Regional distribution of somatostatin-like immunoreactivity in the human brain

The regional distribution and chromatographic characteristics of somatostatin-like immunoreactivity (SLI) were studied in autopsy specimens of 8 human brains. SLI concentrations in 8 different regions of these brains ranged from (mean +/- S.E.) 756.3 +/- 363.4 pmol/g tissue in anterior hypothalamus to 1.6 +/- pmol/g in cerebellum. Chromatography of the extracts of human brain cortex, anterior hypothalamus, thalamus and amygdala on Sephadex G-50 disclosed one major peak which corresponded to the elution peak of synthetic somatostatin. Since the human brains were obtained at autopsy 11-36 h after death, the effects of temperature and time lapse between death and tissue extraction on SLI concentration and chromatographic pattern in rat brain were examined. After storage at 4 degrees C or 23 degrees C for 2 h and 8 h, a significant increase in SLI concentration was noted, although by 24 h in increase was no longer observed. A gradual loss in the eluting forms of SLI on gel chromatography was observed with storage at 4 degrees C or 23 degrees C.     

Vasoactive intestinal peptide and gastrin-releasing peptide play distinct roles in the suprachiasmatic nucleus

Vasoactive intestinal peptide and gastrin-releasing peptide levels were measured in the punched-out suprachiasmatic nucleus tissue from rats kept under a prolonged dim light (in vivo). Vasoactive intestinal peptide content increased from 4 to 8 h, returned to the baseline level at 12 to 16 h, and then increased again until 36 h after the light was switched off (dim light). Gastrin-releasing peptide level, in contrast, showed no significant changes, but a slight decrease from 1 to 4 h was detected under the dim light. In suprachiasmatic nucleus explant-slice culture, in vitro, Arg-vasopressin release increased transiently or showed a decrease at 30 min after exposure to vasoactive intestinal peptide or gastrin-releasing peptide, respectively. Treatment with anti-vasoactive intestinal peptide and anti-gastrin-releasing peptide antibodies reversed these effects. These findings suggest reciprocal roles of vasoactive intestinal peptide and gastrin-releasing peptide in Arg-vasopressin release.     

Beacon immunoreactivity in the rat hypothalamus

Beacon (BC) is a peptide of 73 amino acids, whose gene expression was first reported in the hypothalamus of Psammomys obesus (or Israeli sand rat). To appreciate better the functional role of BC in normal rats and sand rats, the distribution of BC immunoreactivity (irBC) and its subcellular localization were studied in the brain of Sprague-Dawley rats. In the hypothalamus, intense staining was present in neurons of the supraoptic (SO), paraventricular (PVH), and accessory neurosecretory nuclei and in cell processes of median eminence. Double labeling of the hypothalamic sections with mouse monoclonal oxytocin (OT) antibody and rabbit polyclonal BC antiserum revealed that nearly all OT-immunoreactive cells from SO, PVH, and accessory neurosecretory nuclei were irBC. Double labeling of the sections with guinea pig vasopressin (VP) antiserum and BC antiserum showed that a population of VP-immunoreactive neurons was irBC. By immunoelectron microscopy, immunoreactive product was associated with mitochondrial membranes or appeared as electron-dense bodies in many PVH and SO neurons. Most of the neurosecretory granules were unstained for BC. Taken together, our results indicate the presence of beacon in the OT-containing neurons and a population of VP-containing neurons, mostly associated with mitochondrial membrane. Insofar as the amino acids sequence of beacon is identical to that of ubiquitin-like 5, it is possible that the distribution of BC immunoreactivity noted in our study is that of ubiquitin-like 5 peptide in the rat hypothalamus.     

Effects of bombesin and gastrin-releasing peptide on memory processing

We have previously shown that feeding mice immediately following training enhances memory retention and that one of the gastrointestinal hormones released during a meal, cholecystokinin, also enhances retention after peripheral administration. In the studies reported here we demonstrate that another gastrointestinal peptide, gastrin-releasing peptide (GRP), enhances retention after peripheral administration, as does its amphibian counterpart, bombesin. GRP_14–27 had the same effect as the intact peptide, while GRP_1–16 was ineffective at enhancing retention. The dose-response curves showed a characteristic inverted U-shape with high doses of both GRP and bombesin being amnestic. The effect of both peptides was time-dependent and both reversed amnesia induced by the anticholinergic, scopolamine. I.c.v. administration of the peptides required higher doses to produce an effect on memory retention. suggesting that the effect was mediated predominantly through a peripheral mechanism. Doses of the peptides that enhanced memory retention after peripheral administration failed to increase serum glucose, suggesting that glucose modulation was not the mechanism by which GRP and bombesin modulate memory processing. Vagotomy inhibited the memory-enhancing effects of both GRP and bombesin, suggesting that these peptides produced their effect by stimulating ascending vagal pathways. These studies, together with our previous study with cholecystokinin, suggest the existence of a gastrointestinal hormonal system, which is activated by the passage of food through the intestine, that enhances memory retention.     

Substance P-like immunoreactivity and somatostatin-like immunoreactivity in the ventricular fluid of patients with chronic pain syndromes

Summary Substance P-like and somatostatin-like immunoreactivities (SPLI and SLI) were determined in ventricular fluid of patients with chronic pain syndromes and in a comparison group with multiple sclerosis, essential tremor, epilepsy and postanoxic myoclonus. Concentrations of SPLI and SLI were non-significantly decreased by 40% and 33% in chronic pain patients as compared with control patients without pain. There were no differences apparent between subgroups of pain patients (deaferentation pain, neoplasia-induced pain, thalamic pain). High pressure liquid chromatography combined with radioimmunoassay showed marked heterogeneity of SPLI and SLI.     

Effects of lesions in the amygdala and periventricular hypothalamus on striatal somatostatin-like immunoreactivity

Somatostatin has been found in substantial amounts in the basal ganglia by radioimmunoassay and has been demonstrated in both neurons and nerve terminals. Since the levels of somatostatin have been shown to vary in Huntington's and Alzheimer's disease it was of interest to see whether such changes could be produced experimentally. Lesions of the periventricular nucleus of the hypothalamus and knife cuts adjacent to this nucleus had no effect on striatal somatostatin-like immunoreactivity (SLI). Similarly lesions of mediodorsal frontal cortex, and those isolating pyriform cortex or the olfactory bulb had no effect on striatal SLI. Removal of tge amygdala resulted in significant increases in SLI in the ipsilateral striatum and nucleus accumbens, suggesting loss of an inhibitory interaction. Stria terminalis lesions failed to reproduce this effect suggesting that it is mediated via amygdalo-striatal projections traveling in the dorsal longitudinal bundle. Other findings support a somatostatin projection to the amygdala from the bed nucleus of the stria terminalis and one from the amygdala to the ventromedial hypothalamus.     

Regional expressions of Fos-like immunoreactivity in rat cerebral cortex after stress; restraint and intraperitoneal lipopolysaccharide

To demonstrate regional activation in the rat cerebral cortex related to stress-evoked neuroendocrine response, Fos expression in both the cerebral cortex and hypothalamic paraventricular nucleus (PVN) was immunohistochemically examined in two experimental groups; a lipopolysaccharide (LPS) intraperitoneally injected group for inflammatory stress and a restraint group for emotional stress. The LPS injection (100 μg/100 g b.w.) and restraint (for 30 min) had similar effect on Fos-like immunoreactivity (Fos-LI) in PVN with regard to the number of immunoreactive nuclei and their distribution pattern, while the times to maximize Fos-LI were different. Numerical analysis of cortical Fos-LI in untreated rats showed a distinct region-specific pattern. Statistical analysis revealed no significant increase in Fos-LI density in any cortical regions in the LPS group, but restraint resulted in a dramatic and region-specific increase. A significant increase was detected in the prefrontal cortex (the cingulate, orbital and agranular insular cortex), the frontal area 2, the agranular retrosplenial cortex, the parietal cortex, and the medial and lateral occipital area 2. These results indicate that cortical activation relevant to specific functions may be involved in stress-specific neural circuitry.     

The distribution of progesterone receptor immunoreactivity and mRNA in the preoptic area and hypothalamus of the ewe: upregulation of progesterone receptor mRNA in the mediobasal hypothalamus by oestrogen

The distribution of progesterone receptors (PR) was mapped in the hypothalamus of the ewe using immunocytochemistry. These results were confirmed using in situ hybridization with a sheep-specific 35S-labelled riboprobe. In addition, the effect of oestrogen on the level of PR mRNA in the hypothalamus was examined in ovariectomized (OVX) ewes following treatment with an oestrogen implant or without treatment. PR immunoreactive (-ir) cells were readily detected in OVX animals. Labelled cells were observed in four main hypothalamic regions: the preoptic area (POA), including the organum vasculosum of the lamina terminalis, periventricular nucleus (PeVN), ventromedial nucleus (VMN) and the arcuate nucleus (ARC) (including the region ventral to the mamillary recess). In addition, lightly stained PR-ir cells were observed in the supraoptic nucleus and a few PR-ir cells were also found in the diagonal band of Broca. No PR-ir cells were found in the brainstem. PR mRNA-containing cells were found in the same hypothalamic regions as the PR-ir cells. Image analysis of emulsion-dipped slides following in situ hybridization indicated that oestrogen treatment increased (P<0.01) the mean number of silver grains/cell and the density of labelled cells in the VMN and ARC but had no effect on the level of PR mRNA expression in the POA or PeN. The distribution of PR-containing cells in the hypothalamus is similar to that described in other species and all cells were located in nuclei that contain large populations of oestrogen receptor-containing cells. These include regions implicated in the regulation of reproductive neuroendocrine function, and reproductive behaviour. Oestrogen and progesterone synergize to inhibit GnRH secretion and the present results suggest that these functions may involve cells of the VMN and ARC, with oestrogen acting to upregulate PR.     

The distribution of somatostatin-like immunoreactivity in the monkey hippocampal formation

The distribution of somatostatin-like immunoreactivity was studied in the hippocampal formation of the Old World (Macaca fascicularis) and New World (Saimiri sciureus) monkeys. Series of coronal sections were processed by the unlabeled second antiserum method using primary antisera which recognize somatostatin-28 (S309) or somatostatin-28(1-12) (S320). Neuronal cell bodies were more readily stained with antiserum S309 and were observed throughout the hippocampal formation. The most prominent accumulations of stained neurons occur in the hilar region of the dentate gyrus, in strata oriens and pyramidale of regio inferior of the hippocampus, and in the deep layers of the entorhinal cortex. Both antisera demonstrated extensive fiber systems which varied in density regionally in the hippocampal formation. Stained fibers were most prominent in the outer two-thirds of the molecular layer of the dentate gyrus, in stratum lacunosum-moleculare of the hippocampus, in layer I of the presubiculum and in layers I, III, and V of the entorhinal cortex.     

Oxytocin and vasopressin immunoreactivity in rabbit hypothalamus during estrus, late pregnancy, and postpartum

Mother rabbits construct an elaborate maternal nest before parturition and display a single, brief, daily nursing bout throughout lactation. These features present a unique model for investigating the relevance of changes in neuroendocrine secretion associated with pregnancy and parturition for the regulation of maternal behavior. In the present study we analyzed changes in the location, somal size, and number of oxytocin (OT)and arginine vasopressin (AVP)-immunoreactive (IR) neurons in the hypothalamus of rabbits in estrus, late pregnancy (day 29), and postpartum day 1. From estrus to late pregnancy, the number of OT-IR neurons increased in the scattered cell groups located in the lateral hypothalamic area (LHA), but not in the magnocellular nuclei, i.e., paraventricular nucleus (PVN) and supraoptic nucleus (SON). On postpartum day 1 the increase in the number of OT-IR neurons was sustained in the LHA and became apparent also in the main body of the PVN, in which the number of OT-IR neurons doubled. Increases in the somal size of OT-IR cells were seen in all three nuclei only on postpartum day 1. No OT-IR cells were found in the suprachiasmatic nucleus (SCN). From late pregnancy and into postpartum day 1 increases in the somal size of AVP-IR neurons were detected in the PVN, SON, and LHA but not in the SCN. The number of AVP-IR neurons increased between late pregnancy and postpartum day 1 in the SON only. The changes observed in OT and AVP expression in specific hypothalamic nuclei may be related to specific somatic and behavioral events occurring around the time of parturition, e.g., nest-building, maintenance of homeothermy, elevation of blood volume, and nursing in mother rabbits.     

A discrete lesion of ventral hypothalamus and optic chiasm that disturbed the daily temperature rhythm

Summary A patient with a discrete metastasis in the ventral hypothalamus and optic chiasm is reported, who developed an abnormal daily rhythm of oral temperature without alteration of the 24-h mean temperature. This region, its afferents, and its efferents appear to be important in the neural regulation of human circadian rhythmicity.Presented in part at the 36th annual meeting of the American Academy of Neurology, Boston, Massachusetts, April 1984     

Immunohistochemical distribution of somatostatin in the infant hypothalamus

Somatostatin (SS)-containing neurons were mapped in the normal infant hypothalamus with immunohistochemistry, using the peroxidase anti-peroxidase technique. Neurons displaying SS immunoreactivity show a widespread distribution throughout the hypothalamic region. Principal SS-immunoreactive like (SS-IL) perikarya are located in the paraventricular, infundibular and posterior nuclei and in the preoptic region. High SS innervation is also found in the ventromedial and in the lateral mammillary nuclei, and in the median eminence. In general this distribution of SS-IL agrees well with that reported for rat. Compared to the immunohistochemical distribution of SS in human adult hypothalamus, this mapping in the infant hypothalamus is grossly similar. However some differences may be underlined: the presence of a moderately dense group of SS-IL perikarya in the tuberal and posterior nuclei, and a dense innervation of the ventromedial nucleus and in the median eminence. This first detailed distribution of SS immunoreactivity in infant hypothalamus can provide basic knowledge for further studies of infant neuropathology.     

Effects of leptin on spinally projecting neurons in the PVN of the hypothalamus

The study examined whether intravenous (i.v.) leptin increased Fos-production in spinally projecting neurons in the hypothalamic paraventricular nucleus (PVN). We combined (i) rhodamine tagged microspheres injected into the upper thoracic spinal cord and (ii) Fos (marker of neuronal activation) immunohistochemistry. Effects of recombinant murine leptin were compared to vehicle (containing lipopolysaccharide a contaminant present in the leptin solution). Following leptin, 10% of the spinally projecting neurons contained Fos. However, vehicle elicited similar effects and there was no significant difference between the groups.     

Alz-50 immunoreactivity in the hypothalamus of the normal and Alzheimer human and the rat

Alz-50 is a monoclonal antibody recognizing a 68 kilodalton protein that is abundant in Alzheimer's disease (AD) but not detectable by immunoblotting methods in normal brains. When used for immunohistochemistry in AD cortex, Alz-50 recognizes large numbers of neurofibrillary tangles (NFT), neuritic plaques, and some neurons that show no evidence of neurofibrillary degeneration by conventional histopathological staining methods. Alz-50 immunoreactivity is described at the light and electron microscopic levels in the hypothalamus of brains obtained at autopsy from normal and AD subjects. Alz-50 immunoreactivity in the rat hypothalamus is also described. A well-defined population of Alz-50 immunoreactive hypothalamic neurons was identified in both the normal human and rat. At the light microscopic level in the normal human, immunoreactive neurons were most concentrated in the periventricular region, but were also scattered throughout the arcuate nucleus (ARC), lateral hypothalamic area, and tuberal region. Immunoreactive fibers were seen in the periventricular region, dorsal division of the ventromedial nucleus (VMNd), ARC, and external layer of the median eminence (ME). In the rat, reactive neurons were seen only in the periventricular region, and reactive fibers were seen in the periventricular zone, medial preoptic nuclear complex, suprachiasmatic nucleus, VMNd, ARC, and external layer of the ME. Ultrastructurally, all immunoreactivity in the normal human and rat hypothalamus was associated with intraneuronal vesicles. In the AD hypothalamus, Alz-50 identified numerous senile plaques and NFT in addition to the cells and fibers that were stained in the normal brains. Immunoreactive plaques and NFT were most numerous in regions previously reported to undergo neurofibrillary degeneration. At the ultrastructural level, the immunoreactivity in the AD hypothalamus was associated with filaments as well as vesicles. The significance of the selective staining of a specific population of vesicles by Alz-50 is unknown; however, the present results suggest that it is independent of AD pathology.     

Regional distribution of immunoreactive neurotensin in monkey brain

KATAOKA, K., N. MIZUNO AND L. A. FROHMAN. Regional distribution of immunoreactive neurotensin in monkeybrain. BRAIN RES. BULL. 4(1) 57–60, 1979.—The regional distribution of immunoreactive neurotensin (IRNT) in 66 different areas of the monkey brain and spinal cord was studied by radioimmunoassay using specific rabbit antisera to synthetic neurotensin. Each area was found to contain IRNT in uneven concentrations. Highest concentrations were present in the hypothalamic areas and the interpeduncular nucleus, values reaching around 80 pg IRNT/mg wet tissue weight. The habenula, subthalamus and preoptic area were also rich in IRNT. A relatively selective distribution of IRNT was found in the thalamic regions. Low values were obtained in the cerebral and cerebellar cortex, striatum, substantia nigra and white matter areas. Among the fiber bundle studied, the olfactory tract contained a significant amount of IRNT. The present finding of a selective distribution of IRNT suggests a specific neuronal action of neurotensin in primate central nervous system.     

Increase in somatostatin immunoreactivity in the suprachiasmatic nucleus of aged Wistar rats

Decreased immunoreactivity has been reported for several neuropeptides in the aged suprachiasmatic nucleus (SCN). We compared somatostatin (SS) and substance P (SP) immunoreactivity (ir) in aged (20-26 months) and young (6 months) Wistar rats. The old rat SCN revealed a significant increase in SSir (2.6-fold) and SPir. The results show that not all SCN-neuropeptidergic systems decline with age, and suggest a specific age-related role for SS in the SCN.     

Effects of somatostatin antiserum on growth hormone levels in rats with periventricular lesions in the anterior hypothalamus

To determine whether residual inhibitory control of growth hormone (GH) secretion in rats with lesions of the periventricular nucleus (PVN) and depleted median eminence content of somatostatin (SRIF) is due to SRIF, PVN-lesioned rats were treated with SRIF antiserum. Such treatment, in contrast to normal sheep serum, caused increased (P less than 0.0001) plasma GH in lesioned and control groups. These results indicate that biologically important amounts of SRIF are available in the PVN-lesioned rat.     

Male mice lacking the gastrin-releasing peptide receptor (GRP-R) display elevated preference for conspecific odors and increased social investigatory behaviors

Previously, we generated gastrin-releasing peptide receptor null mutant mice (GRP-R-deficient mice), and found that these animals displayed increased non-aggressive social responses in an ordinary social interaction test using a resident-intruder method. In the present study, we examined in more detail the social behaviors of GRP-R-deficient male mice. In social interaction tests, GRP-R-deficient mice showed more social responses, such as sniffing and nosing, relative to wild-type mice, and similar results were obtained whether GRP-R-deficient mice served as intruders or residents. In the same way, they showed more contact behaviors toward an anesthetized conspecific, and less locomotor activity than wild-type mice in a social investigation test toward an anesthetized male mouse. Since olfactory systems play important roles in the social behavior of rodents, olfactory preference tests were conducted in order to evaluate the olfactory properties of GRP-R-deficient mice. The results suggest that no differences exist between wild-type mice and GRP-R-deficient mice in the preference between a novel sawdust odor and their own odor, or that of other male mice. However, GRP-R-deficient mice preferred the odor of other male mice to their own, in contrast to wild-type mice. Furthermore, the preferences of GRP-R-deficient and wild-type mice were not disrupted by intraperitoneal infusion of diazepam (1.5 mg/kg). These results indicate that neither the motion, nor the behavior of conspecifics, nor reduced anxiety lead to the increased non-aggressive social responses and/or social investigatory behaviors in GRP-R-deficient mice. Rather, these latter behaviors may be a consequence of altered cognition of conspecific odors in the mutant mice.     

Colocalization of atriopeptin-like immunoreactivity with choline acetyltransferase-and substance P-like immunoreactivity in the pedunculopontine and laterodorsal tegmental nuclei in the rat

Atriopeptin, the atrial natriuretic peptide, is a circulating hormone that plays an important role in the regulation of fluid and electrolyte balance. Immunohistochemical studies have shown that large, multipolar atriopeptin-like immunoreactive (APir) neurons are present in areas of the midbrain corresponding to the large neurons of the pedunculopontine tegmental (PPT) and lateral dorsal tegmental (TLD) nuclei, all of which can be stained immunohistochemically for choline acetyltransferase-like immunoreactivity (ChATir). A subpopulation of these cholinergic PPT and TLD neurons are also known to contain substance P-like immunoreactivity (SPir). Using an immunofluorescent technique that allows simultaneous localization of two antigens, we have studied the relationship between APir, SPir and ChATir in the pontine tegmentum of the rat. We have found that the large multipolar APir neurons of the pontine tegmentum are identical to the ChATir neurons of the PT and TLD nuclei and a subpopulation of the APir neurons in PPT and TLD neurons are also SPir.     

Regional distribution of kassinin-like immunoreactivity in rat central and peripheral tissues and the effect of capsaicin

The regional distribution of kassinin-like immunoreactivity in rat central and peripheral tissues was investigated by radioimmunoassay and found to resemble closely that of substance P-like immunoreactivity. Neonatal capsaicin treatment caused a similar decrease to both kassinin-like and substance P-like immunoreactivity in primary sensory areas. These results suggest that more than one member of the tachykinin family of neuropeptides exist within the same neuron.