Bogusław Sawicki

Cannabinoids enhance gastric X/A - like cells activity

It has been reported that cannabinoids may cause overeating in humans and in laboratory animals. Although, endogenous cannabinoids and their receptors (CB1) have been found in the hypothalamus, and recently also in gastrointestinal tract, the precise mechanism of appetite control by cannabinoids remains unknown. Recently, ghrelin--a hormone secreted mainly from the stomach X/A-like cells was proposed to be an appetite stimulating agent. The aim of this study was the evaluation of the influence of a single ip injection of a stable analogue of endogenous cannabinoid--anandamide, R-(+)-methanandamide (2.5 mg/kg) and CP 55,940 (0.25 mg/kg), an exogenous agonist of CB1 receptors, on ghrelin plasma concentration and on ghrelin immunoreactivity in the gastric mucosa of male Wistar rats. Four hours after a single injection of both cannabinoids or vehicle, the animals were anaesthetized and blood was taken from the abdominal aorta to determinate plasma ghrelin concentration by RIA. Subsequently, the animals underwent resection of distal part of stomach. Immunohistochemical study of gastric mucosa, using the EnVision method and specific monoclonal antibodies against ghrelin was performed. The intensity of ghrelin immunoreactivity in X/A-like cells was analyzed using Olympus Cell D image analysis system. The attenuation of ghrelin-immunoreactivity of gastric mucosa, after a single injection of R-(+)-methanandamide and CP 55,940 was accompanied by a significant increase of ghrelin plasma concentration. These results indicate that stimulation of appetite exerted by cannabinoids may be connected with an increase of ghrelin secretion from gastric X/A-like cells.

IMMUNOCYTOCHEMICAL STUDY OF PARAFOLLICULAR CELLS OF THE THYROID AND ULTIMOBRANCHIAL REMNANTS OF THE EUROPEAN BISON

The aim of the present study was to compare parafollicular cells in the bison thyroid and its ultimobranchial remnants. The thyroid of 26 European bisons was fixed in Bouins fluid, 5 microns thick paraffin sections were stained with hematoxylin and eosin. Azan or silver Grimelius methods. For immunocytochemical analysis specific rabbit antisera were used against human calcitonin (CT), human calcitonin gene-related peptide (CGRP), bovine (b) or rat (r) neuron-specific enolase (NSE), human synthetic somatostatin (ST), and porcine chromogranin. Strongly positive reactions in the majority of parafollicular cells were observed after application of antisera against CT, CGRP, bNSE and rNSE only. ST-immunopositive cells were found in small numbers. Immunopositive parafollicular cells were also present outside typical structures of the thyroid within persistent ultimobranchial remnants. In persistent ultimobranchial bodies, parafollicular cells were frequently observed in groups between ultimobranchial follicles in form of solid cell nests. Many of these cells did not react with any of the antisera used and showed features of immature cells. It is concluded that histomorphologic analysis and immunocytochemical examination reveals a heterogeneous population of parafollicular cells in the bison thyroid, and this heterogeneity was particularly clear in persistent ultimobranchial bodies.

Immunocytochemical study of parafollicular (C) cells of the thyroid in some wild rodents

Studies were done on 3 wild species of rodents: field voles (Microtus agrestis, Linnaeus 1761), bank voles (Clethrionomys glareolus, Schreber 1780), and forest mice (Apodemus flavicollis, Melchior 1834). Immunocytochemical reactions were used to detect calcitonin (CT), calcitonin gene-related peptide (CGRP), neuron-specific enolase (NSE), chromogranin A (CgA) in the thyroid parafollicular (C) cells in all species examined. Antisera to human CT, rat CGRP, bovine CgA, rat NSE and human NSE give probably a positive reaction in all C cells in the rodents examined. However, in the NSE determining reaction, much feebler positive C cells were observed. Individual variation in respect of shape and distribution of C cells was observed in all species. In forest mice the C cells resembled in shape the C cells previously described in mouse and rat. In field voles and bank voles the C cells were assembled into small groups more often than in forest mice. Antibodies (anti-human CT, anti-rat CGRP, anti-bovine CgA, anti-rat NSE and anti-human NSE) used by us were good markers of C cells in the thyroids of the 3 species of free-living rodents examined.

Estimation of gastric ghrelin-positive cells activity in hyperthyroid rats

Ghrelin is a peptide of 28 amino acids that transmits appetite related signals from peripheral organs to the brain. The main source of ghrelin is stomach. The regulation of ghrelin secretion is still unknown. The finding that fasting and food intake, respectively increase and decrease the secretion of ghrelin suggests that this hormone may be a bridge connecting somatic growth with energy metabolism and appears to play an important role in the alteration of energy homeostasis and body weight in pathophisiological conditions. The purpose of this study was the evaluation of gastric ghrelin immunoreactivity and ghrelin plasma concentration in male Wistar rats with hyperthyroidism. Experimental model of hyperthyroidism was induced by intraperitoneal injection of levothyroxine at the dose of 80 microg/kg daily over 21 days. At the end of experiment the animals were anaesthetized, blood was taken from abdominal aorta to determinate plasma ghrelin concentration by RIA and then the animals underwent resection of distal part of stomach. Immunohistochemical study were performed using monoclonal specific antybodies against ghrelin. Hyperthyroidism was a reason of increase of gastric mucosal ghrelin - immunoreactivity, accompanied by a significant decreased of ghrelin plasma concentration. Those observations may indicate, that chronic administration of L-thyroxine cause the change of ghrelin plasma concentration in rats, probably via direct influence on gastric X/A-like cells, but this effect is not responsible for hyperphagia associated with hyperthyroidism.

C-Zellen in der Schilddrüse und in den Resten des Ultimobranchialkörpers des Wisents

Summary C cells of the thyroid gland of 85 European bison, 43 males and 42 females, of age ranging from 1 month to 25 years were examined. For the identification of the C cells by Grimelius silver impregnation, toluidine blue, pseudoisocyanin and azur A stainings were employed. In the thyroid of the bison the C cells in the central region were distributed denser than in the peripheral region. They may particularly appear in great numbers in the interior of ultimobranchial (UB) remnants; in these cases, however, a great variety of changes has been observed. In part of the adults animals individual differences of the hyperplasia C cells in the preserved UB structures were observed: C cells have not been detected in the internal parathyroid glands. In the discussion, the meaning of paracrine has been described in autoregulatory mechanisms controlling the thyroid activity.