The telencephalic projections of the A8 dopamine cell group of the rat were assessed using both anterograde and combined retrograde-immunohistochemical methods. The projections of the A8 neurons onto the forebrain were more extensive than hitherto realized, and encompassed striatal, limbic, and allocortical regions. The A8 neurons were shown to contribute to the dopaminergic innervation of the striatum, nucleus accumbens, olfactory tubercles, amygdala, and bed nucleus of the stria terminalis, and also innervate the pyriform and entorhinal cortices. In addition, projections within the midbrain were observed, and suggested that there may be direct interconnections between the dopaminergic neurons of the A8, A9, and A10 cell groups. These data therefore suggest that the A8 dopamine cell group is uniquely situated to modulate functional activity within both nigrostriatal and mesocorticolimbic regions, and further suggests that heterogeneities of the midbrain dopamine neurons are embedded within a larger homogeneous mesotelencephalic dopamine system.
The discrete anatomical distribution of arginine vasopressin and corticotropin releasing factor (CRF) immunoreactivity in the paraventricular nucleus (PVN) of the rat hypothalamus is altered after adrenalectomy. Not only is the immunostaining of both peptides enhanced, but vasopressin immunoreactivity, normally confined to the magnocellular subdivision, becomes clear in a large percentage of CRF neurones in the parvocellular subdivision. These changes in immunoreactivity may reflect changes in post-translational events, peptide metabolism or genomic activity that lead indirectly or directly to the enhanced expression of vasopressin. Here we report that levels of transcripts homologous to vasopressin messenger RNA increase in the PVN after adrenalectomy, in parallel with increases in vasopressin immunoreactivity. In fact, after adrenalectomy, vasopressin mRNA can be detected in CRF-immunoreactive neurones. These results indicate that a considerable degree of plasticity is retained by the adult neuronal genome of the rat and that this plasticity may be modulated by the endocrine environment.
Immunocytochemical studies have shown that adrenalectomy produces changes in the content and distribution of [arginine-8]vasopressin (AVP) immunoreactivity in the paraventricular nucleus of the hypothalamus. The purpose of this study was to determine whether manipulation of adrenal hormones affects the levels of AVP mRNA. In situ hybridization assays with highly specific synthetic oligodeoxyribonucleotide probes and immunocytochemistry were used to detect the distribution of AVP mRNA and AVP-immunoreactive perikarya. AVP mRNA is codistributed with AVP immunoreactivity in the posterior magnocellular subdivision of the paraventricular nucleus and its accessory nuclei, the supraoptic nucleus and the suprachiasmatic nucleus. In adrenalectomized rats, the density and distribution of the hybridization signal were increased in the paraventricular nucleus; a 2-fold increase in the area comprising the signal was observed. At the cellular level, silver grains were detected in corticotropin-releasing-factor-immunoreactive neurons throughout the medial parvocellular subdivision of the paraventricular nucleus. No changes were seen in the distribution of AVP mRNA in the supraoptic or suprachiasmatic nuclei. Treatment with dexamethasone prevented the increase in AVP mRNA produced by adrenalectomy. In contrast, adrenalectomy did not alter the hybridization signal obtained with a probe for a-tubulin mRNA. These results suggest, at the cellular level, that adrenalectomy induces a glucocorticoid-sensitive stimulation of AVP mRNA synthesis in the central nervous system. Thus, considerable plasticity in gene expression is retained in the hypothalamus of the adult rat.The discrete anatomical organization of hypothalamic neurosecretory neurons within the paraventricular nucleus (PVN) has established this region as a unique site to study the neurohypophyseal secretory system. [arginine-8]Vasopressin (AVP) and oxytocin are contained within neurons of the PVN and the supraoptic nucleus (SON) (1, 2) and are involved in both autonomic and neurosecretory functions (3, 4). The axons emanating from these perikarya form the hypothalamoneurohypophyseal tracts, which project to the pars nervosa where AVP is released from their terminal endings (5,6). However, AVP has been detected in axon terminals within the external zone of the median eminence (7), suggesting that it may be involved, to some extent, in regulating anterior pituitary functions. Unlike the neurohypophyseal projection, the majority of AVP-immunoreactive axon terminals in the median eminence originate from neurons within the medial parvocellular subdivision of the PVN (8). Biochemical studies have confirmed quantitatively that adrenalectomy increases AVP immunoreactivity (7, 9). After adrenalectomy, corticotropin-releasing factor (CRF) and AVP were reported to be colocalized within neurons in the medial parvocellular region of the PVN (10, 12). This finding is of particular interest because immunocytochemical studies have shown that adrenalectomy produces a dexamethasonesen...
The distribution of cholecystokinin (CCK) mRNA in the rat brain was determined by means of in situ hybridization histochemistry. Our results demonstrate a widespread distribution of neurons containing CCK mRNA throughout the rat brain. Hybridization-positive neurons were distributed throughout the neocortex, olfactory bulb, claustrum, amygdala, the dentate gyrus and hippocampus proper, and several subnuclei of the thalamus and the hypothalamus. The most abundant and most heavily labeled neurons were found in the endopiriform/piriform cortex, tenia tecta, and the ventral tegmental area. The distribution of neurons positive for CCK mRNA paralleled that of CCK-like immunoreactive neurons. These results detail the distribution of CCK mRNA and clearly identify the existence of CCK-synthesizing neurons in regions such as the paraventricular and supraoptic nuclei of the hypothalamus, where the presence of CCK cell bodies was previously uncertain.
Individual neurons containing prosomatostatin mRNA were identified with in situ hybridization histochemistry. Our results demonstrate a widespread distribution of prosomatostatin mRNA in several regions of the rat central nervous system. Neurons containing this transcript were most abundant in the anterior olfactory nucleus, hypothalamus, hippocampus, and amygdala as well as in all regions of the cerebral cortex. Moreover, the distribution of mRNA-containing perikarya was coextensive with the location of neurons containing somatostatin-like immunoreactivity in all areas of the brain examined. Somatostatin neurons varied in their morphology and amount of hybridization signal from region to region. The widespread distribution and regional variations in neuronal morphology and the amount of hybridization signal are consistent with a neurotransmitter and/or a neuromodulator role for somatostatin in addition to its well-established neuroendocrine role. These results demonstrate that both the peptide and its mRNA are found in perikarya in the same areas and that they are therefore the sites of synthesis for somatostatin.
Messenger RNAs (mRNA) coding for vasoactive intestinal polypeptide (VIP), peptide histidine isoleucine (PHI), somatostatin and vasopressin were localized in the suprachiasmatic nucleus (SCN) of the rat hypothalamus using in situ hybridization histochemistry. Specific mRNA coding for each of these peptides was distributed in areas coextensive with the immunohistochemical localization of the appropriate peptide. The autoradiographic signal produced with probes to VIP and PHI created dense concentrations of silver grains over neuronal perikarya in the ventrolateral SCN, and the coextensive distribution of both VIP- and PHI-mRNAs suggests that both peptides are synthesized within the same neurons. The distribution of somatostatin-mRNA was distinct from the of VIP and PHI. Labeled neurons are observed at the interface of the two SCN subdivisions and the distribution of these neurons is identical to those shown to contain somatostatin immunoreactivity. Vasopressin-mRNA is also differentially concentrated within neurons in the dorsomedial subdivision of the SCN in an area that is coextensive with vasopressin-immunoreactive perikarya. The discrete pattern of hybridization for each of these mRNAs indicates that each of these peptides are synthesized in SCN neurons and reaffirms the differential distribution of each of these chemically defined cell populations within cytoarchitecturally distinct subdivisions of the nucleus.
Radioactively labeled RNA probes in conjunction with in situ hybridization histochemistry have become a useful method for studying gene expression in the central nervous system. We used digoxigenin-labeled uridine triphosphate to synthesize cRNA probes for localization of nerve growth factor receptor (NGFR) mRNA in the rat basal forebrain. Detection of cells containing &goxigethn-labeled NGFR mRNA was accomplished using a digoxigenin antibody conjugated with alkaline phosphatase. NGFR mRNA-positive cells were distributed in three major cell groups in the basal
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