An immunohistochemical study of the endocrine cells within the pancreas, intestine, and stomach of the gar (Lepisosteus osseus L.)

The distribution and identity of the various endocrine cell types were examined in the pancreas, stomach, and anterior intestine of the phylogenetically ancient actinopterygian, the gar (Lepisosteus osseus L.), using immunohistochemistry. Antisera used were directed against several insulins (INSs) and somatostatins (SSTs), and members of the pancreatic polypeptide (PP, aPY, NPY) and glucagon (GLUC, GLP) families. In the gar pancreas the most pronounced aggregation of islet tissue is among the exocrine acini near the union of extrahepatic common bile duct with the gastrointestinal junction. Four immunoreactive cell types were identified within well-defined islets (A, B, D, and F cells) but immunoreactive cell types were also seen isolated among the exocrine acini. Centrally located B cells were immunoreactive with mammalian and lamprey INS antisera whereas the widely dispersed D cells immunostained with anti-SST-14, -25, and -34. SST was also localized in the epithelium of the pancreatic ducts. There was a colocalization of immunoreactivity for each member of the PP and GLU families at the periphery of each islet to identify F and A cells, respectively. However, colocalization of peptides from both families is suspected for at least some cells. Although the gastric and intestinal mucosae showed a similar pattern of immunoreactivity to GLP and not GLU, they had contrasting immunoreactivity with the two INS antisera. SST immunoreactivity was restricted to the stomach, whereas three of the four PP-family peptides were only immunoreactive in the intestine. Immunoreactivity to the various antisera used in the study imply that there may be an organ-specific processing of preproinsulin, that the gar SST profile may be more similar to agnathan and bowfin rather than either elasmobranch or teleost SSTs, and that only the GLP portion of the preproglucagon gene is expressed in the gastrointestinal mucosa. Our results are consistent with other recent endocrine studies showing that the gar is a widely distinct actinopterygian.

Isolation, localization, and cardiovascular activity of tachykinins from the stomach of the bowfin Amia calva.

The bowfin is an extant representative of an ancient group of ray-finned fish with evolutionary connections to modern teleosts. A peptide with substance P-like immunoreactivity was isolated from an extract of bowfin stomach and its primary structure was established as Ser-Lys-Ser-His-Gln-Phe-Tyr-Gly-Leu-Met-NH2. This amino acid sequence resembles mammalian substance P only in the COOH-terminal region of the peptide. A second tachykinin with neurokinin A-like immunoreactivity isolated from the extract comprises 23 amino acid residues and shows limited structural similarity to mammalian neuropeptide-gamma. A randomly distributed population of cells in the gastric glands of the bowfin were immunostained with an antiserum raised against substance P, but no immunopositive structures were identified in the surface epithelium, lamina propria, or the nerve plexuses of the submucosa. Bolus injections of synthetic bowfin substance P (0.1-10 nmol/kg) into the bulbus arteriosus of unanesthetized bowfin resulted in a significant and dose-dependent rise in vascular resistance and arterial blood pressure (P < 0.01) and a fall in cardiac output (P < 0.05) without change in heart rate. After 5-10 min, arterial pressure and vascular resistance returned to preinjection levels, but cardiac output significantly (P < 0.05) increased over baseline values. The response to the peptide was unaffected by pretreatment of the animals with phentolamine. The study has shown that the stomach of the bowfin synthesizes tachykinins with novel structural features that display cardiovascular activity in this species.

The corpuscles of Stannius in arawana (Osteoglossum bicirrhosum), an ancient teleost.

The corpuscles of Stannius of arawana (Osteoglossum bicirrhosum), an ancient teleost, were examined by routine light and electron microscopy and following their immunoreactivity to salmon and trout stanniocalcin antisera. Periodic acid-Schiff positive cells of the corpuscles of Stannius had a follicular arrangement and demonstrated a strong immunohistochemical reaction with both stanniocalcin antisera. Fine structural analysis of the paired, posteriorly located, and perirenal ovoid glands revealed two morphologically distinct cell types the basal laminae of which were ramified by nerve terminals. Immunocytochemistry demonstrated that osmiophilic secretory granules in both cell types were immunoreactive to the stanniocalcin antisera. When extracts of arawana corpuscles of Stannius were subjected to sodium dodecyl sulphate electrophoresis and Western blot analysis a diffuse molecular weight band was evident ( approximately 68 kDa) in the non-reduced condition. In all cases, immunoreactivity was abolished by preabsorption of the antisera with salmon stanniocalcin or with a crude extract of arawana corpuscles of Stannius. The corpuscles of Stannius of arawana are similar to those in more recent teleosts with respect to cell structure and their anatomical distribution but their stanniocalcin is more similar in molecular weight to that present in at least one other non-teleost actinopterygian (the gar) which has an ancient lineage.