A subpopulation of dopaminergic neurons in rat ventral mesencephalon contains both neurotensin and cholecystokinin

The coexistence of the neuropeptides neurotensin and cholecystokinin and the catecholamine-synthesizing enzyme tyrosine hydroxylase within neurons of the ventral mesencephalon was analyzed using an immunofluorescence triple-labeling technique. Virtually all of the neurotensin-positive cell bodies in the ventral tegmental area, medial substantia nigra pars compacta, retrorubral field, and rostral and caudal linear raphe nuclei were found to contain both cholecystokinin and tyrosine hydroxylase immunoreactivities. The degree of colocalization was lower and more variable in other regions including the ventral and central periaqueductal grey matter and dorsal raphe nucleus. It appeared that immunoreactivities for these 3 neuroactive substances were not contained within the same axonal-like fibers and terminals in the ventral midbrain. These results demonstrate that a subpopulation of dopaminergic neurons, which presumably comprise part of the ascending mesotelencephalic system, contains the two peptides neurotensin and cholecystokinin. Thus, the data suggest a morphological basis for some of the reported functional interactions of these 3 putative neurotransmitters/neuromodulators within this system.     

Dopaminergic neurons in rat ventral midbrain express brain-derived neurotrophic factor and neurotrophin-3 mRNAs

Studies of the trophic activities of brain-derived neurotrophic factor and neurotrophin-3 indicate that both molecules support the survival of a number of different embryonic cell types in culture. We have shown that mRNAs for brain-derived neurotrophic factor and neurotrophin-3 are localized to specific ventral mesencephalic regions containing dopaminergic cell bodies, including the substantia nigra and ventral tegmental area. In the present study, in situ hybridization with ³⁵S-labeled cRNA probes for the neurotrophin mRNAs was combined with neurotoxin lesions or with immunocytochemistry for the catecholamine-synthesizing enzyme tyrosine hydroxylase to determine whether the dopaminergic neurons, themselves, synthesize the neurotrophins in adult rat midbrain. Following unilateral destruction of the midbrain dopamine cells with 6-hydroxydopamine, a substantial, but incomplete, depletion of brain-derived neurotrophic factor and neurotrophin-3 mRNA-containing cells was observed in the ipsilateral substantia nigra pars compacta and ventral tegmental area. In other rats, combined in situ hybridization and tyrosine hydroxylase immunocytochemistry demonstrated that the vast majority of the neurotrophin mRNA-containing neurons in the substantia nigra and ventral tegmental area were tyrosine hydroxylase immunoreactive. Of the total population of tyrosine hydroxylase-positive cells, double-labeled neurons constituted 25–50% in the ventral tegmental area and 10–30% in the substantia nigra pars compacta, with the proportion being greater in medial pars compacta. In addition, tyrosine hydroxylase/neurotrophin mRNA coexistence was observed in neurons in other mesencephalic regions including the retrorubral field, interfascicular nucleus, rostral and central linear nuclei, dorsal raphe nucleus, and supramammillary region. The present results demonstrate brain-derived neurotrophic factor and neurotrophin-3 expression by adult midbrain dopamine neurons and support the suggestion that these neurotrophins influence dopamine neurons via autocrine or paracrine mechanisms. These data raise the additional possibility that inappropriate expression of the neurotrophins by dopaminergic neurons could contribute to the neuropathology of disease states such as Parkinson's disease and schizophrenia. © 1994 Wiley-Liss, Inc.     

Forebrain projections from cholecystokininlike-immunoreactive neurons in the rat midbrain

The purpose of the present study was to analyze the distribution of cholecystokininlike-immunoreactive (CCK-I) neurons within the rat ventral mesencephalon which project to several forebrain areas. The peroxidase-antiperoxidase immunocytochemical technique was used to examine the anatomical localization of CCK-I within the ventral midbrain and in the following forebrain regions: caudate-putamen, nucleus accumbens, olfactory tubercle, bed nucleus of the stria terminalis, septum, amygdala, and prefrontal, anterior cingulate, and piriform cortices. CCK-I perikarya were distributed throughout the substantia nigra, ventral tegmental area, and several midline raphe nuclei to a greater extent than previously reported, particularly in the substantia nigra pars compacta. Terminallike immunoreactivity for CCK was observed in all of the above forebrain sites. In addition, infrequent CCK-I cell bodies were localized in the caudate-putamen, nucleus accumbens, olfactory tubercle, septum, and bed nucleus of the stria terminalis. To analyze forebrain projections of the ventral midbrain CCK-I neurons, indirect immunofluorescence was combined with fluorescence retrograde tracing. CCK-I neurons of the substantia nigra and/or ventral tegmental area were found to project, to varying extents, to all of the above CCK-I forebrain terminal fields. The nucleus accumbens, olfactory tubercle, and septal and prefrontal cortical projections arose primarily from CCK-I perikarya in the ventral tegmental area whereas the projections to the caudate-putamen and anterior cingulate cortex arose predominantly from immunoreactive neurons in the substantia nigra pars compacta. The amygdala received innervation mainly from CCK-I cell bodies located in the substantia nigra pars lateralis. CCK-I afferents to the bed nucleus of the stria terminalis and piriform cortex originated from perikarya distributed approximately equally across the ventral tegmental area and substantia nigra pars compacta. The general topography of CCK-I forebrain innervation observed in this study is similar to that previously reported for the ascending dopaminergic projections from ventral mesencephalic neurons. CCK-I neurons of the midline raphe nuclei were found to provide relatively minor afferents to the caudate-putamen, bed nucleus of the stria terminalis, septum, and prefrontal cortex and more substantial projections to the amygdala. The results of this study demonstrate that CCK-I neurons of the ventral midbrain supply a much broader innervation of forebrain regions than previously appreciated.(ABSTRACT TRUNCATED AT 400 WORDS)     

Synaptic innervation of midbrain dopaminergic neurons by glutamate-enriched terminals in the squirrel monkey

The excitatory amino acid, glutamate, has long been thought to be a transmitter that plays a major role in the control of the firing pattern of midbrain dopaminergic neurons. The present study was aimed at elucidating the anatomical substrate that underlies the functional interaction between glutamatergic afferents and midbrain dopaminergic neurons in the squirrel monkey. To do this, we combined preembedding immunocytochemistry for tyrosine hydroxylase and calbindin D-28k with postembedding immunostaining for glutamate. On the basis of their ultrastructural features, three types (so-called types I, II, and III) of glutamate-enriched terminals were found to form asymmetric synapses with dendrites and perikarya of midbrain dopaminergic neurons. The type I terminals accounted for more than 70% of the total population of glutamate-enriched boutons in contact with dopaminergic cells in the dorsal and ventral tiers of the substantia nigra pars compacta as well as in the ventral tegmental area, whereas 5–20% of the glutamatergic synapses with dopaminergic neurons involved the two other types of terminals. The major finding of our study is that the glutamate-enriched boutons were involved in 70% of the axodendritic synapses in the ventral tegmental area. In contrast, less than 40% of the boutons in contact with dopaminergic dendrites were immunoreactive for glutamate in the dorsal and ventral tiers of the substantia nigra pars compacta. Approximately 50% of the terminals in contact with the perikarya of the different populations of midbrain dopaminergic neurons displayed glutamate immunoreactivity. In conclusion, our findings provide the first evidence that glutamate-enriched terminals form synapses with midbrain dopaminergic neurons in primates. The fact that the proportion of glutamatergic boutons in contact with dopaminergic cells is higher in the ventral tegmental area than in the substantia nigra pars compacta suggests that the different groups of midbrain dopaminergic neurons are modulated differently by extrinsic glutamatergic afferents in primates. © 1996 Wiley-Liss, Inc.     

Neurotensin terminals form synapses primarily with neurons lacking detectable tyrosine hydroxylase immunoreactivity in the rat substantia nigra and ventral tegmental area.

A light and electron microscopic double antigen localization technique was employed to examine the fine structural relationship between neurotensin-containing axon terminals and dopaminergic neurons in the substantia nigra and ventral tegmental area of the rat. At the light microscopic level, neurotensin-immunoreactive terminals were densely distributed throughout the substantia nigra pars compacta and ventral tegmental area in close proximity to tyrosine hydroxylase-immunoreactive somata and dendrites. On electron microscopic examination, direct synaptic connections were identified between neurotensin-immunoreactive axon terminals and tyrosine hydroxylase-immunopositive perikarya and dendrites. However, only 8.2% and 8.8% of the neurotensin-immunoreactive axonal profiles detected in the substantia nigra and ventral tegmental area, respectively, were found in direct apposition with tyrosine hydroxylase-immunostained elements. In turn, only 9.3% and 10.0% of tyrosine hydroxylase immunoreactive dendrites sampled from the substantia nigra and ventral tegmental area, respectively, were seen in contact with neurotensin immunopositive axon terminals. However, neurotensin-immunoreactive and tyrosine hydroxylase-immunolabelled elements were frequently identified in close anatomical proximity (less than 5 microns) to one another. These results are interpreted in light of the selective association of neurotensin receptors with dopaminergic neurons in the substantia nigra and ventral tegmental area to suggest a predominantly parasynaptic mechanism of action for neurotensin in the ventral midbrain.     

Subpopulations of mesencephalic dopaminergic neurons express different levels of tyrosine hydroxylase messenger RNA.

Subpopulations of mesencephalic dopamine containing neurons possess different electrophysiological, pharmacological, biochemical, and anatomical properties. In order to determine whether such differences are related to the regulation of tyrosine hydroxylase, the rate limiting enzyme in the synthesis of catecholamines, the regional distribution of tyrosine hydroxylase messenger RNA in these neurons was examined using in situ hybridization histochemistry. In the mouse, labelling for tyrosine hydroxylase messenger RNA associated with individual neurons was significantly less in the lateral substantia nigra pars compacta than in the medial substantia nigra pars compacta and the ventral tegmental area. A similar pattern of labelling was observed in the rat. Labelling for tyrosine hydroxylase messenger RNA was significantly less in the lateral substantia nigra pars compacta than in medial pars compacta (a densely cellular region), the area dorsal to the medial substantia nigra pars compacta (a less cell dense region), and the ventral tegmental area. Differences in levels of labelling for messenger RNA in mesencephalic dopamine neurons were not related to differences in cell size as measured in sections processed for tyrosine hydroxylase immunohistochemistry. The results suggest that tyrosine hydroxylase messenger RNA is differentially regulated in subpopulations of mesencephalic dopamine neurons, supporting the view that these neurons are physiologically distinct.     

Correspondence between high affinity 125I-neurotensin binding sites and dopaminergic neurons in the rat substantia nigra and ventral tegmental area: a combined radioautographic and immunohistochemical light microscopic study

Several lines of anatomical, biochemical, and pharmacological evidence have suggested that specific high affinity neurotensin binding sites are associated with dopamine-containing neurons in the rat ventral tegmentum. In the present study we confirmed and quantified the extent of this association by combining monoiodinated neurotensin radioautography and tyrosine hydroxylase immunohistochemistry on adjacent 5-10 microns-thick midbrain sections. We found that 95-100% tyrosine hydroxylase-immunoreactive neurons detected in all subdivisions of the substantia nigra (pars compacta, pars lateralis, and pars reticulata) exhibited intense 125I-neurotensin labeling in adjacent light microscopic radioautographs. Tyrosine hydroxylase-positive dendrites radiating downward from compacta neurons were also heavily labeled throughout the pars reticulata. In the paranigral subdivision of the ventral tegmental area, silver grains were evenly distributed over neuropil and perikarya and therefore could not be readily attributed to any given tyrosine hydroxylase-positive element. In contrast, within the parabrachial pigmentous subdivision of the ventral tegmental area, 80-90% of the tyrosine hydroxylase-immunoreactive somata and proximal processes were clearly in register with 125I-neurotensin labeled cells. Finally, all tightly packed TH-positive neurons in the interfascicular nucleus showed intense 125I-neurotensin labeling. The vast majority of the neurotensin binding sites observed in the ventral midbrain tegmentum were of the high affinity, physiologically active receptor type since levocabastine, a selective blocker of the low affinity neurotensin binding component, had minimal effect on the binding density in any of the midbrain regions examined. The present results demonstrate an extensive overlap between specific, high affinity neurotensin binding sites and dopamine perikarya and dendrites in the rat ventral tegmentum, and thereby provide a direct anatomical substrate for observed neurotensin-dopamine interactions in the mesocorticolimbic and nigrostriatal projection systems.     

Efferent projections of the retrorubral nucleus to the substantia nigra and ventral tegmental area in cats as shown by anterograde tracing

The aim of the present study was to determine whether the retrorubral nucleus projects to the dopaminergic nuclei in the ventral midbrain of the cat. For this purpose, injections of biotinylated dextran-amine or Phaseolus vulgaris-leucoagglutinin were placed into the retrorubral nucleus under stereotaxic guidance. The tracers were visualized by means of (immuno) histochemical procedures. In addition, tyrosine hydroxylase immunohistochemistry was used to evaluate the location of the injection sites and the distribution of the anterogradely labeled fibers. Both tracers reveal the same topography of labeled fibers in the ventral mesencephalon. Labeled fibers with varicosities were found ipsilaterally in the substantia nigra pars compacta, the substantia nigra pars lateralis, the ventral tegmental area and, contralaterally, in the substantia nigra pars compacta, the ventral tegmental area, and the retrorubral nucleus. A considerable number of labeled axons with varicosities were observed to be wrapped around the dendrites and perikarya of tyrosine hydroxylase-positive neurons in these areas. The present results are discussed in view of the possible role of the A8 dopaminergic cell group in the coordination of A9 nigrostriatal and A10 mesolimbic systems, as well as in the progressive pathology seen in patients suffering from Parkinson's disease.     

A catecholaminergic projection from the ventral tegmental area to the diagonal band of broca: Modulation by neurotensin

The ventral tegmental area contains a high density of dopaminergic perikarya having ascending projections to a number of limbic forebrain regions. In this study, we use combined retrograde labeling with horseradish peroxidase (HRP) and immunohistochemical staining for tyrosine hydroxylase to examine the catecholaminergic projection from the ventral tegmental area to the diagonal band of Broca. When injection of HRP was restricted to the diagonal band, only neurons in the nucleus linearis, nucleus interfascicularis and ventromedial portion of the nucleus paranigralis were labeled. In contrast, HRP injection into the adjacent nucleus accumbens labeled neurons throughout these nuclei, plus the nucleus parabrachialis pigmentosus, nucleus retroruber and substantia nigra, pars compacta. Approximately 60% of neurons in the ventral tegmental area labeled from the diagonal band contained tyrosine hydroxylase, compared with 79% of the neurons labeled from the nucleus accumbens. Neurotensin is a tridecapeptide found in the ventral tegmental area which has been shown to activate dopamine neurons projecting to the nucleus accumbens. In this study, microinjection of neurotensin into ventral tegmental nuclei which contained neurons retrogradely labeled from the diagonal band significantly elevated the levels of dopamine metabolites, 3,4-dihydroxyphenylacetic acid and homovanillic acid, in the diagonal band. The results of this study demonstrate that a catecholaminergic projection exists from the ventral tegmental area to the diagonal band of Broca, and that this pathway can be stimulated by intra-ventral tegmental injection with neurotensin.     

Opioid receptors in midbrain dopaminergic regions of the rat I. MU receptor autoradiography

Summary Several lines of evidence indicate that an interaction exists between opioid peptides and midbrain dopaminergic neurons. The purpose of this study was to map and quantify the density of the mu opioid receptor subtype relative to the location of the dopaminergic (DA) neurons in the retrorubral field (nucleus A8), substantia nigra (nucleus A9), and ventral tegmental area and related nuclei (nucleus A10) in the rat. Sections through the rostral-caudal extent of the midbrain were stained with an antibody against tyrosine hydroxylase, as a DA cell marker, and comparable sections were processed for in vitro receptor autoradiography using the mu-selective ligand,³H-Tyr-D-Ala-N-MePhe-Gyl-ol enkephalin. In the nucleus A8 region, there were low levels of mu binding. In the rostral portion of nucleus A9, there was prominent mu binding both in the ventral pars compacta, which contains numerous DA neurons, and in regions that correspond to the location of the DA dendrites which project ventrally into the underlying substantia nigra pars reticulata. In the caudal portion of nucleus A9, mu binding was greatest in the substantia nigra pars reticulata, but also in the same region that contains DA neurons. In nucleus A10, mu receptor densities differed depending upon the nucleus A10 subdivision, and the rostral-caudal position in the nucleus. Low receptor densities were observed in rostral portions of the ventral tegmental area and interfascicular nucleus, and there was negligible binding in the parabrachial pigmented nucleus and paranigral nucleus at the level of the interpeduncular nucleus; all regions where there are high densities of DA somata. Mu binding was relatively high in the central linear nucleus, and in the dorsal and medial divisions of the medial terminal nucleus of the accessory optic system, which has been shown to contain DA dendrites. These data indicate that mu opioid receptors are located in certain regions occupied by all three midbrain DA nuclei, but in a highly heterogeneous fashion.     

Midbrain dopaminergic neurons (nuclei A8, A9, and A10): Three-dimensional reconstruction in the rat

The dopaminergic neurons in the midbrain of the rat are located in three cell groups: nucleus A8 cells in the retrourbal field, nucleus A9 cells in the substantia nigra, and nucleus A10 cells in the ventral tegmental area and related nuclei. The purpose of the present study was to map and quantify the midbrain dopaminergic neurons in two and three dimensions in the rat brain, using immunohistochemical staining and computer imaging techniques. The cells were identified with an antibody against tyrosine hydroxylase, and counted in six midbrain nuclei: the retrorubral field, substantia nigra pars compacta, substantia nigra pars reticulata, central linear nucleus, ventral tegmental area, and interfascicular nucleus. Outlines were traced around the perimeters of the coronal tissue sections, and the locations of all immunoreactive ventral midbrain cells were mapped. On one side of the brain there are approximately 1,300 nucleus A8 cells, 10,500 nucleus A9 cells, and 10,200 nucleus A10 cells. The 2- and 3-dimensional reconstructions illustrate the region-specific density of dopaminergic neurons throughout the midbrain cell complex, and provide a visual appreciation of the location and distribution of the three dopaminergic cell groups in relation to their position in the midbrain. Information about the number and location of midbrain dopaminergic neurons will be useful in conjunction with future studies that characterize these cells more specifically, for example, in terms of their co-transmitters, and afferent and efferent projections. © 1993 Wiley-Liss, Inc.     

Estrogen effects on tyrosine hydroxylase-immunoreactive cells in the ventral mesencephalon of the female rat: further evidence for the two cell hypothesis of dopamine function

The present study was undertaken to examine the differential effect of estrogen (E) on the expression of tyrosine hydroxylase (TH) in the substantia nigra compacta (SNc) and in two subdivisions of the ventral tegmental area in ovariectomized (ovx) and ovx plus estradiol benzoate (ovx+E)-treated female rats. Cell counting of TH-immunoreactive perikarya of the SNc, paranigral (PN) and interfascicular (IF) nucleus was performed and compared. Our findings demonstrate that E eliminated TH immunoreactivity from a number of midbrain neurons, while it seemingly did not affect it in others. This signifies a differential effect of E on ventral mesencephalic dopaminergic neurons.     

Nigrotectal projection to the inferior colliculus: horseradish peroxidase transport and tyrosine hydroxylase immunohistochemical studies in rats, cats, and bats

The present study investigated descending projections from the substantia nigra to the auditory tectum. Small (0.02-0.05 microliters) injections of a 30-60% aqueous solution of horseradish peroxidase (HRP) were made unilaterally into the inferior colliculus in rats, cats, and bats (Eptesicus fuscus). Tissue blocks including the substantia nigra, superior colliculus, and inferior colliculus were removed, sectioned, and processed for visualization of HRP. Results show that the substantia nigra, pars lateralis, projects to the inferior colliculus ipsilaterally. In addition, retrogradely labeled cells are found dorsal to the pars lateralis, in a column within the lateral tegmental area of the midbrain. Analysis of injection sites suggests that the principal target of this nigral projection is the dorsal and rostral pericentral region of the inferior colliculus. Immunohistochemical studies with an antibody to tyrosine hydroxylase demonstrate catecholaminergic neurons within the pars lateralis and lateral tegmentum that are similar in location and morphology to one class of HRP retrogradely labeled cells within these structures. These immunohistochemical studies also demonstrate a plexus of fine, varicose tyrosine hydroxylase-positive axons in the rostral pericentral region of the colliculus. The presence of this nigrotectal projection to the inferior colliculus is discussed in relation to its possible role in the control of acousticomotor behavior.     

The weaver gene continues to target late-generated dopaminergic neurons in midbrain areas at P90

To determine if lethal action of the weaver gene is more intense in late-generated dopaminergic neurons in midbrain areas on postnatal day (P) 90 [3H] thymidine autoradiography and tyrosine hydroxylase immunohistochemistry were combined in the same tissue section in homozygous weaver mice and normal controls. The experimental animals were the offspring of pregnant dams injected with [3H] thymidine on embryonic days (E) 11-12, E12-13, E13-14 and E14-15. Neurogenetic timetables of dopaminergic neurons were different between wild type and homozygous weavers in all midbrain areas analyzed. A substantial number of late-generated neurons in the substantia nigra pars compacta and in the ventral tegmental area are missing at P90, in these dopaminergic areas the loss is greater than at P20 indicating that neuronal loss is progressive. The greatest loss is in the substantia nigra pars compacta, confirming the report of Bayer et al. [Exp. Brain Res. 105 (1995) 200] at P20, while in the retrorubral field and the interfascicular nucleus late-generated neuron loss was less severe. These results furnish more evidence that dopaminergic neuron loss in homozygous weaver midbrain is a phenomenon linked to development.     

Subsets of midbrain dopaminergic neurons in monkeys are distinguished by different levels of mRNA for the dopamine transporter: Comparison with the mRNA for the D2 receptor,tyrosine hydroxylase and calbindin immunoreactivity

The midbrain dopamine system can be divided into two groups of cells based on chemical characteristics and connectivity. The dorsal tier neurons, which include the dorsal pars compacta and the ventral tegmental area, are calbindin positive, and project to the shell of the nucleus accumbens. The ventral tier neurons are calbindin-negative and project to the sensonmotor striatum. This study examined the distribution of the mRNAs for the dopamine transporter molecule (DAT) and the D₂ receptor in the midbrain of monkeys by using in situ hybridization. The distribution patterns were compared to that of tyrosine hydroxylase and calbindin immunohistochemistry. The results show that high levels of hybridization for DAT and the D₂ receptor mRNA are found in the ventral tier, calbindin-negative neurons and relatively low levels are found in the dorsal, calbindin-positive tier. Within the dorsal tier, the dorsal substantia nigra pars compacta has the least amount of both messages. These results show that in monkeys, the ventral tegmental area and the dorsal pars compacta form a dorsal continuum of dopamine neurons which express lower levels of mRNA for DAT and D₂ receptor than the ventral tier. DAT has been shown to be involved in the selective neurotoxicity of N-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). Different levels of DAT mRNA and calbindin may explain the differential effects of MPTP neurotoxicity. © 1995 Wiley-Liss Inc.     

Heterogeneous distribution of cytochrome oxidase activity in the rat substantia nigra: correlation with tyrosine hydroxylase and dynorphin immunoreactivities

Cytochrome oxidase (COase) activity, an endogenous marker of neuronal activity, was examined in the substantia nigra of the adult rat at the light-microscopic level. In addition, the pattern of histochemical staining observed for COase activity was correlated with immunohistochemistry for tyrosine hydroxylase (a marker of dopaminergic neurons) and for dynorphin (a peptide present in afferents from the striatum). Differential oxidative metabolic activity was revealed in subregions of the substantia nigra by COase histochemistry. Neurons of the substantia nigra pars compacta (SNc), ventral tegmental area, and ventrally displaced dopaminergic neurons were characterized by little or no staining for COase. In contrast, the substantia nigra pars reticulata (SNr) possessed a heterogeneous distribution of COase activity that was characterized by denser staining in the ventrolateral than the dorsomedial part of the nucleus throughout its rostrocaudal extent, with the exception of the most rostral levels. This pattern of COase activity was inversely correlated with the density of ventrally descending tyrosine hydroxylase-positive dendrites arising from the medial portion of the SNc, as well as with the density of dynorphin immunoreactivity. The results suggest that the SNc and SNr possess distinct levels of oxidative metabolic activity. Furthermore, within the SNr itself, different levels of COase activity characterize subpopulations of neurons which may be differentially regulated by both striatal and dopaminergic influences.     

Dopaminergic neurons expressing calbindin in normal and parkinsonian monkeys.

In cynomolgus monkeys, midbrain neurons immunoreactive (IR) for the calcium-binding protein calbindin D-28k (CaBP) occur principally in the dorsal tier of substantia nigra pars compacta (SNc) and in the ventral tegmental area (VTA), and most of these neurons co-express tyrosine hydroxylase (TH). In monkeys rendered parkinsonian (PD) after MPTP injections, CaBP-IR neurons are much less severely affected than TH-IR neurons in SNc and in VTA, and most spared neurons in SNc/VTA display both CaBP and TH immunoreactivity. These results reveal that, in contrast to the situation in other neurodegenerative diseases, CaBP may be used as a marker for a specific neuronal population that is less prone to degeneration in Parkinson's disease.     

Organization of midbrain catecholamine-containing nuclei and their projections to the striatum in the North American opossum, Didelphis virginiana.

Presumptive catecholamine (CA) neurons in the opossum midbrain were identified by tyrosine hydroxylase immunohistochemistry. In the midline, small to moderate number of CA cells were present in the rostral third of the nucleus raphe dorsalis and throughout the nucleus linearis. Ventrolaterally, such cells were observed in the deep tegmental reticular formation, in all subnuclei of the ventral tegmental area, and in the three subdivisions of the substantia nigra. The CA cells in these areas conform to the dopamine cell groups, A8, A9, and A10 as described in the rat. In several areas there appeared to be no separation between the CA neurons belonging to cytoarchitecturally different nuclei. In order to determine which CA neurons gave rise to striatal projections, the neostriatum was injected with True Blue (TB), and sections through the midbrain were processed for tyrosine hydroxylase (TH) and visualized by immunofluorescence. Neurons containing both TB and TH were observed in each of the CA cell groups mentioned above. The distribution of these cells confirmed organizational features that may be unique to the opossum's substantia nigra. In addition, different patterns of labeling resulted from caudate versus putamen injections, suggesting a rudimentary medial to lateral topography in the organization of nigrostriatal projections. Although our results suggest that the organization of midbrain CA neurons in the opossum is similar to that in placental mammals, it is clear that differences exist.     

外源性kynurenic acid对大鼠黑质多巴胺能神经元损伤的保护作用研究

目的观察评价预先应用谷氨酸(Glu)受体拮抗剂kynurenic acid(KYNA)对黑质多巴胺(DA)能神经元及神经传导纤维损伤的保护性作用. 方法雌性SD大鼠40只,随机分为4组,每组10只,应用江湾I型C立体定向仪,在单侧黑质致密部及中脑被盖腹侧部, A组注射生理盐水,B组注射KYNA,C组注射KYNA和6-羟基多巴胺(6-OHDA), KYNA先于6-OHDA 30 min, D组注射6-OHDA.注射药物3 d后,进行症状观察,4周后处死大鼠.切片HE染色观察黑质细胞的形态特点,冰冻切片免疫组化特殊染色观察酪氨酸羟化酶(TH)阳性细胞及TH阳性纤维着色情况.结果正常黑质细胞体形较大,富含黑色素颗粒,可见尼氏体.TH着色结果提示B组与A组之间无显著差异,P>0.05.实验组C与A、B、D组比较均有显著性差异,P<0.01.结论外源性Glu受体拮抗剂KYNA通过阻滞Glu受体一定时间阶段内能减轻6-OHDA诱导的黑质DA能神经元毒性损害.     

Lateromedial Gradient of the Susceptibility of Midbrain Dopaminergic Neurons to Neonatal 6-Hydroxydopamine Toxicity

The topography-dependent vulnerability of midbrain dopaminergic neurons to neonatal intracranial exposure to 6-hydroxydopamine (6-OHDA) was investigated at adult age by the quantitative analysis of cell counts of tyrosine hydroxylase-immunopositive neurons. In all cases of intracisternal 6-OHDA treatment, A9 dopaminergic neurons in the substantia nigra (SN) were much more vulnerable to death than more medially located A10 dopaminergic neurons. Moreover, within each cell group, there were also lateromedial topographic gradients. In the A9 neuronal group, cells located in the pars lateralis of the SN and the lateral part of the pars compacta of the SN were more susceptible to 6-OHDA toxicity than those located more medially. In the A10 neuronal group, cells located in the medial part of the ventral tegmental area were more resistant to toxicity than those located more laterally, and dopaminergic cells in the midline midbrain areas (interfascicular nucleus and rostral linear nucleus of raphe) were completely spared from 6-OHDA toxicity. These findings revealed that 6-OHDA is not equally toxic to all midbrain dopaminergic neurons in neonates and that the lateromedial vulnerability pattern shows similarities to those reported in Parkinson's disease.