title: Developmental regulation of two distinct neuronal phenotypes in rat dorsal root ganglia.
abstract: In a previous study we described two distinct neuronal phenotypes in rat dorsal root ganglia based on immunocytochemical assays for the neuronal intermediate filament proteins, peripherin and low-molecular-weight neurofilaments [Goldstein M. E. et al. (1991) J. Neurosci. Res. 30, 92-104]. In this paper we have extended this classification by using in situ hybridization to localize and evaluate the levels of various cytoskeletal and neuropeptide messenger RNAs within the peripherin-immunoreactive and peripherin-immunoreactive-negative neurons found in embryonic day 15 and 20, postnatal day 2 and adult dorsal root ganglia. We found in postnatal and adult dorsal root ganglia in vivo that the large, peripherin-immunoreactive-negative neurons, which are intensely stained by low-molecular-weight neurofilament antibodies, also contain high levels of low, medium and high-molecular-weight neurofilament messenger RNAs, whereas the smaller peripherin-immunoreactive neurons do not. On the other hand, both cell types contained comparable levels of peripherin and alpha-tubulin messenger RNA. The presence of peripherin messenger RNA but no peripherin immunoreactivity in the large cells suggested either a translational or post-translational regulation of this polypeptide, or rapid clearance of this protein from the perikaryon into the axon. In adult dorsal root ganglia, more than 50% of the peripherin-immunoreactive neurons also contained high levels of substance P and/or calcitonin gene-related peptide messenger RNAs, while less than 20% of the large peripherin-immunoreactive-negative neurons did. The attainment of these phenotypic characteristics during development in vivo was studied by northern blot and in situ hybridization histochemistry. In early embryonic stages (embryonic days 15-16), virtually all neurons were peripherin-immunoreactive and were positive for peripherin, alpha-tubulin and low-molecular-weight neuro-filament messenger RNAs, suggesting a homogeneous population. By embryonic day 20, the two adult phenotypes became clearly evident, and were fully established by postnatal day 2. In cultures of embryonic day 15 dorsal root ganglion neurons grown in the presence of nerve growth factor, peripherin and low-molecular-weight neurofilament messenger RNAs were expressed in all neurons, even after nine days in vitro, similar to embryonic dorsal root ganglia in vivo. Nerve growth factor supplemented by skeletal and heart muscle extracts did up-regulate neurofilament gene expression, but not to the extent characteristic of the peripherin-immunoreactive-negative adult phenotype. These results suggest that development of the mature phenotype of dorsal root ganglion neurons occurs by postnatal day 2 in vivo and is dependent upon target contact and/or target-derived factors.
authors.affiliation: Laboratory of Neurochemistry, NINDS, NIH, Bethesda, MD 20892-4130, USA.
authors.forename: M E
authors.initials: ME
authors.lastname: Goldstein
authors.forename: P
authors.initials: P
authors.lastname: Grant
authors.forename: S B
authors.initials: SB
authors.lastname: House
authors.forename: D B
authors.initials: DB
authors.lastname: Henken
authors.forename: H
authors.initials: H
authors.lastname: Gainer
mesh_terms: D000818:Animals; D015152:Blotting, Northern; D005136:Eye Proteins / Q000378:metabolism; D005727:Ganglia, Spinal / Q000166:cytology* / Q000254:growth & development*; D018507:Gene Expression Regulation, Developmental / Q000502:physiology*; D006321:Heart / Q000502:physiology; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D007381:Intermediate Filament Proteins / Q000378:metabolism; D008562:Membrane Glycoproteins; D008970:Molecular Weight; D018482:Muscle, Skeletal / Q000502:physiology; D009419:Nerve Tissue Proteins; D016900:Neurofilament Proteins / Q000096:biosynthesis / Q000235:genetics; D009474:Neurons / Q000502:physiology*; D009479:Neuropeptides / Q000096:biosynthesis / Q000378:metabolism; D015345:Oligonucleotide Probes; D064531:Peripherins; D010641:Phenotype; D015347:RNA Probes; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D051381:Rats; D014020:Tissue Extracts / Q000494:pharmacology
chemical_list: D005136:Eye Proteins; D007381:Intermediate Filament Proteins; D008562:Membrane Glycoproteins; D009419:Nerve Tissue Proteins; D016900:Neurofilament Proteins; D009479:Neuropeptides; D015345:Oligonucleotide Probes; D064531:Peripherins; D015347:RNA Probes; D012333:RNA, Messenger; D014020:Tissue Extracts