Skip to main content
SpringerLink
Log in

Motor function, graft survival and gliosis in rats with 6-OHDA lesions and foetal ventral mesencephalic grafts chronically treated with L-DOPA and carbidopa

  • Regular Papers
  • Published:
Experimental Brain Research Aims and scope Submit manuscript

Summary

In rats with a unilateral 6-OHDA lesion of the nigrostriatal pathway, foetal ventral mesencephalic grafts implanted into the 6-OHDA-lesioned striatum produced a reduction in apomorphine-induced contralateral rotation, and complete abolition of (+)-amphetamine-induced ipsilateral rotation. The graft-induced reduction of apomorphine and (+)-amphetamine-induced rotation was not affected by chronic 27 week administration of L-DOPA and carbidopa to rats receiving foetal grafts.

TH-immunohistochemistry revealed >96% loss of dopamine cells in the substantia nigra ipsilateral to the 6-OHDA lesion in all animals, but cell loss in the ipsilateral ventral tegmental area was more variable (21–46% of the intact side). TH-positive cells in the intact substantia nigra and ventral tegmental area were not affected by chronic treatment with L-DOPA and carbidopa. In the lesioned striatum of rats receiving sham grafts, no TH-positive cells or fibres were seen. In the 6-OHDA-lesioned striatum of animals receiving foetal grafts, many TH-positive cells were seen in the grafts and chronic treatment with L-DOPA and carbidopa did not reduce cell survival.

GFA-P immunohistochemistry revealed that a unilateral 6-OHDA lesion followed by a sham graft was not associated with a reactive gliosis reaction in the striatum at the time of study (38 weeks after lesion surgery and 30 weeks after sham-graft), and treatment of such rats with L-DOPA and carbidopa was also without effect on glia. In contrast there was a marked gliosis in the striatum surrounding foetal grafts which was unaffected by chronic treatment with L-DOPA and carbidopa. The grafts themselves were surrounded by a rim of glial cells, and the glial density within the grafts was higher in animals receiving chronic L-DOPA and carbidopa treatment. However, there was no obvious relationship between the number of TH-positive cells within the grafts, or graft volume, and glial cell density within the grafts.

These results suggest that long-term treatment with L-DOPA and carbidopa does not impair either the behavioural recovery produced by foetal ventral mesencephalic grafts in rats or the long-term survival of grafts as revealed by TH-immunohistochemistry. The presence of a foetal graft is associated with a reactive gliosis in the implanted striatum, which was not altered by long-term treatment with L-DOPA and carbidopa. However such treatment did result in an increase in glial density within the grafts themselves.

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

Similar content being viewed by others

Abbreviations

BBB:

blood brain barrier

CNS:

central nervous system

GFAP:

glial fibrillary acidic protein

6-OHDA:

6-hydroxydopamine

IHC:

immunohistochemistry

L-DOPA:

L-3,4-dihydroxyphenylalanine

PET:

positron emission tomography

SN:

substantia nigra

TH:

tyrosine hydroxylase

VTA:

ventral tegmental area

References

  • Abrous N, Guy J, Vigny A, Calas A, Le Moal M, Herman JP (1988) Development of intracerebral dopaminergic grafts: a combined immunohistochemical and autoradiographic study of its time course and environmental influences. J Comp Neurol 273:26–41

    Article  PubMed  Google Scholar 

  • Azmitia EC, Whitaker PM (1983) Formation of a glial scar following microinjection of fetal neurons into the hippocampus or midbrain of the adult rat: an immunocytochemical study. Neurosci Lett 38:145–150

    Article  PubMed  Google Scholar 

  • Bailey RC, Jackson DM, Bracs PU (1979) Long-term L-DOPA pretreatment of mice: central receptor subsensitivity or supersensitivity? Psychopharmacology 66:55–61

    Article  PubMed  Google Scholar 

  • Bignami A, Dahl D (1976) The astroglial response to stabbing. Immunofluorescence studies with antibodies to astrocyte-specific protein (GFA) in mammalian and submammalian species. Neuropath Appl Neurobiol 2:99–110

    Google Scholar 

  • Björklund A, Dunnett SB, Stenevi U, Lewis ME, Iversen SD (1980) Reinnervation of the denervated striatum by substantia nigra transplants: functional consequences as revealed by pharmacological and sensorimotor testing. Brain Res 199:307–333

    Article  PubMed  Google Scholar 

  • Björklund A, Stenevi U, Schmidt RH, Dunnett SB, Gage FH (1983a) Intracerebral grafting of neuronal suspensions II. Survival and growth of nigral cell suspensions implanted in different brain sites. Acta Phys Scand [Suppl] 522:9–18

    Google Scholar 

  • Björklund A, Stenevi U, Schmidt RH, Dunnett SB, Gage FH (1983b) Intracerebral grafting of neuronal cell suspensions I. Introduction and general methods of preparation. Acta Physiol Scand [Suppl] 522:1–7

    Google Scholar 

  • Blin J, Bonnet A, Agid Y (1988) Does levodopa aggravate Parkinson's disease? Neurology 38:1410–1416

    PubMed  Google Scholar 

  • Blunt SB, Jenner P, Marsden CD (1991a) The effect of L-DOPA and carbidopa treatment on the survival of rat foetal dopamine grafts assessed by tyrosine hydroxylase immunohistochemistry and 3H-mazindol autoradiography. Neuroscience 43:95–110

    Article  PubMed  Google Scholar 

  • Blunt SB, Jenner P, Marsden CD (1991b) The effect of chronic L-DOPA treatment on motor function in 6-OHDA-lesioned rats with ventral mesencephalic grafts. Neuroscience 40:453–464

    Article  PubMed  Google Scholar 

  • Brundin P, Isacson O, Björklund A (1985) Monitoring of cell viability in suspensions of embryonic CNS tissue and its use as a criterion for intracerebral graft survival. Brain Res 331:251–259

    Article  PubMed  Google Scholar 

  • Brundin P, Barbin G, Strecker RE, Isacson O, Prochiantz A, Björklund A (1988) Survival and function of dissociated rat dopamine neurones grafted at different developmental stages or after being cultured in vitro. Dev Brain Res 39:233–243

    Article  Google Scholar 

  • Cavanagh JB (1970) The proliferation of astrocytes around a needle wound in the rat brain. J Anat 106:471–487

    PubMed  Google Scholar 

  • David S, Bouchard C, Tsa Tas O, Giftochristos N (1990) Macrophages can modify the non permissive nature of the adult mammalian central nervous system. Neuron 5:463–469

    Article  PubMed  Google Scholar 

  • Denis-Donini S, Glowinski J, Prochiantz A (1984) Glial heterogeneity may define the 3-dimensional shape of mouse mesencephalic dopaminergic neurones. Nature 307:641–643

    Article  PubMed  Google Scholar 

  • Dunnett SB, Björklund A, Stenevi U, Iversen SD (1981) Behavioural recovery following transplantation of substantia nigra in rats subjected to 6-OHDA lesions of the nigrostriatal pathway. I. Unilateral lesions. Brain Res 215:147–161

    Article  PubMed  Google Scholar 

  • Dunnett SB, Björklund A, Schmidt RH, Stenevi U, Iversen SD (1983) Intracerebral grafting of neuronal suspensions IV. Behavioural recovery in rats with unilateral 6-OHDA lesions following implantation of nigral cell suspensions in different forebrain sites. Acta Physiol Scand [Suppl] 522:29–37

    Google Scholar 

  • Dunnett SB, Hernandez TD, Summerfield A, Jones GH, Arbuthnott G (1988) Graft-derived recovery from 6-OHDA lesions: specificity of ventral mesencephalic graft tissue. Exp Brain Res 71:411–424

    Article  PubMed  Google Scholar 

  • Fishman PS, Nilaver G, Kelly J (1983) Astrogliosis limits the integration of peripheral nerve grafts into the spinal cord. Brain Res 277:175–180

    Article  PubMed  Google Scholar 

  • Freed CR, Breeze RE, Rosenberg NL, Schnek S, Wells T, Barrett JN, Grafton ST, Huang SC, Eidelberg D, Rottenberg DA (1990) Transplantation of human fetal dopamine cells for Parkinson's disease. Arch Neurol 47:505–512

    PubMed  Google Scholar 

  • Gaudin DP, Rioux L, Bedard PJ (1990) Fetal dopamine neuron transplants prevent behavioural supersensitivity induced by repeated administration of L-DOPA in the rat. Brain Res 506:166–168

    Article  PubMed  Google Scholar 

  • Graham DG, Tiffany SM, Bell WR (1978) Autooxidation versus covalent binding of quinones as the mechanism of toxicity of dopamine, 6-OHDA and related compounds towards C1300 neuroblastoma cells in vitro. Mol Pharmacol 14:644–653

    PubMed  Google Scholar 

  • Hatten ME, Liem RKH (1981) Astroglial cells provide a template for the positioning of developing cerebellar neurons in vitro. J Cell Biol 90:622–630

    Article  PubMed  Google Scholar 

  • Hefti F, Melamed E, Bhawan J, Wurtman RJ (1981) Long-term administration of L-DOPA does not damage dopaminergic neurons in the mouse. Neurology 31:1194–1195

    Google Scholar 

  • Hitchcock ER, Clough C, Hughes R, Kenny B (1988) Embryos and Parkinson's disease. Lancet I:1274

    Article  Google Scholar 

  • Hsu SM, Raine L, Fanger H (1981) Use of avidin-biotin-peroxidase complex (ABC) in immunoperoxidase techniques: a comparison between ABC and unlabelled antibody (PAP) procedures. J Histochem Cytochem 29:577–580

    PubMed  Google Scholar 

  • Jackson DM, Jenkins OF, Malor R, Christie MJ, Gregory P (1983) Chronic L-DOPA treatment of rats and mice does not change the sensitivity of post-synaptic dopamine receptors. Arch Pharmacol 324:271–274

    Article  Google Scholar 

  • Jaeger CB (1985) Cytoarchitectonics of substantia nigra grafts: a light and electron microscopic study of immunocytochemically identified dopaminergic neurons and fibrous astrocytes. J Comp Neurol 231:121–135

    Article  PubMed  Google Scholar 

  • Jarvis MF, Wagner GC (1985) Age-dependent effects of MPTP. Neuropharmacology 24:581–583

    Article  PubMed  Google Scholar 

  • Klawans HL, Goetz C, Nauseida P, Weiner W (1977) Levodopa-induced dopamine receptor hypersensitivity. Ann Neurol 2:125–129

    Article  Google Scholar 

  • Konigsmark BW (1970) Methods for the counting of neurons. In: Nauta WJH, Ebbesson SOE (eds) Contemporary research methods in neuroanatomy. Springer, Berlin Heidelberg New York pp 315–340

    Google Scholar 

  • Krum JM, Rosenstein JM (1989) The fine structure of vascularastroglial relations in transplanted fetal neocortex. Exp Neurol 103:203–212

    Article  PubMed  Google Scholar 

  • Levitt P, Pintar JE, Breakfield XO (1982) Immunocytochemical demonstration of monoamine-oxidase B in brain astrocytes and serotonergic neurons. Proc Natl Acad Sci USA 79: 6385–6389

    PubMed  Google Scholar 

  • Lindvall O, Rehncrona S, Brundin P, Gustavii B, Astedt B, Widner H, Lindholm T, Björklund A, Leenders K, Rothwell J, Frackowiak R, Marsden CD, Johnels B, Steg G, Freedman R, Hoffer BJ, Seiger A, Bygdeman M, Stromberg I, Olson L (1989) Human fetal dopamine neurons grafteed into the striatum in two patients with severe Parkinson's disease. Arch Neurol 46: 615–631

    PubMed  Google Scholar 

  • Lindvall O, Brundin P, Widner H (1990) Grafts of fetal dopamine neurons survive and improve motor function in Parkinson's disease. Science 247: 574–577

    PubMed  Google Scholar 

  • Madrazo I (1988) Transplantation of fetal substantia nigra and adrenal medulla to the caudate nucleus in two patients with Parkinson's disease. N Engl J Med 318: 51

    Google Scholar 

  • Nieto-Sampedro M, Saneto RP, de Vellis J, Cotman CW (1985) The control of glial populations in brain: changes in astrocyte mitogenic and morphogenic factors in response to injury. Brain Res 343: 320–328

    Article  PubMed  Google Scholar 

  • Nishi K, Kondon T, Narabayashi H (1989) Difference in recovery of striatal dopamine content, tyrosine hydroxylase activity and total biopterin content after MPTP administration: a comparison of young and older mice. Brain Res 489: 157–162

    Article  PubMed  Google Scholar 

  • O'Callaghan JP, Miller DB, Reinhard JF (1990a) Characterisation of the origins of astrocyte response to injury using the dopaminergic neurotoxicant, MPTP. Brain Res 521: 73–80

    Article  PubMed  Google Scholar 

  • O'Callaghan JP, Miller DB, Reinhard JF (1990b) MPTP-induced damage of striatal dopaminergic fibers attenuates subsequent astrocyte response to MPTP. Neurosci Lett 117: 228–233

    Article  PubMed  Google Scholar 

  • Ogawa M, Araki M, Nagatsu I, Yoshida M (1989) Astroglial cell alteration caused by neurotoxins: immunohistochemical observations with antibodies to glial fibrillary acidic protein, laminin and tyrosine hydroxylase. Exp Neurol 106: 187–196

    Article  PubMed  Google Scholar 

  • Olney JW, Zorumski CF, Stewart GR, Price MT, Wang G, Labruyere J (1990) Excitotoxicity of L-DOPA and 6-OHDA: implications for Parkinson's and Huntington's diseases. Exp Neurol 108: 269–272

    Article  PubMed  Google Scholar 

  • Parenti M, Flauto C, Parati E, Vescoci A, Groppetti (1986) Differential effect of repeated treatment with L-DOPA on dopamine D-1 or D-2 receptors. Neuropharmacology 25: 331–334

    Article  PubMed  Google Scholar 

  • Paxinos G, Watson G (1982) The rat brain in stereotaxic coordinates. Academic Press, New York

    Google Scholar 

  • Pellegrino LJ, Pellegrino AS, Cushhman AJ (1979) A stereotaxic atlas of the rat brain, 2nd edn. Plenum Press, New York

    Google Scholar 

  • Perry TL, Young VW, Masatoshi I, Foulks JG, Wall RA, Godin DV, Clavier RM (1984) Nigrostriatal dopaminergic neurons remain undamaged in rats given high doses of L-DOPA and carbidopa chronically. J Neurochem 43: 990–993

    PubMed  Google Scholar 

  • Quinn N, Parkes D, Janota I, Marsden CD (1986) Preservation of the substantia nigra and locus coeruleus in a patient receiving L-DOPA (2 kg) plus decarboxylase inhibitor over a 4 year period. Movement Disorders 1: 65–68

    Article  PubMed  Google Scholar 

  • Redmond DE, Spencer D, Naftolin F, Leranth C, Robbins RJ, Vollmer TL, Roth RH, Bunney BS, Kim JH (1989) Cryopreserved human fetal neural tissue remains viable four months after transplantation into human caudate nucleus. Soc Neurosci Abstr 15: 123

    Google Scholar 

  • Redmond DE, Leranth C, Spencer DD, Robbins R, Vollmer T, Kim JH and Roth RH (1990) Fetal neural graft survival. Lancet II: 820–822

    Article  Google Scholar 

  • Sahakian BJ, Carlson KA, De Girolami U, Bhawan J (1980) Functional and structural consequences of long-term dietary L-DOPA treatment in mice. Commun Psycopharmacol 4: 169–176

    Google Scholar 

  • Schmidt RH, Björklund A, Stenevi U, Dunnett SB, Gage FH (1983) Intracerebral grafting of neuronal cell suspensions. III. Activity of intrastriatal nigral suspension implants as assessed by measurements of dopamine synthesis and metabolism. Acta Physiol Scand [Suppl] 522: 19–28

    Google Scholar 

  • Shetty AK, Banerjee R, Gopinath G, Tandon PH (1991) Fetal nigral grafts in the anterior eye chamber of adult rats: a long term morphological study. Exp Neurol 111: 106–114

    Article  PubMed  Google Scholar 

  • Solberg Y, Pollack Y, Silverman WF (1989) Expression of tyrosine hydroxylase mRNA in transplanted foetal dopamine neurons. Neurosci Lett 106: 36–42

    Article  PubMed  Google Scholar 

  • Steece-Collier K, Collier TJ, Sladek CD, Sladek JR (1990) Chronic levodopa impairs morphological development of grafted embryonic dopamine neurons. Exp Neurol 110: 201–208

    Article  PubMed  Google Scholar 

  • Sternberger LA (1986) Immunocytochemistry Wiley, New York, pp 90–208

    Google Scholar 

  • Stromberg I, Björklund H, Dahl D, Jonsson G, Sundstrom E, Olson L. (1986) Astrocyte responses to dopaminergic denervations by 6-hydroxydopamine and 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine as evidenced by glial fibrillary acidic protein immunohistochemistry. Brain Res Bull 17: 225–236

    Article  PubMed  Google Scholar 

  • Takada M, Li ZK, Hattori T (1990) Astroglial ablation prevents MPTP-induced nigrostriatal neuronal death. Brain Res 509: 55–61

    Article  PubMed  Google Scholar 

  • Yahr MD, Wolf A, Antunes JL, Miyoshi K, Duffy P (1972) Autopsy findings in parkinsonism following treatment with L-DOPA. Neurology 22 [Suppl]: 56–66

    PubMed  Google Scholar 

  • Zhou FC, Buchwald N, Hull C, Towle A (1989) Neuronal and glial elements of fetal neostriatal grafts in the adult neostriatum. Neuroscience 30: 19–31

    PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Parkinson's Disease Society Experimental Research Laboratories, Pharmacology Group, Biomedical Sciences Division, King's College London, Manresa Road, SW3 6LX, London, UK

    S. B. Blunt & P. Jenner

  2. Department of Clinical Neurology, Institute of Neurology, Queen Square, WC1N 3BG, London, UK

    S. B. Blunt & C. D. Marsden

Authors
  1. S. B. Blunt
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. P. Jenner
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. C. D. Marsden
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Blunt, S.B., Jenner, P. & Marsden, C.D. Motor function, graft survival and gliosis in rats with 6-OHDA lesions and foetal ventral mesencephalic grafts chronically treated with L-DOPA and carbidopa. Exp Brain Res 88, 326–340 (1992). https://doi.org/10.1007/BF02259108

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02259108

Key words

Search

Navigation