title: Ciliary neurotrophic factor overexpression in neural progenitor cells (ST14A) increases proliferation, metabolic activity, and resistance to stress during differentiation.
abstract: Neurotrophic factors exert considerable neuroprotective and neurorestorative effects in neurodegenerative diseases. Because neuronal progenitor cells have, at least in part, the potency to restore degenerated neuronal networks, transgenic high-dosage expression of neurotrophins by these cells in neurotransplantation may be advantageous. In the present study, a retroviral vector containing the gene of rat ciliary neurotrophic factor (rCNTF) was permanently transfected into a striatal neuronal progenitor cell line. Qualitative and quantitative analyses demonstrated a sustained expression of the transgene; i.e., rCNTF was present at the mRNA level and protein level. Moreover, cocultivation in separate chambers of transgenic CNTF-ST14A cells and CNTF-dependent TF1 cells exerted typical biological effects, such as increased proliferation and differentiation of the TF1 cells, indicating the functional integrity of the secreted recombinant neurotrophin. The CNTF-ST14A cells displayed improved stress response compared with native ST14A cells under differentiation conditions, i.e., at the nonpermissive temperature of 39 degrees C and after staurosporine exposure, respectively. This effect coincided with a relatively reduced apoptosis rate and a raised metabolic activity of CNTF-ST14A cells at 39 degrees C. Neurotransplantation of CNTF-ST14A cells in the rat quinolinic acid model of Huntington's disease showed a significant and sustained decline in pathological apomorphine-induced rotations compared with parental ST14A cells. We conclude that sustained functional transgene CNTF production improves stress response as well as metabolic activity, making CNTF-ST14A cells a promising tool for neurotransplantation in the quinolinic acid model of Huntington's disease.
authors.affiliation: Neurobiological Laboratory, Department of Neurology, University of Rostock, Rostock, Germany.
authors.forename: Sabine
authors.initials: S
authors.lastname: Weinelt
authors.forename: Sabine
authors.initials: S
authors.lastname: Peters
authors.forename: Peter
authors.initials: P
authors.lastname: Bauer
authors.forename: Eilhard
authors.initials: E
authors.lastname: Mix
authors.forename: Stefan J P
authors.initials: SJ
authors.lastname: Haas
authors.forename: Aline
authors.initials: A
authors.lastname: Dittmann
authors.forename: Stanislov
authors.initials: S
authors.lastname: Petrov
authors.forename: Andreas
authors.initials: A
authors.lastname: Wree
authors.forename: Elena
authors.initials: E
authors.lastname: Cattaneo
authors.forename: Rupert
authors.initials: R
authors.lastname: Knoblich
authors.forename: Ulf
authors.initials: U
authors.lastname: Strauss
authors.forename: Arndt
authors.initials: A
authors.lastname: Rolfs
mesh_terms: D000818:Animals; D001058:Apomorphine / Q000633:toxicity; D017209:Apoptosis / Q000502:physiology; D015153:Blotting, Western; D002454:Cell Differentiation / Q000502:physiology*; D002455:Cell Division / Q000502:physiology; D002478:Cells, Cultured; D020934:Ciliary Neurotrophic Factor / Q000378:metabolism* / Q000502:physiology; D018920:Coculture Techniques; D003342:Corpus Striatum / Q000293:injuries; D016895:Culture Media, Serum-Free / Q000494:pharmacology; D004195:Disease Models, Animal; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005347:Fibroblasts / Q000378:metabolism; D015870:Gene Expression; D020287:In Situ Nick-End Labeling; D017378:Quinolinic Acid / Q000633:toxicity; D012333:RNA, Messenger / Q000096:biosynthesis; D051381:Rats; D017208:Rats, Wistar; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012399:Rotation; D019311:Staurosporine / Q000494:pharmacology; D013234:Stem Cells / Q000502:physiology*; D013312:Stress, Physiological; D013778:Tetrazolium Salts; D013697:Thermosensing; D013936:Thymidine / Q000096:biosynthesis; D013997:Time Factors; D014162:Transfection; D014316:Tritium
chemical_list: C110656:2-(4-iodophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium; D020934:Ciliary Neurotrophic Factor; D016895:Culture Media, Serum-Free; D004791:Enzyme Inhibitors; D012333:RNA, Messenger; D013778:Tetrazolium Salts; D014316:Tritium; D017378:Quinolinic Acid; D019311:Staurosporine; D001058:Apomorphine; D013936:Thymidine