Generation of functional human serotonergic neurons from fibroblasts

Mol Psychiatry. 2016 Jan;21(1):49-61. doi: 10.1038/mp.2015.161. Epub 2015 Oct 27.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Astrocytes / physiology
  • Basic Helix-Loop-Helix Transcription Factors / genetics
  • Basic Helix-Loop-Helix Transcription Factors / metabolism
  • Cell Line
  • Cell Transdifferentiation / physiology
  • Cytological Techniques / methods*
  • DNA-Binding Proteins / genetics
  • DNA-Binding Proteins / metabolism
  • Fibroblasts / physiology*
  • GATA2 Transcription Factor / genetics
  • GATA2 Transcription Factor / metabolism
  • Genetic Vectors
  • Homeodomain Proteins / genetics
  • Homeodomain Proteins / metabolism
  • Human Embryonic Stem Cells / physiology
  • Humans
  • LIM-Homeodomain Proteins / genetics
  • LIM-Homeodomain Proteins / metabolism
  • Lentivirus / genetics
  • Mice
  • Nerve Tissue Proteins / genetics
  • Nerve Tissue Proteins / metabolism
  • Nuclear Proteins / genetics
  • Nuclear Proteins / metabolism
  • Serotonergic Neurons / physiology*
  • Transcription Factors / genetics
  • Transcription Factors / metabolism
  • Transcriptome
  • Zebrafish Proteins

Substances

  • ASCL1 protein, human
  • Basic Helix-Loop-Helix Transcription Factors
  • DNA-Binding Proteins
  • FEV protein, human
  • GATA2 Transcription Factor
  • GATA2 protein, human
  • Homeodomain Proteins
  • LIM homeobox transcription factor 1 beta
  • LIM-Homeodomain Proteins
  • NEUROG2 protein, human
  • Nerve Tissue Proteins
  • Nkx2.2 protein
  • Nuclear Proteins
  • Transcription Factors
  • Zebrafish Proteins