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Neurochemical Research

, Volume 21, Issue 4, pp 403–409 | Cite as

Expression of a unique globo-series glycolipid in cultured rat dorsal root ganglion neurons: Relationship with neuronal development

  • Shubhro Pal
  • John W. Bigbee
  • Megumi Saito
  • Toshio Ariga
  • Robert K. Yu
Original Articles

Abstract

Previous studies from this laboratory demonstrated the presence of a UDP-galactose:Gb3Cer α1-3galactosyltansferase activity responsible for the synthesis of a unique glycosphingolipid (GSL), Galα1-3Gb3Cer, in cultured PC12 pheochromocytoma cells (21). In this investigation, we examined the presence of this enzyme activity in isolated rat embryonic dorsal root ganglion neurons (DRGN), which, like pheochromocytoma cells, originate from the neural crest cells. DRGN exhibited the α-galactosyltransferase activity and the activity was comparable to that of the PC12 cells while several other rat tissues, with the exception of kidney, showed minimal activity. In order to define the spatial and temporal expression of Galα1-3Gb3Cer in DRGN, we examined the expression of Galα1-3Gb3Cer in cultured DRGN derived from embryonic day 16 rat embryos. Using a polyclonal antibody raised against Galα1-3Gb3Cer, we examined the localization of this glycolipid in DRGN cells after, 5, 8, 12, and 15 days in culture. Immunostaining was restricted to the neurons while Schwann cells were negative. At day 5, the immunostaining was weak and confined to the cell body of the DRGN, though neurites were present at this stage. The period between days 5 and 15 represented a period of rapid neuritic growth and continued enlargement of the cell bodies. Immunoreactivity in the cell bodies increased dramatically by day 8. By day 12, immunoreactivity was present in neurites, and by day 15, was strong in both cell bodies and neurites. The expression of Galα1-3Gb3Cer in vivo was confirmed by immunostaining of frozen sections of dorsal root ganglia. Our present studies which demonstrate neuron-specific expression of Galα1-3Gb3Cer during neurotigenesis combined with previous observations for its expression in PC12 cells, strongly implicates this GSL in neuronal development.

Key words

Glycosphingolipid dorsal rest ganglion neuronal development 

List of Abbreviations

GSL

Glycosphingolipid

HPTLC

Highperformance thin-layer chromatography

Cer

Ceramide

NGF

Nerve growth factor

DRG

Dorsal root ganglion

UDP

Uridine 5′-diphosphate

CDP

Cytidine 5′-diphosphate

DEAE

Diethylaminoethyl

Gb3Cer

Galα1-4Galβ1-4Glcβ1-1’Cer

GalGb3Cer

Galα1-3Galα1-4Galβ1-4Glcβ1-1’Cer

GalGalGb3Cer

Galα1-3Galα1-3Galα1-4Galβ1-4Glcβ1-1’Cer

PBS

Phosphate buffered saline

GFAP

Glial fibrillary acidic protein

ELISA

Enzyme linked immunosorbent assay

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Copyright information

© Plenum Publishing Corporation 1996

Authors and Affiliations

  • Shubhro Pal
    • 1
  • John W. Bigbee
    • 2
  • Megumi Saito
    • 1
  • Toshio Ariga
    • 1
  • Robert K. Yu
    • 1
  1. 1.Department of Biochemistry and Molecular BiophysicsVirginia Commonwealth UniversityRichmond
  2. 2.Department of Anatomy, Medical College of VirginiaVirginia Commonwealth UniversityRichmond

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