Central Medial Thalamic Nucleus Research Articles

Mapping of somatostatin-28 (1–12) in the alpaca diencephalon

Using an immunocytochemical technique, we report for the first time the distribution of immunoreactive cell bodies and fibers containing somatostatin-28 (1–12) in the alpaca diencephalon. Somatostatin-28 (1–12)-immunoreactive cell bodies were only observed in the hypothalamus (lateral hypothalamic area, arcuate nucleus and ventromedial hypothalamic nucleus). However, immunoreactive fibers were widely distributed throughout the thalamus and hypothalamus. A high density of such fibers was observed in the central medial thalamic nucleus, laterodorsal thalamic nucleus, lateral habenular nucleus, mediodorsal thalamic nucleus, paraventricular thalamic nucleus, reuniens thalamic nucleus, rhomboid thalamic nucleus, subparafascicular thalamic nucleus, anterior hypothalamic area, arcuate nucleus, dorsal hypothalamic area, around the fornix, lateral hypothalamic area, lateral mammilary nucleus, posterior hypothalamic nucleus, paraventricular hypothalamic nucleus, suprachiasmatic nucleus, supraoptic hypothalamic nucleus, and in the ventromedial hypothalamic nucleus. The widespread distribution of somatostatin-28 (1–12) in the thalamus and hypothalamus of the alpaca suggests that the neuropeptide could be involved in many physiological actions.

Neonatal maternal separation enhances central sensitivity to noxious colorectal distention in rat

Psychological stress experienced in early life plays an important role in the development of visceral hyperalgesia in irritable bowel syndrome (IBS). Neonatal maternal separation has been shown to trigger a long-term alternation in stress-induced responses to visceral nociceptive stimuli in rats. The aim of the present study was to show a direct evidence of stress-induced alteration in central neuronal responses to colorectal distention (CRD) in rats by a quantitative study of c-fos expression in relevant brain structures. Male Wistar rat pups were subjected to 180-min daily neonatal maternal separation (NMS) for 13 consecutive days (from PND 2 to PND 14). The expression of c-fos was examined by using immunohistochemistry. Increased c-fos expression was observed, for the first time, in the cingulate cortex (3-fold) in NMS rats in comparison with the control group at basal condition. At noxious CRD (80 mm Hg), c-fos expression was induced in the supraspinal centers and in both the superficial (laminae I–II) and the deeper laminae (laminae V–VI and X) of the spinal cord in rats. Significantly more Fos-IR nuclei were found in the laminae I and II, and laminae V–VI of the lumbarsacral spinal cord, the paraventricular thalamic nucleus, the cingulate cortex, the amygdaloid central nucleus in NMS rats, but not in the solitary tract, the central medial thalamic nucleus, the ventromedial hypothalamic nucleus, and the periaquaductal gray. The present results indicate that NMS has sensitized the cingulate cortex and upregulated the activity of the ascending pathway at spinal level as well as the thalamo-cortico-amydala pathway to CRD. The upregulation and sensitization of these pathways may be responsible for the development of visceral hypersensitivity in IBS.

Up‐regulation of NMDA receptor subunit and post‐synaptic density protein expression in the thalamus of elderly patients with schizophrenia

Numerous studies have described structural and functional abnormalities of the thalamus in schizophrenia, but surprisingly few studies have examined neurochemical abnormalities that accompany these pathological changes. We previously identified abnormalities of multiple molecules associated with glutamatergic neurotransmission, including changes in NMDA receptor subunit transcripts and binding sites and NMDA receptor-associated post-synaptic density (PSD) protein transcripts in the thalamus of elderly patients with schizophrenia. In the present study, we performed western blot analysis to determine whether protein levels of NMDA receptor subunits (NR1, NR2A, NR2B) and associated PSD proteins (NF-L, PSD95, SAP102) are altered in schizophrenia. Thalamic tissue from each subject was grossly dissected into two regions: a dorsomedial region containing limbic-associated dorsomedial, anterior and central medial thalamic nuclei; and a ventral thalamus region that primarily consisted of the ventral lateral nucleus. We observed increased protein expression of the NR2B NMDA receptor subunit and its associated intracellular protein, PSD95, in the dorsomedial thalamus of patients with schizophrenia, but the other molecules were unchanged, and we found no changes in the ventral thalamus. These data provide additional evidence of thalamic neurochemical abnormalities, particularly in thalamic nuclei which project to limbic regions of the brain. Further, these findings provide additional evidence of NMDA receptor alterations in schizophrenia, which may play an important role in the neurobiology of the illness.

Comparison of thalamic populations in mammals and birds: expression of ErbB4 mRNA

The expression of ErbB4 mRNA was examined in dorsal thalamic regions of chicks and rats. In rats, ErbB4 expression was observed in the habenular nuclei, the paraventricular nucleus, intermediodorsal nucleus, the central medial thalamic nucleus, the posterior nucleus, the parafascicular nucleus, the subparafascicular nucleus, the suprageniculate nucleus, the posterior limitans nucleus, the medial part of the medial geniculate nucleus, the peripeduncular nucleus, the posterior intralaminar nucleus, the lateral subparafascicular nucleus, the lateral posterior nucleus, and all ventral thalamic nuclei. In chicks, expression was observed in the subhabenular nucleus, the dorsomedialis posterior nucleus, the dorsointermedius posterior nucleus, the nucleus of the septomesencephalic tract, and areas surrounding the rotundus and ovoidalis nuclei, including the subrotundal and suprarotundal areas, and all ventral thalamic nuclei. Most cells within ovoidalis and rotundus were not labeled. The similar pattern of afferent and efferent projections originating from ErbB4-expressing regions of the mammalian and bird dorsal thalamus suggests that ErbB4 hybridizing cells may be derived from a single anlage that migrates into multiple thalamic regions. Most neurons in the rotundus and ovoidalis nuclei of chick may be homologous to unlabeled clusters of neurons intermingled with ErbB4-expressing cells of the mammalian posterior/intralaminar region.

Relationships between thalamostriatal neurons and pedunculopontine projections to the thalamus: a neuroanatomical tract-tracing study in the rat.

The present study aimed to investigate whether the pedunculopontine projection to the thalamus overlaps with identified thalamostriatal neurons. These projections were studied using a dual tract-tracing procedure combining anterogradely transported biotinylated dextran amine (pedunculopontine projections) and retrogradely transported Fluoro-Gold (thalamostriatal projections). Overlapping thalamic territories between thalamostriatal neurons and the axon terminals arising from the pedunculopontine tegmental nucleus were observed in the midline (paraventricular) and in the intralaminar (centrolateral, central medial, paracentral and parafascicular) thalamic nuclei. Other thalamic nuclei, such as the ethmoid, intermediodorsal, mediodorsal, paratenial, posteromedian, ventromedian, ventrolateral and rhomboid thalamic nuclei, displayed a lesser degree of overlap. These observations suggest the existence of presumptive contacts between thalamostriatal neurons and axons emerging from the pedunculopontine tegmental nucleus, therefore supporting the possible existence of feedback circuits in the rat basal ganglia in which the tegmento-thalamic projection would play a major role.

Differential projections to the intralaminar and gustatory thalamus from the parabrachial area: A PHA-L study in the rat

The organization of projections from the parabrachial (PB) area to the ventral posterior parvicellular (VPpc) "gustatory" and intralaminar nuclei of the thalamus was studied in the rat by using microinjections of Phaseolus vulgaris leucoagglutinin (PHA-L), into subregions of the PB area. The present study is a follow-up of three former studies (Bernard et al. [1993] J. Comp. Neurol. 329:201-229; Aldén et al. [1994] J. Comp. Neurol. 341:289-314; Bester et al. [1997a] J. Comp. Neurol. 383:245-281) that examined PB projections onto the amygdala, the bed nucleus of the stria terminalis, and the hypothalamus. Our data showed that (1) the region centered in the internal lateral PB subnucleus projects densely with a bilateral and symmetric pattern to the caudal portion of the paracentral and, to a lesser extent, to the adjacent portion of the central and parafascicular medial thalamic nuclei; (2) the mesencephalic PB region centered in the ventral lateral subnucleus and scattered neurons in the subjacent brachium conjunctivum project primarily, although diffusely, to the central medial thalamic nucleus. The third region includes two subgroups: (3a) the medial subgroup, including the medial, the waist area, and the ventral lateral subnuclei of the pontine PB area, projects bilaterally but with a weak ipsilateral predominance to the VPpc, terminals bearing large varicosities. Additionally, a diffuse projection with small varicosities spreads in the area between the two VPpc nuclei and the central medial nucleus. (3b) The lateral subgroup, centered in the external medial subnucleus, projects with a contralateral predominance in the periphery of the VPpc nuclei, most terminals being located around the dorsomedial tip. It is suggested that the PB projections to the intralaminar nucleus could be involved in cortical limbic arousal processing in relation with nociceptive, (somatic, visceral, and intraoral) and gustatory aversive stimuli. The projection with large varicosities inside the VPpc could process gustatory discrimination.

Differential projections to the intralaminar and gustatory thalamus from the parabrachial area: A PHA‐L study in the rat

The organization of projections from the parabrachial (PB) area to the ventral posterior parvicellular (VPpc) “gustatory” and intralaminar nuclei of the thalamus was studied in the rat by using microinjections of Phaseolus vulgaris leucoagglutinin (PHA-L), into subregions of the PB area. The present study is a follow-up of three former studies (Bernard et al. [1993] J. Comp. Neurol. 329:201–229; Aldén et al. [1994] J. Comp. Neurol. 341:289–314; Bester et al. [1997a] J. Comp. Neurol. 383:245–281) that examined PB projections onto the amygdala, the bed nucleus of the stria terminalis, and the hypothalamus. Our data showed that (1) the region centered in the internal lateral PB subnucleus projects densely with a bilateral and symmetric pattern to the caudal portion of the paracentral and, to a lesser extent, to the adjacent portion of the central and parafascicular medial thalamic nuclei; (2) the mesencephalic PB region centered in the ventral lateral subnucleus and scattered neurons in the subjacent brachium conjunctivum project primarily, although diffusely, to the central medial thalamic nucleus. The third region includes two subgroups: (3a) the medial subgroup, including the medial, the waist area, and the ventral lateral subnuclei of the pontine PB area, projects bilaterally but with a weak ipsilateral predominance to the VPpc, terminals bearing large varicosities. Additionally, a diffuse projection with small varicosities spreads in the area between the two VPpc nuclei and the central medial nucleus. (3b) The lateral subgroup, centered in the external medial subnucleus, projects with a contralateral predominance in the periphery of the VPpc nuclei, most terminals being located around the dorsomedial tip. It is suggested that the PB projections to the intralaminar nucleus could be involved in cortical limbic arousal processing in relation with nociceptive, (somatic, visceral, and intraoral) and gustatory aversive stimuli. The projection with large varicosities inside the VPpc could process gustatory discrimination. J. Comp. Neurol. 405:421–449, 1999. © 1999 Wiley-Liss, Inc.

Daily rhythms in Fos activity in the rat ventrolateral preoptic area and midline thalamic nuclei.

The present experiment investigated the expression of the nuclear phosphoprotein Fos over the 24-h light-dark cycle in regions of the rat brain related to sleep and vigilance, including the ventrolateral preoptic area (VLPO), the paraventricular thalamic nucleus (PVT), and the central medial thalamic nucleus (CMT). Immunocytochemistry for Fos, an immediate-early gene product used as an index of neuronal activity, was carried out on brain sections from rats perfused at zeitgeber time (ZT) 1, ZT 5, ZT 12.5, and ZT 17 (lights on ZT 0-ZT 12). The number of Fos-immunopositive (Fos+) cells in the VLPO was elevated at ZT 5 and 12.5 (i.e., during or just after the rest phase of the cycle). Fos+ cell number increased at ZT 17 and ZT 1 in the PVT and CMT, 180 degrees out of phase with the VLPO. A positive correlation was found between the numbers of Fos+ cells in the PVT and CMT, and Fos expression in each thalamic nucleus was negatively correlated with VLPO Fos+ cell number. The VLPO, PVT, and CMT may integrate circadian and homeostatic influences to regulate the sleep-wake cycle.

Behavioral changes and expression of heat shock protein hsp-70 mRNA, brain-derived neurotrophic factor mRNA, and cyclooxygenase-2 mRNA in rat brain following seizures induced by systemic administration of kainic acid

Kainic acid-induced seizures in rats represent an established animal model for human temporal lobe epilepsy. However, it is well-known that behavioral responses to the systemic administration of kainic acid are inconsistent between animals. In this study, we examined the relationship between expression of genes, neuropathological damage, and behavioral changes (seizure intensity and body temperature) in rats after systemic administration of kainic acid. The considerable differences in the response to kainic acid-induced seizures were observed in rats after a single administration of kainic acid (12 mg/kg i.p.). There was no detection of the expression of heat shock protein hsp-70 mRNA and HSP-70 protein in brain of vehicle-treated controls and in animals exhibiting weak behavioral changes (stage 1–2). A moderate expression of hsp-70 mRNA was detected throughout all regions (the pyramidal cell layers of CA1–3 and dentate gyrus) of the hippocampus, the basolateral, lateral, central and medial amygdala, the piriform cortex, and the central medial thalamic nucleus of rats that developed moderate seizures (stage 3–4). Marked expression of hsp-70 mRNA was detected in the all regions (cingulate, parietal, somatosensory, insular, entorhinal, piriform cortices) of cerebral cortex and all regions of hippocampus, and the central medial thalamic nucleus of the rats that developed severe seizures (stage 4–5). In addition, marked HSP-70 immunoreactivity was detected in the pyramidal cell layers of CA1 and CA3 regions of hippocampus, all regions (cingulate, parietal, somatosensory, insular, piriform cortices) of cerebral cortex, and the striatum of rats that developed severe seizures (stage 4–5). Furthermore, a marked expression of cyclooxygenase-2 (COX-2) mRNA and brain-derived neurotrophic factor (BDNF) mRNA levels by kainic acid-induced behavioral seizures (stage 3–4 or stage 4–5) was detected in all hippocampal pyramidal cell layers, granule layers of dentate gyrus, piriform cortex, neocortex, and amygdala. The present study suggest that the behavioral changes (seizure intensity and body temperature) and neuropathological damage after systemic administration of kainic acid are inconsistent between animals, and that these behavioral changes (severity of kainic acid-induced limbic seizures) might be correlated with gene expression of hsp-70 mRNA, COX-2 mRNA, and BDNF mRNA in rat brain.

Generalized Tonic Seizures Induced by a Unilateral Microinjection of Kainic Acid into the Pontine Reticular Formation in Rats

Purpose: Following Penfield and Jasper's proposal of the centrencephalic theory, the brainstem reticular formation was believed to be involved in the expression of generalized seizures and the generalization of partial seizures. In our study, an experimental animal model of generalized tonic seizures was produced by the unilateral microinjection of kainic acid (KA) into the pontine reticular formation (PRF) of Wistar rats. In addition, local cerebral glucose utilization (LCGU) was measured during the acute phase of the seizure by autoradiography by using [14C]2‐deoxyglucose to determine the extent of the hypermetabolic area in the brain induced by the seizure. Methods: Under general anesthesia with sodium pentobarbital, adult male Wistar rats weighing 280–320 g were implanted stereotaxially with a chemitrode in the left PRF and a monopolar electrode in the central medial thalamic nucleus. Stainless‐steel screw electrodes for recording the electrocorticogram were placed bilaterally on the dura over the sensorimotor cortex. After ≥1 week after surgery, 0.8 kg of KA was injected into the left PRF via the chemitrode. After the injection, the electroencephalographic and behavioral consequences were observed over a period of 7 days. For the LCGU study, the rats were unrestrained for 7 days after recovery from stereotaxic surgery. They were anesthetized with 1.5% halothane, and catheters were inserted into the femoral artery and vein. They were immobilized from the waist down by means of a loosefitting plaster cast and allowed to recover from anesthesia for ≥2 h. Then KA or a phosphate buffer solution was administered into the PRF via an implanted cannula. The LCGU study described by Sokoloff et al. was started 75 min later. Results: At an average of 16.9 min after the injection, the rats began to run and jump intermittently in the recording chamber (wild running). At an average of 22.5 min after the injection, they began to assume a tonic posture in which the upper body was strongly contorted to the left or backward and the four limbs were fully extended. About 2 h after the injection, wild running ceased, but the tonic seizure status persisted. Occasionally, clonic seizures were detected in the face or limbs during tonic seizures. The tonic seizures peaked at 3–4 h after the injection and then gradually recovered, completely disappearing within 24 h. On the EEG, at an average of 50.9 min after the KA injection, a high‐amplitude spike discharge appeared in the PRF and simultaneously spread bilaterally to the sensorimotor cortex. Synchronous interictal discharges persisted for 24 h after the injection but disappeared 3 days after the KA injection. LCGU was significantly augmented bilaterally in the PRF, mesencephalic reticular formation, periaqueductal gray, superior colliculus, raphe nucleus, locus ceruleus, zona incerta, medial forebrain bundle, CA1 and CA3 fields of the hippocampus, and amygdala. LCGU was also markedly increased in the lateral and medial geniculate nuclei, lateral septa1 nucleus, ventromedial nucleus of the hypothalamus, and lateral preoptic area on the side of the KA injection. The degree of LCGU in the entorhinal cortex and piriform cortex varied greatly. Conclusions: These results suggest that the PRF plays an important role in the primary and secondary generalization of tonic seizure. The epileptic discharge passed through the medial forebrain bundle from the PRF, entered the hypothalamus and basal forebrain, and affected the limbic system. When the epileptic discharge reaches the cerebral cortex from the PRF through this route, clonic seizures may occur.

Widespread expression of Fos protein induced by acute haloperidol administration in the rat brain.

The effect of acute haloperidol administration on Fos protein expression was examined immunohistochemically in discrete regions of the rat brain. Male Wistar rats were injected subcutaneously (s.c.) with 0.1, 0.25, or 1.0 mg/kg of haloperidol. Two h after the injection, the rats were perfused, and the numbers of Fos immunoreactive neurons were counted in 24 brain regions. In contrast to the limited changes in Fos immunoreactivity at the low dose of haloperidol (0.1 mg/kg), the moderate dose (0.25 mg/kg) induced widespread increases in Fos-positive neurons in the rat brain. Large increases were produced in the caudate-putamen, nucleus accumbens, central amygdaloid nucleus, dorsomedial hypothalamic nucleus, hippocampus CA1 and substantia nigra pars compacta. Moderate increases were observed in the entorhinal cortex, lateral septum, lateral habenula, lateral amygdaloid nucleus, dentate gyrus, and mesencephalic central grey. Mild increases were induced in the anterior cingulate, temporal, occipital and perirhinal cortex, and central medial thalamic nucleus. The distribution of changes in Fos immunoreactivity at the high dose of haloperidol (1.0 mg/kg) were comparable to their distribution at the moderate dose. These findings indicate that the effect of acute haloperidol on Fos expression is widely distributed in the rat brain beyond the previously known dopamine-rich areas at the dose which produces plasma levels equivalent to those within the therapeutic range used clinically in humans. Further studies on the effects of chronic antipsychotic treatment are needed in order to identify the sites of the therapeutic action of antipsychotic drugs.

Calretinin Immunoreactivity in the Developing Thalamus of the Rat: A Maker of Early Generated Thalamic Cells

The present work was aimed to study the immunocytochemical localization of the calcium-binding protein, calretinin, in the rat thalamus from embryonic day 14 to the third postnatal week. In the adult rat thalamus, calretinin immunoreactivity is intensely expressed in some intralaminar and midline nuclei, as well as in selected regions of the reticular nucleus. At embryonic day 14, calretinin was expressed by immature and migrating neurons and fibres laterally to the neuroepithelium of the diencephalic vesicle in the region identified as reticular neuroepithelium. At embryonic day 16, immunoreactive neurons were present in the primordium of the reticular nucleus and in the region of the reticular thalamic migration, where neurons showed the morphology of migratory cells. At the end of embryonic development and in the first postnatal week, calretinin-positive neurons were observed in selected region of the reticular nucleus and it was intensely expressed in some intralaminar and midline nuclei. Bands of immunopositive fibres were also observed crossing the thalamus. During the second postnatal week, the immunolabelling in the reuniens, rhomboid, paraventricular and central medial thalamic nuclei remains very intense while a decrease of immunoreactivity in mediodorsal, centrolateral and laterodorsal nuclei was observed. The immunostaining of fibres, particularly evident in the perinatal period, progressively decreased and it was no longer visible by the end of the second postnatal week when the distribution and intensity of calretinin immunostaining was similar to that observed in the adult rat thalamus. The present findings indicate that the immunolocalization of calretinin can be used to identify subsets of thalamic neuronal population during pre- and postnatal maturation allowing also the detection of the migratory pattern of early generated reticular thalamic neurons.

Quantitative Autoradiography on [(35)S]TBPS Binding Sites of Gamma- Aminobutyric Acid(A) Receptors in Discrete Brain Regions of High- Alcohol-Drinking and Low-Alcohol- Drinking Rats Selectively Bred forHigh- and Low-Alcohol Preference.

It has been documented that ethanol can potentiate brain gamma-aminobutyric acid (GABA)ergic function, and there is a close link between the GABA(A) receptor complex and effects of ethanol, including reinforcement of alcohol which is a fundamental element of alcohol preference. However, it is unknown in what discrete brain regions GABA(A) receptors might be associated with alcohol preference. In the present study, [(35)S]t-butylbicyclophosphorothionate ([(35)S]TBPS) was used to localize GABA(A) receptors in high-alcohol-drinking (HAD) rats and low-alcohol-drinking (LAD) rats which were selectively bred for high and low alcohol preference, respectively. Initial qualitative observations indicated that [(35)S]TBPS binding sites were abundant in many brain areas including the cerebral cortex, hypothalamus and amygdala of HAD and LAD rats. Furthermore, the quantitative autoradiographic analysis revealed fewer [(35)S]TBPS binding sites of GABA(A) receptors in the amygdaloid complex, central medial thalamic nucleus, lateral hypothalamic nucleus and anterior hypothalamic nucleus of HAD rats than LAD rats. Collectively, this study has indicated that HAD rats selectively bred for high alcohol preference possess lower [(35)S]TBPS binding in the brain. Since lower TBPS binding has been proposed to reflect enhanced GABAergic function, as evidenced in rats with seizure or under alcohol withdrawal, the results from the present study suggest that HAD rats might have an enhanced GABAergic function. It is thus likely that enhanced GABAergic function in the brain might be related to high alcohol preference which is characteristic in HAD rats. In addition, the present result showing no difference of [(35)S]TBPS binding in the nucleus accumbens is also in agreement with a notion that [(35)S]TBPS binding may represent only a small spectrum of the GABA(A) receptor complex which is constituted of a sophisticated subunit combination whose functional compositions are still unknown. In conclusion, the present study supports the working hypothesis that GABA(A) receptors are involved in alcohol preference in HAD rats.

A comparison of the ultrastructure of synapses in the cerebello-rubral and cerebello-thalamic pathways in the rat

The aim of this study was to compare the ultrastructure of anterogradely labelled cerebellar terminals in the red nucleus (RN), ventrolateral (VL), parafascicular (PF) and central medial (CM) thalamic nuclei, as well as in the zona incerta (ZI). No differences were found in the morphology of synapses in any of the nuclei. Terminals in RN and VL were larger than those in PF, CM and ZI and synapsed proximally. In contrast, synapses in PF, CM and ZI formed mainly on distal dendrites. These findings indicate that cerebellar output neurones (a) form morphologically similar synapses (Gray's type I) on neurones in functionally different nuclei, and (b) form larger, more proximal synapses in RN and VL than in PF, CM and ZI.

Effect of the tetrapeptide tuftsin on the bioelectrical activity of brain structures in cats with different functional states of the central nervous system

The tetrapeptide tuftsin (Thr-Lys-Pro-Arg) injected intraperitoneally into cats altered the bioelectrical activity of sensory, motor, and limbicoreticular structures in the wakeful state, during two different phases of sleep, and also when the caudate nucleus was electrically stimulated. In wakefulness and during REM sleep, absolute values of the electroencephalographic power spectrum were increased in all the brain structures under study, while during slow-wave sleep they were increased in the sensorimotor cortical areas and decreased in the central medial thalamic nuclei and hippocampus. Tuftsin facilitated the genesis of slow waves in subcortical structures and the expression of theta waves in the hippocampus. It also increased the amplitude of evoked potentials and elongated spindles when the caudate nucleus was stimulated.

The efferent projections of the periaqueductal gray in the rat: A Phaseolus vulgaris‐leucoagglutinin study. I. Ascending projections

This study has examined the ascending projections of the periaqueductal gray in the rat. Injections of Phaseolus vulgaris-leucoagglutinin were placed in the dorsolateral or ventrolateral subregions, at rostral or caudal sites. From either region, fibers ascended via two bundles. The periventricular bundle ascended in the periaqueductal and periventricular gray matter. At the posterior commissure level, this bundle divided into a dorsal component that terminated in the intralaminar and midline thalamic nuclei, and a ventral component that supplied the hypothalamus. The ventral bundle formed in the deep mesencephalic reticular formation and supplied the ventral tegmental area, substantia nigra pars compacta, and the retrorubral field. The remaining fibers were incorporated into the medial forebrain bundle. These supplied the lateral hypothalamus and forebrain structures, including the preoptic area, the nuclei of the diagonal band, and the lateral division of the bed nucleus of the stria terminalis. The dorsolateral subregion preferentially innervated the centrolateral and paraventricular thalamic nuclei and the anterior hypothalamic area. The ventrolateral subregion preferentially innervated the parafascicular and central medial thalamic nuclei, the lateral hypothalamic area, and the lateral division of the bed nucleus of the stria terminalis. Although the dorsolateral and ventrolateral subregions gave rise to differential projections, the projections from both the rostral and caudal parts of either subregion were similar. This suggests that the dorsolateral and ventrolateral subregions are organized into longitudinal columns that extend throughout the length of the periaqueductal gray. These columns may correspond to those demonstrated in recent physiological studies.

A Novel Rat Serotonin Receptor: Primary Structure, Pharmacology, and Expression Pattern in Distinct Brain Regions

Serotonin mediates various central and peripheral functions by its ability to bind to and activate a variety of receptors that belong to four pharmacologically defined classes termed 5-HT1 to 5-HT4. Using a polymerase chain reaction-derived probe, a clone, GPRFO, was isolated from a rat brain cDNA library. This cDNA encodes a putative G protein-coupled receptor that exhibits highest similarity to receptors for biogenic amines. RNA blot analysis indicated that the corresponding gene is expressed in the diencephalon. Consistent with the RNA blot data, GPRFO mRNA has been detected by in situ hybridization in the centrolateral, central medial, and intermediodorsal thalamic nuclei. Highest amounts of GPRFO mRNA, however, have been observed in a small area, i.e., the hippocampal rudiments and the stria longitudinalis. COS-7 cells transiently transfected with the GPRFO cDNA acquire saturable high-affinity binding sites for [3H]serotonin (KD = 41 nM). The pharmacological properties of the receptor differ from those of the known serotonin receptor subtypes, suggesting that the GPRFO cDNA encodes a novel serotonin receptor that is expressed in distinct rat brain regions.

Ontogeny of bombesin/gastrin-releasing peptide binding sites in rat brain

The development of bombesin/gastrin-releasing peptide (BN/GRP) binding sites was determined in the rat brain. Using rat brain homogenate, the density of radio-labeled peptide binding sites increased dramatically around birth, remained constant during the first post-natal week, and then increased again between Postnatal (P) Days 7 and 10; the density of binding sites was similar in P10 and adult animals. Rat brain homogenates derived from P1, P7, or P10 animals bound ( 125I-Tyr 4)BN with high affinity ( K d = 2–4 n M) to a single class of sites ( B max = 11, 17, and 51 pmol/mg protein, respectively). BN, GRP, and GRP 18–27 inhibited specific radiolabeled peptide binding to rat brain homogenate with high affinity using P1, P3, P7, or P14 animals, whereas GRP 1–16 was inactive. These data indicate that the C-terminal of BN or GRP is essential for high-affinity binding activity in the rat brain during development. In vitro autoradiography revealed high ( 125I-Tyr 4)BN grain densities in the anterior olfactory nucleus, olfactory tubercle, nucleus accumbens, central medial thalamic nucleus, and hippocampus. Moderate grain densities were observed in the parietal cortex, periventricular hypothalamic nucleus, central gray, inferior colliculus, pontine reticular, and gigantocellular reticular nuclei in P1, P3, and P7 rat brains. In P14 and adult animals, relative to P1, P3, or P7 rat brains, the density of binding sites increased significantly in the parietal cortex. The ( 125I-Tyr 4)BN grain density was not significantly changed in the anterior olfactory nucleus, periventricular nucleus of the hypothalamus, olfactory tubercle, hippocampus, dentate gyrus, central medial thalamic nucleus, and nucleus accumbens but decreased significantly in the central grey, inferior colliculus, and pontine reticular nucleus. These data indicate that there is a redistribution of BN/GRP binding sites in the rat brain during development.

A PHA‐L analysis of ascending projections of the dorsal raphe nucleus in the rat

Ascending projections from the dorsal raphe nucleus (DR) were examined in the rat by using the anterograde anatomical tracer, Phaseolus vulgaris leucoagglutinin (PHA-L). The majority of labeled fibers from the DR ascended through the forebrain within the medial forebrain bundle. DR fibers were found to terminate heavily in several subcortical as well as cortical sites. The following subcortical nuclei receive dense projections from the DR: ventral regions of the midbrain central gray including the 'supraoculomotor central gray' region, the ventral tegmental area, the substantia nigra-pars compacta, midline and intralaminar nuclei of the thalamus including the posterior paraventricular, the parafascicular, reuniens, rhomboid, intermediodorsal/mediodorsal, and central medial thalamic nuclei, the central, lateral and basolateral nuclei of the amygdala, posteromedial regions of the striatum, the bed nucleus of the stria terminalis, the lateral septal nucleus, the lateral preoptic area, the substantia innominata, the magnocellular preoptic nucleus, the endopiriform nucleus, and the ventral pallidum. The following subcortical nuclei receive moderately dense projections from the DR: the median raphe nucleus, the midbrain reticular formation, the cuneiform/pedunculopontine tegmental area, the retrorubral nucleus, the supramammillary nucleus, the lateral hypothalamus, the paracentral and central lateral intralaminar nuclei of the thalamus, the globus pallidus, the medial preoptic area, the vertical and horizontal limbs of the diagonal band nuclei, the claustrum, the nucleus accumbens, and the olfactory tubercle. The piriform, insular and frontal cortices receive dense projections from the DR; the occipital, entorhinal, perirhinal, frontal orbital, anterior cingulate, and infralimbic cortices, as well as the hippocampal formation, receive moderately dense projections from the DR. Some notable differences were observed in projections from the caudal DR and the rostral DR. For example, the hippocampal formation receives moderately dense projections from the caudal DR and essentially none from the rostral DR. On the other hand, virtually all neocortical regions receive significantly denser projections from the rostral than from the caudal DR. The present results demonstrate that dorsal raphe fibers project significantly throughout widespread regions of the midbrain and forebrain.

Fibroblast growth factor receptor-bearing neurons in the CNS: identification by receptor-mediated retrograde transport.

Neurons that internalize and retrogradely accumulate acidic (aFGF) or basic (bFGF) fibroblast growth factor were identified by autoradiography after injections of 125 I-aFGF or 125I-bFGF into the adult rat central nervous system (CNS). Neuronal cell bodies within the lateral hypothalamus, pedunculpontine tegmental nucleus, laterodorsal tegmental nucleus, and the paracentral dorsal tegmental nucleus accumulated 125I-aFGF. Neurons in the hippocampus, subiculum, the centrolateral, paracentral, central medial, and parafascicular thalamic nuclei, the supramammillary nucleus, and substantia nigra compacta accumulated 125I-bFGF. The pattern of neuronal labeling with 125I-bFGF in adult rats was similar to that observed in newborn guinea pigs. No 125I-FGF labeling was observed in nerve growth factor (NGF) receptor-bearing neurons, including the basal forebrain cholinergic neurons. Time-course studies indicate that 125I-FGF was internalized at the terminals and retrogradely transported to the neuronal cell bodies. Neurons were retrogradely labeled either by injection of 125I-bFGF into the lateral ventricle or by injection into innervated target tissues. Co-injection of a 250-fold excess of unlabeled FGF with the 125I-FGF abolished the neuronal labeling. Co-injection of wheat germ agglutinin (WGA), which nonspecifically blocks binding of 125I-bFGF to its receptor, also prevented neuronal labeling. These studies demonstrate that specific neuronal populations within the CNS express functional receptors for aFGF and/or bFGF; in these neurons, aFGF and/or bFGF bind specifically to these receptors, are internalized and retrogradely transported to the neuronal soma in a manner analogous to NGF. The data indicate that FGF can provide trophic support to CNS neurons by both direct and indirect mechanisms.