Immunoreactive Endocrine Cells Research Articles

Effects of glucagon-like peptide-2 on TNF-alpha/actinomycin D-induced intestinal epithelial injury: a scanning electron microscopic and immunohistochemical study

Glucagon-like peptide-2 (GLP-2) is a peptide hormone with intestinotrophic activity, which is released from the intestinal endocrine cells. The aim of this study was to investigate the effects observed by scanning electron microscope of GLP-2 in intestinal epithelial injury induced by tumor necrosis factor-alpha (TNF-α)/actinomycin D (Act D). In addition, it aimed to elucidate whether the action mechanism of GLP-2 might be mediated by gastrointestinal hormones such as cholecystokinin and somatostatin . The intestinal epithelial injury was induced by intraperitoneal administration of 15 µg/kg TNF-α and 800 µg/kg Act D per mouse. Animals were injected subcutaneously 200 µg/kg GLP-2 analogue every 12 hr for 10 consecutive days prior to the administration of TNF-α and Act D. Scanning electron microscopic examination revealed severe epithelial damage in the small intestine of TNF-α/Act D-administered mice. The administration of TNF-α/Act D was significantly increased the number of somatostatin -immunoreactive endocrine cells, but did not affect the number of cholecystokinin -immunoreactive endocrine cells in the intestinal mucosa . On the other hand, GLP-2 analogue pretreatment prevented TNF-α/Act D-induced intestinal epithelial injury by causing a marked decrease in the degenerative changes and the number of somatostatin -immunoreactive endocrine cells, and a significant increase in the number of cholecystokinin -immunoreactive endocrine cells. As a result, the present study indicates that GLP-2 has a protective effect against TNF-α/Act D-induced intestinal epithelial injury. Morever protective effect of GLP-2 might be related to somatostatin and cholecystokinin.

The Small Intestine of the Adult New Hampshire Chicken: an Immunohistochemical Study

The presence and distribution of glucose-dependent insulinotropic polypeptide or gastric inhibitory polypeptide (GIP), gastric-releasing peptide (GRP) and glucagon immunoreactivity were studied in the small intestine of the New Hampshire chicken using immunohistochemistry. This is the first report of the presence of GIP-immunoreactive (ir) cells in avian small intestine. GIP, GRP and glucagon immunoreactivity was localized in the epithelium of the villi and crypts of the duodenum, jejunum and ileum. In particular, both in the duodenum and in the jejunum immunoreactive endocrine cells to GIP, GRP and glucagon were observed. In the ileum, we noticed GIP-ir and glucagon-ir cells. GRP-ir was found in nerve fibres of all three segments of the small intestine. The distribution of these bioactive agents in the intestinal tract of the chicken suggests that GIP and glucagon may play a role in the enteropancreatic axis in which intestinal peptides modulate pancreas secretion.

Apoptosis, cell proliferation and serotonin immunoreactivity in gut of Liza aurata from natural heavy metal polluted environments: preliminary observations

In the present paper, the effect of natural environment non-lethal heavy metal concentration on cell renewal of Liza aurata intestinal epithelium, was studied by the TUNEL (terminal deoxynucleotidyltransferase-mediated dUTP nick end labelling) method and anti-PCNA (proliferating cell nuclear antigen) immunohistochemistry, in order to detect, respectively, apoptosis and cell proliferation. In addition, the presence and distribution of the cell renewal regulator, serotonin, was immunohistochemically investigated. In order to reduce variability, only immature specimens were considered. The results indicated that in the control specimens from non-polluted areas, the PCNA immunoreactive nuclei of the proximal intestinal epithelium were only located at the bottom of the intestinal folds, together with a few TUNEL-positive nuclei, and goblet mucous differentiated cells. In the specimens from polluted areas, the number of PCNA immunoreactive cells was greatly enhanced, and they extended along the mid portion of the intestinal folds; the number of TUNEL-positive nuclei was enhanced as well, but they were almost exclusively detected in the third apical portion of the intestinal folds. Serotonin immunoreactive nerve elements were more frequently detected in the intestinal wall of L. aurata specimens from polluted areas, and besides that, some serotonin immunoreactive endocrine cells were also present. Variations in distribution and frequency of TUNEL-positive nuclei, PCNA immunoreactive nuclei, and serotonin immunoreactivity put in evidence an alteration of cell renewal with an enhancement of cell proliferation, probably leading to morphological intestinal fold changes.

The distribution and frequency of endocrine cells in the splenic lobe of grass lizard (Takydromus wolteri): An immunohistochemical study

The regional distribution and frequency of the pancreatic endocrine cells in the splenic lobe of grass lizard, Takydromus wolteri, were studied by immunohistochemical (PAP) method using six types of specific mammalian antisera against bovine Sp-1/chromogranin (bCG), serotonin, insulin, glucagon, somatostatin and human pancreatic polypeptide (hPP). The pancreas was subdivided into two regions--islet kike and exocrine regions. The frequency of each immunoreactive (IR) endocrine cells was calculated as mean number/total 100 islet cells and as mean number/total 1,000 cells (including exocrine and endocrine cells) using automated image analysis process. In addition, the percentage of each IR cell was also calculated. All of six endocrine cells were demonstrated. They were dispersed in the whole pancreatic parenchyma between exocrine acinar cells, or they were also observed as islet like clusters. In islet-like regions, bCG-, insulin- and glucagon-IR cells were detected as one or two cell layer cords and they were located between this cell-cords with 14.30+/-5.62, 61.50+/-9.76 and 26.50+/-9.31/100 cells frequencies, respectively. However, somatostatin-IR cells were mainly located in the peripheral parts not in cell-cords with 12.40+/-4.86/100 cells, and no serotonin- and hPP-IR cells were demonstrated. In exocrine regions, all of bCG-, serotonin-, insulin-, glucagon-, somatostatin- and hPP-IR cells were detected and they occurred mainly among the exocrine parenchyma as solitary cells with 10.30+/-2.54, 0.80+/-0.63, 15.50+/-5.30, 5.80+/-2.66, 3.10+/-1.29 and 11.00+/-3.33/1000 cells frequencies, respectively. In addition, serotonin-IR cells were mainly located between epithelia and connective tissue of pancreatic duct. Overall, there were 0.58+/-0.49% serotonin-, 56.44+/-9.35% insulin-, 23.73+/-8.22% glucagon-, 11.28+/-3.03% somatostatin- and 7.97+/-2.02% hPP-IR cells.

Altered Hypothalamic-Pituitary-Adrenal Axis Activity in Patients with Chronic Heart Failure

Neuroendocrine factors play an important role in the pathogenesis of chronic heart failure. Despite numerous clinical and experimental studies, the role of the hypothalamic-pituitary-adrenal axis and glucocorticoid hormones is not fully characterised. Here we present a study of plasma cortisol concentration in 74 chronic heart failure patients, divided into four groups based on NYHA functional classes I-IV, and in 17 control subjects. In parallel, we performed morphological analysis of the hypothalamic-pituitary-adrenal axis components from 8 male patients who had died from chronic heart failure, and 9 male controls. In our study we applied immunohistochemical method and quantitative analysis to investigate an expression of hypothalamic neurohormones (corticotropin-releasing hormone, vasopressin) and adrenocorticotropin hormone in the pituitary, as well as performed general histological examination of the adrenal cortex. Measurement of morning cortisol concentration in plasma of chronic heart failure patients revealed neither difference compared to controls nor with the severity of the disease. Despite this, a two-fold increase in the density of corticotropin-releasing hormone-immunoreactive neurons as well as a two-fold increase in the number of corticotropin-releasing hormone neurons co-expressing vasopressin in the hypothalamic paraventricular nucleus were found. In the anterior pituitary the density of adrenocorticotropin hormone-immunoreactive cells was significantly increased. General histological analysis of the adrenal cortex revealed a drastic thinning of the zona fasciculata and dystrophic changes in corticocytes. Structural changes, observed in the adrenal cortex, suggest a relative glucocorticoid deficiency, which may contribute to corticotropin-releasing hormone and adrenocorticotropin hormone upregulation in hypothalamus and pituitary of chronic heart failure patients.

Distribution of Neuropeptide F-Like Immunoreactivity in the Eastern Subterranean Termite,Reticulitermes flavipes

The nervous system and gut of worker, soldier and alate castes of the eastern subterranean termite, Reticulitermes flavipes Kollar (Isoptera: Rhinotermitidae) were examined for immunoreactivity to an antiserum to Helicoverpa zea (Boddie) (Leipidoptera: Noctuidae) MP-I (QAARPRF-NH2), a truncated form of neuropeptide F. More than 145 immunostained axons and cell bodies were seen in the brain and all ganglia of the ventral nerve cord. Immunoreactive axons exiting the brain projected anteriorly to the frontal ganglion and posteriorly to the corpora cardiaca and corpora allata. In the stomatogastric nervous system, immunoreactive axons were observed over the surface of the foregut, salivary glands, midgut and rectum. These axons originated in the brain and from 15–25 neurosecretory cells on the foregut. Staining patterns were consistent between castes, with the exception of immunostaining observed in the optic lobes of alates. At least 600 immunoreactive endocrine cells were evenly distributed in the midguts of all castes with higher numbers present in the worker caste. Immunostaining of cells in the nervous system and midgut was blocked by preabsorption of the antiserum with Hez MP-I but not by a peptide having only the RF-NH2 in common. This distribution suggests NPF-like peptides coordinate feeding and digestion in all castes of this termite species.

Immunocytochemical study of gastrintestinal endocrine cells in insectivorous bats (Mammalia: Chiroptera)

The regional distribution and relative frequency of endocrine cells in the stomach and intestine of Phyllostomidae: Lonchorhina aurita and Molossidae: Molossus molossus bats were studied immunohistochemically. Three types of immunoreactive (IR) endocrine cells--to serotonin (5-HT), gastrin (GAS) and enteroglucagon (GLUC)--were found in the gastric mucosa and four types of IR cells were identified in the intestinal mucosa. This study showed an interespecfic difference in the regional distribution and relative frequency of endocrine cells in the Chiropteran alimentary tract.

The characteristic change in the distribution of S-100 immunoreactive folliculostellate cells in rat anterior pituitary upon long-term estrogen treatment is prevented by concomitant progesterone treatment

The presence of folliculostellate cells in the anterior pituitary was described 49 years ago. These cells give about 10% of the whole cell population and through their long processes they provide intrahypophyseal communication. The folliculostellate cells contain S-100 protein. Its immunostaining was used to identify these cells. It was previously found that the diethylstilbestrol treatment basically influences the morphology and function of the trophic hormone secreting as well as the folliculostellate cells. In the present experiment, we have studied whether a concomitant progesterone treatment can prevent or attenuate changes caused by diethylstilbestrol treatment in the distribution of folliculostellate, prolactin, and GH cells. Diethylstilbestrol alone induced the appearance of prolactinomas. Inside the prolactinomas, folliculostellate cells were scattered but outside the prolactinomas they formed a demarcation line. Inside the prolactinomas, there were only a few growth hormone immunoreactive cells but they surrounded the prolactinomas in a ring-like pattern. When diethylstilbestrol was implanted with progesterone, the changes being characteristic for diethylstilbestrol treatment, could not develop. Concomitant progesterone influence prevented morphological changes in the anterior pituitary. Progesterone alone had no effect. In accordance with the formation of prolactinomas, the plasma prolactin level was very high in diethylstilbestrol treated rats. Concomitant progesterone treatment prevented the effect of diethylstilbestrol. Progesterone alone did not influence the prolactin level. GH levels did not significantly differ in any groups.

An immunohistochemical study of endocrine cells in the iodoacetamine-induced rat's gastritis

The gastric epithelium is a complex stucture formed by tubular gastric glands. The gastric glands contain a wide variety of cell types according to the secretion of hydrochloric acid, proteases, mucus, pepsinogen and hormones. Endocrine and local paracrine mediators are of crucial importance for maintaining structural and functional integrity of the epithelium. Therefore our aim was to investigate the precise localization and density of the endocrine cells and their contacts in iodoacetamine-induced rat's gastritis. Methods: somatostatin (SOM), calcitonin gene-related peptide CGRP), substance P (SP), vasoactive intestinal polypeptide (VIP), neuropeptide Y (NPY) and galanin immunoreactive (IR) endocrine cells and nerve elements were detected by ABC immunohistochemistry. Results: The quantitative analysis of the density of the different neuropeptide containing endocrine cells and nerve fibres showed that the (IR) nerve fibres were found with different density. The number of SOM and CGRP IR endocrine cells increased significantly in the gastritis. Both endocrine cells possessed long cytoplasmic extensions and no cell-to-cell contact between parietal and endocrine cells was found. The number of SP, VIP, NPY IR nerve fibres also increased significantly in the gastritis, however, the others showed no alteration. The number of the immunocompetent cells was also increased (being lymphocytes, plasma cells and mast cells) and 16,8% of the immunocytes were IR for SP and 9, 4% of them were IR for NPY. In several cases close contacts were detected between IR nerve fibres and the immunocytes. Conclusions: The increased number of SOM and CGRP endocrine cells and SP IR nerve fibres might play a crucial role in the regulation of gastric acid secretion, including gastric ulcer disease and autoimmune gastritis. The main inhibitor of acid secretion is somatostatin. CGRP activates the mucin biosynthesis, both SOM and CGRP have a preventive action on gastric mucosal injury. The changes in the density of nerve fibres and immunocytes might be the consequences of the inflammation.

Cyclooxygenase inhibitors decrease the number of gonadotrophin-releasing hormone-immunoreactive cells in the hypothalamus in ovariectomized estrogen-primed rats.

Cyclooxygenase inhibitors decrease the number of gonadotrophin-releasing hormone-immunoreactive cells in the hypothalamus in ovariectomized estrogen-primed rats.

Cholinergic afferents to gonadotropin-releasing hormone neurons of the rat

Gonadotropin-releasing hormone-synthesizing neurons represent the final common pathway in the hypothalamic regulation of reproduction and their secretory activity is influenced by a variety of neurotransmitters and neuromodulators acting centrally in synaptic afferents to gonadotropin-releasing hormone neurons. The present study examined the anatomical relationship of cholinergic neuronal pathways and gonadotropin-releasing hormone neurons of the preoptic area. The immunocytochemical detection of choline acetyltransferase or vesicular acetylcholine transporter revealed a fine network of cholinergic fibers in this region. At the light microscopic level, the cholinergic axons formed appositions to the gonadotropin-releasing hormone immunoreactive cell bodies and dendrites. Results of electron microscopic studies confirmed the absence of glial interpositions in many of these neuronal contacts. Classical cholinergic synapses, which belonged to the asymmetric category, were only observed rarely on gonadotropin-releasing hormone neurons. The lack of synaptic density in most contacts corroborates previous observations on the cholinergic system elsewhere in the brain. Further, it suggests a dominantly non-synaptic route also in this cholinergic neuronal communication. This study provides direct neuromorphological evidence for the involvement of the cholinergic system in the afferent neuronal regulation of gonadotropin-releasing hormone neurons. The sources of cholinergic afferents and the receptorial mechanisms underlying this interaction will require further clarification.

E-box regulation of gonadotropin-releasing hormone (GnRH) receptor expression in immortalized gonadotrope cells

The pituitary gland's ability to respond to the hypothalamic hormone GnRH (gonadotropin-releasing hormone) depends directly on the gonadotrope-specific expression of the GnRH receptor (GnRHR), a G-protein coupled transmembrane protein coded by the GnRHR gene. In the present study, we have investigated the potential regulatory role of seven noncanonical E-box enhancer sequences within the 856 bp proximal 5′-flanking region of the mGnRHR gene in regulating transcription. These sequences are known to mediate the action of clock gene proteins on the expression of a diverse array of genes both central and peripheral. In the present studies the expression of all of the cognate clock genes was identified in the αT3-1 gonadotrope cell line. Additionally, luteinizing hormone-immunoreactive cells in the adult rodent pituitary gland were also shown to co-express the PERIOD-1 protein. By means of chromatin immunoprecipitation of αT3-1 nuclear extracts we were able to capture promoter fragments of the GnRHR and Period-1 genes, indicating that E-boxes in these promoters bind the CLOCK protein. RNA interference experiments with αT3-1 cells in which Bmal1 expression was attenuated also confirmed the involvement of E-boxes in transcriptional regulation of the mGnRHR gene. Subsequent luciferase reporter assay experiments with GnRHR constructs possessing intact or mutated E-boxes confirmed the use of these sequences for the regulation of mGnRH-R/ luc expression. Transient overexpression of the dominant negative E-box-binding factor CLOCK-Δ19, or the inhibitory clock protein mPER1, markedly reduced CLOCK/BMAL1-driven mGnRH-R/ luc expression in a dose-dependent fashion. Our data implicate the clock genes as important factors controlling GnRHR expression in murine gonadotrope cells.

The galaninergic enteric nervous system of pleuronectiformes (Pisces, Osteichthyes): An immunohistochemical and confocal laser scanning immunofluorescence study

The galaninergic enteric nervous system of three flatfishes was studied using immunohistochemical and immunofluorescence methods. Galanin-like immunoreactivity was mainly detected within the enteric intramural neurons of the stomachs and the proximal intestines. The sole, Solea solea L. and the flounder, Platichthys flesus L. showed a similar occurrence and relative distribution of galaninergic intramural neurons. Rare nervous fibre immunoreactive to the anti-galanin serum were observed in the muscular layers of the oesophagus of the turbot, Psetta maxima L. The presence and relative abundance of galanin-like immunoreactive neurons in the remaining organs of the alimentary canal of the turbot showed a different pattern in comparison to those observed in the sole and the flounder. A galanin-like peptide was detected in nerve fibres running through the exocrine parenchyma of the pancreas of all three species. It is conceivable that the galaninergic system in these species plays a role in regulating gut muscle activity, and in controlling pancreatic secretion. Galanin and choline acetyltransferase were co-localized within the same neurons of the stomach and intestine. The result confirms the hypothesis that galanin in the gut of fish functions as a cholinergic modulator. Differently from other fish species, immunoreactive endocrine cells were not detected in the studied pleuronectiformes.

Development of the Chick Pancreas with Regard to Estimation of the Relative Occurrence and Growth of Endocrine Tissue

Endocrine cells in chick pancreas were observed to map their distribution during development and to perform morphometric studies starting on embryonic day 5. The ratio of exocrine to endocrine tissues first prevailed in favour of the endocrine ones, and changed abruptly after day 9 when rapid growth of exocrine tissue began. Endocrine tissue was formed of two types of islets. The 'light' (or B) islets were composed of insulin-immunoreactive cells, completed perhaps by a few somatostatin-immunoreactive cells occurring on the periphery. The majority of the somatostatin- and glucagon-immunoreactive cells were present in the 'dark' (or A) islets. Endocrine elements were also scattered as single cells over the pancreas. Sporadically, the endocrine cells established contacts with exocrine ducts. In morphometric analysis, volume density of insulin-, glucagon-, and somatostatin-immunoreactive cells was measured, and ratios were calculated between particular components. The volume density of endocrine cells and their ratio appeared stable in individual lobes but varied significantly between each other. Increase of the glucagon volume density is exponential, whereas insulin increases almost linearly especially in splenic lobe. The process results in the increase of the hormone-immunoreactive cell volume density in favour of glucagon-immunoreactive cells typical for birds.

Age‐related changes in growth hormone‐immunoreactive cells in the anterior pituitary gland of Jcl: Wistar‐TgN (ARGHGEN) 1Nts rats (Mini rats)

Rats of the Jcl: Wistar-TgN (ARGHGEN) 1Nts strain (Mini rats) are transgenic animals carrying an antisense RNA transgene for rat growth hormone (GH); they show poor somatic growth and a low blood GH level compared to age-matched wild-type Wistar (non-Mini) rats. The purpose of the present study was to investigate age-related changes in growth hormone-immunoreactive (GH-IR) cells in the anterior pituitary gland (AP) of Mini rats at four, six, and eight weeks of age. The body weight and size of the GH-IR cells of Mini rats was significantly lower than that of non-Mini rats at six and eight weeks of age; however, this difference was not observed at four weeks of age. The AP volume and the number of GH-IR cells in Mini rats were significantly smaller than those of the age-matched non-Mini rats at the three ages. These results suggest that the abnormal development of GH-IR cells in the AP induced by the GH antisense RNA transgene is responsible for the poor somatic growth and the low blood GH levels in Mini rats.

A comparative immunohistochemical study on a galanin‐like peptide in the neuroendocrine system of the alimentary canal of three species of siluriform catfishes

Immunohistochemical examination of the galaninergic and the diffuse endocrine systems of the alimentary canal of three catfish species, Ameiurus melas, Silurus glanis and Clarias gariepinus showed the presence of galanin‐like immunoreactive endocrine cells in the stomachs and the proximal intestines and a galanin‐like immunoreactivity within the enteric intramural neurons. The three species differed notably in the relative abundance of galaninergic neurons of the digestive system. A galanin‐like peptide was detected in a high number of nerve bundles and nerve fibres running through the exocrine parenchyma and the endocrine islets of the pancreas of all three species. It is conceivable that the galaninergic system in these species plays a role in regulating gut muscle activity, in modulating mucosal secretive and absorptive processes, and in controlling pancreatic hormone secretion.

Prolactin-releasing peptide is a potent mediator of stress responses in the brain through the hypothalamic paraventricular nucleus

The effects of i.c.v. administration of prolactin-releasing peptide on neurons in the paraventricular nucleus of rats and plasma corticosterone levels were examined by measuring changes in Fos-like immunoreactivity, c- fos mRNA using in situ hybridization histochemistry, and plasma corticosterone using a specific radioimmunoassay. Approximately 80% of corticotropin-releasing hormone immunoreactive cells exhibited Fos-like immunoreactivity in the parvocellular division of the paraventricular nucleus 90 min after i.c.v. administration of prolactin-releasing peptide. The greatest induction of the c-fos mRNA expression in the paraventricular nucleus was observed 30 min after administration of prolactin-releasing peptide, and occurred in a dose-related manner. Plasma corticosterone levels were also significantly increased 30 min after administration of prolactin-releasing peptide. Next, the effects of restraint stress, nociceptive stimulus and acute inflammatory stress on the expression of the prolactin-releasing peptide mRNA in the dorsomedial hypothalamic nucleus, nucleus of the solitary tract and ventrolateral medulla were examined using in situ hybridization histochemistry for prolactin-releasing peptide mRNA. Restraint stress and acute inflammatory stress upregulated the prolactin-releasing peptide mRNA expression in the nucleus of the solitary tract and ventrolateral medulla. Nociceptive stimulus upregulated the prolactin-releasing peptide mRNA expression in the ventrolateral medulla. Finally, we observed that pretreatment (i.c.v. administration) with an anti-prolactin-releasing peptide antibody significantly attenuated nociceptive stimulus-induced c- fos mRNA expression in the paraventricular nucleus. These results suggest that prolactin-releasing peptide is a potent and important mediator of the stress response in the brain through the hypothalamic paraventricular nucleus.

Changes in the neuromodulators of the diffuse endocrine system of the alimentary canal of farmed rainbow trout, Oncorhynchus mykiss (Walbaum), naturally infected with Eubothrium crassum (Cestoda)

A histopathological and immunohistochemical study on the intestines of 45 specimens of farmed rainbow trout, Oncorhynchus mykiss (Walbaum), from Loch Awe, Scotland, revealed a number of cellular deviations in individuals naturally infected with the pseudophyllidean cestode Eubothrium crassum (Bloch, 1779). Twenty-five individuals (55.5%) were infected with an average worm burden of 18.84 +/- 4.06 (mean +/- SE) cestodes per host (range, 2-80 worms; total 471 worms). The cestodes, measuring an average 8.23 +/- 1.10 cm (mean +/- SE; range, 5.3-13.0 cm) in length, were found attached by their scolices to the mucosal lining of the distal portion of the pyloric caeca. Within the caeca, the strobila evoked a mild catarrhal enteritis, namely an enhanced mucus production with epithelial cellular desquamation, a leucocytic infiltration of the lamina propria-submucosa and vacuolization of the intestinal epithelial cells. Eosinophilic granular cells of the stratum granulosum exhibited granular depletion, while within the catarrh, the presence of a high number of rodlet cells was noticed. Immunohistochemically, the occurrence of E. crassum caused a significant reduction in the number of bombesin-, gastrin-releasing peptide and glucagon-like immunoreactive endocrine cells, but an increase in the relative densities of endocrine cells containing cholecystokinin-8- and gastrin-like substances. There were, however, no significant differences in the number of endocrine cells that were immunoreactive to secretin, neuropeptide Y and peptide histidine-isoleucine antisera in the digestive tracts of either the infected or non-infected O. mykiss.

Immunostaining for allatotropin and allatostatin-A and -C in the mosquitoes Aedes aegypti and Anopheles albimanus

Confocal laser-scanning microscopy was used to carry out a comparative study of the immunostaining for three families of neuropeptides, viz., allatostatin-A (AS-A), allatostatin-C (AS-C) and allatotropin (AT), in adult female mosquitoes of Aedes aegypti and Anopheles albimanus. The specific patterns of immunostaining for each of the three peptides were similar in both species. The antisera raised against AT, AS-A, and AS-C revealed intense immunoreactivity in the cells of each protocerebral lobe of the brain and stained cells in each of the ventral ganglia and neuronal projections innervating various thoracic and abdominal tissues. Only the AS-A antiserum labeled immunoreactive endocrine cells in the midgut. The distribution of the peptides supports the concept that they play multiple regulatory roles in both species.

Neuronal distribution of melanin-concentrating hormone, cocaine- and amphetamine-regulated transcript and orexin B in the brain of the Djungarian hamster ( Phodopus sungorus)

The distribution of melanin-concentrating hormone-, cocaine- and amphetamine-regulated transcript- and orexin B-immunoreactive elements as well as their morphological relationships in selected brain structures harbouring the neuroendocrine pathways controlling energy balance and circadian rhythmicity in the Djungarian hamster ( Phodopus sungorus) were studied. Cocaine- and amphetamine-regulated transcript-(55–102)-immunoreactive perikarya co-expressed melanin-concentrating hormone-immunoreactivity in the lateral hypothalamic area, dorsomedial hypothalamic nucleus, zona incerta and posterior hypothalamic area. In addition, arcuate nucleus, hypothalamic periventricular nucleus, Edinger–Westphal nucleus, and the rostral aspect of the dorsal raphe nucleus contained cocaine- and amphetamine-regulated transcript-immunoreactive cell profiles. Orexin B-immunoreactive perikarya were distributed in the lateral hypothalamic area, dorsomedial hypothalamic nucleus and retrochiasmatic area. Cells immunoreactive for orexin B did not co-express melanin-concentrating hormone-immunoreactivity, but orexin B-immunoreactive fibers had close apposition to many melanin-concentrating hormone-immunoreactive cells. Whereas immunoreactivity for all examined peptides was absent in the suprachiasmatic nucleus, dense and large orexin B-immunoreactive fibers and to a lesser extent melanin-concentrating hormone- and cocaine- and amphetamine-regulated transcript-immunoreactive fibers of smaller size were present in the intergeniculate leaflet and raphe nucleus. These observations in Djungarian hamsters indicate that the neuronal distribution of the examined peptides is strongly conserved between species. In addition, the presence of fibers within the neuronal components of the circadian timing system suggests that they may indirectly influence circadian rhythms.