Differential Expression of Nasal Embryonic LHRH Factor (NELF) Variants in Immortalized GnRH Neuronal Cell Lines
| Autor: | Durkadin Demir, Richard S. Cameron, Lynn P. Chorich, Eunkyung Ko, Lindsey Y. Goldberg, Lawrence C. Layman, Samuel D. Quaynor, Hyung Goo Kim, Robert K. Stanley |
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| Jazyk: | angličtina |
| Rok vydání: | 2013 |
| Předmět: |
Nuclear Localization Signals
Gonadotropin-releasing hormone Biology Biochemistry Article Gonadotropin-Releasing Hormone Mice Endocrinology Species Specificity Cell Movement medicine NLS Animals Humans Protein Isoforms RNA Messenger Molecular Biology Cellular localization Cell Line Transformed Regulation of gene expression Cell Nucleus Neurons Alternative splicing NASAL EMBRYONIC LHRH FACTOR Molecular biology Cell nucleus Alternative Splicing medicine.anatomical_structure Gene Expression Regulation Nuclear localization sequence Signal Transduction Transcription Factors |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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