Functional and anatomical reconstruction of the 6-hydroxydopamine lesioned nigrostriatal system of the adult rat

doi:10.1016/0306-4522(95)00509-9

Neuroscience

Volume 71, Issue 4, April 1996, Pages 913-925

https://doi.org/10.1016/0306-4522(95)00509-9 Get rights and content

Abstract

In an attempt to reconstruct the 6-hydroxydopamine lesioned nigrostriatal system of the adult rat we have combined homotopic grafting of embryonic ventral mesencephalon suspensions with the implantation of long oblique “bridge” grafts of fibroblast growth factor-4-transfected RN-22 schwannoma cells stretching from the site of the neuronal grafts to the striatum. At seven weeks after receiving both grafts, animals were killed and processed for immunohistochemistry against tyrosine hydroxylase. Tyrosine hydroxylase-immunoreactive axons were seen to extend from the nigral grafts, along the bridge graft to the striatum where terminal arborizations could be seen. The retrograde tracer Fluoro-gold was injected intrastriatally in some of the experimental animals and was taken up by grafted neurons confirming their projection to the striatum. In parallel to the anatomical reconstruction of the system, a decrease in amphetamine-induced rotation was demonstrated in those animals receiving both grafts which had received > 98% complete lesions. This decrease was greatest in those animals with the most tyrosine hydroxylase-immunoreactive axons in their bridge grafts. The presence of the bridge graft also led to an increase in neuronal graft survival with twice as many tyrosine hydroxylase-immunoreactive neurons being found in the grafts of those animals that had received both grafts compared to those that had received a neuronal graft but no bridge graft.

Reference (72)

AgidY.
Parkinson's disease: pathophysiology
Lancet
(1991)
AhlskogJ.E.
Cerebral transplantation for Parkinson's disease: current progress and future prospects
BjörklundA. et al.
Reconstruction of the nigrostriatal pathway by intracerebral nigral transplants
Brain Res.
(1979)
BrundinP. et al.
Monitoring of cell viability in suspensions of embryonic CNS tissue and its use as a criterion for intracerebral graft survival
Brain Res.
(1985)
CarmanL.S. et al.
Partial lesion of the substantia nigra: relation between extent of lesion and rotational behaviour
Brain Res.
(1991)
ChomczynskiP. et al.
Single-step method of RNA isolation by acid guanidium thiocyanate-phenol-chloroform extraction
Analyt. Biochem.
(1987)
CossuM. et al.
Axonal elongation into peripheral nerve grafts between thalamus and somatosensory cortex of the rat. An experimental model
Brain Res.
(1987)
DoucetG. et al.
Host afferents into intrastriatal transplants of fetal ventral mesencephalon
Expl Neurol.
(1989)
EwingS.E. et al.
Recovery in hemiparkinsonian rats following intrastriatal implantation of activated leukocytes
Brain Res.
(1992)
FawcettJ.W. et al.
The growth of axons in three-dimensional astrocyte cultures
Devl Biol.
(1989)

HeftiF. et al.

Circling behaviour in rats with partial, unilateral nigro-striatal lesions: effect of amphetamine, apomorphine, and DOPA

Pharmac. Biochem. Behav.

(1980)

HudsonJ.L. et al.

Correlation of apomorphine- and amphetamine-induced turning with nigrostriatal dopamine content in unilateral 6-hydroxydopamine lesioned rats

Brain Res.

(1993)

JerussiT.P. et al.

Amphetamine-induced rotation in rats without lesions

Neuropharmacology

(1974)

KromerL.F. et al.

Regeneration of the septohippocampal pathways in adult rats is promoted by utilizing embryonic hippocampal implants as bridges

Brain Res.

(1981)

LindvallO.

Prospects of transplantation in human neurodegenerative diseases

Trends Neurosci.

(1991)

LoughlinS.E. et al.

Mesostriatal projections from ventral tegmentum and dorsal raphe: cells project ipsilaterally or contralaterally but not bilaterally

Neurosci. Lett.

(1982)

MayerE. et al.

Basic fibroblast growth factor promotes the survival of embryonic ventral mesencephalic dopaminergic neurons—II. Effects on nigral transplants in vivo

Neuroscience

(1993)

PritzelM. et al.

Behavioural and neuronal reorganisation after unilateral substantia nigra lesions: Evidence for increased interhemispheric nigrostriatal projections

Neuroscience

(1983)

ReisD.J. et al.

Reversible changes in the activities and amounts of tyrosine hydroxylase in dopamine neurons of the substantia nigra in response to axonal injury as studied by immunochemical and immunocytochemical methods

Brain Res.

(1978)

RobertsonH.A.

Dopamine receptor interactions: some implications for the treatment of Parkinson's disease

Trends Neurosci.

(1992)

SoK.-F. et al.

Lengthy regrowth of cut axons from retinal ganglion cells after peripheral nerve transplantation into the retina of adult rats

Brain Res.

(1985)

SteinbuschH.W.M. et al.

Basic fibroblast growth factor enhances survival and sprouting of fetal dopaminergic cells implanted in the denervated rat caudate-putamen: preliminary observations

Prog. Brain Res.

(1990)

UngerstedtU. et al.

Quantitative recording of rotational behaviour in rats after 6-hydroxydopamine lesions of the nigrostriatal dopamine system

Brain Res.

(1970)

UnsickerK. et al.

Comparison of the effects of laminin and the polyornithine-binding neurite promoting factor from RN22 schwannoma cells on neurite regeneration from cultured newborn and adult rat dorsal root ganglion neurons

Devl Brain Res.

(1985)

ZhouF.C.

Connectivities of the striatal grafts and laminin guiding

Prog. Brain Res.

(1990)

ZhouF.C. et al.

Excitochemical-induced trophic bridging directs axonal growth of transplanted neurons to distant target

Cell Transplant.

(1995)

AbercrombieM.

Estimation of nuclear population from microtome sections

Anat. Rec.

(1946)

BjörklundA. et al.

Dopamine containing systems in the CNS

BjörklundA. et al.

Intracerebral grafting of neuronal suspensions I. Introduction and general methods of preparation

Acta physiol. scand.

(1983)

BjörklundA. et al.

Intracerebral grafting of neuronal cell suspensions II. Survival and growth of nigral cell suspensions implanted in different brain sites

Acta physiol. scand.

(1983)

BohnM.C. et al.

Adrenal medulla grafts enhance recovery of striatal dopaminergic fibres

Science

(1987)

BungeR.P.

The role of the Schwann cell in trophic support and regeneration

J. Neurol.

(1994)

ChenA. et al.

Methylprednisolone administration improves axonal regeneration into Schwann cell (SC) grafts in thoracic rat spinal cord

Soc. Neurosci. Abstr.

(1994)

ClarkeD.J. et al.

Human fetal dopamine neurons grafted in a rat model of Parkinson's disease: ultrastructural evidence for synapse formation using tyrosine hydroxylase immunocytochemistry

Expl Brain Res.

(1988)

DavidS. et al.

Axonal elongation into peripheral nervous system “bridges” after central nervous system injury in adult rats

Science

(1981)

DavisG.E. et al.

Human amnion membrane serves as a substratum for growing axons in vitro and in vivo

Science

(1987)

Cited by (48)

Mechanisms and use of neural transplants for brain repair
2017, Progress in Brain Research
Citation Excerpt :
A far more effective cell source for the bridges, as predicted by the studies in spinal cord outlined earlier, was the use of cultured Schwann cells, in particular when modified to express specific trophic factors on which DA neuron survival is dependent, FGF and GDNF. These engineered Schwann cell bridges promoted extensive outgrowth of TH-positive axons from dopamine neurons within the intranigral VM grafts over 10–15 mm to the 6-OHDA-lesioned striatum (Brecknell et al., 1996b; Wilby et al., 1999). In a related study, in which the lesions were made not by the toxin 6-OHDA (which kills the nigral neurons), but by a knife cut lesion (which transects the axons leaving the nigral neurons themselves largely intact) the oblique Schwann cell bridge was seen to stimulate axon regeneration of the endogenous axotomized host nigral cells to enable a more extensive reinnervation of the denervated striatum (Brecknell et al., 1996a).
Under appropriate conditions, neural tissues transplanted into the adult mammalian brain can survive, integrate, and function so as to influence the behavior of the host, opening the prospect of repairing neuronal damage, and alleviating symptoms associated with neuronal injury or neurodegenerative disease. Alternative mechanisms of action have been postulated: nonspecific effects of surgery; neurotrophic and neuroprotective influences on disease progression and host plasticity; diffuse or locally regulated pharmacological delivery of deficient neurochemicals, neurotransmitters, or neurohormones; restitution of the neuronal and glial environment necessary for proper host neuronal support and processing; promoting local and long-distance host and graft axon growth; formation of reciprocal connections and reconstruction of local circuits within the host brain; and up to full integration and reconstruction of fully functional host neuronal networks. Analysis of neural transplants in a broad range of anatomical systems and disease models, on simple and complex classes of behavioral function and information processing, have indicated that all of these alternative mechanisms are likely to contribute in different circumstances. Thus, there is not a single or typical mode of graft function; rather grafts can and do function in multiple ways, specific to each particular context. Consequently, to develop an effective cell-based therapy, multiple dimensions must be considered: the target disease pathogenesis; the neurodegenerative basis of each type of physiological dysfunction or behavioral symptom; the nature of the repair required to alleviate or remediate the functional impairments of particular clinical relevance; and identification of a suitable cell source or delivery system, along with the site and method of implantation, that can achieve the sought for repair and recovery.
Rebuilding the nigrostriatal dopamine pathway: 30 years and counting
2014, Experimental Neurology
Survival, differentiation, and connectivity of ventral mesencephalic dopamine neurons following transplantation
2012, Progress in Brain Research
Citation Excerpt :
This appeared to be the result of a restrictive host environment rather than a limitation of the growth capacity of the grafted neurons. Experiments using so-called bridge grafts using pieces of peripheral nerve or Schwann cells to provide a permissive growth substrate between the midbrain and striatum showed that the grafted DA neurons had the intrinsic potential to extend axons from the midbrain in order to innervate the host striatum (Aguayo et al., 1984; Brecknell et al., 1996; Gage et al., 1985; Wilby et al., 1999). Xenografting studies also showed that DA neurons from human or porcine VM placed into the midbrain of 6-OHDA-lesioned rats could innervate appropriate forebrain targets, including the striatum (Isacson et al., 1995; Wictorin et al., 1992) leading to the hypothesis that this was the result of a failure of outgrowing axons to recognize species-specific growth-inhibitory cues.
The reconstruction of midbrain dopamine (DA) circuitry through intracerebral transplantation of new DA neurons contained in embryonic ventral mesencephalon (VM) is a promising therapeutic approach for Parkinson's disease (PD). Although some of the early open-label trials have provided proof-of-principal that VM grafts can provide sustained improvement of motor function in some patients, subsequent trials showed that the functional response can be highly variable. This chapter reviews an extensive body of basic and clinical research on the survival, differentiation, and connectivity of DA neurons in VM grafts, and also looks at how these parameters are affected by certain host- and donor-specific variables. We also review how technical advances in the tools available to study the integration of grafted DA neurons, such as transgenic reporter mice, have made significant contributions to our understanding of the capacity of different DA neuronal subtypes for target-directed growth and innervation of appropriate host brain structures. Our established and on-going understanding of the capacity of grafted DA neurons to structurally and functionally integrate following transplantation forms an important basis for the refinement and optimization of VM grafting procedures, and also the development of new procedures based on the use of stem cells.
Nigral grafts in animal models of Parkinson's disease. Is recovery beyond motor function possible?
2012, Progress in Brain Research
Citation Excerpt :
The belief that fetal nigral tissues positioned in the adult midbrain did not retain a capacity to reinnervate remote targets in the striatum (Björklund et al., 1983) then requires specific cotransplantation strategies to bridge the distance between nigra and striatum with a cell or tissue that can both stimulate and direct long-distance axon growth to the appropriate target (Aguayo et al., 1984; Dunnett et al., 1989; Zhou et al., 1996). Although such bridges have been developed sufficient to restore significant axon growth from nigra to striatum and sufficient to yield recovery in simple and sensitive behavioral measures, such as rotation (Brecknell et al., 1996; Wilby et al., 1999), the density of projections required to alleviate deficits in skilled reaching tests has never been achieved. Moreover, whereas the specific difficulties of demonstrating recovery in skilled reaching from ectopic graft placements have been replicated many times (Abrous et al., 1993a,b; Cenci et al., 1994; Döbrössy et al., 2000; Dunnett et al., 1987; Montoya et al., 1990; Olsson et al., 1995; Torres et al., 2008), several other studies have suggested that combined nigral and striatal placements of nigral grafts or particular training and testing parameters can reveal a limited degree of recovery on the staircase test of skilled reaching (Cordeiro et al., 2010; Nikkhah et al., 1995a,b).
Parkinson's disease (PD) has long been considered predominantly to be a “movement disorder,” and it is only relatively recently that nonmotor symptoms of PD have been recognized to be a major concern to patients. Consequently, there has been surprisingly little investigation into the feasibility of utilizing cell replacement therapies to ameliorate any of the nonmotor dysfunctions of PD. In this chapter, we identify nonmotor impairments associated predominately with dopaminergic dysmodulation, evaluate the few emerging studies that have identified a role for dopamine and nigral transplantation in nonmotor performance, and consider a number of outstanding questions and considerations dominating the field of nigral transplantation today. Preliminary results obtained from rodent models of PD, despite being limited in number, give clear indications of graft effects on striatal processing beyond the simple activation of motor output and promise a major, exciting, and fruitful new avenue of research for the next decade. We can now consider the prospect of rewriting the opportunities for treating patients, with new stem cell sources to be complemented by new targets for therapeutic benefit.
Olfactory enseathing glia enhances reentry of axons into the brain from peripheral nerve grafts bridging the substantia nigra with the striatum
2011, Neuroscience Letters
Reconstruction of lost axonal pathways in the central nervous system (CNS) is possible with the use of peripheral nerve grafts (PNG). However, these permit the entry of axons, while their reentry back into the CNS is compromised. Olfactory enseathing glia (OEG) may permit this reentry of axons if cografted with PNG. We compared the number of tyrosine-hydroxylase positive (TH+) fibers reinnervating PNGs and crossing the graft–striatum interface in PNG placed between the substantia nigra and the striatum in rats receiving both PNG and OEGs and animals receiving PNG only. More TH fibers were seen inside the grafts when OEG was cografted. Although the number of fibers decreases along the graft's length, this effect is less severe when OEG is present. TH+ fibers are seen crossing the PNG–striatum interface in the OEG group. This is correlated with a higher synaptic density at the striatum near the graft when OEG is co-grafted. While these results must be replicated in animal models of Parkinsonism, their implications may apply both to the treatment of Parkinson's disease and to other pathologies, such as spinal cord lesions, where regeneration of long axonal pathways is necessary.
Transgenic reporter mice as tools for studies of transplantability and connectivity of dopamine neuron precursors in fetal tissue grafts
2009, Progress in Brain Research
Citation Excerpt :
Earlier studies using so-called “bridge grafts” have demonstrated that mDA neurons grafted into the midbrain at least bare the intrinsic capacity for axonal extension over the long distances required to reach forebrain targets. In these experiments, growth-permissive substrates, such as peripheral nerve (Aguayo et al., 1984; Gage et al., 1985) or Schwann cells (Brecknell et al., 1996; Wilby et al., 1999), placed between the striatum and the intra-nigral grafts allow for the extension of TH-positive axons along the substrate which can then innervate the striatum. Other encouraging findings come from xenografting studies, which show that when pig or human VM tissue is grafted into the midbrain of 6-OHDA-lesioned adult rats the dopamine neurons can extend axons along the nigro-striatal pathway in order to innervate appropriate forebrain targets, including the striatum (Wictorin et al., 1992; Isacson et al., 1995).
Cell therapy for Parkinson's disease (PD) is based on the idea that new midbrain dopamine (mDA) neurons, implanted directly into the brain of the patient, can structurally and functionally replace those lost to the disease. Clinical trials have provided proof-of-principle that the grafted mDA neurons can survive and function after implantation in order to provide sustained improvement in motor function for some patients. Nonetheless, there are a number of issues limiting the application of this approach as mainstream therapy, including: the use of human fetal tissue as the only safe and reliable source of transplantable mDA neurons, and variability in the therapeutic outcome. Here we review recent progress in this area from investigations using rodent models of PD, paying particular attention to the use of transgenic reporter mice as tools for neural transplantation studies. Cell type-specific expression of reporter genes, such as green fluorescent protein, affords valuable technical advantages in transplantation experiments, such as the ability to selectively isolate specific cell fractions from mixed populations prior to grafting, and the unambiguous visualization of graft-derived dopamine neuron fiber patterns after transplantation. The results from these investigations have given new insights into the transplantability of mDA precursors as well as their connectivity after grafting and have interesting implications for the development of stem cell based approaches for the treatment of PD.

View all citing articles on Scopus

View full text

Functional and anatomical reconstruction of the 6-hydroxydopamine lesioned nigrostriatal system of the adult rat

Abstract

Lancet

Brain Res.

Brain Res.

Brain Res.

Analyt. Biochem.

Brain Res.

Expl Neurol.

Brain Res.

Devl Biol.

Pharmac. Biochem. Behav.

Brain Res.

Neuropharmacology

Brain Res.

Trends Neurosci.

Neurosci. Lett.

Neuroscience

Neuroscience

Brain Res.

Trends Neurosci.

Brain Res.

Prog. Brain Res.

Brain Res.

Devl Brain Res.

Prog. Brain Res.

Cell Transplant.

Estimation of nuclear population from microtome sections

Anat. Rec.

Dopamine containing systems in the CNS

Intracerebral grafting of neuronal suspensions I. Introduction and general methods of preparation

Acta physiol. scand.

Intracerebral grafting of neuronal cell suspensions II. Survival and growth of nigral cell suspensions implanted in different brain sites

Acta physiol. scand.

Adrenal medulla grafts enhance recovery of striatal dopaminergic fibres

Science

The role of the Schwann cell in trophic support and regeneration

J. Neurol.

Methylprednisolone administration improves axonal regeneration into Schwann cell (SC) grafts in thoracic rat spinal cord

Soc. Neurosci. Abstr.

Human fetal dopamine neurons grafted in a rat model of Parkinson's disease: ultrastructural evidence for synapse formation using tyrosine hydroxylase immunocytochemistry

Expl Brain Res.

Axonal elongation into peripheral nervous system “bridges” after central nervous system injury in adult rats

Science

Human amnion membrane serves as a substratum for growing axons in vitro and in vivo

Science