S. Morgan

MPTP susceptibility in the mouse: behavioral, neurochemical, and histological analysis of gender and strain differences.

To investigate the impact of strain and sex in the 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) animal model of Parkinsons disease, C57BL/6 and BALB/c mice were treated with either systemic MPTP-HCl (4 × 15 mg/kg) or saline and were examined in a number of behavioral tests. Furthermore, neostriatal and ventral striatal monoamine contents were determined, and the numbers of tyrosine hydroxylase-immunostained cells were counted in the substantia nigra and ventral tegmental area. Open-field testing showed that locomotor activity was drastically reduced as an acute effect of MPTP in both strains; however, subsequent recovery to control levels was faster in BALB/c mice than in C57BL/6. Nest building also indicated strain-dependent effects, since it was delayed only in C57BL/6 mice treated with MPTP. The other tests (grip test, pole test, rotarod, elevated plus-maze), although partly sensitive for overall strain or gender differences, turned out not to be useful to compare MPTP effects in these two strains. Neurochemically, MPTP led to more severe neostriatal dopamine depletions in C57BL/6 (−85%) than in BALB/c mice (−58%). Histologically, a loss of tyrosine hydroxylase immunoreactivity (−25%) was observed only in the substantia nigra of C57BL/6 animals. Thus, our analysis consistently showed that the C57BL/6 mouse strain is more susceptible to MPTP than the BALB/c strain. Sex differences in MPTP sensitivity were not observed in our mice. The implications of these findings for the search for genes related to susceptibility to neurodegeneration are discussed.

Interhemispheric nigrostriatal projections in the rat: Bifurcating nigral projections and loci of crossing in the diencephalon

The organization of interhemispheric nigrostriatal projections in the rat was studied with (1) a double labeling technique combined with a histofluorescence method for monoamines and with (2) horseradish-peroxidase (HRP) as a tracer. Each animal received an injection of Fast Blue (FB) into the caudate nucleus (NC) of one hemisphere and an injection of Nuclear Yellow (NY) into the NC of the other hemisphere. Brains were processed according to the sucrose-potassium phosphate-glyoxylic acid (SPG) method in order to identify monoaminergic neurons. About 5% of the neurons which were labeled by either of the tracers in the substantia nigra (SN) ipsilateral to the injection site were also labeled in the contralateral SN. Most of these interhemispherically projecting SN-neurons were monoaminergic. A small number of neurons in either SN was labeled with both fluorescent tracers. This suggests that bifurcating, monoaminergic neurons in the SN project to both NC. In the second experiment, rats received a unilateral injection of HRP into NC. After a survival time of several hours the animals were perfused and the brains were processed using benzidine-dihydrochloride. Fibers crossing the hemisphere were found within the massa intermedia of the thalamus and within the inferior thalamic peduncle. The results are discussed with respect to a possible functional role of interhemispheric nigrostriatal connections.

The basal ganglia-orofacial system: Studies on neurobehavioral plasticity and sensory-motor tuning

We have employed the unilateral removal of the vibrissae as a tool to examine ensuing behavioral changes in relation to concomitant changes in the central nervous system. In this paper we review a series of studies showing that unilateral removal of the vibrissae leads to behavioral asymmetries (e.g., in thigmotactic scanning) from which rats recover over time. Time-related to these behavioral changes we found neuronal alterations in striatal afferents, that is, in uncrossed and crossed projections from the substantia nigra and the tuberomammillary nucleus. The involvement of dopaminergic mechanisms was indicated by results showing that dopaminergic agonists can induce asymmetries in thigmotactic scanning and turning; the direction of these asymmetries was also dependent on time after vibrissae removal. Furthermore, it was shown that endogenous preferential use of one vibrissae side in thigmotactic scanning interacts with the expression of spontaneous and drug-induced behavioral asymmetries exhibited after unilateral vibrissae removal. Neurochemical studies indicated that both unilateral vibrissae removal and unilateral perioral stimulation can have lateralized effects on biogenic amines in the brain. Finally, using electrical stimulation of the substantia nigra, evidence was found for a lateralized and bidirectional interaction between basal ganglia and the orofacial systems, indicating an involvement in mechanisms of motivation and particular stimulation. These results are important from several perspectives. One, they indicate functional links between the orofacial systems and the basal ganglia. Two, they raise the possibility that unilateral removal of the vibrissae can serve as a model (a) to investigate the dynamics of recovery of function after CNS insults, in general, and specifically, (b) to study neuronal plasticity in the nigrostriatal and tuberomammillary-striatal pathways, and (c) to investigate the neuropharmacology of catecholamine systems in the brain.

Apomorphine reverses direction of asymmetry in facial scanning after 10 days of unilateral vibrissae removal in rat vibrissotomy induced denervation supersensitivity

Sensorimotor asymmetries of hemivibrissotomized rats were investigated in an open field. To examine possible behavioral plasticity after unilateral removal of the vibrissae, we compared two groups of rats which had experienced the sensory imbalance for different durations. One group was shaved on one side of the face daily for 10 days before behavioral testing (group 10-Days). The other group was first shaved 4 h prior to the behavioral testing and was sham-shaved for the preceding 10 days (group 4-H). On day 11, the undrugged behavior was examined, and then, one hour later, the behavior induced by apomorphine (0.75 mg/kg, s.c.) was analyzed. In the undrugged state both groups tended to scan the walls of the enclosure with the vibrissae side of the face. After injection of apomorphine, the 4-H group also scanned the walls preferentially with the vibrissae side. In contrast, the 10-Days group reversed the scanning asymmetry under the influence of this drug. That is, animals in this latter group now preferred to scan the walls with the shaved side of the face. These findings are suggestive of a plasticity in dopamine transmission subsequent to unilateral removal of vibrissae. The reversal in direction of asymmetry under apomorphine of the 10-Days group is reminiscent of the apomorphine-induced reversal in sensorimotor asymmetries found after unilateral 6-hydroxydopamine lesion of the substantia nigra, which has been attributed to dopamine receptor supersensitivity.

Dissociation of crossed and uncrossed nigrostriatal projections with respect to site of origin in the rat

In the present study we have investigated the relative rostrocaudal position of the neurons in the substantia nigra, which project to the ipsilateral or contralateral neostriatum. The retrograde tract tracer horseradish peroxidase was implanted into the striatum on one side. The substantia nigra, ventral tegmental area and retrorubral area ipsilateral and contralateral to the site of implantation were examined for labeled cells. The distributions of the cells which give rise to the crossed and uncrossed nigrostriatal projections were found to be inverse. More labeled cells were found in the rostral than the caudal part of the substantia nigra ipsilateral to the site of horseradish peroxidase implantation. In contrast, there was a greater likelihood of finding labeled cells in the middle and caudal parts of the contralateral substantia nigra than in the rostral part. Sparse projections from the ipsilateral and contralateral retrorubral area and ventral tegmental area were also found.

Evidence for a Dissociation between MPTP Toxicity and Tyrosinase Activity Based on Congenic Mouse Strain Susceptibility

The neurotoxicity induced by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) is one of the most valuable available models for investigating critical aspects of human Parkinsons disease. In order to analyze the relevance of pigmentation for MPTP sensitivity, we compared C57Bl/6 wild-type mice with the albino mutant C57Bl/6J-Tyr(c-2J) of the same strain. These animals were treated either with systemic MPTP or with saline and were examined in behavioral tests. Seven days after treatment, the contents of dopamine and other monoamines were determined postmortem in the neostriatum and ventral striatum. Furthermore, the numbers of tyrosine hydroxylase-positive cells were counted in the substantia nigra and ventral tegmental area. Open field testing showed that rearing activity was drastically reduced as an acute effect of MPTP in both wild type and mutants; however, subsequent recovery to control levels was faster in wild-type mice. Nest building also indicated strain-dependent effects, since it was delayed only in mutants treated with MPTP. Neurochemically, MPTP led to severe neostriatal dopamine depletions, which did not differ significantly between wild-type (72.9%) and mutant mice (82.1%). Less severe dopamine depletions were also found in the ventral striatum. Histologically, a loss of tyrosine hydroxylase-labeled cells was observed only in the substantia nigra of both wild-type and mutant mice (13.3 and 21.3%, respectively), but not in the ventral tegmental area. Together, our data do not provide evidence that tyrosinase-deficient mice are less affected by MPTP treatment than the comparable wild type, thus arguing strongly against the hypothesis that enhanced MPTP sensitivity in pigmented mouse strains is caused by tyrosinase activity.

Evidence for resistance to MPTP in C57BL/6 x BALB/c F1 hybrids as compared with their progenitor strains.

The neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) is capable of producing a syndrome in mice which shares major characteristics with human Parkinsons disease. There is evidence for a genetic influence on the degree of damage exerted by MPTP, since different strains of mice can dramatically differ in their response to MPTP. We produced reciprocal F1 hybrids by crossbreeding the MPTP-susceptible C57BL/6 strain with resistant BALB/c. These hybrids were compared to the parental strains using neural and behavioral measures in order to characterize the genetic transmission of MPTP-susceptibility. The F1 generation as a whole had a lower depletion of neostriatal dopamine levels than even found in BALB/c. Furthermore, there was no significant loss of tyrosine hydroxylase-positive cells in the substantia nigra and quick recovery from deficits in motor behavior in F1, herein resembling BALB/c. We suggest that several loci are involved in susceptibility to MPTP, and that the trait is under control of recessive susceptibility and/or dominant resistance alleles, which interact in F1, leading to extremely low susceptibility.

Neuronal Plasticity in the Nigrostriatal System of the Rat after Unilateral Removal of Vibrissae

The vibrissae of rats were shaved from one side of the face daily for 10 days. To see whether or not this treatment had an effect on crossed and uncrossed striatal afferent connections from the midbrain, the tract tracer horseradish peroxidase was applied to the caudate-putamen on day 11. When the tracer was deposited on the side opposite the vibrissae removal, more labeled cells were found in the contralateral substantia nigra than when it was applied on the same side as the vibrissae removal, or placed in animals with intact vibrissae. Unilateral removal of vibrissae did not affect uptake of the tracer by the cells which give rise to the homolateral nigrostriatal projections. These changes in HRP labeling in the crossed projection from the substantia nigra were seen after 10 days of unilateral removal of the vibrissae; i.e., at a time when the animals have had opportunity to learn to compensate for vibrissotomy-induced behavioral asymmetries.

Asymmetries in crossed and uncrossed nigrostriatal projections dependent on duration of unilateral removal of vibrissae in rats

SummaryThe influence of unilateral removal of vibrissae on the crossed and uncrossed nigrostriatal projections was examined with the horseradish peroxidase tract tracing technique. Hemivibrissotomy mainly affected the projections arising from the rostral part of the substantia nigra. One to three days after clipping the vibrissae, rats were found to have more labeled neurons in the crossed projection to the caudate-putamen (CPU) on the same side as vibrissae removal than in the crossed projection to the CPU opposite to vibrissae removal. A reversed asymmetry was seen in rats examined 4–20 days after vibrissae removal. These animals had more labeled cells in the crossed and uncrossed projections terminating in the CPU opposite to the shaved side, i.e. in the hemisphere deprived of vibrissal sensory input. This time-course of neural alterations is similar to that of the recovery from behavioral asymmetries seen after hemivibrissotomy. Similar time-dependent alterations in the nigrostriatal projection had been found after unilateral injection of 6-OHDA into the substantia nigra.

Parallels in Behavioral and Neural Plasticity Induced by Unilateral Vibrissae Removal and Unilateral Lesion of the Substantia Nigra

A prominent facial feature of mammals is the presence of long tactile hairs referred to as “vibrissae” or “whiskers”. In fact, such tactile sense organs are found in almost all mammals except Homo sapiens (Ahl 1986). They are employed in a great variety of behaviors (see Gustafson and Felbain-Keramidas 1977 and Ahl 1986 for reviews of vibrissae function). For example, in a novel environment rats whisk their vibrissae over surfaces and edges in a repetitive manner (Vincent 1912; Welker 1964). This behavior is thought to serve for sampling tactile stimuli (Welker 1964; Zucker and Welker 1969; Shipley 1974). There is evidence that such tactile information from the vibrissae is analyzed in a complex form, which may even allow “shape” or “pattern” perception (Shipley 1974). The importance of the vibrissae system for the rat is also reflected by its large cortical representation. About 20% of the somatosensory cortex has been found to respond to stimulation of the vibrissae region of the face (Welker 1971).