Arrestins and two receptor kinases are upregulated in Parkinson's disease with dementia.

Arrestins and G proteins-coupled receptor kinases (GRKs) regulate signaling and trafficking of G protein-coupled receptors. We investigated changes in the expression of arrestins and GRKs in the striatum of patients with Parkinson's disease without (PD) or with dementia (PDD) at postmortem using Western blotting and ribonuclease protection assay. Both PD and PDD groups had similar degree of dopamine depletion in all striatal regions. Arrestin proteins and mRNAs were increased in the PDD group throughout striatum. Protein and mRNA of GRK5, the major subtype in the human striatum, and GRK3 were also upregulated, whereas GRK2 and 6 were mostly unchanged. The PD group had lower concentration of arrestins and GRKs than the PDD group. There was no statistical link between the load of Alzheimer's pathology and the expression of these signaling proteins. Upregulation of arrestins and GRK in PDD may confer resistance to the therapeutic effects of levodopa often observed in these patients. In addition, increased arrestin and GRK concentrations may lead to dementia via perturbation of multiple signaling mechanisms.

PIB is a non-specific imaging marker of amyloid-beta (Abeta) peptide-related cerebral amyloidosis.

The in vivo imaging probe [11C]-PIB (Pittsburgh Compound B, N-methyl[11C]2-(4'-methylaminophenyl-6-hydroxybenzathiazole) is under evaluation as a key imaging tool in Alzheimer's disease (AD) and to date has been assumed to bind with high affinity and specificity to the amyloid structures associated with classical plaques (CPs), one of the pathological hallmarks of the disease. However, no studies have systematically investigated PIB binding to human neuropathological brain specimens at the tracer concentrations achieved during in vivo imaging scans. Using a combination of autoradiography and histochemical techniques, we demonstrate that PIB, in addition to binding CPs clearly delineates diffuse plaques and cerebrovascular amyloid angiopathy (CAA). The interaction of PIB with CAA was not fully displaceable and this may be linked to the apolipoprotein E-epsilon4 allele. PIB was also found to label neurofibrillary tangles, although the overall intensity of this binding was markedly lower than that associated with the amyloid-beta (Abeta) pathology. The data provide a molecular explanation for PIB's limited specificity in diagnosing and monitoring disease progression in AD and instead indicate that the ligand is primarily a non-specific marker of Abeta-peptide related cerebral amyloidosis.

Age-related changes in nigrostriatal dopaminergic function are accentuated in +/- brain-derived neurotrophic factor mice.

The effects of a deletion for the brain derived neurotrophic factor (BDNF) allele (+/- BDNF) upon age-related changes in nigrostriatal dopaminergic (NSDA) function were assessed. Behavioral (beam crossing and spontaneous activity) and neurochemical (potassium-stimulated dopamine release from superfused striatum) measures were compared among Young (4-5 month), Middle (11-13 month) and Aged (19-21 month) +/- BDNF and their wild type littermate control (+/+ BDNF) mice. No statistically significant differences were obtained between +/+ and +/- BDNF mice at the Young age sampling period for any of the behavioral or neurochemical measures. Behavioral and neurochemical responses indices of NSDA function begin to diverge between +/+ and +/- Middle age BDNF mice and maximal differences were observed at the Aged period. For both movement and stereotypy times, scores obtained from +/+ mice were significantly decreased compared with +/- BDNF mice at the Aged period and center time scores of +/+ mice were decreased at both the Middle and Aged periods compared with +/- BDNF mice. Neurochemically, potassium-stimulated DA release of +/+ mice was significantly greater than +/- BDNF mice with maximal differences obtained at the Aged period. These results demonstrate marked differences in age-related changes of NSDA function between +/+ and +/- BDNF mice and suggest that the deletion of one allele for BDNF may make these mice more susceptible to age-related declines in NSDA function.

Loss of response to levodopa in Parkinson's disease and co-occurrence with dementia: role of D3 and not D2 receptors.

Previous data suggest a relationship between the loss of response to levodopa in Parkinson's disease (PD) patients with the co-occurrence of dementia, but the role of alterations in the dopamine system has not been explored. We measured the extent of striatal DA loss and changes in striatal DA D(2) and D(3) receptors in postmortem striatum of PD patients who historically had or had not lost their clinical response to dopaminergic drugs and/or had an additional diagnosis of dementia. Clinical evaluation and retrospective chart reviews for PD and dementia, and neuropathological diagnoses were obtained. All PD cases (+/-dementia), regardless of response to dopaminergic drugs, exhibited a significant and similar degree and pattern of loss of tyrosine hydroxylase immunocytochemistry and DA transporter binding in striatum, and loss of tyrosine hydroxylase-immunoreactive neurons and brain-derived neurotrophic-immunoreactive neurons from the ventral midbrain. D(2) receptor concentrations were modestly elevated in the rostral striatum of all the PD cases (+/-dementia), whether or not they continued to respond to dopaminergic drugs. In contrast, loss of D(3) receptor concentration correlated with loss of response to dopaminergic drugs, independent of the presence or absence of dementia. A maintained response to dopaminergic drugs correlated with an elevation of D(3) receptors. Dementia with PD was highly correlated with a loss of response to dopaminergic drugs, and was also correlated with reduced D(3) receptors. The alterations in D(3) receptor concentrations were greatest in the nucleus accumbens, caudal striatum, and globus pallidus. Thus, loss of dopamine D(3) receptors may be a more important contributing factor to a loss of response to dopaminergic drugs than changes in the D(2) receptor.

Dopamine D3 receptor as a therapeutic target for antipsychotic and antiparkinsonian drugs.

The cloning of the gene for the D3 receptor and subsequent identification of its distribution in brain and pharmacology allowed for serious consideration of the possibility that it might be a target for drugs used to treat schizophrenia and Parkinson's disease (PD). That is because it is highly expressed in limbic regions of the brain, exhibits low expression in motor divisions, and has pharmacologic similarity to the D2 receptor. Thus, antipsychotics that were presumed to block D2 receptors also had high affinity for the D3 receptor. Dopamine agonists used to treat the clinical symptoms of PD also have high affinity for the D3 receptor, and two D3 receptor-preferring agonists were found to be effective for treatment of PD. Many compounds achieving high potency and selectivity are now available, but few have reached clinical testing. Recent findings with respect to the anatomy of this receptor in human brain, altered expression in schizophrenia and PD, and biological models to study its function support the proposal that it is a target for development of drugs to alleviate symptoms in neuropsychiatric and neurologic disorders. Because of distinct aspects of regulation of the D3 receptor, it represents a unique target for therapeutic intervention in schizophrenia without high potential for unintended side effects such as tardive dyskinesia. It may also be that D3 receptor agonists can provide neuroprotective effects in PD and can modify clinical symptoms that D2 receptor-preferring agonists cannot provide.

Thalamo-cortical afferents control transient expression of the dopamine D(3) receptor in the rat somatosensory cortex.

The D(3) dopamine receptor (D(3)R) is selectively and transiently expressed in the barrel neurons of the somatosensory cortex (SI) between the first and second postnatal weeks. The D(3)R expression starts after the initial ingrowth of thalamocortical afferents (TCAs) into the barrel cortex and could be induced or controlled by them. We show that unilateral electrolytic lesion of the thalamic ventrobasal complex immediately after birth leads to a decrease in the D(3)R mRNA concentration in the lesioned SI 7 days after the lesion, whereas the D(3)R binding is little affected. Fourteen days after the neonatal thalamic lesion, the D(3)R binding and mRNA are drastically reduced and the barrel-like pattern of the D(3)R is absent. Elevation of the D(3) binding normally seen between the first and second postnatal weeks does not occur. Thalamic lesion on P6 differentially affects the D(3)R expression. One day after the lesion, the D(3) binding and mRNA are down-regulated, but the effect is transient. Five days after the lesion the concentration of D(3) mRNA in the lesioned hemisphere returns to the control level. The typical barrel-like pattern of D(3)R expression is evident in the lesioned SI, although TCAs are completely absent. Quantitative analysis demonstrated elevated cellular levels of the D(3) mRNA in barrel neurons 5 days after the lesion. These higher levels are needed, perhaps, to support the increased production of the D(3)R protein appropriate for this age. Age-related dynamics of the D(3)R binding is retained in the lesioned SI, although the concentration of D(3)R sites remains reduced. These data demonstrate that intact thalamic input is essential for the formation of mechanisms responsible for developmental regulation of the D(3)R expression in the SI.

Ventral striatal D(3) receptors and Parkinson's Disease.

Antiparkinsonian drugs are thought to act largely through the D2 receptor family that includes the D(2) and D(3) receptors. D(2) and D(3) receptors exhibit both complementary and overlapping expression at the macro and cellular level. The D(3) receptor appears to be a primary target of the mesolimbic dopamine system, is highly enriched in expression within the "limbic" striato-pallidal-thalamic loop, and is recognized as being regulated by dopaminergic activity in distinctly different ways from the D(2) receptor. In Parkinson's Disease it has been determined that loss of dopaminergic innervation results in elevation of the D(2) receptor but reduced levels of the D(3) receptor. In many late-stage Parkinson's Disease patients there is a loss of antiparkinsonian response to L-dopa and other antiparkinsonian drugs that is often correlated with clinical signs for dementia. We have determined that the reduction of D(3) receptor, and not that of the D(2) receptor, is associated with the loss of response to L-dopa and other antiparkinsonian drugs. The reduction of D(3) receptor is also related to the presence of dementia. An elevation of D(3) receptors was evident in those Parkinson's Disease cases with continued good response to L-dopa. Thus, we believe that reduced D(3) receptor number is correlated with certain subgroups of Parkinson's Disease and may also be related to a further diminishment in the mesolimbic DA system.

D2 but not D3 receptors are elevated after 9 or 11 months chronic haloperidol treatment: influence of withdrawal period.

Previous postmortem studies have identified divergent alterations in D2 and D3 receptors in schizophrenia but those results cannot be interpreted without further understanding of whether antipsychotic regulation of the D3 receptor is different from that of the D2 receptor. Depot parenteral administration of haloperidol decanoate was utilized to achieve consistent high levels in rat brain for 9 months with 2-month withdrawal or 11 months with 48-h withdrawal and compared to vehicle control and acute haloperidol (48-h) treatment groups. Autoradiographic means for measuring levels of D2 ([(3)H]-spiperone) and D3 receptors ([(125)I]trans 7-OH-PIPAT) and of D3 mRNA by in situ hybridization histochemistry in rat caudate-putamen, nucleus accumbens, islands of Calleja, and olfactory tubercle determined that there were significant group differences for regulation of D2 receptor. Chronic haloperidol for 9 or 11 months elevated D2 but not D3 receptors or D3 mRNA in all regions measured. Acute haloperidol treatment had no significant effects for any measure. Treatment for 9 months with a 2-month withdrawal resulted in a persistent increase in D2 receptors that was greater than that observed in the 11 months with 48-h withdrawal. This effect was most noticeable in the olfactory tubercle. These data confirm previous findings that short- or long-term haloperidol treatment leads to elevations in D2 but not D3 receptors or D3 mRNA, and long-term withdrawal from chronic haloperidol does not lead to elevations in D3 receptors or D3 mRNA. This suggests that an elevation in D3 receptors identified at postmortem in schizophrenics withdrawn from antipsychotics is not the result of the previous drug history [Gurevich et al. (1997) Arch Gen Psychiatry 54:225-232].

The zitter mutant rat exhibits loss of D3 receptors with degeneration of the dopamine system.

Experimental studies of the regulation of the dopamine (DA) D3 receptor have utilized techniques causing massive depletions of the DA system. Zitter mutant rats exhibiting a heterogeneous loss of striatal DA innervation were examined for DA transporter (DAT) binding and DA D3 receptor number by autoradiography and compared with Sprague-Dawley rats. A significant loss of DAT binding was observed with the greatest in the lateral caudate-putamen (72% loss) and the least in the nucleus accumbens (45% loss). DA D3 receptors were reduced by > 70% in all regions, even in the nucleus accumbens that exhibited considerably smaller depletions of DAT binding, indicating that loss of D3 receptors may occur prior to massive loss of DA innervation.

Dopamine D(3) receptor is selectively and transiently expressed in the developing whisker barrel cortex of the rat.

The rodent primary somatosensory cortex (SI) contains a map of the body surface, the most conspicuous part of which are "barrels," neuronal aggregates in layer IV that receive somatotopic projections from whiskers on the rodent's snout. We report that the D(3) dopamine receptor (D(3)R) is selectively and transiently expressed in SI during the first 2 weeks of postnatal development. D(3)R binding sites and mRNA overlap completely and are limited to layer IV of SI. D(3)R/mRNA are organized in a pattern corresponding to somatotopic representations of the body (e.g., whiskers, jaws, paws, etc.) with the highest expression in the barrel field. D(3) mRNA is first detected at postnatal day (P)4, increases rapidly until P7-10, and sharply decreases after P14. D(3)R binding sites are detectable at P6, peak at P14, and decline afterwards. D(1), D(2), D(4), or D(5) mRNAs display dissimilar expression pattern. D(1) mRNA is mostly confined to infragranular layers throughout the cortex. D(4) mRNA expression in layer IV rises by 4 weeks postnatal, when D(3)R expression is virtually undetectable. Quantitative analysis of D(3) mRNA expression demonstrates that the proportion of D(3) mRNA-positive cells decreases between P7 and P14, whereas mRNA concentration per cell remains stable. Moreover, D(3)R number continues to rise, whereas mRNA levels begin to decline. Thus, a process limiting D(3)R expression to fewer cells may occur that also induces changes in post-transcriptional regulation of D(3)R expression in remaining cells. These findings indicate that dopamine acting via D(3)R may play an important role in the development or function of the SI.

Ontogeny of the dopamine D2 receptor mRNA expressing cells in the human hippocampal formation and temporal neocortex.

The study details the cellular expression of the dopamine D2 receptor mRNA in the human temporal lobe during prenatal development. At 13 embryonic weeks (E13) D2 mRNA was widely expressed in the temporal lobe. At this time point in the dentate gyrus D2 mRNA positive cells first appeared at the outer border of the granular layer and their number increased with development. The CA1 exhibited the highest level of D2 mRNA expression. By E19-25 the hippocampal formation underwent rapid morphological maturation. D2 mRNA expression became more uniform and dense in the ammonic subfield. At all ages the subiculum appeared more mature morphologically but less intensely stained for D2 mRNA than the ammonic fields. In the entorhinal cortex D2 mRNA expression was most conspicuous in the future layer II at all ages. In the temporal neocortex D2 mRNA-positive cells were detected in the subplate and cortical plate. Differentiation of the cortical plate was accompanied by concentration of D2 mRNA-positive cells in layer V. The most conspicuous cells expressing D2 mRNA were found in the marginal zone of all regions and resembled Cajal-Retzius cells in morphology and location. Density of putative Cajal-Retzius cells expressing D2 mRNA decreased with development. They all but disappeared from the hippocampal areas by mid gestation, but in the temporal neocortex occasional cells were seen even at term. Early and widespread but region and cell type specific expression of D2 receptor mRNA suggests an important role of this DA receptor subtype in prenatal development of the human temporal lobe.

D3 receptors and the actions of neuroleptics in the ventral striatopallidal system of schizophrenics.

The mesolimbic dopamine (DA) system and an important target receptor, the D3 receptor, have been implicated in schizophrenia. We have identified, using non-radioactive in situ hybridization histochemistry, that D3 mRNA-positive neurons are highly concentrated in the ventral striatum, efferents of the ventral striatum (globus pallidus internal, ventral palladium, substantia nigra pars reticulata), and in regions projecting to the ventral striatum (medial dorsal thalamus, nucleus basalis, extended amygdala). D3 receptors are also highly enriched in the "limbic" striatal-pallidal-thalamic loop, exhibiting segregation from the D2 receptor-enriched "motor loop." This supports data developed in rats showing that the D3 receptor is a target of the mesolimbic DA system that can modulate the limbic striatal-palladial-thalamic loop. However, D2 and D3 receptors and their mRNAs are co-localized in many sensory regions (lateral and medial geniculate nuclei, basolateral and basomedial amygdala, regions of thalamus), suggesting mechanisms of cross-talk. We have also demonstrated that there are 45% elevations in D3 receptor number in ventral striatal neurons and their striatopalladial targets in schizophrenics that is reduced by concurrent antipsychotic treatment. Chronic haloperidol treatment to rats for 6 months with a 2-month withdrawal does not result in elevated D3 receptor number. We hypothesize that antipsychotic treatment via D3 receptors returns balance to limbic efferents of the ventral striatum. We established that early neonatal damage to the nigrostriatal DA system in rats produces characteristic adaptations in the pre- and post-synaptic components of the mesolimbic DA system that can provide a model to explore regulation by antipsychotics. This includes elevated release of DA from the mesolimbic DA terminals, elevated D3 receptor mRNA in the Islands of Calleja and nucleus accumbens, and enhanced behavioral response to psychostimulants.

Dopamine transporter-immunoreactive neurons decrease with age in the human substantia nigra.

Unbiased disector stereologic cell counting was applied to sections from the human substantia nigra that were immunostained by using a monoclonal antibody against the dopamine transporter (DAT). This antibody was found to penetrate the full thickness of the stained section. Quantification of the number of DAT immunostained neurons was performed in human cases stratified into three age groups, young (ages 0-49 years), middle aged (ages 50-69 years), and aged (ages 70-85 years). The number of DAT-immunoreactive nigral neurons was normalized for each case by constructing a ratio of the number of DAT-containing neurons to total number of neuromelanin-containing cells in each subject's sample. Three types of DAT nigral neurons were seen: type 1, intensely stained; type 2, lightly stained; and type 3, DAT-immunonegative neuromelanin-containing perikarya. By 50 years of age, the number of type 1 neurons decreased significantly (P < 0.0001), whereas the number of type 2 neurons increased with age (P < 0.0001). Type 3 neurons also increased with age (P < 0.01), although less robustly than type 2 neurons. Type 1 neurons decreased by 11.2% per decade, and the total number of nigral neurons (types 1-3) decreased by 6.7% per decade. Relative to the young group, there were 75% and 88% reductions in type 1 neurons in the middle-aged and aged groups, respectively. This contrasts with the 35% and 41% reductions in total number of neuromelanin-containing neurons seen in middle-aged and aged groups, respectively. The young group had significantly more type 1 neurons and fewer type 2 neurons compared with middle-aged and aged participants. Post-hoc analyses indicated that the young group had significantly fewer type 3 neurons compared with middle-aged and aged participants. These findings demonstrate an age-related reduction in the number of substantia nigra DAT-immunoreactive neurons. Therefore, insight into the mechanisms regulating the rate of DAT synthesis may aid in our understanding of the decline of DATs with aging and its functional significance.

Timing: A critical determinant of the functional consequences of neonatal 6-OHDA lesions.

Previous data have indicated that intrastriatal (IS) lesions of the dopamine (DA) system early in development result in a selective effect on D1 receptor expression and sensitivity, which is not seen with adult lesions or lesions made later in development. The purpose of the present study was to test the hypothesis that the timing of the lesion is a critical determinant of the consequences of DA depletion during development. Rats received IS injections of 6-hydroxydopamine (6-OHDA) on day of birth/postnatal day 1 (P0/1) or P7, which resulted in similar decreases in the number of DA uptake sites (> or =70% loss), a measure of DA terminal density. As adults, lesioned rats were challenged with DA receptor agonists to examine the functional sensitivity of D1 and D2 receptors. In adulthood, P0/1-lesioned rats exhibited increases in oral dyskinesias and rearing behavior following treatment with the partial D1 receptor agonists, SKF38393 and SKF77434, whereas rats lesioned on P7 exhibited increases in grooming. P7-lesioned rats also exhibited increases in gnawing, explosive jumping, and self-biting behavior following treatment with the full D1 receptor agonist SKF82958, which were not observed in the other groups. The results support the hypothesis that the timing of DA denervation is of paramount importance for governing the functional consequences of neonatal lesions, as measured by the incidence of DA agonist-induced behaviors in adulthood.

Developmental regulation of expression of the D3 dopamine receptor in rat nucleus accumbens and islands of Calleja.

The dopamine D3 receptor (D3R) belongs to the D2 subfamily and is expressed in the rat brain in targets of the mesolimbic dopaminergic system. Little is known about its normal development and control by dopaminergic innervation. We studied developmental expression of D3R in the rat nucleus accumbens (NAC) and islands of Calleja (ISC). At postnatal day (P) 7, D3 binding sites and mRNA were low in both areas. By P14, D3R and mRNA concentrations were close to adult levels in the ISC, whereas, in the NAC, binding increased until 3 months after birth. Cellular concentrations of D3 mRNA in the ISC increased with age in conjunction with a decrease in the number of D3 positive cells. In the NAC, the number of positive cells increased, whereas cellular levels of expression remained unchanged. Neonatal 6-hydroxydopamine lesion caused age-dependent changes in D3R expression. D3 binding sites did not change at P7 or P14, but there was a reduction in the number of D3 mRNA positive neurons accompanied by an increase in cellular levels of D3 mRNA at P14, suggesting that changes occurred in a subset of neurons. Up-regulation of D3 binding sites in NAC and ISC occurred 1 month after the lesion (P35) concomitant with a decrease in cellular levels of D3 mRNA and the number of D3 mRNA positive cells. At 3 months (P90) after the lesion, an increase in D3 mRNA occurred with no change in D3 binding sites. D3R shows region-specific dynamics in receptor/mRNA expression during development and is sensitive to loss of dopamine in early postnatal development.

Distribution of dopamine D3 receptor expressing neurons in the human forebrain: comparison with D2 receptor expressing neurons.

The dopamine D2 and D3 receptors are members of the D2 subfamily that includes the D2, D3 and D4 receptor. In the rat, the D3 receptor exhibits a distribution restricted to mesolimbic regions with little overlap with the D2 receptor. Receptor binding and nonisotopic in situ hybridization were used to study the distribution of the D3 receptors and neurons positive for D3 mRNA in comparison to the D2 receptor/mRNA in subcortical regions of the human brain. D2 binding sites were detected in all brain areas studied, with the highest concentration found in the striatum followed by the nucleus accumbens, external segment of the globus pallidus, substantia nigra and ventral tegmental area, medial preoptic area and tuberomammillary nucleus of the hypothalamus. In most areas the presence of D2 receptor sites coincided with the presence of neurons positive for its mRNA. D3 binding sites and D3 mRNA positive neurons were most abundant in the limbic striatum and efferent structures, such as the nucleus accumbens, ventral striatum, substantia nigra, internal segment of the globus pallidus, anteroventral nucleus of the thalamus, and rostral pars reticulata of the substantia nigra. One important difference from the rat is that D3 receptors were virtually absent in the ventral tegmental area. D3 receptor and D3 mRNA positive neurons were observed in sensory, hormonal, and association regions such as the nucleus basalis, anteroventral, mediodorsal, and geniculate nuclei of the thalamus, mammillary nuclei, the basolateral, basomedial, and cortical nuclei of the amygdala. As revealed by simultaneous labeling for D3 and D2 mRNA, D3 mRNA was often expressed in D2 mRNA positive neurons. Neurons that solely expressed D2 mRNA were numerous and regionally widespread, whereas only occasional D3-positive-D2-negative cells were observed. The regions of relatively higher expression of the D3 receptor and its mRNA appeared linked through functional circuits, but co-expression of D2 and D3 mRNA suggests a functional convergence in many regions of the signals mediated by the two receptor subtypes.

Loss of dopamine D2 receptors in Alzheimer's disease with parkinsonism but not Parkinson's or Alzheimer's disease.

A significant proportion of patients with Alzheimer's disease (AD) exhibit extrapyramidal features that are referred to as parkinsonism (AD/Park) to distinguish the clinical and pathological features that differ from Parkinson's disease (PD). Previous results from this laboratory have shown that, although the presynaptic components of the dopamine (DA) system are markedly affected in AD/Park, the pathology is not similar to PD (Murray et al. 1995; Joyce et al. 1997). In the present study, we determined whether the parkinsonian symptoms in AD/Park might also reflect changes in numbers of postsynaptic DA receptors. We analyzed the binding of [125I]epidepride biding to DA D2/D3 receptors and [3H]SCH 23390 to D1 receptors by autoradiography in the striatum of six patients with PD, nine patients with AD, seven patients with AD/Park, and 14 neurologically intact control subjects. D2 receptors were reduced in the caudate and putamen of the AD/Park group (by 42 and 27% of controls, respectively) but not reduced in AD or PD. D1 receptors were elevated by 36% in the putamen of the PD group. Dopamine receptor changes are, therefore, not similar in PD, AD, and AD/Park. The elevation in D1 receptors in PD may contribute to the unwanted side effects of L-dopa treatment. The loss of D2 receptors in AD/Park, not observed in AD lacking overt parkinsonian symptomatology, may contribute to the presence of parkinsonian features and lack of responsiveness to L-dopa.

There is a limited critical period for dopamine's effects on D1 receptor expression in the developing rat neostriatum.

Neonatal lesions of the dopamine (DA) system have different behavioral and neurochemical effects than lesions made in adulthood. Previous data from this laboratory have indicated that in the early postnatal period, lesions to the DA system induced by instrastriatal 6-hydroxydopamine (6-OHDA) result in a rapid and permanent loss of striatal D1 binding sites, but D2 receptor binding is unaffected. The length of the postnatal period within which neonatal instrastriatal 6-OHDA administration is effective in modulating D1 receptor binding is not known. To determine when D1 and D2 receptors are vulnerable to lesions of the DA system, we administered 6-OHDA intrastriatally to damage the DA innervation at different ages in the early postnatal period, at day of birth/postnatal day 1 (P0/1), P7 or P15 and examined DA receptor binding at P90 with quantitative autoradiography. Using [3H]mazindol binding to DA transporters (DAT) to verify the extent of the lesion, we then quantified the number of D1 binding sites using [3H]SCH23390 and D2 sites with [3H]spiroperidol. There were significant reductions in DAT sites at P0/1 (78 to 88%) and P7 (67 to 81%) but less significant changes at P15 (34 to 50% losses). The lesions were most effective for the dorsal caudate-putamen than more ventrally or in the nucleus accumbens. Our results demonstrate a significant reduction in D1 sites in all regions of the neostriatum following lesions at P0/1. The dorsal caudate-putamen was affected the most (51% loss, and the nucleus accumbens (41%) and ventral caudate-putamen less so (31%). No significant changes in D1 receptors were found at P7 or P15 and D2 receptors were unaffected with lesions in any of the age groups. The results indicate that there is a critical period for affecting expression of D1 receptors and this effect may, in addition, be related to the pattern of DA loss. Additionally, regulation of D2 receptors by this degree of loss of DA innervation does not occur during the first two weeks postnatally.

Dopamine D3 receptor is decreased and D2 receptor is elevated in the striatum of Parkinson's disease.

The mesolimbic dopamine (DA) system preferentially innervates the D3 receptor, whereas the D2 receptor is, in addition, a target of the nigrostriatal DA system. In human brain D3 receptors and D3 mRNA-expressing neurons are largely segregated to brain regions that are the targets of the mesolimbic DA system and the efferents of the "limbic striatum." Thus, D3 receptors may regulate effects of DA on the "limbic" cortico-striatal-pallidal-thalamic-cortical loop. The nigrostriatal DA system is considerably more damaged in Parkinson's disease (PD) than the mesolimbic DA system. We report here, using radioligands selective for the D2 and D3 receptor, that these receptors are independently changed in PD. Tissue collected at autopsy from nine subjects with a diagnosis of PD and eight age-matched subjects with no evidence of a neurologic disorder was processed for [125I]epidepride binding to D2 receptors, [125I] trans-7-OH-PIPAT binding to D3 receptors, [125I]RTI-55 for the DA transporter (DAT), and immunoautoradiography for tyrosine hydroxylase (TH) using autoradiographic methods. Dopaminergic innervation to the caudal putamen was profoundly reduced and to a lesser extent in the rostral putamen in PD. DAT sites but not TH protein levels were reduced in the nucleus accumbens (NAS) in PD compared with age-matched control subjects. This is consistent with a loss of dopaminergic innervation from the mesolimbic DA system but elevation in TH production. D3 receptors were significantly reduced in PD by 40-45% particularly in the NAS and putamen. D2 receptors were elevated in PD in the dorsal putamen by 15%. The reduction in D3 receptor number was not observed in PD cases with a diagnosis of less than 10 years. The changes in DA D3 receptor number is interesting in light of the development of antiparkinsonian agents that are D3-preferring agonists.

Dopamine D2 receptor bands in normal human temporal cortex are absent in Alzheimer's disease.

A modular organization of bands enriched in high concentrations of D2 receptors are observed throughout the rostral to caudal aspects of the temporal cortex of the normal human at postmortem, but are most frequently observed in the inferior and superior temporal cortices [S. Goldsmith, J.N. Joyce, Dopamine D2 receptors are organized in bands in normal human temporal cortex, Neuroscience 74 (1996) 435-451]. In the tissue derived at postmortem from Alzheimer's disease cases (AD), these D2 receptor-enriched modules were found to be largely absent at rostral and mid-levels of the temporal cortex. Regions exhibiting this loss of receptor binding also showed a marked reduction in the number of pyramidal neurons stained for D2 mRNA. In addition, the AD material exhibited numerous thioflavin-positive plaques and tangle-filled extraneuronal (ghost) pyramidal neurons that were D2 mRNA-negative. Regions that are the earliest affected and most susceptible to classical AD pathology are also most sensitive to the loss of D2 receptors. These results, along with our previous data [J.N. Joyce, C. Kaeger, H. Ryoo, S. Goldsmith, Dopamine D2 receptors in the hippocampus and amygdala in Alzheimer's disease, Neurosci. Lett. 154 (1993) 171-174; H. Ryoo, J. N. Joyce, The loss of dopamine D2 receptors varies along the rostrocaudal axis of the hippocampal complex in Alzheimer's disease, J. Comp. Neurol. 348 (1994) 94-110], indicate that specific pathways enriched with D2 receptors, including that within modules of higher order association cortices of the temporal lobe and continued through segregated pathways within the parahippocampus and hippocampus, are particularly susceptible to the loss in AD. These dopamine D2 receptor-enriched modules may play an important role in the reciprocal activity of large groups of neurons in these high-order association cortical regions. Hence, the loss of the D2 receptor-enriched modules in Alzheimer's disease contributes to disturbances in information processing in these high-order association cortices, and may promote the cognitive and non-cognitive impairments observed in Alzheimer's disease.