Skip to main content

Advertisement

SpringerLink
Log in

Targeting of neural stem cells in the hippocampus of adult rats by custom-made Ad vectors

  • Original Article
  • Published:
Brain Structure and Function Aims and scope Submit manuscript

Abstract

Adult hippocampal neural stem cells (NSC) are an intriguing source for cell replacement or could serve as delivery vehicles for therapeutic genes. We recently reported selective transduction of adult mouse NSC in the DG by in vivo injection of GFP encoding adenoviral (Ad) vectors engineered to bind NSC-specific peptides. Here, we investigated the specificity of these peptide-tagged vectors in the adult rat DG, and whether they can be used to follow differentiation of infected cells over time. The virus-containing solution was injected into the DG by stereotaxic surgery. Specific transduction of NSC was demonstrated by the radial glia-like morphology of GFP-expressing type-1 cells and co-labeling with nestin or glial fibrillary acidic protein. Three days post-injection more than 82% of GFP-containing cells were nestin-immunoreactive, as revealed by unbiased stereology and no GFP-expressing neurons were observed. However, 30 days after injection, the amount of GFP and nestin-containing cells declined (56%), whereas now neurons that contained NeuN or possessed the typical granular nerve cell morphology expressed GFP, indicating that they were derived from initially transduced NSC. Importantly, still more than 20% of nestin-immunoreactive NSC was found to be GFP-positive 90 days after infection, but unfortunately at this time point no GFP-containing neurons were detectable. Our results demonstrate that Ad vectors tagged with NSC-specific ligands can be used to target type-1 NSC, the low-proliferating cell population, in the rat hippocampus. They are a valuable tool to monitor the differentiation of their descendants, at least over short time periods.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  • Alemany R, Curiel DT (2001) CAR-binding ablation does not change biodistribution and toxicity of adenoviral vectors. Gene Ther 8:1347–1353

    Article  CAS  PubMed  Google Scholar 

  • Altman J, Das GD (1965) Autoradiographic and histological evidence of postnatal hippocampal neurogenesis in rats. J Comp Neurol 124:319–335

    Article  CAS  PubMed  Google Scholar 

  • Alvarez-Buylla A, Seri B, Doetsch F (2002) Identification of neural stem cells in the adult vertebrate brain. Brain Res Bull 57:751–758

    Article  PubMed  Google Scholar 

  • Anthony TE, Klein C, Fishell G, Heintz N (2004) Radial glia serve as neuronal progenitors in all regions of the central nervous system. Neuron 41:881–890

    Article  CAS  PubMed  Google Scholar 

  • Biebl M, Cooper CM, Winkler J, Kuhn HG (2000) Analysis of neurogenesis and programmed cell death reveals a self-renewing capacity in the adult rat brain. Neurosci Lett 291:17–20

    Article  CAS  PubMed  Google Scholar 

  • Chen LW, Wei LC, Qiu Y, Liu HL, Rao ZR, Ju G, Chan YS (2002) Significant up-regulation of nestin protein in the neostriatum of MPTP-treated mice. Are the striatal astrocytes regionally activated after systemic MPTP administration? Brain Res 925:9–17

    Article  CAS  PubMed  Google Scholar 

  • Curtis MA, Penney EB, Pearson AG, Van Roon-Mom WM, Butterworth NJ, Dragunow M, Connor B, Faull RL (2003) Increased cell proliferation and neurogenesis in the adult human Huntington’s disease brain. Proc Natl Acad Sci USA 100:9023–9027

    Article  CAS  PubMed  Google Scholar 

  • Dahlstrand J, Zimmerman LB, Mckay RD, Lendahl U (1992) Characterization of the human nestin gene reveals a close evolutionary relationship to neurofilaments. J Cell Sci 103:589–597

    CAS  PubMed  Google Scholar 

  • Dahlstrand J, Lardelli M, Lendahl U (1995) Nestin mRNA expression correlates with the central nervous system progenitor cell state in many, but not all, regions of developing central nervous system. Brain Res Dev Brain Res 84:109–129

    Article  CAS  PubMed  Google Scholar 

  • Dayer AG, Ford AA, Cleaver KM, Yassaee M, Cameron HA (2003) Short-term and long-term survival of new neurons in the rat dentate gyrus. J Comp Neurol 460:563–572

    Article  PubMed  Google Scholar 

  • Dolbeare F (1995) Bromodeoxyuridine: a diagnostic tool in biology and medicine, part II: oncology, chemotherapy and carcinogenesis. Histochem J 27:923–964

    CAS  PubMed  Google Scholar 

  • Duggal N, Schmidt-Kastner R, Hakim AM (1997) Nestin expression in reactive astrocytes following focal cerebral ischemia in rats. Brain Res 768:1–9

    Article  CAS  PubMed  Google Scholar 

  • Eriksson PS, Perfilieva E, Bjork-Eriksson T, Alborn AM, Nordborg C, Peterson DA, Gage FH (1998) Neurogenesis in the adult human hippocampus. Nat Med 4:1313–1317

    Article  CAS  PubMed  Google Scholar 

  • Ge S, Yang CH, Hsu KS, Ming GL, Song H (2007) A critical period for enhanced synaptic plasticity in newly generated neurons of the adult brain. Neuron 54:559–566

    Article  CAS  PubMed  Google Scholar 

  • Geraerts M, Eggermont K, Hernandez-Acosta P, Garcia-Verdugo JM, Baekelandt V, Debyser Z (2006) Lentiviral vectors mediate efficient and stable gene transfer in adult neural stem cells in vivo. Hum Gene Ther 17:635–650

    Article  CAS  PubMed  Google Scholar 

  • Graham FL, Prevec L (1995) Methods for construction of adenovirus vectors. Mol Biotechnol 3:207–220

    Article  CAS  PubMed  Google Scholar 

  • Gubert F, Zaverucha-Do-Valle C, Pimentel-Coelho PM, Mendez-Otero R, Santiago MF (2009) Radial glia-like cells persist in the adult rat brain. Brain Res 1258:43–52

    Article  CAS  PubMed  Google Scholar 

  • Gundersen HJ, Bagger P, Bendtsen TF, Evans SM, Korbo L, Marcussen N, Moller A, Nielsen K, Nyengaard JR, Pakkenberg B (1988) The new stereological tools: disector, fractionator, nucleator and point sampled intercepts and their use in pathological research and diagnosis. Apmis 96:857–881

    Article  CAS  PubMed  Google Scholar 

  • Kempermann G (2006) Adult neurogenesis: stem cells and neuronal development in the adult brain. Oxford University Press, New York

    Google Scholar 

  • Kempermann G, Kuhn HG, Gage FH (1998) Experience-induced neurogenesis in the senescent dentate gyrus. J Neurosci 18:3206–3212

    CAS  PubMed  Google Scholar 

  • Kempermann G, Jessberger S, Steiner B, Kronenberg G (2004) Milestones of neuronal development in the adult hippocampus. Trends Neurosci 27:447–452

    Article  CAS  PubMed  Google Scholar 

  • Kirik D, Rosenblad C, Björklund A, Mandel RJ (2000) Long-term, rAAV-mediated gene transfer of GDNF in the rat Parkinson’s model: intrastriatal but not intranigral transduction promotes functional regeneration in the lesioned nigrostriatal system. J Neurosci 20:4686–4700

    CAS  PubMed  Google Scholar 

  • Kronenberg G, Reuter K, Steiner B, Brandt MD, Jessberger S, Yamaguchi M, Kempermann G (2003) Subpopulations of proliferating cells of the adult hippocampus respond differently to physiologic neurogenic stimuli. J Comp Neurol 467:455–463

    Article  PubMed  Google Scholar 

  • Malatesta P, Hack MA, Hartfuss E, Kettenmann H, Klinkert W, Kirchhoff F, Gotz M (2003) Neuronal or glial progeny: regional differences in radial glia fate. Neuron 37:751–764

    Article  CAS  PubMed  Google Scholar 

  • Ming GL, Song H (2005) Adult neurogenesis in the mammalian central nervous system. Annu Rev Neurosci 28:223–250

    Article  CAS  PubMed  Google Scholar 

  • O’Riordan CR, Lachapelle A, Delgado C, Parkes V, Wadsworth SC, Smith AE, Francis GE (1999) PEGylation of adenovirus with retention of infectivity and protection from neutralizing antibody in vitro and in vivo. Hum Gene Ther 10:1349–1358

    Article  PubMed  Google Scholar 

  • Overstreet LS, Hentges ST, Bumaschny VF, de Souza FSJ, Smart JL, Santangelo AM, Low MJ, Westbrook GL, Rubinstein M (2004) A transgenic marker for newly born granule cells in dentate gyrus. J Neurosci 23:3251–3259

    Article  Google Scholar 

  • Paxinos G (1986) The rat brain in stereotaxic coordinates, 2nd edn. Academic Press, London

    Google Scholar 

  • Pützer BM, Gnauck J, Kirch HC, Brockmann D, Esche H (1997) A cis-acting element 7 bp upstream of the ESF-1-binding motif is involved in E1A 13S autoregulation of the adenovirus 12 TS2 promoter. J Gen Virol 78:879–891

    PubMed  Google Scholar 

  • Pützer BM, Stiewe T, Parssanedjad K, Rega S, Esche H (2000) E1A is sufficient by itself to induce apoptosis independent of p53 and other adenoviral gene products. Cell Death Differ 7:177–188

    Article  PubMed  Google Scholar 

  • Romanczuk H, Galer CE, Zabner J, Barsomian G, Wadsworth SC, O’Riordan CR (1999) Modification of an adenoviral vector with biologically selected peptides: a novel strategy for gene delivery to cells of choice. Hum Gene Ther 10:2615–2626

    Article  CAS  PubMed  Google Scholar 

  • Schmidt A, Haas SJ, Hildebrandt S, Scheibe J, Eckhoff B, Racek T, Kempermann G, Wree A, Pützer BM (2007) Selective targeting of adenoviral vectors to neural precursor cells in the hippocampus of adult mice: new prospects for in situ gene therapy. Stem Cells 25:2910–2918

    Article  CAS  PubMed  Google Scholar 

  • Schmidt-Hieber C, Jonas P, Bischofberger J (2004) Enhanced synaptic plasticity in newly generated granule cells of the adult hippocampus. Nature 429:184–187

    Article  CAS  PubMed  Google Scholar 

  • Schmitt O, Eggers R (1997) Systematic investigations of the contrast results of histochemical stainings of neurons and glial cells in the human brain by means of image analysis. Micron 28:197–215

    Article  CAS  PubMed  Google Scholar 

  • Seki T, Arai Y (1999) Temporal and spacial relationships between PSA-NCAM-expressing, newly generated granule cells, and radial glia-like cells in the adult dentate gyrus. J Comp Neurol 410:503–513

    Article  CAS  PubMed  Google Scholar 

  • Seri B, Garcia-Verdugo JM, Mcewen BS, Alvarez-Buylla A (2001) Astrocytes give rise to new neurons in the adult mammalian hippocampus. J Neurosci 21:7153–7160

    CAS  PubMed  Google Scholar 

  • Shapiro LA, Korn MJ, Shan Z, Ribak CE (2005) GFAP-expressing radial glia-like cell bodies are involved in a one-to-one relationship with doublecortin-immunolabeled newborn neurons in the adult dentate gyrus. Brain Res 1040:81–91

    Article  CAS  PubMed  Google Scholar 

  • Slomianka L, West MJ (2005) Estimators of the precision of stereological estimates: an example based on the CA1 pyramidal cell layer of rats. Neuroscience 136:757–767

    Article  CAS  PubMed  Google Scholar 

  • Song H, Kempermann G, Overstreet Wadiche L, Zhao C, Schinder AF, Bischofberger J (2005) New neurons in the adult mammalian brain: synaptogenesis and functional integration. J Neurosci 25:10366–10368

    Article  CAS  PubMed  Google Scholar 

  • Steiner B, Klempin F, Wang L, Kott M, Kettenmann H, Kempermann G (2006) Type-2 cells as link between glial and neuronal lineage in adult hippocampal neurogenesis. Glia 54:805–814

    Article  PubMed  Google Scholar 

  • Thomas CE, Abordo-Abesida E, Maleniak TC, Stone D, Gerdes CA, Lowenstein PR (2001) Gene transfer into rat brain using adenoviral vectors. Curr Prot Neurosci Chapter 4:Unit 4.24

  • van Praag H, Kempermann G, Gage FH (2000) Neural consequences of environmental enrichment. Nat Rev Neurosci 1:191–198

    Article  PubMed  Google Scholar 

  • van Praag H, Schinder AF, Christie BR, Toni N, Palmer TD, Gage FH (2002) Functional neurogenesis in the adult hippocampus. Nature 415:1030–1034

    Article  PubMed  Google Scholar 

  • West MJ (1993) New stereological methods for counting neurons. Neurobiol Aging 14:275–285

    Article  CAS  PubMed  Google Scholar 

  • West MJ, Slomianka L, Gundersen HJ (1991) Unbiased stereological estimation of the total number of neurons in the subdivisions of the rat hippocampus using the optical fractionator. Anat Rec 231:482–497

    Article  CAS  PubMed  Google Scholar 

  • Yoo YM, Lee U, Kim YJ (2005) Apoptosis and nestin expression in the cortex and cultured astrocytes following 6-OHDA administration. Neurosci Lett 382:88–92

    Article  CAS  PubMed  Google Scholar 

  • Zhao C, Teng EM, Summers RG Jr, Ming GL, Gage FH (2006) Distinct morphological stages of dentate granule neuron maturation in the adult mouse hippocampus. J Neurosci 26:3–11

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

We thank Frauke Winzer and Susann Lehmann for skillful technical assistance. We are grateful to Ramon Alemany for providing AdTLY477RAE vector. Grant sponsor: Bundesminister für Bildung und Forschung (to B.M.P.); Grant number: 01ZZ0108; Grant sponsor: Deutsche Forschungsgemeinschaft (to B.M.P. and A.W.); Grant numbers: PU188/8-1 and WR20/2-1; Grant sponsor: FORUN program Medical Faculty of Rostock (to A.S. and S.J.-P.H); Grant numbers: 889048 and 889011.

Author information

Authors and Affiliations

  1. Institute of Anatomy, Medical Faculty, University of Rostock, Gertrudenstr. 9, 18057, Rostock, Germany

    Steve Hildebrandt, Oliver Schmitt, Andreas Wree & Stefan J.-P. Haas

  2. Department of Vectorology and Experimental Gene Therapy, Biomedical Research Center, Rostock, Germany

    Anke Schmidt, Anja Stoll & Brigitte M. Pützer

  3. Institute of Physiology, Medical Faculty, University of Rostock, Rostock, Germany

    Rüdiger Köhling

Authors
  1. Steve Hildebrandt
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Anke Schmidt
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Anja Stoll
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Oliver Schmitt
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Rüdiger Köhling
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Andreas Wree
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Stefan J.-P. Haas
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Brigitte M. Pützer
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Stefan J.-P. Haas.

Additional information

S. Hildebrandt and A. Schmidt contributed equally to this work.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Hildebrandt, S., Schmidt, A., Stoll, A. et al. Targeting of neural stem cells in the hippocampus of adult rats by custom-made Ad vectors. Brain Struct Funct 215, 105–113 (2010). https://doi.org/10.1007/s00429-010-0275-8

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00429-010-0275-8

Keywords

Search

Navigation