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Biomedical subjects

J E Brinn

Publications and source records attributed to J E Brinn.

15 recordsLinked to original sources

Effects of combined secretagogues and extracellular calcium on neonatal insulin release.

The insulin response of 3-day old neonatal rat islets was evaluated following a 1 h incubation with glucose alone and in the presence of 30 nM sulfated cholecystokinin octapeptide (CCK) and/or 20 microM carbachol (CCh). Insulin secretion was found to be incrementally increased from the lowest glucose concentration and enhanced several fold in the presence of CCK and/or CCh. In combination, CCK and CCh increased glucose-stimulated insulin secretion by an amount equivalent to the sum of their individual increases. The presence of either CCK alone or CCK plus CCh increased phosphoinositide hydrolysis by the same relative amount that they increased insulin secretion when compared to 8.3 mM glucose. Glucose-stimulated insulin secretion was totally inhibited when calcium was omitted from the incubation buffer; this effect was partially negated by CCK alone and more so by CCK combined with CCh. Insulin secretion in response to 8.3 mM glucose alone was unchanged when calcium in the incubation buffer was increased from 1 to 5 mM; however, the insulin response to 16.7 mM glucose alone and 8.3 mM glucose in the presence of CCK and/or CCh was increased under this condition. Thus, we have shown that, even at 3 days postpartum, insulin secretion from isolated islets is a complex response capable of being molded by several secretagogues at once and ultimately determined by interplay of different signaling systems activated.

Animals

Catecholamine release by catecholamines in the eel does not require the presence of brain or anterior spinal cord.

The catecholamine-producing chromaffin cells of the American eel are strongly innervated by fibers, which, by ultrastructural criteria, seem to be cholinergic. However, neither removal of the brain nor removal of the brain combined with extirpation of the anterior spinal cord prevents the release of catecholamines into the circulation by catecholamines. It appears that the chromaffin cells are controlled by both nervous and humoral stimuli, and that at least some of the latter do not require the presence of "preganglionic" innervation.

Anguilla

Age-dependent stimulation of neonatal insulin release and inositol phosphate accumulation by CCK-8 and carbachol.

Development of a robust insulin secretory response to glucose occurs during the early neonatal period. To determine if neuroendocrine agents play a role during this time, we studied the effects of selected peptides and neurotransmitters on insulin release and polyphosphoinositide metabolism in islets isolated from 1- and 3-day neonatal rats. Vasoactive intestinal peptide had no effect on glucose-stimulated release in either islet population. In contrast, sulfated cholecystokinin octapeptide (CCK-8) significantly enhanced glucose-induced insulin release in both islet groups. One-day islets were stimulated only by a concentration of 300 nM, whereas 3-day islets were responsive at 3 nM. Similar to CCK-8, there were clear differences in responses to carbachol between 1- and 3-day islets. One-day islets required a concentration of 200 microM for insulin release to be significantly greater than with glucose alone; 3-day islet insulin release was significant at 2 microM carbachol. Both agonists stimulated inositol phosphate accumulation in 3-day islets, but only CCK-8 caused a significant increase over glucose-induced levels in 1-day islets. These results indicate that islet responsiveness to CCK-8 and carbachol develops in parallel during the early neonatal period. This development may be linked to the maturation of a critical step of stimulus-secretion coupling through which these agents act.

Age Factors

Mongolian gerbil tympanic membrane. Normal and with induced otitis media.

Otoscopic observation of the tympanic membrane is the most effective noninvasive means of diagnosing and following the course of experimental otitis media in animal models of the disease in both short-term and in longitudinal studies. Because of the difficulty of viewing the entire tympanic membrane of the gerbil model of otitis media without manipulation, we present methods of visualizing and color illustrations of the normal tympanic membrane of the Mongolian gerbil, Meriones unguiculatus. These are compared with illustrations of animals with acute experimental otitis media. An artist's rendition of the entire normal gerbil tympanic membrane is presented. These methods and illustrations will enable investigators to more rapidly and easily adopt the Mongolian gerbil as an animal model for studies of otitis media.

Acute Disease

Vasoactive intestinal polypeptide enhances hormone content and insulin release in cultured fetal rat islets.

The effect of porcine vasoactive intestinal polypeptide (VIP) on development of the biphasic insulin release response in cultured fetal rat islets was investigated. Fetal islets, 21.5 days gestational age, were cultured for 7 days in RPMI 1640 culture medium containing either 2.8 or 11.1 mM glucose adn subsequently challenged with 16.7 mM glucose in a perfusion system. Islets were exposed to VIP at a final concentration of 13.2 nM by adding the peptide to the perifusion buffer (acute exposure) or by adding it to the culture medium throughout the culture period (chronic exposure). Islet hormone and DNA contents were also quantitated at the end of the culture period. Acute exposure to VIP resulted in no alterations of the insulin release pattern after culture in the presence of either glucose concentration. However, chronic treatment of islets with 13.2 nM VIP in the presence of 2.8 mM glucose resulted in significant increases in the maximum rate of insulin release during the first phase and the total amount of insulin release during both phases. Similarly, islets cultured in the presence of 11.1 mM glucose and 13.2 nM VIP demonstrated enhanced biphasic insulin release patterns with increased maximum rate and total amount of release during both phases. The presence of VIP and 2.8 mM glucose increased islet glucagon and somatostatin contents, but islet DNA and insulin contents remained unchanged. These findings indicate that VIP plays a significant role in the in vitro development of the biphasic insulin release pattern and may be a factor controlling the maturation of the fetal islet in vivo.

Animals

Longitudinal studies of experimental otitis media with Haemophilus influenzae in the gerbil.

Haemophilus influenzae non-typable strain 119 was found to cause severe otitis media with sequellae when inoculated into the middle ear cavities of the Mongolian gerbil, Meriones unguiculatus. Acute inflammation was followed by the development of highly vascular granulation tissue and formation of new bone within the middle ear bulla. These changes persisted throughout the 14-week study in the untreated animals. The gerbil was variably susceptible to otitis media caused by inoculating 30-3000 bacterial cells and 100% susceptible to greater than 3000 cells. The susceptibility of the gerbil to common etiological agents of otitis media allows its use as an appropriate model of the disease.

Acute Disease

Experimental pneumococcal otitis media: longitudinal studies in the gerbil model.

In order to ascertain that the Mongolian gerbil, Meriones unguiculatus, is an acceptable model for studying the development of chronic from acute otitis media, we used previously published methods for experimental otitis media in a longitudinal study of the acute disease and sequellae. The gerbil was found to be susceptible to as few as 30 viable Streptococcus pneumoniae type 3 cells, indicating that only a few viable pathogens are able to cause the disease. Untreated experimental infections with S. pneumoniae type 23 resulted in a mild, self-limiting disease with little permanent sequellae, while S. pneumoniae type 3 produced severe disease characterized by an acute phase of from 2 to 3 weeks, followed by the development of new bone formation and a vascularized granulation tissue which persisted throughout the 13-week study. Viable pneumococci could be recovered from the middle ears for only two weeks. We conclude that the gerbil is a useful model for otitis media.

Animals

Innervation of the cultured fetal rat pancreas.

The fetal rat pancreas, explanted at 18 days of gestation and cultured up to ten days, contains numerous acetylcholinesterase-positive neurons. These nerves usually appear in small ganglia although single nerve cells are encountered. The axons of these intrapancreatic nerves appear to terminate only in the islet tissue and not on any exocrine components of the explant. It is concluded that the fetal rat pancreas contains an islet-specific group of cholinergic neurons.

Animals

New types of islet cells in a cyclostome, Petromyzon marinus L.

Four types of acidophilic granular cells, in addition to B-cells, are identified in the islet organ of anadromous specimens of two subspecies of Petromyzon marinus by light and electron microscopy. Three of these acidophils (PI, PII and PIV-cells) occur in both the cranial and hepatic islets while a fourth type (PIII-cell) has only been found in the hepatic islet of some animals. The granules of the PI-cells stain with ponceau de xylidine, give a distinct tryptophan reaction and in ultrastructural examination show large, dense granules. The PII-cells contain unusual crystals and appear to be a non-secretory stage of the PI. The PIII-cells stain deep-red and acid fuchsin. They contain very large, dense granules and some lysosomes. PIV-cells stain selectively with phosphotungstic acid-hematoxylin and ultrastructurally, contain small, more or less dense granules. It appears that PI- and PIV-cells develop directly from B-cells, while the PIII-cells derive from PI-cells. despite their direct or indirect origin from B-cells, the PI-, PIII- and PIV-cells show characteristic features of functionally independent endocrine cells. Petromyzon marinus may be an ideal model for the understanding of phylogenetic and pathological interrelationships between islet and gastrointestinal hormones. It is clear that the interpretation of the islet organ of the cyclostomes, which has been generally considered a source of insulin only, requires a revaluation.

Animals

Pancreatic islet cytology of Ictaluridac (Teleostei).

The endocrine pancreas of the bullhead catfish, Ictalurus nebulosus, and the channel catfish, I. punctatas was studied by light and electron microscopy. In addition to the usual A, B and D cells, a fourth endocrine cell type was consistently observed in the electron microscope. All endocrine cell types were innervated. The vesicles of most of the nerve endings were ultrastructurally different from typical adrenergic and cholinergic vesicles, strongly suggesting the possibility of a third autonomic neurotransmitter serving as a regulator of catfish islet secretion.

Animals

Stains for A, B, and D cells in fetal rat islets.

Ten techniques often used for identification of A, B, and D cells in adult islets of Langerhans were applied to fetal rat pancreas. Modifications were tried with many of these techniques. Two indole methods (xanthydrol and postocoupled benxylidene reactions) and a cryostat technique using o-phthaladehyde failed to stain fetal islets. Phosphotungstic acid hematoxylin and lead hematoxylin lightly stained fetal A cell granules in Helly's fixed tissue. The Grimelius silver nitrate technique stains adult rat A cells but failed to stain fetal cells. A modification of this technique stained fetal A cells and a possible 4th cell type. The specificity of this method was confirmed by restaining stained cells with a fluorescent antibody technique and with pseudoisocyanin. B cells, as previously reported, were readily stained by the aldehyde fuchsin technique. Fetal D cells were not stained by the Hellerstrom-Hellman alcoholic silver nitrate method, nor did they display pseudoisocyanin metachromasia after acid hydrolysis; they did fluoresce brightly with this technique when viewed with UV light. It was thus possible to distinguish the three usual cell types, plus a possible fourth type, in the fetal rat pancreas.

Animals

Ontogeny of four cell types in fetal rat islets using histochemical techniques.

The pancreas of the fetal rat was collected from the first appearance of the pancreatic bud at 11 days of gestation, and every day thereafter until birth. After birth the neonatal pancreas was collected every day for one week, and at intervals thereafter. Fetal B-cells were stained with Gomori's aldehyde fuchsin at 16 1/2 days, and with the immunofluorescent technique for insulin at 14 days. The A-cells were stained as early as 13 days using the fluorescent antibody technique for glucagon. The D-cells first stained at 17 days with pseudoisocyanin. A 4th cell type was found which stained black with silver nitrate, using a method derived from the Grimelius technique for A-cells. This 4th cell type appeared at 15 days in the fetus, reaching its greatest abundance around 19 days, and then declined in numbers after birth until adulthood, when occasionally one or two cells were found.

Age Factors

Effects of cholecystokinin octapeptide and carbachol on neonatal insulin secretory dynamics.

The effects of glucose, sulfated cholecystokinin-octapeptide (CCK-8), or carbachol on insulin secretory dynamics were studied in pancreatic islets isolated from 1- and 3-day-old neonatal rats. When challenged with glucose, 1-day islets responded with a definite first phase and elevated secretion during the latter part of the stimulation period; 3-day islets had a first phase and a rising, sustained second phase. The presence of stimulatory concentrations of CCK-8 or carbachol in addition to glucose caused dramatic changes in the release pattern in both islet populations. In 1-day islets, carbachol stimulated mainly first phase secretion whereas CCK-8 enhanced first phase release and produced a definite second phase response. The two secretagogues increased significantly both phases of release in 3-day islets with no differences between the two agents in their effects. These results indicate that CCK-8 and carbachol differentially stimulate neonatal insulin secretion, possibly through different steps in the stimulus-secretion pathway. They also suggest that the cellular mechanism for second phase release is present in 1-day islets and can be activated by CCK-8.

Animals