Hans-Jürgen Hahn

Calcium and pancreatic β-cell function: 6. glucose and intracellular 45Ca distribution

Glucose stimulates the uptake of 45Ca into beta-cell-rich pancreatic islets isolated from ob/ob-mice. The distribution of the incorporated radioactivity was analysed by labelling the organelles with 45Ca in their cellular environment. The radioactive content of the organelles was measured after homogenization and fractionation of the islets under conditions preventing 45Ca redistribution. The 45Ca taken up in response to glucose appeared essentially in the secretory granule fraction and in that enriched in mitochondria. Modification of the 45Ca loading procedure, involving reduction of the oxygen tension and incubation volume, resulted in the disappearance of the glucose effect on the mitochondrial fraction whereas part of the stimulatory effect on the secretory granules persisted. Buffering of calcium by the secretory granules and mitochondria may be important for regulating the cytoplasmic Ca2+ involved in stimulus-secretion coupling.

Regional distribution of glucagon-like immunoreactive material in the brain of rats and sand rats. An immunohistochemical investigation

Glucagon-like immunoreactivity was demonstrated in different parts of rat and sand rat central nervous system by the indirect immunofluorescence method. In hyperglycemic animals the level of immunoreactive material was reduced to a great extent.

An immunofluorescent reaction appears to insulin-antiserum in different CNS regions of two rat species

Using immunofluorescence technique a positive reaction to insulin antiserum could be revealed in different parts of the CNS of Wistar rats and sand rats.

C-peptide immunoreactive neurons in human brain

C-peptide/C-peptide-like immunoreactivity was shown to be present in the cytoplasm of the soma and the proximal part of apical dendrites of some pyramidal cells in the Neocortex (Gyrus precentralis) and Hippocampus of man. C-peptide (connecting peptide) is a metabolic product in insulin biosynthesis and its localization in neurons is a proof for extrapancreatic insulin production.

Immunoreactive glucagon in neurons of various parts of the human brain. Demonstration by immunofluorescence technique

The presence of glucagon-like immunoreactivity in nerve cells of different parts of the human brain was demonstrated by the indirect immunofluorescence technique. A bright fluorescent reaction was observed in the pyramidal cells of lamina V of the Neocortex. Less prominent concentrations of the glucagon-like material were detected in a few pyramidal cells of the Hippo-campus and in some neurons of the Presubiculum and Subiculum. Within the Corpus amygdaloideum, only a few magnocellular neurons showed a positive reaction. The Hypothalamus was evidenced by a moderate, but widely distributed, reaction in magnocellular and medium-sized nerve cells in different nuclei (especially Nuc. ventromedialis and Nuc. arcuatus). A strong immunofluorescence was localized to some neurocytes in the Nuc. amibigus, and Nuc. n. hypoglassi. The Purkinje cells of the cerebellar cortex were free from immunoreactive material, but fluorescence occurred in some very small nerve cells of the Cerebellum (probably granular cells). A dependence of the strength of immunofluorescence of the time delay between autopsy and death is shown.

Investigations on isolated islets of langerhans in vitro: 16. Modification of the glucose-dependent inhibition of glucagon secretion☆

Abstract Investigation of glucagon secretion in isolated Wistar rat islets was carried out to elucidate further the regulatory function of glucose and arginine on pancreatic A-cells. The suppressive effect of D -glucose could also be demonstrated with L -glucose, D -mannose, D -fructose, D -galactose, D -glyceraldehyde and DL -dihyroxyacetone, but not in the presence of 3- O -methylglucose or mannitol. Sugars other than D -glucose inhibited glucagon secretion only at much higher concentrations than those at which D -glucose was effective. Furthermore, although 7.5 mM D -glucose caused up to 80% inhibition, the effects of other sugars appeared to level off at only 50–60% inhibition. The inhibitory action of D -glucose or D -glyceraldehyde on glucagon secretion could not be overcome by L -arginine, but 3- O -methylglucose, mannoheptulose, 2-deoxy- D -glucose, iodoacetamide, theophylline, epinephrine and acetylcholine were effective. The insulin secretion in response to glucose was inhibited by the metabolic inhibitors used, whereas the B-cell response in the presence of glyceraldehyde was diminished by iodoacetamide only. Like D -glucose, a variety of other sugars markedly reduced the stimulatory effect of L -arginine in glucagon release. The results show that the suppression of glucagon secretion is not specific for D -glucose and not strongly connected on a stimulated insulin secretion.

Neuron-specific enolase (NSE) as a neuroendocrine cell marker in the human fetal pancreas

Using the PAP technique, we investigated the presence of neuron-specific enolase in the human fetal pancreas of 10, 12, and 14 weeks of gestational age. Neuron-specific enolase is present in the islet cells in the 10th week. Positive cells are situated mainly in duct epithelium. The number of cells with a positive reaction increases from the 12th to the 14th week. In the 14th week, they are clustered either near the ducts or between the acini. The numbers and localizations of the cells correspond to those obtained in previous studies with 4 basic islet cell types in the same material. The present results are a further proof that islet cells are biologically active during early fetal development.

Are certain peculiarities in the distribution of somatostatin-like immunoreactivity (SLI) in the brain of sand rats (Psammomys obesus) possibly connected with an altered carbohydrate metabolism?

The hypothalamic and extrahypothalamic distribution of somatostatin-like immunoreactivity (SLI) was investigated in fed adult sand rats (Psammomys obesus) of both sexes using the immunofluorescence technique. Blood glucose and insulin concentrations were determined in these animals prior to decapitation. In a blind study, the amount of immunofluorescence of certain CNS areas and the glucose: Insulin ratio were compared and found to be connected: High amounts of SLI were detected in the dorsal hippocampus, in some hypothalamic nuclei and the median eminence of rats displaying low glucose levels. Sand rats with high blood glucose values did not show SLI in the hippocampal formation and the immunofluorescence of the circumventricular (hypothalamic) regions and the Eminentia mediana was drastically reduced. A possible correlation of SLI with insulin concentrations in the blood could not be revealed. The data obtained are discussed as a possible expression of a CNS influence on glucoregulation in this species.

Polypeptides in the human brain

Summary Using an indirect fluorescence technique and the unlabelled enzyme technique of Sternberger (PAP-techniques) we were able to demonstrate a widespread distribution of insulin, C-peptide and glucagon throughout the human brain.

Detection of proinsulin, C-peptide, insulin-A-chain, and Glicentin in pancreatic islet cells of early human fetogenesis

The presence of C-peptide, proinsulin, insulin-A-chain, and glicentin in human fetal pancreatic cells by using the PAP-technique was investigated and the results obtained compared with the occurrence of insulin or glucagon immunoreactive cells. In pancreatic sections obtained from 10 weeks old human fetuses we could identify cells reacting with antibodies directed against C-peptide, proinsulin, and insulin-A-chain. The majority of the cells were found in the duct epithelium and their number increased from the 10th to 14th week forming clusters near the ducts. The number and localization of the cells correspond exactly to the insulin positive cells. The presence of proinsulin and insulin-A-chains is a further proof of biological activity already in an early step of fetal development. The presence of glicentin-positive cells in the 10th week of gestational age as well as cells reacting with glucagon antibodies provide evidence for active glucagon biosynthesis. The number of these cells increased markedly in the 14th week of gestational age.