Abstract
Neural stem cell lines represent a homogeneous source of cells for genetic, developmental, and gene transfer and repair studies in the nervous system. Since both gene transfer of neurotrophic factors and cell replacement strategies are of immediate interest for therapeutical purposes, we have generated BDNF-secreting neural stem cell lines and investigated to what extent different endogenous levels of BDNF expression affect in vitro survival, proliferation and differentiation of these cells. Also, we have investigated the in vivo effects of such BDNF gene transfer procedure in the rat neostriatum. Hippocampus- and cerebellum-derived cell lines reacted differently to manipulations aimed at varying their levels of BDNF production. Over-expression of BDNF enhanced survival of both cell types, in a serum-deprivation assay. Conversely, and ruling out unspecific effects, expression of an antisense version of BDNF resulted in compromised survival of cerebellum-derived cells, and in a lethal phenotype in hippocampal progenitors. These data indicate that endogenous BDNF level strongly influences the in vitro survival of these cells. These effects are more pronounced for hippocampus- than for cerebellum-derived progenitors. Hippocampus-derived BDNF overproducers showed no major change in their capacity to differentiate towards a neuronal phenotype in vitro. In contrast, cerebellar progenitors overproducing BDNF did not differentiate into neurons, whereas cells expressing the antisense BDNF construct generated cells with morphological features of neurons and expressing immunological neuronal markers. Taken together, these results provide evidence that BDNF controls both the in vitro survival and differentiation of neural stem cells. After in vivo transplantation of BDNF-overproducing cells to the rat neostriatum, these survived better than the control ones, and induced the expected neurotrophic effects on cholinergic neurons. However, long-term (3 months) administration of BDNF resulted in detrimental effects, at this location. These findings may be of importance for the understanding of brain development, for the design of therapeutic neuro-regenerative strategies, and for cell replacement and gene therapy studies.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$259.00 per year
only $21.58 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Gage FH, Ray J, Fisher LJ . Isolation, characterization, and use of stem cells from the CNS Annu Rev Neurosci 1995 18: 159–192
Morrison SJ, Shah NM, Anderson DJ . Regulatory mechanisms in stem cell biology Cell 1997 88: 287–298
Birren SJ, Anderson DJ . v-myc immortalized sympathoadrenal progenitor cell line in which neuronal differentiation is initiated by FGF but not NGF Neuron 1990 4: 189–201
Birren SJ, Verdi JM, Anderson DJ . Membrane depolarization induces p140trk and NGF responsiveness, but not p75LNGFR, in MAH cells Science 1992 257: 395–397
Ip NY et al. CNTF, FGF, and NGF collaborate to drive the terminal differentiation of MAH cells into postmitotic neurons Neuron 1994 13: 443–455
Snyder EY . Grafting immortalized neurons to the CNS Curr Op Neurobiol 1994 4: 742–751
Snyder EY, Macklis JD . Multipotent neural progenitor or stem-like cells may be uniquely suited for therapy for some neurodegenerative conditions Clin Neurosci 1996 3: 310–316
Martínez-Serrano A, Björklund A . Gene transfer to the mammalian brain using neural stem cells: a focus on trophic factors, neuroregeneration and cholinerergic neuron systems Clin Neurosci 1996 3: 301–309
Martínez-Serrano A, Björklund A . Immortalized neural progenitor cells for CNS gene transfer and repair Trends Neurosci 1997 20: 530–538
Martínez-Serrano A, Björklund A . Progress towards gene therapy for nervous system diseases. In: Friedmann T (ed) . The Development of Human Gene Therapy Cold Spring Harbor Laboratory Press: Cold Spring Harbor 1999 607–642
Fisher LJ . Neural precursor cells: applications for the study and repair of the central nervous system Neurobiol Dis 1997 4: 1–22
Temple S, Qian X . bFGF, neurotrophins, and the control of cortical neurogenesis Neuron 1995 15: 249–252
Temple S, Qian X . Vertebrate neural progenitor cells: subtypes and regulation Curr Op Neurobiol 1996 6: 11–17
Stemple DL, Mahanthappa NK . Neural stem cells are blasting off Neuron 1997 18: 1–4
McKay R . Stem cells in the central nervous system Science 1997 276: 66–71
Martínez-Serrano A, Hantzopoulos P, Björklund A . Ex vivo gene transfer of brain-derived neurotrophic factor to the intact rat forebrain: neurotrophic effects on cholinergic neurons Eur J Neurosci 1996 8: 727–735
Renfranz PJ, Cunningham MG, McKay RDG . Region-specific differentiation of the hippocampal stem cell line HiB5 upon implantation into the developing mammalian brain Cell 1991 66: 713–729
Ryder EF, Snyder EY, Cepko CL . Establishment and characterization of multipotent neural cell lines using retrovirus vector-mediated oncogene transfer J Neurobiol 1990 21: 356–375
Snyder EY et al. Multipotent neural cell lines can engraft and participate in development of mouse cerebellum Cell 1992 68: 33–51
Snyder EY, Taylor RM, Wolfe JH . Neural progenitor cell engraftment corrects lysosomal storage throughout the MPS VII mouse brain Nature 1995 374: 367–370
Snyder EY, Yoon CH, Flax JD, Macklis JD . Multipotent neural precursors can differentiate toward replacement of neurons undergoing targeted apoptotic degeneration in adult mouse neocortex Proc Natl Acad Sci USA 1997 94: 11645–11650
Martínez-Serrano A, Snyder EY . Neural stem cell lines for brain repair. In: Tuszynsky M (ed) . CNS Regeneration Academic Press: San Diego 1999 203–250
Ip NY, Li Y, Yancopoulos GD, Lindsay RM . Cultured hippocampal neurons show responses to BDNF, NT-3 and NT-4, but not NGF J Neurosci 1993 13: 3394–3405
Vicario-Abejón C et al. Functions of basic fibroblast growth factor and neurotrophins in the differentiation of hippocampal neurons Neuron 1995 151: 105–114
Snider WD . Functions of the neruotrophins during nervous system develoment: what the knockouts are teaching us Cell 1994 77: 627–638
Henderson CE . Role of neurotrophic factors in neuronal development Curr Op Neurobiol 1996 6: 64–70
Jungbluth S, Koentges G, Lumsden A . Coordination of early neural tube development by BDNF/trkB Development 1997 124: 1877–1885
Frade JM et al. Control of early cell death by BDNF in the chick retina Development 1997 124: 3313–3320
Ghosh A, Carnahan J, Greenberg ME . Requirement for BDNF in activity-dependent survival of cortical neurons Science 1994 263: 1618–1623
Acheson A et al. A BDNF autocrine loop in adult sensory neurons prevents cell death Nature 1995 374: 450–453
Eaton MJ, Whittemore SR . Autocrine BDNF secretion enhances the survival and serotonergic differentiation of raphe neuronal precursor cells grafted into the adult rat CNS Exp Neurol 140: 105–114
Sah DWY, Ray J, Gage FH . Bipotent progenitor cell lines from the human CNS Nat Biotech 1997 15: 574–580
Schwartz PM et al. Abnormal cerebellar development and foliation in BDNF −/− mice reveals a role for neurotrophins in CNS patterning Neuron 1997 19: 269–281
Alcántara S et al. TrkB signalling is required for postnatal survival of CNS neurons and protects hippocampal and motor neurons from axotomy-induced cell death J Neurosci 1997 17: 3623–3633
Johe KK et al. Single factors direct the differentiation of stem cells from the fetal and adult central nervous system Genes Dev 1996 10: 3129–3140
Lowenstein DH, Arsenault L . The effects of growth factors on the survival and differentiation of cultured dentate gyrus neurons J Neurosci 1996 16: 1759–1769
Kirschenbaum B, Goldman SA . Brain-derived neurotrophic factor promotes the survival of neurons arising from the adult rat forebrain subependymal zone Proc Natl Acad Sci USA 1995 92: 210–214
Schinstine M, Iacovitti L . 5-azacytidine and BDNF enhance the maturation of neurons derived from EGF-generated neural stem cells Exp Neurol 1997 144: 315–325
Lachyankar MB et al. Embryonic precursor cells that express trk receptors: induction of different cell fates by NGF, BDNF, NT-3 and CNTF Exp Neurol 1997 144: 350–360
Ahmed S, Reynolds BA, Weiss S . BDNF enhances the differentiation but not the survival of CNS stem-cell derived neuronal precursors J Neurosci 1995 15: 5765–5778
Hoshimaru M, Ray J, Sah DW, Gage FH . Differentiation of the immortalized adult neuronal progenitor cell line HC2S2 into neurons by regulatable suppression of the v-myc oncogene Proc Natl Acad Sci USA 1996 93: 1518–1523
Falkenberg T et al. Transgenic mice overexpressing BDNF in neural stem cells Soc Neurosci Abstracts 1994 20: 451.6
Ringstedt T et al. BDNF regulates Reelin expression and Cajal-Retzius cell development in the cerebral cortex Neuron 1998 21: 305–315
Anton R et al. Neural-targeted gene therapy for rodent and primate hemiparkinsonism Exp Neurol 1994 127: 207–218
Martínez-Serrano A, Björklund A . Protection of the neostriatum against excitotoxic damage by neurotrophin-producing, genetically modified neural stem cells J Neurosci 1996 16: 4604–4616
Martínez-Serrano A, Fischer W, Björklund A . Reversal of age-dependent cognitive impairments and cholinergic neuron atrophy by NGF-secreting neural progenitors grafted to the basal forebrain Neuron 1995 15: 473–484
Martínez-Serrano A et al. Long-term functional recovery from age-induced spatial memory impairments by nerve growth factor (NGF) gene transfer to the rat basal forebrain Proc Natl Acad Sci USA 1996 93: 6355–6360
Martínez-Serrano A, Björklund A . Ex vivo nerve growth factor gene transfer to the basal forebrain in presymptomatic middle-aged rats prevents the development of cholinergic neuron atrophy and cognitive impairment during aging Proc Natl Acad Sci USA 1998 95: 1858–1863
Andsberg G et al. Amelioration of ischemia-induced neuronal death in the rat striatum by NGF-secreting neural stem cells Eur J Neurosci 1998 10: 2026–2036
Snyder EY, Fisher LJ . Gene therapy in neurology Curr Op Pediatr 1996 8: 558–568
Lindsay RM, Wiegand SJ, Altar CA, DiStefano PS . Neurotrophic factors: from molecule to man Trends Neurosci 1994 17: 182–190
Lindvall O et al. Neurotrophins and brain insults Trends Neurosci 1994 17: 490–496
Lindvall O, Odin P . Clinical application of cell transplantation and neurotrophic factors in CNS disorders Curr Op Neurobiol 1994 4: 752–757
Yan Q et al. Distribution of intracerebral ventricularly administered neurotrophins in rat brain and its correlation with Trk receptor expression Exp Neurol 1994 127: 23–36
Anderson KD et al. The differential distribution of exogenous BDNF, NGF, and NT-3 in the brain corresponds to the relative abundance and distribution of high-affinity and low-affinity neurotrophin receptors J Comp Neurol 1995 357: 296–317
Gurtu V, Yan G, Zhang G . IRES bicistronic expression vectors for efficient creation of stable mammalian cell lines Biochem Biophys Res Comm 1996 229: 295–298
Friedmann T . Overcoming the obstacles to gene therapy Sci Am 1997 276: 96–101
Craig CG et al. In vivo growth factor expansion of endogenous subependymal neural precursor cell populations in the adult mouse brain J Neurosci 1996 16: 2649–2658
Holmin S, Almqvist P, Lendahl U, Mathiesen T . Adult nestin-expressing subependymal cells differentiate to astrocytes in response to brain injury Eur J Neurosci 1997 9: 65–75
Eriksson PS et al. Neurogenesis in the adult human hippocampus Nature Med 1998 4: 1313–1317
Kuhn HG, Dickinson-Anson H, Gage FH . Neurogenesis in the dentate gyrus of the adult rat: age-related decrease of neuronal progenitor proliferation J Neurosci 1996 16: 2027–2033
Lundberg C et al. Survival, integration and differentiation of neural stem cell lines after transplantation to the adult rat striatum Exp Neurol 1997 145: 342–360
Kokaia Z et al. Focal cerebral ischemia in rats induces expression of p75 neurotrophin receptor in resistant striatal cholinergic neurons Neuroscience 1998 84: 1113–1125
Li XY et al. Enhancement of neurotransmitter release induced by brain-derived neurotrophic factor in cultured hippocampal neurons J Neurosci 1998 18: 10231–10240
Takei N et al. Brain-derived neurotrophic factor induces rapid and transient release of glutamate through the non-exocytotic pathway from cortical neurons J Biol Chem 1998 273: 27620–27624
Rutherford LC, Nelson SB, Turrigiano GG . BDNF has opposite effects on the quantal amplitude of pyramidal neuron and interneuron excitatory synapses Neuron 1998 21: 521–530
Frederiksen K et al. Immortalization of precursor cells from the mammalian CNS Neuron 1988 1: 439–448
Villalba M et al. The role of pyruvate in neuronal calcium homeostasis. Effects on intracellular calcium pools J Biol Chem 1994 269: 2468–2476
Ruiz F et al. Protection by pyruvate and malate against glutamate-mediated neurotoxicity Neuroreport 1998 9: 1277–1282
Sambrook J, Fritsch EF, Maniatis T (eds) . Molecular Cloning: A Laboratory Manual, 2nd edn Cold Spring Harbor Laboratory Press: Cold Spring Harbor 1989
Honma M, Glancey G, Mizusaw H . Identity testing In: Cell and Tissue Culture: A Laboratory Manual Wiley: Chichester 1994 pp 9A5.1–9A5.13.
Nawa H, Carnahan J, Gall C . BDNF protein measured by a novel enzyme immunoassay in normal brain and after seizure: partial disaccordance with mRNA levels Eur J Neurosci 1995 7: 1527–1535
Kokaia Z et al. Regional brain-derived neurotrophic factor mRNA and protein levels following transient forebrain ischemia in the rat Mol Brain Res 1996 38: 139–144
Elmér E et al. Dynamic changes of BDNF protein levels in the rat forebrain after single and recurring kindling-induced seizures Neuroscience 1998 83: 351–362
Brewer GJ, Cotman CW . Survival and growth of hippocampal neurons in defined medium at low density: advantages of a sandwich culture technique or low oxygen Brain Res 1989 494: 65–74
Paxinos G, Watson C . The Rat Brain in Stereotaxic Coordinates Academic Press: New York 1986
Acknowledgements
We wish to thank the excellent technical assistance of B Sesé. This work was supported by grants from Comisión Interministerial de Ciencia y Tecnología and Boehringer-Ingelheim SA (JS), NeuroSearch (AMS), Swedish Medical Research Council, Segerfalk, Kock and Wiberg Foundations (OL), National Institutes of Health (NS34247) (EYS). The institutional grant from Ramón Areces Foundation to the CBMSO is also gratefully acknowledged.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Rubio, F., Kokaia, Z., del Arco, A. et al. BDNF gene transfer to the mammalian brain using CNS-derived neural precursors. Gene Ther 6, 1851–1866 (1999). https://doi.org/10.1038/sj.gt.3301028
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/sj.gt.3301028
Keywords
This article is cited by
-
Current Challenges for the Advancement of Neural Stem Cell Biology and Transplantation Research
Stem Cell Reviews and Reports (2012)
-
BDNF regulation under GFAP promoter provides engineered astrocytes as a new approach for long-term protection in Huntington's disease
Gene Therapy (2010)
-
Neural stem cells display an inherent mechanism for rescuing dysfunctional neurons
Nature Biotechnology (2002)
-
Global gene and cell replacement strategies via stem cells
Gene Therapy (2002)