Distribution of somatostatinlike immunoreactivity in the brain of the little brown bat (Myotis lucifugus)

The distribution of somatostatinlike immunoreactive (SLI) perikarya, axons, and terminals was mapped in subcortical areas of the brain of the little brown bat, Myotis lucifugus, using light microscopic immunocytochemistry. A preponderance of immunoreactivity was localized in reticular, limbic, and hypothalamic areas including: 1) in the forebrain: the bed nucleus of the stria terminalis; lateral preoptic, dorsal, anterior, lateral and posterior hypothalamic areas; amygdaloid, periventricular, arcuate, supraoptic, suprachiasmatic, ventromedial, dorsomedial, paraventricular, lateral and medial mammillary, and lateral septal nuclei; the nucleus of the diagonal band of Broca and nucleus accumbens septi; 2) in the midbrain: the periaqueductal gray, interpeduncular, dorsal and ventral tegmental, pretectal, and Edinger-Westphal nuclei; and 3) in the hindbrain: the superior central and parabrachial nuclei, nucleus incertus, locus coeruleus, and nucleus reticularis gigantocellularis. Other areas containing SLI included the striatum (caudate nucleus and putamen), zona incerta, infundibulum, supramammillary and premammillary nuclei, medial and dorsal lateral geniculate nuclei, entopeduncular nucleus, lateral habenular nucleus, central medial thalamic nucleus, central tegmental field, linear and dorsal raphe nuclei, nucleus of Darkschewitsch, superior and inferior colliculi, nucleus ruber, substantia nigra, mesencephalic nucleus of V, inferior olivary nucleus, inferior central nucleus, nucleus prepositus, and deep cerebellar nuclei. While these results were similar in some respects to those previously reported in rodents, they also provided interesting contrasts.     

The efferent connections of the lateral septal nucleus in the guinea pig: projections to the diencephalon and brainstem

Summary The anterograde Phaseolus vulgaris-leucoagglutinin (PHA-L) tracing technique was used to determine the distribution of efferent fibers originating in the lateral septal nucleus of the guinea pig. For complementary detection of the chemical identity of the target neurons, double-labeling immunocytochemistry was performed with antibodies to PHA-L and to vasopressin, oxytocin, vasoactive intestinal polypeptide, serotonin or dopamine -hydroxylase, respectively. The hypothalamus received the majority of the PHA-L-stained septofugal fibers. Here, a specific topography was observed. (1) The medial and lateral preoptic area, (2) the anterior, lateral, dorsal, posterior hypothalamic and retrochiasmatic area, (3) the supraoptic, paraventricular, suprachiasmatic, dorsomedial, caudal ventromedial and arcuate nuclei, and (4) the tuberomammillary, medial and lateral supramammillary, dorsal and ventral premammillary nuclei always contained PHA-L-labeled fibers. The rostral portion of the ventromedial nucleus and the medial and lateral mammillary nucleus only occasionally showed weak terminal labeling. In other diencephalic areas, termination of PHA-L-labeled fibers was observed in the epithalamus and the nuclei of the midline region of the thalamus. In the mesencephalon, terminal varicosities occurred in the ventral tegmental area, interfascicular and interpeduncular nucleus, and periaqueductal gray. In addition, the dorsal and medial raphe nuclei of the metencephalon, together with the locus coeruleus and the dorsal tegmental nucleus, received lateral septal efferents.     

Pattern of afferents to the lateral septum in the guinea pig

Summary The septal region represents an important telencephalic center integrating neuronal activity of cortical areas with autonomous processes. To support the functional analysis of this brain area in the guinea pig, the afferent connections to the lateral septal nucleus were investigated by the use of iontophoretically applied horseradish peroxidase (HRP). Retrogradely labeled perikarya were located in telencephalic, diencephalic, mesencephalic and metencephalic sites. The subnuclei of the lateral septum (pars dorsalis, intermedia, ventralis, posterior) receive afferents from the (i) medial septal nucleus, diagonal band of Broca (pars horizontalis and pars ventralis), and the principal nucleus of the stria terminalis, the hippocampus, and amygdala (nucleus medialis); (ii) the medial habenular nucleus, and the para- (peri-) ventricular, parataenial and reuniens nuclei of the thalamus; the anterior, lateral and posterior hypothalamic areas in particular, the medial and lateral preoptic, suprachiasmatic, periventricular, paraventricular, arcuate, premammillary, and supramammillary nuclei; (iii) the periaquaeductal grey, ventral tegmental area, nucleus interfascicularis, nucleus reticularis linearis, central linear nucleus, interpeduncular nucleus; (iv) dorsal and medial raphe complex, and locus coeruleus. Each subnucleus of the lateral septum displays an individual, differing pattern of afferents from the above-described regions. Based on a double-labeling method, the vasopressinergic and serotonergic afferents to the lateral septum were found to originate in the nucleus paraventricularis hypothalami and the raphe nuclei, respectively.Abbreviations ARC arcuate nucleus - BNST bed nucleus of the stria terminalis - CL central linear nucleus - DBBh diagonal band of Broca (pars horizontalis) - DBBv diagonal band of Broca (pars ventralis) - DR dorsal raphe nucleus - HC hippocampus - IF interfascicular nucleus - IP interpeduncular nucleus - LC locus coeruleus - LDT laterodorsal tegmental nucleus - LHA lateral hypothalamic area - LPO lateral preoptic area - LSN lateral septal nucleus - MA medial amygdaloid nucleus - MH medial habenular nucleus - MPO medial preoptic region - MR medial raphe nucleus - MSN medial septal nucleus - PAG periaquaeductal grey - PEN periventricular nucleus - PHA posterior hypothalamic area - PMd premammillary region (pars dorsalis) - PMv premammillary region (pars ventralis) - PT parataenial nucleus - PVN paraventricular hypothalamic nucleus - PVT paraventricular thalamic nucleus - RE nucl. reuniens - RL nucl. reticularis linearis - SCN suprachiasmatic nucleus - SMl supramammillary region (pars lateralis) - SMm supramammillary region (pars medialis) - SUB subiculum - TS triangular septal nucleus - VTA ventral tegmental area - ac anterior commissure - bc brachium conjunctivum - bp brachium pontis - cc corpus callosum - fr fasciculus retroflexus - fx fornix - ml medial lemniscus - mlf fasciculus longitudinalis medialis - mp mammillary peduncle - mt mammillary tract - oc optic chiasm - on optic nerve - pc posterior commissure - pt pyramidal tract - sm stria medullaris - st stria terminalis - vhc ventral hippocampal commissure Supported by the Deutsche Forschungsgemeinschaft (Nu 36/2-1)     

Blood supply of the rat hypothalamus. VI. Posterior region of the hypothalamus (nucleus hypothalamicus posterior, nuclei praemammillares, nucleus supramammilaris, mammilary body).

It was shown by the double ink-filling technique that the arteries of the rat premammillary region and mammillary body arise from the a. communicans posterior while these areas are drained by the anterior interpeduncular vein. Disregarding some minor overlaps and anastomoses, the blood supplies of the two territories are independent of each other and from the neighbouring areas of the hypothalamus, diencephalon and mesencephalon. Arteries of the premammillary region arise from the premammillary artery, except for some branches of the posterior tuberal and interpeduncular arteries. The mammillary body is supplied by three mammillary arteries (anterior, posterior and lateral). The premammillary region drains into the anterior and posterior premammillary veins. Venous blood of the mammillary body is collected by the anterior and posterior mammillary veins which end in the anterior interpeduncular vein. The circulation of individual premammillary and mammillary nuclei is described in detail.     

A horseradish peroxidase study on the mammillothalamic tract in the rat

The mammillary projections to the anterior thalamic nuclei were investigated in the rat, using the horseradish peroxidase (HRP) method. Pars centralis of the medial mammillary nucleus projects to the medial portion of the ateromedial nucleus (AM). Pars medialis (Mm) of the medial mammillary nucleus sends fibers to the ipsilateral AM and sparsely to the medial portion of the contralateral side. The ventral and dorsal portions of Mm project to the anterior and posterior portions of AM, respectively. The pars latralis (Ml) and pars posterior (Mp) of the medial mammillary nucleus send fibers predominantly to the ipsilateral anteroventral nucleus and sparsely to the contralateral side. A slight difference between Ml and Mp projections was observed. The lateral mammillary nucleus projects bilaterally to the anterodorsal nucleus.     

Cholecystokinin receptor binding levels in the genetically obese rat brain

Cholecystokinin (CCK) receptor binding levels were compared between groups of genetically obese (fa/fa) and non-obese (Fa/-) Zucker rats of both sexes. The radioligand used was the iodinated octapeptide (CCK-8). Binding was measured in eight brain regions. The relative distribution among different brain regions of specifically bound CCK per mg protein was similar in all groups of animals. High binding levels were present in the olfactory bulb, cortex and caudate nucleus. Moderate levels were seen in hippocampus and hypothalamus, and low levels were observed in hindbrain, midbrain and thalamus. Obese animals of both sexes had significantly higher CCK receptor binding levels in the hippocampus and in the midbrain in comparison to lean controls. The male obese animals also had significantly elevated binding levels in the thalamic sample. These results demonstrate a correlation between genetic obesity and elevated CCK receptor binding levels in specific brain regions.     

Neuropeptide Y Y1 receptors in the rat forebrain: Autoradiographic demonstration of [125I][Leu31,Pro34]-NPY binding sites and neurons expressing Y1 receptor mRNA

Abstract

Using the specific monoiodinated NPY analog [Leu³¹,Pro³⁴]-NPY we have localized NPY binding sites of the Y₁ type in forebrain areas of the rat. The resulting receptor autoradiograms were compared with the regional distribution and cellular localization of the mRNA encoding Y₁ receptor as demonstrated by in situ hybridization histochemistry. High densities of Y₁ binding sites were present in the cerebral cortex, the claustrum, the thalamus and the medial mammillary nucleus, while moderate densities of Y₁ binding sites were observed in the amygdalahippocampal complex. Lower binding densities were observed in septal nuclei, most hypothalamic nuclei and the circumventricular organs. High levels of Y₁ mRNA were observed in the granula cell layer of the hippocampal dentate gyrus, several thalamic nuclei and the hypothalamic arcuate nucleus, while moderate levels of Y₁ mRNA were seen in the frontoparietal cortex, several thalamic nuclei, the hippocampal pyramidal layers, the subiculum, the olfactory tubercle, the claustrum and a number of hypothalamic nuclei. Using the hypothalamic arcuate nucleus as an example, the distribution of immunoreactive NPY, Y₁ mRNA and Y₁ binding sites was compared, and possible implications of Y₁ mediated actions within this nucleus are discussed. The present study further enlightens the anatomical distribution of NPY binding sites of the Y₁ type within the central nervous system of the rat, and extends the understanding of central actions of NPY mediated via this type of receptor.     

Corticotropin-releasing factor (CRF)-immunoreactive neurons in the mammillary body of the rat

Summary The presence and distribution of CRF-immunoreactive cells and nerve fibers were studied in the mammillary body of the rat, 12 days after placing various types of lesions within the hypothalamus. Anterior and anteriolateral cuts, placed in the midhypothalamus immediately behind the paraventricular nuclei resulted in an almost complete disappearance of CRF-immunoreactive fibers from the median eminence and simultaneous appearance of CRF-containing neurons in the mammillary body. Posterior or postero-lateral hypothalamic cuts carried out in front of the mammillary body caused the accumulation of CRF-immunoreactive material in neurons and neural processes located behind the cut-line. This type of intervention had no effect on the quantity of CRF fibers in the median eminence. A cut running through the central part of the mammillary body in the frontal plane resulted in appearance of CRF neurons only in the posterior half of the mammillary region. Placing a cut behind and over the mammillary body, CRF-immunoreactive neurons became detectable below the superior cut-line. No immunoreactive neurons were observed in the mammillary body when the frontal cut reached the base of the brain at the posterior border of the nucleus, leaving intact its anterior and superior connections. In all these cases when the mammillo-thalamic tract was transected, CRF neurons became detectable in the mammillary body.     

Correlation between morphology and function in the rat hippocampus and hypothalamus

Correlation between morphology and function in the hippocampus and hypothalamus was studied by electrophysiological and morphological techniques. Single unit responses were recorded extracellularly in the arcuate and medial preoptic nuclei of the hypothalamus to application of single stimuli to the hippocampus. Phasic responses and primary inhibition predominated in the arcuate nucleus, whereas both phasic and tonic responses were observed in the medial preoptic nucleus. In the morphological experiments horseradish peroxidase was injected into the same region of the hippocampus. Stained cells were found in the nuclei of the mammillary body, mediobasal hypothalamus, and medial preoptic nucleus. Groups of stained neurons were discovered at the periphery of the ventro- and dorsomedial and also in the lateral and mammillary nuclei of the hypothalamus. Besides fusiform and triangular neurons, reticular neurons also were found in all structures except the medial mammillary nucleus. The results are discussed from the standpoint of interaction between hypothalamus and hippocampus.A. A. Zhdanov State University, Leningrad. Translated from Neirofiziologiya, Vol. 11, No. 5, pp. 427–434, September–October, 1979.     

Chondroitin sulfate proteoglycans in the rat thalamus: expression during postnatal development and correlation with calcium-binding proteins in adults

Postnatal expression of chondroitin sulfate proteoglycans was studied in the rat thalamus by immunocytochemistry and Western immunoblotting techniques with monoclonal antibodies that recognize carbohydrate epitopes (clones CS-56, 1-B-5, 2-B-6). The complex of the results shows that these antibodies recognize mostly nonoverlapping molecules whose expression is regulated during postnatal development. Chondroitin sulfate proteoglycans, recognized by antibody CS-56, and hyaluronan, identified by antibody 1-B-5 after hyaluronidase digestion, are abundant in the neuropil of most thalamic nuclei at the perinatal stage and progressively decrease during the second week of life, attaining levels barely detectable by immunocytochemistry at the end of the third week. In adult thalamus, chondroitin sulfate proteoglycans of high molecular mass, bearing glycosaminoglycans unsulfated in the linking region, and recognized by antibody 1-B-5 are confined to perineuronal nets around neurons chiefly localized in thalamic reticular nucleus. The immunoreactvity for antibody 2-B-6, specific for chondroitin-4-sulfate, is low at the perinatal stage and is not detectable in adult thalamus. Double-immunolabeling has shown that, along the rostrocaudal extension of reticular nucleus, the most developed perineuronal nets are associated with a subset of neurons expressing calretinin, and not with parvalbumin-positive neurons, which represent the largest neuronal population of the nucleus. The distribution of perineuronal nets supports the presence, in thalamic reticular nucleus, of neuronal subpopulations with different morphological and physiological features.     

Histamine-immunoreactive neurons in the hypothalamus of cats

The localization of histaminergic neurons in the cat brain was determined immunohistochemically with an antibody against histamine. We found that histamine-immunoreactive neurons are observed exclusively in the posterior hypothalamus of colchicine treated cats. The larger group of neurons was found in the ventrolateral part of the posterior hypothalamus, including the tuberomammillary nucleus. Histamine-positive neurons were also observed in the supramammillary area and adjacent posterior hypothalamic area, as well as in the peri- and premammillary regions. In addition, numerous histamine immunoreactive fibers were detected, not only in the posterior hypothalamus, but also in other brain areas, such as the preoptic area of the anterior hypothalamus.     

Induction of CCK mRNA levels in the limbic-hypothalamic circuit: Time course and site-specific effects of estrogen

Estrogenic regulation of cholecystokinin (CCK) and its receptors is correlated with the initiation and termination of lordosis behavior. To understand the effect of circulating estrogen concentration on the temporal aspects of CCK mRNA expression in the posterodorsal medial amygdaloid nucleus (MeApd) and the central part of the medial preoptic nucleus (MPNc) of the limbic-hypothalamic circuit, ovariectomized female rats were treated with a 10 mm Silastic™ capsule filled with estradiol, a bolus injection of 50 μg estradiol benzoate or 2 μg estradiol benzoate every 4 days for five “cycles.” In situ hybridization was used to compare the relative changes of CCK mRNA levels at 0 h to levels measured at 6, 12, 24, 48, 72, or 96 h after estrogen administration. In the MPNc and the MeApd, the 10-mm capsule significantly increased and maintained CCK mRNA levels from 6 to 96 h. The range of the increase was 3.0–5.1-fold in the MPNc and 2.8–5.0 in the MeApd. The 50-μg injections significantly increased and maintained CCK mRNA levels in the MPNc from 12 to 96 h (range of the increase 2.4–4.1-fold) and in the MeApd from 24 to 96 h (range of the increase 2.2–2.8-fold). The repeated administration of 2 μg estrogen induced a significant increase of message levels in the MPNc at 12 and 24 h that were 4.2- and 4.7-fold, respectively. In the MeApd this estrogen treatment did not significantly increase CCK mRNA. These studies demonstrate that small doses (2 μg) of estrogen that mimic the pattern and circulating levels of estrogen dramatically stimulate CCK mRNA levels in the limbic-hypothalamic circuit. To further study this steroid stimulation, ovariectomized female rats were implanted with estradiol-filled cannulae into the bed nucleus of the stria terminalis or MeA. Estrogen elevated CCK mRNA levels locally in each nucleus. Implants in the bed nucleus also elevated CCK mRNA levels in the MeApd indicating that physiologic estrogen stimulation of CCK in the MeApd is the result of both local and distal transsynaptic elevation of CCK mRNA levels. The site-specific induction of CCK mRNA levels within the limbic-hypothalamic nuclei provides another important facet of estrogenic modulation of CCK induction. © 1996 John Wiley & Sons, Inc.     

Branching-pattern analysis of the dendritic arborization in the thalamic nuclei of the rat brain

We investigated the dendritic patterns of rapid Golgi-impregnated, highly similar multipolar neurons from two functionally different thalamic regions of the rat brain: two dorsal nuclei (the nucleus laterodorsalis thalami, pars dorsomedialis and the nucleus laterodorsalis thalami, pars ventrolateralis), and two ventral nuclei (the nucleus ventrolateralis thalami and the nucleus ventromedialis thalami). The analysis involved conventional morphometric parameters (height and size) and a new parameter derived from graph theory, the relative imbalance (RI), derived from the branching patterns of the dendrites, which permits quantitative characterization of the dendritic arborization of a neuron. On this basis, neurons can be grouped into three fundamentally different types: type A, or highly-polarized (imbalanced) neurons (RI values close to 1); type B, or medium-polarized neurons (RI values around 0.5); and type C, or balanced neurons with low polarization (RI values close to 0). The orientations of the dendritic arbor, and thus the receptive fields, of the dorsal and ventral thalamic neurons, were mutually perpendicular. The H and S values indicated that the neurons in the dorsal and ventral thalamic nuclei differed significantly. However, their RI values demonstrated that they were similar neurons of type B. Our data reveal that 1 ) the dendritic arbor cannot be reliably characterized purely on the basis of height and size, and 2) RI is a valuable morphometric parameter that identifies the true nature of the dendritic arborization.     

Somatostatin-like immunoreactivity in the amygdala of the pig

The distribution and morphology of neurons containing somatostatin (SOM) was investigated in the amygdala (CA) of the pig. The SOM-immunoreactive (SOM-IR) cell bodies and fibres were present in all subdivisions of the porcine CA, however, their number and density varied depending on the nucleus studied. The highest density of SOM-positive somata was observed in the layer III of the cortical nuclei, in the anterior (magnocellular) part of the basomedial nucleus and in the caudal (large-celled) part of the lateral nucleus. Moderate to high numbers of SOM-IR cells were also observed in the medial and basolateral nuclei. Many labeled neurons were also consistently observed in the lateral part of the central nucleus. In the remaining CA regions, the density of SOM-positive cell bodies varied from moderate to low. In any CA region studied SOM-IR neurons formed heterogeneous population consisting of small, rounded or slightly elongated cell bodies, with a few poorly branched smooth dendrites. In general, morphological features of these cells clearly resembled the non-pyramidal Golgi type II interneurons. The routine double-labeling studies with antisera directed against SOM and neuropeptide Y (NPY) demonstrated that a large number of SOM-IR cell bodies and fibers in all studied CA areas contained simultaneously NPY. In contrast, co-localization of SOM and cholecystokinin (CCK) or SOM and vasoactive intestinal polypeptide (VIP) was never seen in cell bodies and fibres in any of nuclei studied. In conclusion, SOM-IR neurons of the porcine amygdala form large and heterogeneous subpopulation of, most probably, interneurons that often contain additionally NPY. On the other hand, CCK- and/or VIP-IR neurons belonged to another, discrete subpopulations of porcine CA neurons.     

Neuronal Types in the Human Anterior Ventral Thalamic Nucleus: A Golgi Study

Neurons in the anterior ventral (AV) thalamic nucleus of human adults were impregnated by Golgi-Kopsch impregnation method. Results showed that at least three morphological types of neurons could be recognized in the human AV thalamic nucleus. Type I neurons were medium to large with rich dendritic arborization. Both tufted and radiating dendritic branching patterns were seen in almost every neuron of this type. Only the initial axonal segments of these cells were impregnated suggesting that these axons were heavily myelinated. Type II neurons were medium in size with poor to moderate dendritic arborization. Many of these cells possess a few dendritic grape-like appendages. Long segments (up to 300 μm) of their axons were impregnated suggesting that these axons were either unmyelinated or thinly myelinated. These axons change their direction and form loops very often. No local branches were seen for these axons suggesting that they could be projection axons. Type III neurons were small with only one or two dendrites with poor arborization. No axons for these cells were seen in this study. The three neuronal types in the human AV thalamic nucleus were compared with neuronal types already described in other thalamic nuclei of human and non-human species. The results of this study might provide a morphological basis for further electrophysiological and / or pathological studies.     

1,25 (OH)2 vitamin D3 sites of action in the brain. An autoradiographic study

After injection of 3H 1,25 (OH)2 vitamin D3 to adult rats and mice, under normal or vitamin D deficient diet, the hormone was found to be accumulated in nuclei of neurons in certain brain regions. Nuclear concentration was prevented or diminished, when excess unlabeled 1,25 (OH)2 vitamin D3 was injected before 3H 1,25 (OH)2 vitamin D3, while excess 25 (OH) vitamin D3 did not prevent nuclear labeling. Highest nuclear concentration of 3H 1,25 (OH)2 vitamin D3 is observed in certain neurons in the nucleus interstitialis striae terminalis, involving its septo-preoptic pars dorsolateralis and its anterior hypothalamic-thalamic portion, and in the nucleus centralis of the amygdala, all constituting a system of target neurons linked by a component of the stria terminalis. Nuclear concentration of 3H 1,25 (OH)2 vitamin D3 is also found in neurons in the periventricular nucleus of the preoptic-hypothalamic region, including its extensions, the parvocellular paraventricular and arcuate nucleus, in the ventromedial nucleus, supramammillary nucleus, reticular nucleus of the thalamus, ventral hippocampus, caudate nucleus, pallium, in the midbrain-pontine central gray, dorsal raphe nucleus, parabrachial nuclei, cranial motor nuclei, substantia gelatinosa of the sensory nucleus of the trigeminus, Golgi type II cells of the cerebellum, and others. The extensive distribution of target neurons suggests that 1,25 (OH)2 vitamin D3 regulates the production of several aminergic and peptidergic messengers, and influences the activity of certain endocrine-autonomic, sensory and motor systems.     

Genetic inactivation of prohormone convertase (PC1) causes a reduction in cholecystokinin (CCK) levels in the hippocampus, amygdala, pons and medulla in mouse brain that correlates with the degree of colocalization of PC1 and CCK mRNA in these structures in rat brain

Prohormone convertase (PC1) is found in endocrine cell lines that express cholecystokinin (CCK) mRNA and process pro CCK to biologically active products. Other studies have demonstrated that PC1 may be a one of the enzymes responsible for the endoproteolytic cleavages that occur in pro CCK during its biosynthesis and processing. Prohormone convertase 1 (PC1) has a distribution that is similar to cholecystokinin (CCK) in rat brain. A moderate to high percentage of CCK mRNA-positive neurons express PC1 mRNA. CCK levels were measured in PC1 knockout and control mice to assess the degree to which loss of PC1 changed CCK content. CCK levels were decreased 62% in hippocampus, 53% in amygdala and 57% in pons-medulla in PC1 knockout mice as compared to controls. These results are highly correlated with the colocalization of CCK and PC1. The majority of CCK mRNA-positive neurons in the pyramidal cell layer of the hippocampus express PC1 mRNA and greater than 50% of CCK mRNA-positive neurons in several nuclei of the amygdala also express PC1. These results demonstrate that PC1 is important for CCK processing. PC2 and PC5 are also widely colocalized with CCK. It may be that PC2, PC5 or another non-PC enzyme are able to substitute for PC1 and sustain production of some amidated CCK. Together these enzymes may represent a redundant system to insure the production of CCK.