Generation of functional human serotonergic neurons from fibroblasts.
The brain's serotonergic system centrally regulates several physiological processes and its dysfunction has been implicated in the pathophysiology of several neuropsychiatric disorders. While in the past our understanding of serotonergic neurotransmission has come mainly from mouse models, the development of pluripotent stem cell and induced fibroblast-to-neuron (iN) transdifferentiation technologies has revolutionized our ability to generate human neurons in vitro. Utilizing these techniques and a novel lentiviral reporter for serotonergic neurons, we identified and overexpressed key transcription factors to successfully generate human serotonergic neurons. We found that overexpressing the transcription factors NKX2.2, FEV, GATA2 and LMX1B in combination with ASCL1 and NGN2 directly and efficiently generated serotonergic neurons from human fibroblasts. Induced serotonergic neurons (iSNs) showed increased expression of specific serotonergic genes that are known to be expressed in raphe nuclei. iSNs displayed spontaneous action potentials, released serotonin in vitro and functionally responded to selective serotonin reuptake inhibitors (SSRIs). Here, we demonstrate the efficient generation of functional human serotonergic neurons from human fibroblasts as a novel tool for studying human serotonergic neurotransmission in health and disease.
Laboratory of Genetics, The Salk Institute for Biological Studies, La Jolla, CA, USA.
K C
KC
Vadodaria
Laboratory of Genetics, The Salk Institute for Biological Studies, La Jolla, CA, USA.
J
J
Mertens
Laboratory of Genetics, The Salk Institute for Biological Studies, La Jolla, CA, USA.
A
A
Paquola
Laboratory of Genetics, The Salk Institute for Biological Studies, La Jolla, CA, USA.
C
C
Bardy
Laboratory of Genetics, The Salk Institute for Biological Studies, La Jolla, CA, USA.|University of California, San Diego (UCSD), La Jolla, CA, USA.
X
X
Li
Laboratory of Genetics, The Salk Institute for Biological Studies, La Jolla, CA, USA.
R
R
Jappelli
Laboratory of Genetics, The Salk Institute for Biological Studies, La Jolla, CA, USA.|University of California, San Diego (UCSD), La Jolla, CA, USA.
L
L
Fung
Laboratory of Genetics, The Salk Institute for Biological Studies, La Jolla, CA, USA.
M C
MC
Marchetto
The Salk Institute for Biological Studies, La Jolla, CA, USA.
M
M
Hamm
Laboratory of Genetics, The Salk Institute for Biological Studies, La Jolla, CA, USA.
M
M
Gorris
Institute of Reconstructive Neurobiology, University of Bonn, Bonn, Germany.
P
P
Koch
Laboratory of Genetics, The Salk Institute for Biological Studies, La Jolla, CA, USA.
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FH
Gage
D000818:Animals; D001253:Astrocytes / Q000502:physiology; D051792:Basic Helix-Loop-Helix Proteins / Q000235:genetics / Q000378:metabolism; D002460:Cell Line; D054338:Cell Transdifferentiation / Q000502:physiology; D003584:Cytological Techniques / Q000379:methods*; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D005347:Fibroblasts / Q000502:physiology*; D050989:GATA2 Transcription Factor / Q000235:genetics / Q000378:metabolism; D005822:Genetic Vectors; D000090322:Homeobox Protein Nkx-2.2; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism; D000066449:Human Embryonic Stem Cells / Q000502:physiology; D006801:Humans; D060850:LIM-Homeodomain Proteins / Q000235:genetics / Q000378:metabolism; D016086:Lentivirus / Q000235:genetics; D051379:Mice; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism; D059326:Serotonergic Neurons / Q000502:physiology*; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D059467:Transcriptome; D029961:Zebrafish Proteins
D051792:Basic Helix-Loop-Helix Proteins; D004268:DNA-Binding Proteins; D050989:GATA2 Transcription Factor; D000090322:Homeobox Protein Nkx-2.2; D018398:Homeodomain Proteins; D060850:LIM-Homeodomain Proteins; D009419:Nerve Tissue Proteins; D009687:Nuclear Proteins; D014157:Transcription Factors; D029961:Zebrafish Proteins; C065500:ASCL1 protein, human; C474280:FEV protein, human; C494711:GATA2 protein, human; C112149:LIM homeobox transcription factor 1 beta; C491183:NEUROG2 protein, human; C000715631:NKX2-2 protein, human; C000715627:Nkx2-2 protein, mouse; C000715630:nkx2.2b protein, zebrafish