title: Differential expression of nasal embryonic LHRH factor (NELF) variants in immortalized GnRH neuronal cell lines.
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.
authors: Section of Reproductive Endocrinology, Infertility, & Genetics, Department of Obstetrics & Gynecology; Institute of Molecular Medicine and Genetics; Georgia Regents University, Augusta, GA 30912, USA.
authors: Samuel D
authors: SD
authors: Quaynor
authors: Department of Internal Medicine, University of Kentucky, Lexington, KY 40536-0284, USA.
authors: Lindsey Y
authors: LY
authors: Goldberg
authors: Section of Reproductive Endocrinology, Infertility, & Genetics, Department of Obstetrics & Gynecology; Institute of Molecular Medicine and Genetics; Georgia Regents University, Augusta, GA 30912, USA.
authors: Eun Kyung
authors: EK
authors: Ko
authors: Department of Chemistry and Biochemistry, University of Notre Dame, South Bend, IN 46556, USA.
authors: Robert K
authors: RK
authors: Stanley
authors: Department of Medical Biology and Genetics, Akdeniz University, Antalya 07058, Turkey.
authors: Durkadin
authors: D
authors: Demir
authors: Section of Reproductive Endocrinology, Infertility, & Genetics, Department of Obstetrics & Gynecology; Institute of Molecular Medicine and Genetics; Georgia Regents University, Augusta, GA 30912, USA.
authors: Hyung-Goo
authors: HG
authors: Kim
authors: Section of Reproductive Endocrinology, Infertility, & Genetics, Department of Obstetrics & Gynecology; Institute of Molecular Medicine and Genetics; Georgia Regents University, Augusta, GA 30912, USA.
authors: Lynn P
authors: LP
authors: Chorich
authors: Department of Medicine; Institute of Molecular Medicine and Genetics; Georgia Regents University, Augusta, GA 30912, USA.
authors: Richard S
authors: RS
authors: Cameron
authors: Section of Reproductive Endocrinology, Infertility, & Genetics, Department of Obstetrics & Gynecology; Institute of Molecular Medicine and Genetics; Georgia Regents University, Augusta, GA 30912, USA. Electronic address: lalayman@gru.edu.
authors: Lawrence C
authors: LC
authors: Layman
mesh_terms: D017398:Alternative Splicing; D000818:Animals; D002461:Cell Line, Transformed; D002465:Cell Movement; D002467:Cell Nucleus / Q000378:metabolism; D005786:Gene Expression Regulation; D007987:Gonadotropin-Releasing Hormone / Q000378:metabolism; D006801:Humans; D051379:Mice; D009474:Neurons / Q000166:cytology / Q000378:metabolism*; D019913:Nuclear Localization Signals; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics* / Q000378:metabolism; D015398:Signal Transduction; D013045:Species Specificity; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism
chemical_list: C582413:NELF protein, mouse; C411985:NSMF protein, human; D019913:Nuclear Localization Signals; D020033:Protein Isoforms; D012333:RNA, Messenger; D014157:Transcription Factors; D007987:Gonadotropin-Releasing Hormone
keywords: GN11 cells; GT1-7 cells; GnRH neurons; NLT cells; NSMF; Nasal embryonic LHRH factor (NELF)