Induction of a serotonergic and neuronal phenotype in thyroid C-cells.
We have investigated whether rat thyroid C-cells can acquire a phenotype similar to serotonergic neurons. C-cells are neural crest derived endocrine cells with some intrinsic neuronal and serotonergic properties. A relatively simple isolation scheme yielded cultures of about 50% initial purity, as measured by fluorescence activated cell sorting. These enriched C-cells could extend neurites up to 550 microns on a laminin-containing substratum in the presence of NGF. The cultured C-cells expressed neurofilaments and this expression was enhanced by NGF treatment. The C-cells also expressed two markers of the sympathoadrenal neural crest lineage, the mammalian achaete scute homolog-1 (MASH-1) transcription factor, and the B2 cell surface antigen. Interestingly, MASH-1 was not detectable after the C-cells were placed in culture, which is consistent with neuronal differentiation, since MASH-1 is only expressed in neuronal progenitors prior to differentiation. We then demonstrated that C-cells possess the fundamental features of serotonergic neurons: synthesis and secretion, uptake, and feedback control. The enriched C-cells, as well as the CA77 C-cell line, showed 5-HT immunostaining, expression of tryptophan hydroxylase mRNA, 5-HT1B autoreceptor mRNA, and 5-HT transporter mRNA and activity. NGF greatly induced 5-HT transporter activity as determined by sensitivity to sertraline, a selective 5-HT reuptake inhibitor. Based on these results, we propose that thyroid C-cells are derived from a vagal sympathoadrenal progenitor, similar to serotonergic enteric neurons, and can undergo neuronal transdifferentiation. Hence, these cells should provide suitable and convenient models for molecular and cellular studies on serotonergic neurons.
Clark
M S
MS
Molecular Biology Program, University of Iowa, Iowa City 52242, USA.
Lanigan
T M
TM
Page
N M
NM
Russo
A F
AF
D000818:Animals; D000954:Antigens, Surface / Q000378:metabolism; D001483:Base Sequence; D051792:Basic Helix-Loop-Helix Proteins; D002352:Carrier Proteins / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism; D005260:Female; D008297:Male; D008562:Membrane Glycoproteins / Q000378:metabolism; D026901:Membrane Transport Proteins; D015335:Molecular Probes / Q000235:genetics; D008969:Molecular Sequence Data; D009414:Nerve Growth Factors / Q000494:pharmacology; D009419:Nerve Tissue Proteins; D016501:Neurites / Q000502:physiology; D016900:Neurofilament Proteins / Q000502:physiology; D009474:Neurons / Q000502:physiology*; D010641:Phenotype; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D011985:Receptors, Serotonin / Q000235:genetics / Q000378:metabolism; D012701:Serotonin / Q000502:physiology*; D050486:Serotonin Plasma Membrane Transport Proteins; D013961:Thyroid Gland / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D014157:Transcription Factors / Q000378:metabolism
D000954:Antigens, Surface; D051792:Basic Helix-Loop-Helix Proteins; D002352:Carrier Proteins; D004268:DNA-Binding Proteins; D008562:Membrane Glycoproteins; D026901:Membrane Transport Proteins; D015335:Molecular Probes; D009414:Nerve Growth Factors; D009419:Nerve Tissue Proteins; D016900:Neurofilament Proteins; D012333:RNA, Messenger; D011985:Receptors, Serotonin; D012701:Serotonin; D050486:Serotonin Plasma Membrane Transport Proteins; D014157:Transcription Factors; C499318:Ascl1 protein, rat; C493232:Slc6a4 protein, rat