Autoradiographic localisation of D1 and D2 dopamine receptors in primordial striatal tissue grafts in rats

doi:10.1016/0304-3940(90)90006-U

Neuroscience Letters

Volume 109, Issue 3, 16 February 1990, Pages 271-276

https://doi.org/10.1016/0304-3940(90)90006-U Get rights and content

Abstract

Using in vitro autoradiography the study examined the binding of the dopamine (DA) D₁ receptor and D₂ receptor radiolabelled ligands, [¹²⁵I]SCH 23982 and [¹²⁵I]sulpiride, respectively, to striatal sections of control rats, those with unilateral ibotenic acid (IA) lesions of the striatum and those with grafts of primordial striatal tissue implanted into the IA-lesioned striatum. In animals with IA-lesions, there was almost total degeneration of striatal neurones and a complete disappearance of both D₁ and D₂ receptors in the lesioned striatum. Within the striatal grafts, dense ‘patches’ of both D₁ and D₂ receptors were observed with densities comparable to those in the intact neostriatum. These data indicate that both D₁ and D₂ receptors are localized on the soma of intrinsic striatal neurones. Moreover, the specific localisation of D₁ and D₂ receptors in the grafts provides the neurochemical basis for the mechanisms by which the host nigrostriatal DA system can regulate the functional capacity of striatal grafts.

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Cited by (29)

Fetal tissue transplants in animal models of Huntington's disease: The effects on damaged neuronal circuitry and behavioral deficits
2000, Progress in Neurobiology
Accumulating evidence indicates that grafts of embryonic neurons achieve the anatomical and functional reconstruction of damaged neuronal circuitry. The restorative capacity of grafted embryonic neural tissue is most illustrated by studies with striatal tissue transplantation in animals with striatal lesions. Striatal neurons implanted into the lesioned striatum receive some of the major striatal afferents such as the nigrostriatal dopaminergic inputs and the gluatmatergic afferents from the neocortex and thalamus. The grafted neurons also send efferents to the primary striatal targets, including the globus pallidus (GP, the rodent homologue of the external segment of the globus pallidus) and the entopeduncular nucleus (EP, the rodent homologue of the internal segment of the globus pallidus). These anatomical connections provide the reversal of the lesion-induced alterations in neuronal activities of primary and secondary striatal targets. Furthermore, intrastriatal striatal grafts improve motor and cognitive deficits seen in animals with striatal lesions. Since the grafts affect motor and cognitive behaviors that are critically dependent on the integrity of neuronal circuits of the basal ganglia, the graft-mediated recovery in these behavioral deficits is most likely attributable to the functional reconstruction of the damaged neuronal circuits. The fact that the extent of the behavioral recovery is positively correlated to the amount of grafted neurons surviving in the striatum encourages this view. Based on the animal studies, embryonic striatal tissue grafting could be a viable strategy to alleviate motor and cognitive disorders seen in patients with Huntington’s disease where massive degeneration of striatal neurons occurs.
Embryonic donor age and dissection influences striatal graft development and functional integration in a rodent model of Huntington's disease
2000, Experimental Neurology
The method of embryonic dissection and the age of the donor material remain areas of controversy in the preparation of striatal tissue for intrastriatal implantation. This study explores the relationship between these two parameters with respect to the morphology, function, and physiological integration of the resultant grafts. Tissue derived from embryos of 14 and 16 days of gestation (CRL 10–11 and 14–15 mm, respectively) was prepared as whole, lateral, and medial ganglionic eminence suspensions (WGE, LGE, and MGE, respectively). The embryonic material was implanted into the excitotoxically lesioned striatum of host rats. Grafts derived from E14 LGE attenuated drug-induced rotational bias whereas grafts from E14 MGE ameliorated contralateral deficits in paw reaching. Six months after grafting retrograde tracing of graft projections to the globus pallidus was performed followed by electrical excitation of cortical afferent fibers. Grafts derived from E14 WGE had the largest volume of striatum-like tissue and more striatal neurons compared to LGE from the same donor age. These results suggest that MGE tissue as well as LGE plays a role in the structural and functional integration of striatal grafts.
The integration and function of striatal grafts
2000, Progress in Brain Research
This chapter discusses striatal transplantation in rodents. Most of the studies investigating the functional activity of striatal transplants have been undertaken in animals with unilateral or bilateral excitotoxic lesions of the striatum. The anatomical, neurochemical, and behavioral sequelae of lesions are at present well characterized, and they provide a straightforward model within which to evaluate the structural and functional repair provided by striatal transplants. The understanding of the connectivity of striatal grafts has been complemented by electron microscopic analysis, in particular by combining degeneration, Golgi, and immunohistochemical methods at the ultrastructural level. Striatal grafts can restore functional deficits in experimental rats in both motor and cognitive domains. Such studies have been informative in developing the present understanding of alternative mechanisms of graft function. In addition, the degree of observed recovery following experimental striatal destruction has provided a major stimulus for considering whether a similar strategy may be of clinical application in human neurological disease, in particular ones involving striatal neurodegeneration, of which the prototypical example is Huntington's disease.
GABA(A) receptor subunit expression in intrastriatal striatal grafts. Comparison between normal developing striatum and developing striatal grafts
1997, Developmental Brain Research
Expression of the α1, α2 and β2/3 GABA_A receptor subunits in maturing cell-suspension striatal grafts and in normal developing striatum was studied by immunocytochemistry. During normal postnatal development, the α1 subunit was present in the striatum only at very low density, while the α2 and β2/3 subunits were present with a patchy distribution, in some patches at high density. Double-staining techniques indicated that DARPP-32 (a marker of striatal projection neurons) was not colocalized with α1, but was present in some β2/3-positive areas and all α2-positive areas. In striatal grafts, α1 immunoreactivity was first detected 2 weeks post-grafting (p.g.), and by 3–10 weeks p.g. the pattern was similar to that observed in mature grafts (1 year p.g.), in which α1-immunopositive patches surrounding DARPP-32-positive (i.e. striatum-like) areas are observed. α2 and β2/3 immunoreactivity was observed within the first week p.g., and by 3–10 weeks p.g. was similar to that observed in mature grafts (i.e. immunoreactivity throughout the graft but with patches of different intensity). During graft maturation there was a marked decline in α2 immunoreactivity in DARPP-32-negative areas, as is observed during normal development of the globus pallidus and ventral pallidum. Interestingly, α1- and β2/3-positive fibers (perhaps mostly dendrites) entered DARPP-32-positive patches from DARPP-32-negative areas. This study indicates that the time course of expression of GABA_A receptor subunits in grafted striatal neurons, closely matches that of morphological maturation of the transplant, that of the development of functional synaptic activity and that of GABA_A receptor subunit immunoreactivity in normal developing striatum. Our results also suggest that there are significant interactions between DARPP-32-positive and DARPP-32-negative areas with respect to the expression of GABA_A receptors, and support the suggestion that miniature `striatopallidal systems' may develop within grafts; such interactions may be important for the functional integration of striatal grafts with the host brain.
The effects of donor stage on the survival and function of embryonic striatal grafts in the adult rat brain. I. Morphological characteristics
1997, Neuroscience
The effects of the stage of donor embryos on the survival of grafts from different neuronal cell types have been well documented. Indeed, this parameter has been shown to be highly important in the survival and function of transplants of various tissues of the CNS. However this question has not been addressed in grafts of embryonic striatal tissue transplanted into animal models of Huntington's disease. In this study, rats which had received a unilateral ibotenic acid lesion in the dorsal striatum received grafts from a standard dissection of embryonic striatal primordium taken from donors of embryonic stage either E14, E16, E17 or E19 days. Three months after transplantation six rats from each group were killed for analysis of graft survival and morphology. The remaining animals in each group were killed between 10 and 14 months after grafting. Graft morphology was detected using a range of markers including: acetylcholinesterase and Cresyl Violet, the 32 000 mol. wt dopamine- and cyclic AMP-regulated phosphoprotein (DARPP-32), tyrosine hydroxylase and striatally-enriched phosphatase. All the grafts from different donor stages survived well at both time-points and Cresyl Violet staining indicated neuronal cell types spread throughout the grafts. The transplants were seen to have a characteristic “patchy” appearance with areas of dense AChE activity and DARPP-32 immunopositivity interspersed with areas of much lighter expression. These areas also co-localized consistently with striatally-enriched phosphatase and tyrosine hydroxylase expression, indicating that they comprised the striatal-like compartment of the graft (the so called P zones, containing cells of the mature striatum), and receiving specific afferent input from the host dopaminergic system.
There was no significant difference in total graft volume, when comparing individual groups at both time-points from grafting. However, when comparing the volume of the P zones, the striatal primordium from the youngest donor stages (E14 and E16) produced grafts with a significantly higher proportion of striatal-like tissue. Therefore, in order to increase the proportion of striatal tissue within these grafts, tissue from younger embryonic donors should be used. This has important implications in the application of this model towards clinical trials in Huntington's disease.
The effects of donor stage on the survival and function of embryonic striatal grafts in the adult rat brain. II. Correlation between positron emission tomography and reaching behaviour
1997, Neuroscience
Grafts of embryonic striatal primordia are able to elicit behavioural recovery in rats which have received an excitotoxic lesion to the striatum, and it is believed that the P zones or striatal-like tissue within the transplants play a crucial role in these functional effects. We performed this study to compare the effects of different donor stage of embryonic tissue on both the morphology (see accompanying paper) and function of striatal transplants. Both the medial and lateral ganglionic eminence was dissected from rat embryos of either 10 mm, 15 mm, 19 mm, or 23 mm crown-rump length, and implanted as a cell suspension into adult rats which had received an ibotenic acid lesion 10 days prior to transplantation. After four months the animals were tested on the “staircase task” of skilled forelimb use. At 10–14 months rats from the groups which had received grafts from 10 mm or 15 mm donor embryos were taken for positron emission tomography scanning in a small diameter postiron emission tomography scanner, using ligands to the dopamine D₁ and D₂ receptors, [¹¹C]SCH 23390 and [¹¹C]raclopride, respectively. A lesion-alone group was also scanned with the same ligands for comparison. Animals which had received transplants from the 10 mm donors showed a significant recovery with their contralateral paw on the “staircase test”. No other groups showed recovery on this task. Similarly, the animals with grafts from the youngest donors showed a significant increase in D₁ and D₂ receptor binding when compared to the lesion-alone group. No increase in signal was observed with either ligand in the group which had received grafts from 15 mm donors. Success in paw reaching showed a strong correlation to both the positron emission tomography signal obtained and the P zone volume of the grafts.
These results suggest that striatal grafts from younger donors (10 mm CRL) give greater behavioural recovery than grafts prepared from older embryos. This recovery is due to both the increased proportion of striatal-like tissue within the grafts and an increase in functional D₁ and D₂ dopamine receptors measured by positron emission tomography, i.e. a more extensive integration of the graft with the host brain.

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Autoradiographic localisation of D1 and D2 dopamine receptors in primordial striatal tissue grafts in rats

Abstract

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Autoradiographic localisation of D₁ and D₂ dopamine receptors in primordial striatal tissue grafts in rats