverley M
BM
Henley
Brian A
BA
Williams
Rahul
R
Srinivasan
Bruce N
BN
Cohen
Cheng
C
Xiao
Elisha D W
ED
Mackey
Barbara J
BJ
Wold
Henry A
HA
Lester
D000818:Animals; D004298:Dopamine / Q000378:metabolism; D059290:Dopaminergic Neurons / Q000378:metabolism*; D005786:Gene Expression Regulation / Q000187:drug effects*; D016678:Genome; D051379:Mice; D008810:Mice, Inbred C57BL; D009538:Nicotine / Q000008:administration & dosage / Q000494:pharmacology*; D013312:Stress, Physiological; D013378:Substantia Nigra / Q000166:cytology; D059467:Transcriptome; D005680:gamma-Aminobutyric Acid / Q000378:metabolism
D005680:gamma-Aminobutyric Acid; D009538:Nicotine; D004298:Dopamine
DAVID; Database for Annotation, Visualization and Integrated Discovery; Dopamine-related genes; ER; FPKM; IPA; Ingenuity Pathway Analysis; Nicotinic acetylcholine receptor; PD; Parkinson's disease; RNA-Seq; SNc; Substantia nigra pars compacta; UPR; Ubiquitin–proteasome pathway; VTA; endoplasmic reticulum; fragments per kilobase per million mapped reads; nAChR; nicotinic acetylcholine receptor; substantia nigra pars compacta; unfolded protein response; ventral tegmental area