Ellagic acid in Emblica officinalis exerts anti-diabetic activity through the action on ß-cells of pancreas.

PURPOSE: The present study was undertaken to explore the possible anti-diabetic mechanism(s) of Emblica officinalis (EO) and its active constituent, ellagic acid (EA), in vitro and in vivo. METHOD: Neonatal streptozotocin-induced non-obese type 2 diabetic rats were treated with a methanolic extract of EO (250 or 500 mg/kg) for 28 days, and blood glucose, serum insulin, and plasma antioxidant status were measured. Insulin and glucagon immunostaining and morphometry were performed in pancreatic section, and liver TBARS and GSH levels were measured. Additionally, EA was tested for glucose-stimulated insulin secretion and glucose tolerance test. RESULTS: Treatment with EO extract resulted in a significant decrease in the fasting blood glucose in a dose- and time-dependent manner in the diabetic rats. It significantly increased serum insulin in the diabetic rats in a dose-dependent manner. Insulin-to-glucose ratio was also increased by EO treatment. Immunostaining of pancreas showed that EO250 increased ß-cell size, but EO500 increased ß-cells number in diabetic rats. EO significantly increased plasma total antioxidants and liver GSH and decreased liver TBARS. EA stimulated glucose-stimulated insulin secretion from isolated islets and decreased glucose intolerance in diabetic rats. CONCLUSION: Ellagic acid in EO exerts anti-diabetic activity through the action on ß-cells of pancreas that stimulates insulin secretion and decreases glucose intolerance.

Brain insulin infusion does not augment the counterregulatory response to hypoglycemia or glucoprivation.

Although high dosages of insulin can cause hypoglycemia, several studies suggest that increased insulin action in the head may paradoxically protect against severe hypoglycemia by augmenting the sympathoadrenal response to hypoglycemia. We hypothesized that a direct infusion of insulin into the third ventricle and/or the mediobasal hypothalamus (MBH) would amplify the sympathoadrenal response to hypoglycemia. Nine-week-old male rats had insulin (15 mU) or artificial cerebrospinal fluid (aCSF, control) infused bilaterally into the MBH or directly into the third ventricle. During the final 2 hours of the brain insulin or aCSF infusions, the counterregulatory response to either a hyperinsulinemic hypoglycemic (approximately 50 mg/dL) clamp or a 600-mg/kg intravenous bolus of 2-deoxyglucose (2DG) was measured. 2-Deoxyglucose was used to induce a glucoprivic response without peripheral insulin infusion. In response to insulin-induced hypoglycemia, epinephrine rose more than 60-fold, norepinephrine rose more than 4-fold, glucagon rose 8-fold, and corticosterone rose almost 2-fold; but these increments were not different in aCSF vs insulin treatment groups with either intracerebroventricular or bilateral MBH insulin protocols. Intracerebroventricular insulin infusion stimulated insulin signaling as noted by a 5-fold increase in AKT phosphorylation. In the absence of systemic insulin infusion, 2DG-induced glucopenia resulted in an equal counterregulatory response with brain aCSF and insulin infusions. Under the conditions studied, although insulin infusion acted to stimulate hypothalamic insulin signaling, neither intrahypothalamic nor intracerebroventricular insulin infusion augmented the counterregulatory response to hypoglycemia or to 2DG-induced glucoprivation. Therefore, it is proposed that the previously noted acute actions of insulin to augment the sympathoadrenal response to hypoglycemia are likely mediated via mechanisms exterior to the central nervous system.

Beneficial effect of chungkukjang on regulating blood glucose and pancreatic beta-cell functions in C75BL/KsJ-db/db mice.

The current study investigated the antidiabetic effect of chungkukjang, a widely used traditional Korean soybean fermentation food, in a type 2 diabetic animal model, C57BL/KsJ-db/db mice. After a 2-week acclimation period, the db/db mice (male, 5 weeks old) were divided into three groups: diabetic control (AIN-76 diet), chungkukjang (5 g/100 g of diet), and rosiglitazone (0.005 g/100 g of diet). The supplementation of chungkukjang induced a significant reduction of blood glucose and glycosylated hemoglobin level, and it improved insulin tolerance compared to the diabetic control group. Plasma and pancreatic insulin levels of the chungkukjang-supplemented group were significantly higher than those of the diabetic control mice, and the plasma glucagon level was also significantly different. The supplementation of chungkukjang and rosiglitazone significantly elevated hepatic glucokinase activity with a simultaneous reduction of glucose-6-phosphatase and phosphoenolpyruvate carboxykinase activity in the db/db mice compared to the diabetic control mice. In addition, the chungkukjang-supplemented group had an increased hepatic glycogen content compared to the diabetic control and rosiglitazone-supplemented groups. Consequently, these results suggest that chungkukjang may be beneficial in improving insulin resistance and hyperglycemia in type 2 diabetic animals that are partly medicated by the regulation of hepatic glucose enzymes and insulin sensitivity in peripheral tissues.

Fructo-oligosaccharide supplementation: effects on metabolic, endocrine and hematological traits in veal calves.

Fructo-oligosaccharides (FOS) are soluble fibres which exert various effects in the gastrointestinal tract, and induce metabolic and endocrine changes. The effects are favourable in diabetes mellitus, and may be favourable in veal calves, which during late periods of fattening often develop hyperglycemia, glucosuria and insulin resistance, especially during high lactose intake. Based on this we have studied metabolic, endocrine and haematological traits in veal calves (Simmental x Red Holstein) fed FOS (10 g/day; group GrF) or no FOS (group GrC). Whole milk and milk replacer in both groups, on a kg body weight basis, were fed in identical amounts. Experiments, lasting for 3 weeks, started when calves were 10 weeks old and weighed 117 kg. During week 3 lactose was supplemented to enhance post-absorptive glucose loads. Feed intakes were similar in both groups, but weight gain tended to be higher in GrF than GrC. The post-prandial increase of glucose concentrations was significantly smaller, of lactate tended to be smaller, and growth hormone peak frequency tended to be lower, whereas maximal insulin concentrations reached post-prandially were significantly higher in GrF than GrC. Eosinophil granulocytes increased during FOS feeding. In conclusion, FOS had basically similar effects on metabolic and endocrine traits in veal calves as in animals and humans with diabetes mellitus, but changes were small, albeit more prominent after lactose loads.

Immunocytochemical characterization of quisqualic acid- and N-methyl-D-aspartate-induced excitotoxicity in the retina of chicks.

A single, large dose of N-methyl-D-aspartate (NMDA) or quisqualic acid (QA) injected into the chick eye has been shown previously to destroy many retinal amacrine cells and to induce excessive ocular growth accompanied by myopia. The purpose of this study was to identify distinct populations of retinal cells, particularly those believed to be involved in regulating ocular growth, that are sensitive to NMDA or QA. Two pmol of NMDA or 0.2 micromol of QA were injected unilaterally into eyes of 7-day-old chicks, and retinas were prepared for observation 1, 3, or 7 days later. Retinal neurons were identified by using immunocytochemistry, and cells containing fragmented DNA were identified by 3'-nick-end labelling in frozen sections. NMDA and QA destroyed many amacrine cells, including those immunoreactive for vasoactive intestinal polypeptide, Met-enkephalin, and choline acetyltransferase, but they had little effect upon tyrosine hydroxylase-immunoreactive cells. Other cells affected by both QA and NMDA included those immunoreactive for glutamic acid decarboxylase, gamma-aminobutyric acid, parvalbumin, serotonin, and aminohydroxy methylisoxazole propionic acid (AMPA) receptor subunits GluR1 and GluR2/3. Cells largely unaffected by QA or NMDA included bipolar cells immunoreactive for protein kinase C (alpha and beta isoforms) and amacrine cells immunoreactive for glucagon. DNA fragmentation was detected maximally in many amacrine cells and in some bipolar cells 1 day after exposure to QA or NMDA. We propose that excitotoxicity caused by QA and NMDA induces apoptosis in specific populations of amacrine cells and that these actions are responsible for the ocular growth-specific effects of QA and NMDA reported elsewhere.

Development of embryonic chick insulin cells in culture: beneficial effects of serum-free medium, raised nutrients, and biomatrix.

A previous finding that insulin cells do not survive or differentiate in explants of embryonic avian pancreas cultured in collagen gel with a serum-containing medium has provided a model system for identification of conditions favorable for development of these cells. To this end, we here modify the substrate and the medium. The epithelial component of dorsal pancreatic buds of 5-d chick embryos was cultured for 7 d on Matrigel in serum-containing and in serum-free medium, the latter incorporating insulin, transferrin, and selenium. Endocrine cell types were distinguished by immunocytochemistry; insulin cell counts were expressed as a proportion of insulin plus glucagon cells. With serum-containing medium, Matrigel stimulated a significant increase in this proportion as compared with collagen gel--3.1% as against 0.2%; the serum-free medium further increased this proportion to 17.3%. Raising the level of essential amino acids approximately fivefold increased the latter figure somewhat (to 18.9%), but it was more than doubled (to 37.4%) by raising the glucose concentration from 10 mM to 20 mM. Raising the levels of amino acids and glucose simultaneously yielded a lesser increase (to 31.8%). Some cultures grown in collagen gel and serum-containing medium for 7 d were transferred to Matrigel and serum-free medium for a further 7 d. Insulin cell development recovered, indicating that progenitor cells had survived and were stimulated to develop by the improved conditions. This study indicates that components of the biomatrix and the medium (in particular, a raised glucose concentration) are important for the survival and differentiation of embryonic insulin cells.

[The vasopressin-, oxytocin- and corticoliberin-synthesizing structures of the hypothalamus in rats with diabetes mellitus under hypoxic exposures]. / Sostoianie vazopressin-, oksitotsin- i kortikoliberinsinteziruiushchikh struktur gipotalamusa u krys s sakharnym diabetom pri gipoksicheskikh vozdeistviiakh.

Hypothalamic mechanisms of neurohormone regulation of endocrine pancreas in diabetes mellitus, adaptation to hypoxia and their combination were studied on Wistar rats. To evaluate the condition of supraoptic nucleus (SON) secretory function, paraventricular subnuclei (PVH) of hypothalamus and endocrine pancreas, we used radioimmunoassay, immunocytochemical, morphometrical and histochemical methods. Hyperglycemia, hypoinsulinemia, glucagon and somatostatin synthesis and secretion intensification in diabetes mellitus is accompanied by marked reorganization of hypothalamic neurohormones (CRF, vasopressin, oxytocin) secretion with corresponding signs of activity increase of synthesizing their hypothalamus nuclei and subnuclei and also ACTH, corticosterone, cortisol rise in blood. Adaptation to hypoxia caused hypoglycemia, activated insulin biosynthesis, changed glucagon and somatostatin synthesis and secretion. CRF concentration, corticosterone and cortisol, ACTH in blood was not changed, vasopressin concentration lowered, oxytocin in median eminence of hypothalamus increased to a higher degree than in diabetes. Adaptation to hypoxia corrected impaired hormone balance and state of Langerhans islets (beta-cells destruction process inhibition, insulin biosynthesis stimulation, glucagon and somatostatin secretion decrease) in diabetes mellitus, hypothalamic neurohormones participating in this process.

Glucagon-like immunoreactivity in hypothalamic neurons of the rat.

Antisera specific for three different regions of pancreatic proglucagon were used to examine the distribution of such immunoreactivity in rat hypothalamus. Neurons in the supraoptic and paraventricular nuclei were immunoreactive with an antiserum against glucagon, but not with antisera directed towards the aminoterminal region of proglucagon (glicentin) or the glucagon-like peptide I sequence in the carboxyl-terminal region of proglucagon. These findings confirm a previous report of glucagon-like immunoreactivity in the supraoptic and paraventricular nuclei, but indicate that, while this material is immunochemically related to glucagon, it is not derived from a proglucagon-like precursor.

Tissue-specific differences in the levels of proglucagon-derived peptides in streptozotocin-induced diabetes.

The synthesis and secretion of proglucagon-derived peptides are regulated in a tissue-specific manner in pancreas, intestine, and brain. We have examined the plasma and tissue levels of these peptides during the first 3 weeks of streptozotocin (STZ)-induced diabetes in the rat. Plasma glucose levels were markedly elevated (P less than 0.0001) within 24 h of STZ administration. The plasma levels of glucagon-like immunoreactive (GLI) peptides were significantly elevated on days 8-22 of diabetes (P less than 0.05-0.001). Ileal GLI peptide concentrations rose in parallel with the plasma levels (r = 0.39; P less than 0.006) to 250% of control levels (P less than 0.001); however, pancreatic GLI peptide content increased only transiently on day 1 (P less than 0.05). No significant differences in the posttranslational processing of proglucagon in normal or diabetic rats could be detected. The increment in ileal GLI peptide content was not associated with increases in intestinal proglucagon mRNA transcripts. In contrast, pancreatic, but not intestinal, somatostatin mRNA levels were increased on day 22 of diabetes. Diabetic rats were found to have small but significant changes in GLI and immunoreactive glucagon peptide content of the hypothalamus and medulla oblongata (P less than 0.05). These observations suggest that STZ-induced diabetes may produce tissue-specific perturbations in the biosynthesis and secretion of the proglucagon-derived peptides.

Effects of basal insulin supplementation on disposition of mixed meal in obese patients with NIDDM.

Basal insulin supplementation has been used as a therapy for patients with non-insulin-dependent diabetes mellitus (NIDDM) who require insulin. To determine whether basal insulin supplementation in addition to lowering postabsorptive plasma glucose concentration also improves the postprandial pattern of glucose disposition, glucose metabolism after ingestion of a solid mixed meal was assessed in obese patients with NIDDM before and after treatment with ultralente and compared with glucose metabolism observed in nondiabetic subjects. Splanchnic uptake of ingested glucose clearance was assessed by including [2-3H]glucose (a tracer that only minimally cycles through glycogen) in a solid mixed meal. Postprandial gluconeogenesis was estimated by measuring the rate of incorporation of carbon dioxide into glucose. Net glucose and lipid oxidation were measured by indirect calorimetry. Both splanchnic uptake of ingested glucose (27 +/- 1 vs. 14 +/- 2 g) and postprandial hepatic glucose release (51 +/- 5 vs. 24 +/- 3 g) were greater (P less than .001) in diabetic than in nondiabetic subjects. Although the percentage of postprandial hepatic glucose release accounted for by glucose synthesis from bicarbonate was similar in the two groups (25 +/- 2 vs. 35 +/- 5%), the absolute rate was greater in the diabetic patients (13 +/- 1 vs. 8 +/- 1 g; P less than .05). Postprandial glucose oxidation and glucose disposal (measured either isotopically or by the forearm-catheterization technique) were similar in both groups. However, total lipid oxidation was increased in the diabetic patients. (P less than .05). Two weeks of basal insulin supplementation lowered fasting glucose concentrations (from 219 +/- 22 to 144 +/- 21 mg/dl; P less than .01) and integrated postprandial glycemic response (from 814 +/- 68 to 621 +/- 72 min.mg.ml-1) but not to normal. Although circulating insulin concentrations were two- to threefold greater (P less than .02) after 3 mo of basal insulin supplementation, the postprandial pattern of glucose metabolism remained essentially the same. Basal insulin supplementation decreased (P less than .05) both splanchnic uptake of ingested glucose and hepatic glucose release. The addition of a preprandial injection of soluble insulin to basal insulin supplementation further suppressed (P less than .05) postprandial hepatic glucose release, thereby further improving postprandial glucose tolerance. These studies indicate that initial splanchnic glucose clearance, hepatic glucose release, and new glucose synthesis, as well as extrahepatic substrate metabolism, are altered in NIDDM after ingestion of a mixed meal.(ABSTRACT TRUNCATED AT 400 WORDS)

Distribution and molecular forms of glucagon-like peptide in the dog.

Using glucagon-like peptide-1 N-terminus and C-terminus directed antisera, we investigated concentration and molecular forms of GLP-1 immunoreactivity (IR) in extracts of various tissues of the dog. GLP-1 IR measured with C-terminus-directed antiserum R2337 (GLP-1 IR-CT) was high in the ileum, appendix, jejunum, colon, and gastric fundus and body. GLP-1 IR measured with N-terminus-directed antiserum R1043 (GLP-1 IR-NT) was high only in the pancreas, and gastric fundus and body. Only GLP-1 IR-CT was found in the hypothalamus, thalamus and medulla oblongata. No immunoreactive materials were detected in the liver, spleen and kidney. Gel-filtration with Sephadex G-50 showed two peaks of both GLP-1 IR-CT and GLP-1 IR-NT, at 10kd and at the position of GLP-1 (1-36 amide) in the pancreatic extract, and one peak at 10kd in the stomach extract. Ileal extracts showed 3 peaks of GLP-1 IR-CT at 10kd, at the position of GLP-1(1-36 amide) and GLP-1(7-36 amide), respectively, but GLP-1 IR-NT was coeluted with GLP-1(1-36 amide). Hypothalamic extracts showed a single peak at the position of GLP-1(7-36 amide). These results suggest that processing of preproglucagon differs in different organs, and that the main GLP-1-related products are a large molecular form and GLP-1(1-36 amide) or GLP-1(1-37) in the pancreas, and GLP-1(7-36 amide) or GLP-1 (7-37) in the ileum and hypothalamus.

Insulin-like growth factor-I, GH, insulin and glucagon concentrations in bovine colostrum and in plasma of dairy cows and neonatal calves around parturition.

1. The IGF-I concentrations in colostrum on days 1 and 2 after parturition were higher than those in cow and neonate plasma. 2. The modest increase in GH concentrations in cow plasma around parturition would not be enough to stimulate IGF-I release by tissues. 3. The concentrations of insulin, GH and glucagon in colostrum were substantially lower than those in plasma.

Oxyntomodulin and glicentin: brain-gut peptides in the rat.

Glucagon-like materials and glucagon have been identified by immunoassay and immunocytochemistry in the mammalian central nervous system. However, the molecular forms relevant to brain glucagon-like immunoreactivity (GLI) have not been precisely defined. In the rat small intestine, more than 90% of GLI is constituted by two peptides: oxyntomodulin (OXM) and glicentin. This work was initiated to characterize and determine the concentrations of these two peptides and glucagon in the rat central nervous system and to compare their relative proportions with those found in the gut. Different regions from the adult rat brain were analyzed by HPLC in association with RIA, using a central glucagon antiserum and an antibody directed toward the C-terminal end of OXM and glicentin. The elution profiles of hypothalamus extracts were constituted by two main peaks, both detected by the two antibodies used and displaying the same retention times as glicentin and OXM, respectively. A third small peak, which coeluted with glucagon, was constantly recorded with the central glucagon antiserum. The percentages of glicentin, OXM, and glucagon in 10 hypothalami were 37 +/- 1%, 55 +/- 1%, and 8 +/- 2%, respectively (n = 8). This distribution was quite similar to that in small intestinal extracts (38 +/- 1%, 59 +/- 1%, and 1.3 +/- 0.1%, respectively; n = 7); however, the peptide concentrations were almost 50-fold greater in intestine than in hypothalamus. In the medulla oblongata, the same peptide ratio was observed, with 10-fold lower concentrations compared to those in hypothalamus. In olfactory bulb, cerebellum, and cortex the concentrations were close the the detection limit, whereas they could be not detected in the pituitary. The combination of HPLC and specific RIAs allowed us to unambiguously characterize OXM and glicentin as the major components of GLI in the rat hypothalamus and medulla oblongata. The same proportion of these two peptides in the central nervous system and the gut indicates that a similar posttranslational processing exists in these rat tissues, another example of the brain-gut axis.

Distribution of glucagonlike peptide I (GLP-I), glucagon, and glicentin in the rat brain: an immunocytochemical study.

Although glucagonlike immunoreactants (GLIs) are present in the central nervous system of several mammalian species, their structural relationship with pancreatic proglucagon is not defined, and their precise anatomical distribution has not been studied extensively. To obtain further information about the structure and biological significance of brain GLIs, the anatomical distribution of three different antigenic determinants of pancreatic proglucagon--glucagonlike peptide I (GLP-I), glucagon, and glicentin--was mapped in the brain of colchicine-treated rats by immunocytochemistry using the avidin-biotin-peroxidase method. Neuronal cell bodies immunoreactive with antisera specific for GLP-I, glucagon, and glicentin were found only in the caudal medulla oblongata. Within the caudal medulla immunostained cell bodies were found at levels from approximately 0.55 mm rostral to the obex to 0.45 mm caudal to the obex, and were located within the nucleus of the solitary tract (NTS) and the dorsal (MdD) and ventral (MdV) parts of the medullary reticular nucleus. The NTS contained three times more immunoreactive cell bodies than the MdD and MdV, and these cell bodies were located in the midline, medial, and lateral subnuclei of the caudal third of the NTS. Immunostaining of the same cell bodies in paired adjacent sections incubated with GLP-I and glucagon antisera or glucagon and glicentin antisera provided evidence for coexistence of the three antigens within the same neurons of the NTS. Nerve fibers and terminals immunoreactive with GLP-I, glucagon, and glicentin antisera were widely distributed throughout the rat brain and there was no discernible difference in the distribution of fibers and terminals immunoreactive with each of the three antisera. The highest densities of immunostained fibers and terminals were observed in the hypothalamus, thalamus, and septal regions, and the lowest in the cortex and hindbrain. The localization of neuronal cell bodies containing GLP-I, glucagon, and glicentin within the NTS and the MdD and MdV, and the extensive distribution of immunoreactive fibers and terminals throughout the rat brain suggest a role for these peptides in the integration of autonomic as well as central nervous system functions.

Pituitary, hypothalamic and pancreatic 7B2 concentrations in rats with streptozotocin-induced diabetes and spontaneously diabetic mice.

The pituitary protein, 7B2 has been demonstrated in the pancreas and is known to be present in very high concentrations in pancreatic endocrine tumors. To investigate whether any changes in 7B2 concentrations might be present in the pancreases affected by different types of diabetes and whether the diabetic state itself might affect pituitary and hypothalamic 7B2 concentrations, streptozotocin (STZ)-treated rats and mice with natural-onset diabetes (obese hyperglycemic, or ob/ob, mice and non-obese diabetic, or NOD, mice) were employed. A significant reduction in pancreatic 7B2 concentrations was found in STZ-treated rats. The pancreatic 7B2 concentration was significantly high in ob/ob mice (p less than 0.05, versus the concentration in their lean littermates, and the decrease observed in older NOD mice, appeared to parallel their insulin reserve. Pituitary and hypothalamic 7B2 concentrations were similar in STZ-treated and control rats. A reduction in pituitary and hypothalamic 7B2 concentrations was observed in older NOD mice (both p less than 0.01 versus respective values in younger mice). In ob/ob mice, a significant reduction was also found in hypothalamic 7B2 concentration (p less than 0.01 versus that in control mice). Gel permeation chromatography showed that 7B2 immunoreactivity comprised two molecular components, and the relative proportion in the pancreas differed from that in other tissues. In the pancreas, a smaller molecular component was predominant (elution coefficient, 0.62).(ABSTRACT TRUNCATED AT 250 WORDS)

Identification and localization of glucagon-like peptide-1 and its receptor in rat brain.

The existence and distribution of glucagon-like peptide-1 (GLP-1) and its receptor in rat brain in relation to that of glucagon were examined. The concentration of GLP-1 immunoreactivity (GLP-1-IR), measured by a specific and sensitive RIA established in this study with anti GLP-1 serum (LMT-01), was found to be highest in the thalamus-hypothalamus, followed by the medulla oblongata. The distribution of glucagon-like immunoreactivity was similar to that of GLP-1-IR. However, appreciable glucagon immunoreactivity was detected only in the thalamus-hypothalamus. Gel filtration analysis showed the presence of GLP-1-IR of various molecular weights in the extract of thalamus-hypothalamus including that eluted at the same position as synthetic GLP-1 (1-37); moreover, HPLC analysis also confirmed the presence of GLP-1-IR, eluted at the exact position as synthetic GLP-1 (1-37). The distribution of receptors for GLP-1 corresponded with that of GLP-1-IR in the rat brain, except in the pituitary gland. The distribution of these receptors was also similar to that of glucagon receptors. The thalamus-hypothalamus, pituitary gland, and medulla oblongata were rich in GLP-1 and glucagon-binding sites. The binding affinities of GLP-1 and glucagon were in the nanomolar range [disocciation constant Kd approximately equal to 4 nM]. The presence of specific, high affinity receptors for GLP-1 was confirmed by demonstrating that GLP-1 stimulated cAMP formation in the thalamus-hypothalamus and the pituitary gland. The concentration of GLP-1 required for half-maximal stimulation of cAMP formation in these regions was about 1 nM. These results suggest that GLP-1 may be synthesized in certain parts of the brain and play a role as a neurosignal transmitter.

Immunohistochemistry of the hypothalamic neuropeptides and anterior pituitary cells in the Japanese quail.

The pars distalis of the avian adenohypophysis consists of well-defined cephalic and caudal lobes which are distinct in their cellular constituents. Immunocytochemical investigations on the pituitary hormones of the pars distalis of the Japanese quail reveal five types of secretory cells, adenocorticotropin (ACTH) cells, prolactin (PRL) cells, thyroid-stimulating hormone (TSH) cells, growth hormone GH (STH) cells, and FSH/LH (gonadotropic) cells. The ACTH cells, TSH cells, and PRL cells are restricted to the cephalic lobe, and GH (STH) cells are confined to the caudal lobe, while FSH/LH cells are distributed throughout the cephalic and caudal lobes. The median eminence of birds has distinct anterior and posterior divisions, each with different neuronal components. The avian hypophysial portal vessels also consists of two groups, anterior and posterior. The peculiar arrangement and distribution of the avian hypophysial portal vessels are possibly related to the distribution of neuropeptides in the two divisions of the median eminence and to the cytological and functional differentiation of two lobes of the pars distalis. The localization of perikarya and fibers containing luteinizing hormone releasing hormone (LHRH), somatostatin, vasotocin, mesotocin, corticotropin-releasing factor (CRF), vasoactive intestinal polypeptide (VIP), glucagon, metenkephalin, and substance P in the hypothalamus and median eminence of the Japanese quail has been investigated by means of immunohistochemistry using antisera against the respective neuropeptides. LHRH-, somatostatin-, VIP-, met-enkephalin-, and substance P-immunoreactive fibers are localized in the external layer of the anterior and posterior divisions of the median eminence, while CRF- and vasotocin-reactive fibers are demonstrated only in the external layer of the anterior division of the median eminence. The metenkephalin fibers are thicker in the anterior median eminence but the substance P fibers are more abundant in the posterior division. Mesotocin fibers occur only in the internal layer of the median eminence and neural lobe.

Neural network of glucose monitoring system.

Glucose-sensitive neural elements exist in the hypothalamus, the nucleus of the solitary tract (NTS) and autonomic afferents from visceral organs such as liver and gastrointestinal tract. Glucose affects neural activity through these central and peripheral chemosensors. Glucose is generally suppressive in the liver, the NTS and the lateral hypothalamic area (LHA), and generally excitatory in the small intestine and ventromedial hypothalamic nucleus (VMH). The hypothalamus is involved in the control of pancreatic hormone secretion through autonomic efferent nerves. Stimulation or lesion of the hypothalamus induces various changes in pancreatic autonomic nerve activity. The VMH, the dorsomedial hypothalamic nucleus and the paraventricular nucleus have inhibitory effects on vagal nerve activity and excitatory effects on splanchnic nerve activity. The LHA is excitatory to the vagal nerve, and both excitatory and inhibitory to the splanchnic nerve. These findings suggest that the neural network of the glucose monitoring system, which also analyzes and integrates information concerning other metabolites and peptides in the blood and cerebrospinal fluid, contributes to regulation of peripheral metabolism and endocrine activity as well as feeding behavior. The physiological function and input-output organization of this network are discussed.

Synaptosomal localization and release of glucagon-like materials in the rat brain.

The subcellular localization of glucagon-like materials in the thalamus-hypothalamus and brain stem of the rat was investigated. Both glucagon immunoreactivity (GI) determined by C-terminal specific antibody and glucagon-like immunoreactivity (GLI) determined by non-specific antibody were enriched in the microsomal and synaptosomal fractions relative to the nuclear, myelin and mitochondrial fractions. Furthermore, the synaptosomal fraction of both the thalamus-hypothalamus and brain stem incubated in Krebs-Ringer bicarbonate buffer with 55 mM K+ at 37 degrees C released GI and GLI in the presence of Ca++. These findings suggested that glucagon-like substances detected in the brain have a role in the synaptic function.