Topographical distribution of [125I]-glial cell line-derived neurotrophic factor in unlesioned and MPTP-lesioned rhesus monkey brain following a bolus intraventricular injection

doi:10.1016/S0006-8993(97)01495-9

Brain Research

Volume 789, Issue 1, 6 April 1998, Pages 9-22

https://doi.org/10.1016/S0006-8993(97)01495-9 Get rights and content

Abstract

The present study determined the topographical distribution profile for [¹²⁵I]-glial cell line-derived neurotrophic factor in unlesioned and MPTP-lesioned (unilateral intracarotid injection) rhesus monkeys following an intraventricular injection. Autoradiographic analysis showed that following a bolus intraventricular injection, there was widespread distribution of [¹²⁵I]-glial cell line-derived neurotrophic factor throughout the ventricular system (walls of lateral, third, and fourth ventricles and aqueduct), with some accumulation at the lateral ventricle injection site, possibly associated with the ependymal cell layer. In both unlesioned and MPTP-lesioned monkeys, there was labelling of the cerebral cortex, substantia nigra/ventral tegmental area and sequestration of [¹²⁵I]-glial cell line-derived neurotrophic factor adjacent to the hippocampal formation, globus pallidus, ventral to and in the substantia nigra. However, [¹²⁵I]-glial cell line-derived neurotrophic factor did not appear to diffuse readily or accumulate in the caudate–putamen even though there was some penetration away from the ventricular walls. Throughout the brain, there was also substantial non-parenchymal labelling of [¹²⁵I]-glial cell line-derived neurotrophic factor, possibly associated with extracellular matrix components, meninges and vasculature due to the heparin binding properties of glial cell line-derived neurotrophic factor. In addition to the extensive loss of tyrosine hydroxylase immunoreactivity within the substantia nigra, there was also decreased accumulation of [¹²⁵I]-glial cell line-derived neurotrophic factor and reduced glial cell line-derived neurotrophic factor immunoreactivity ipsilateral to the lesion. Microscopic analysis showed that glial cell line-derived neurotrophic factor immunoreactivity was associated with upper cortical layers including a high density of immunoreactivity at the surface of the cortex (meningeal, pial layer, vasculature) and around the ventricular walls (with some cellular labelling and labelling of vasculature). Moderate staining was observed in nigral cells contralateral to the MPTP-lesion, whereas only minimal levels of that glial cell line-derived neurotrophic factor immunoreactivity were detected ipsilateral to the lesion. This study shows that intraventricularly injected glial cell line-derived neurotrophic factor accumulates not only around the ventricular walls, but also in specific brain regions in which sub-populations of cells are more readily accessible than others. The presence of cells labelled with [¹²⁵I] and immunopositive for glial cell line-derived neurotrophic factor in the substantia nigra indicates that these cells are a target for the trophic factor following intraventricular administration. Thus, the behavioral improvement observed in MPTP-lesioned monkeys following an intraventricular injection of glial cell line-derived neurotrophic factor is likely the result of activation of nigral cells.

Introduction

The loss of dopaminergic neurons in the nigrostriatal pathway is characteristic of the degeneration that occurs in Parkinson's disease (PD) [21]. The neurodegeneration of nigrostriatal neurons is a complex process involving multiple pathways resulting in a dopaminergic denervation phenomenon that produces severe clinical symptoms including resting tremor, rigidity, bradykinesia and akinesia in PD [6]. Although several pharmacological agents have been used to treat the disease symptomatology, it does not appear that the majority of pharmacological agents currently available reverse or slow the progression of the disease. Recently, attention has focused on preventing neurodegeneration, perhaps by using neurotrophic factor therapy. Of the known neurotrophic factors, glial cell line-derived neurotrophic factor (GDNF) has been shown to be a potent survival and differentiation factor for dopamine (DA) cells in vitro 18, 19. Studies in rodent and primate models of PD have demonstrated that GDNF also is neurotrophic for DA neurons in vivo 4, 14, 15. In rodents, intranigral administration of GDNF increases nigral tyrosine hydroxylase (TH)-immunoreactivity (IR) and DA levels 7, 10. In rats with 6-hydroxydopamine (6-OHDA)-induced lesions of the nigrostriatal pathway, GDNF pretreatment prevents the loss of TH-IR, and in rats with an established lesion, GDNF increases nigral DA levels and TH activity, and reduces apomorphine-induced rotational behavior 1, 2, 5, 8, 10, 13, 15, 16a. GDNF administration produces a long-lasting increase in nigral TH-IR and DA levels in both unlesioned and methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-lesioned primates and in addition, attenuates motor function deficits in the lesioned animals 4, 10, 22. Therefore, GDNF's overall neurotrophic properties for nigrostriatal DA neurons make it a potential candidate as a therapeutic agent for PD 10, 11, 12, 13, 15.

Preclinical studies have recently concentrated on determining whether intraparenchymal or intraventricular (i.c.v.) administration of GDNF improves the behavioral deficits evident in a well-characterized primate model of PD [4]. Although i.c.v. administration of GDNF affects substantia nigra (SN) DA neurons, it is not clear whether these effects are directly related to accumulation of the trophic factor in the caudate–putamen or SN of lesioned primates. Therefore, the main objective of the present study was to determine whether the primate nigrostriatal pathway is a target for i.c.v. administered GDNF. For this, the topographical distribution of [¹²⁵I]-GDNF in the CNS of unlesioned and MPTP-lesioned rhesus monkeys following an i.c.v. injection was assessed and compared to the localization of TH-IR. In addition, in brain regions that accumulated [¹²⁵I]-GDNF, immunocytochemistry with a monoclonal antibody directed against GDNF was done to determine the cellular localization of GDNF.

Section snippets

Animals

The six adult female rhesus monkeys (Macaca mulatta) ranging from 7–20 years of age used in this study were obtained from commercial suppliers (TSI Mason Laboratories, Worcester, MA and HRP, Alice, TX).

GDNF

GDNF was produced at Amgen (Thousand Oaks, CA) and was custom-iodinated using the [¹²⁵I]Bolton Hunter process by Amersham (Buckinghamshire, England). Using controlled trypsin cleavage of [¹²⁵I]-GDNF, the label was shown to be distributed throughout the protein. Moreover, [¹²⁵I]-GDNF retained full

Unlesioned monkeys

The topographical distribution of [¹²⁵I]-GDNF labelling in unlesioned monkey brains 48 and 72 h following an i.c.v. injection is presented in Fig. 1Fig. 2, respectively. Since the monkeys were perfused prior to any tissue processing, the distribution of [¹²⁵I]-GDNF represents `tissue-bound GDNF'. Levels of labelling in the various brain structures are characterized as light to heavy and correlate with a grey scale on autoradiographic film. Thus, very light grey corresponds to little

Discussion

The present study characterized the topographical distribution of [¹²⁵I]-GDNF in the normal unlesioned and MPTP-lesioned primate brain following a single i.c.v. injection. At the 24-, 48- and 72-h time points following [¹²⁵I]-GDNF administration, label was distributed throughout the ventricular system and adjacent structures. There was substantial labelling of the ependymal cell layer, vasculature and limited parenchymal labelling throughout the brain. In addition, accumulation of [¹²⁵I]-GDNF

Conclusion

Overall, this study shows that i.c.v. delivery of GDNF distributes bilaterally throughout the primate brain and accumulates in the SN. Thus, access to the SN by the i.c.v. route of administration results in bilateral stimulation of nigrostriatal dopaminergic tone and may be considered as a potentially viable therapy for PD.

Acknowledgements

We thank P. Miller for assisting with photograph panel production.

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Research reportTopographical distribution of [125I]-glial cell line-derived neurotrophic factor in unlesioned and MPTP-lesioned rhesus monkey brain following a bolus intraventricular injection

Abstract

Introduction

Section snippets

Animals

GDNF

Unlesioned monkeys

Discussion

Conclusion

Acknowledgements

Neurosci. Lett.

Brain Res. Bull.

Brain Res.

Neuroscience

Neurodegeneration

Exp. Neurol.

Neuroscience

Exp. Neurol.

Brain Res.

Neuroscience

Eur. J. Pharmacol.

6-Hydroxydopamine induced the loss of the dopaminergic phenotype in substantia nigra neurons in the rat—a possible mechanism for restoration of the nigrostriatal circuit mediated by glial cell line-derived neurotrophic factor

Exp. Brain Res.

Research report
Topographical distribution of [¹²⁵I]-glial cell line-derived neurotrophic factor in unlesioned and MPTP-lesioned rhesus monkey brain following a bolus intraventricular injection