Steen Lindkœr Jensen

Distribution and molecular forms of peptides containing somatostatin immunodeterminants in extracts from the entire gastrointestinal tract of man and pig

Specimens from human porcine mucosal and muscular tissue from the entire gastrointestinal tract were extracted in acid ethanol, subjected to chromatography and analysed for somatostatin-like immunoreactivity by region-specific radioimmunoassays. The concentration of somatostatin-like immunoreactivity from man and pig ranged from 1.13 +/- 0.37 to 101.15 +/- 33.93 pmol/g wet weight, and from 7.64 to 159.48 +/- 23.79 pmol/g wet weight, respectively. In both species the highest concentrations were found in the jejunum. The immunoreactivity in intestinal mucosal extracts was distributed among four major peaks, two of which were identified by HPLC as somatostatin 1-28 and somatostatin 1-14, respectively. A peak of approx. 10 kDa was resolved by ion exchange plus HPLC into three components, two containing at least part of the somatostatin 1-14 sequence as well as (part of) the somatostatin 1-28(1-14) sequence (but differing in charge), the third containing only the 1-28(1-14) sequence. These peptides probably represent uncleaved and partially cleaved prosomatostatin. The fourth component to be identified by gel filtration was slightly larger than somatostatin 1-14. Extracts from the antrum, the pancreas and from muscular tissues contained almost exclusively somatostatin 1-14, and very little somatostatin 1-28, indicating that the somatostatin precursor is processed differently at these sites. Furthermore, extracts of porcine gastric antrum, analysed for somatostatin 1-28(1-14) immunoreactivity, showed two immunoreactive forms in the mucosa and three major forms in the muscular layers.

Secretin release from the isolated, vascularly perfused pig duodenum.

1. A method for the isolation and vascular perfusion of the porcine pancreas and duodenum was developed. 2. The oxygen consumption of the whole preparation was similar to that of the pancreas alone, and since the duodenal arteriovenous oxygen deficit was similar to that of the total preparation, it was concluded that the duodenum respired adequately. 3. The duodenum rapidly absorbed luminally administered radioactive glucose, and this absorption was strongly inhibited by ouabain and phloridzin. 4. The duodenum secreted secretin rapidly in response to hydrochloric acid, but did not respond to any other luminal stimuli, including lipids, proteins, carbohydrates and bile. Neither was secretin release stimulated by intra‐arterially injected acetylcholine. 5. By gel permeation chromatography the release immunoreactive secretin behaved identically to pure natural secretin, indicating that the tissue form and the circulating form have identical molecular size. 6. It is concluded that this model offers an unique opportunity to study the endocrine secretion of the duodenum.

Effect of Sulfation of CCK-8 on Its Stimulation of the Endocrine and Exocrine Secretion from the Isolated Perfused Porcine Pancreas

The secretory effect of sulfated and nonsulfated cholecystokinin octapeptide (CCK-8) was studied on the isolated perfused porcine pancreas. Both CCKs in concentrations from 10(-11) to 10(-8) mol/l in the presence of glucose (7.5, 5.0 or 3.5 mmol/l) were without effect on insulin and glucagon release. The exocrine secretion was stimulated by both CCKs in a dose-dependent manner, but sulfated CCK-8 was considerably more potent. The study shows that CCK-8, a major constituent of endogenous CCK, does not contribute to the incretin mechanism, irrespective of degree of sulfation. In contrast, CCK-8 is a potent stimulator of exocrine pancreatic secretion. For this effect sulfation is crucial.

Effect of intragastric pH on antral gastrin and somatostatin release in anaesthetised, atropinised duodenal ulcer patients and controls.

The synchronous change in the antral release of gastrin and somatostatin into a vein draining the stomach was studied during acidic and alkaline intragastric pH in six anaesthetised duodenal ulcer patients and six controls after atropinisation. No differences in the basal secretion of gastrin and somatostatin were observed among the two groups. Alkaline as well as acidic intragastric pH had no effect on the antral release of somatostatin in duodenal ulcer patients and controls. In contrast, alkaline intragastric pH was associated with a significantly higher antral gastrin release in duodenal ulcer patients than in controls. Acidic intragastric pH was associated with a significantly smaller inhibition of antral gastrin release in duodenal ulcer patients than in controls. These results suggest that atropinised anaesthetised duodenal patients release gastrin abnormally in the presence of acidic or alkaline intragastric pH and that any inverse relationship between antral gastrin and somatostatin release is uncoupled under these conditions.

Galanin: Distribution and Effect on Contractile Activity and Release of Vasoactive Intestinal Polypeptide from the Isolated Perfused Porcine Ileum

In the pig ileum galanin (GAL)-like immunoreactivity was identified in nerve cell bodies of the submucous plexus and in nerve fibers of the circular and longitudinal muscle layer. Infusion of 5.10(-10)-10(-8) M of GAL into the arterial line of the isolated perfused porcine ileum decreased the frequency of spontaneous phasic contractions in a dose-dependent manner. The frequency of phasic contractions during maximal inhibition by GAL 10(-8) M was 13 +/- 4% (mean +/- SE) of basal frequency (p less than 0.05). The recovery from inhibition by GAL 10(-8) M lasted 16 +/- 1 min. Tonic contractions were not observed in this experimental set-up, neither by standard perfused catheter manometry nor by measurement of cross-sectional area of an intraluminally located balloon. Infusion of GAL 10(-8) M decreased the venous release of vasoactive intestinal polypeptide to 80 +/- 8% of basal release (p less than 0.05). It is concluded that GAL may participate in the regulation of small intestinal motility in the pig.