Differential expression of nasal embryonic LHRH factor (NELF) variants in immortalized GnRH neuronal cell lines

Samuel D. Quaynor, Lindsey Y. Goldberg, Eun Kyung Ko, Robert K. Stanley, Durkadin Demir, Hyung Goo Kim, Lynn P. Chorich, Richard S Cameron, Lawrence C Layman

Research output: Contribution to journalArticle

3 Scopus citations

Abstract

NELF, a protein identified in migratory GnRH neurons, is predominantly nuclear and alternatively spliced. However, specific NELF splice variants expressed in immortalized GnRH neuronal cell lines from mouse and human are not known. RNA from migratory (GN11 and NLT) and postmigratory (GT1-7) cells in mouse, and (FNCB4-hTERT) cells in human was subjected to RT-PCR. RT-PCR products were cloned, electrophoresed on denaturing gradient gels and sequenced. In addition, quantitative RT-PCR was performed using variant-specific primers. Western blot and immunofluorescence using confocal microscopy were performed for selected variants. Nelf variant 2 (v2), which contains a nuclear localization signal (NLS), was the predominant variant in all mouse and human GnRH neurons. Variants without a NLS (v3 in mouse; v4 in human) were identified. In mouse, v2 protein expression was nuclear, while v3 was non-nuclear. In mouse GnRH neurons, six Nelf splice variant transcripts were identified, including three previously unreported variants. In human, four NELF variant transcripts were observed. In both mouse and human, nuclear and non-nuclear variant transcript and protein were identified, explaining variable NELF cellular localization.

Original languageEnglish (US)
Pages (from-to)32-37
Number of pages6
JournalMolecular and Cellular Endocrinology
Volume383
Issue number1
DOIs
StatePublished - Mar 5 2014

Keywords

  • GN11 cells
  • GT1-7 cells
  • GnRH neurons
  • NLT cells
  • NSMF
  • Nasal embryonic LHRH factor (NELF)

ASJC Scopus subject areas

  • Endocrinology
  • Molecular Biology
  • Biochemistry

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