ISSN:
0003-276X
Keywords:
Insulin
;
Islet
;
Isolation
;
Secretion
;
Morphology
;
Rat
;
Immunoassay
;
Life and Medical Sciences
;
Cell & Developmental Biology
Source:
Wiley InterScience Backfile Collection 1832-2000
Topics:
Medicine
Notes:
Laboratories engaged in secretory studies of rat pancreatic islets often encounter high baseline insulin secretion with poor secretory response to secretagogues, such as glucose. The specific morphologic abnormalities that accompany this unregulated release have not been described. We isolated islets comparing two approaches. Both used stationary digestion with collagenase. In method I, we distended the biliary duct extracorporeally with collagenase and minced the pancreas after a 28 min digestion (37°C). In method II, we distended the pancreas intracorporeally and digested for 40 min without mincing. Both methods utilized a similar collagenase concentration (2 μg/ml in Hank's balanced salt solution (HBSS)). Both methods yielded over 300 islets/rat. Islets from both methods appeared intact, when viewed under the dissecting microscope. We found that adequate secretion from incubated islets was evoked with method I, i.e., low basal insulin levels at low glucose (3.3 mM), tripling at 11.0 mM glucose, and nearly quadrupling in response to higher glucose (16.7 mM). In contrast, method II was characterized by high basal levels without response to higher glucose. Ultramicroscopic examination of islet B cells in method I revealed normal cytological features, while B cells in method II showed marked degranulation, profiles of swollen endoplasmic reticulum, and swollen mitochondria. Morphometric analysis of B cells confirmed quantitatively a decrease in secretory granule density and mitochondrial enlargement in method II compared to method I. Anatomic changes, largely confined to the B cells of islets may account for functional alterations of responses. Defects cannot be predicted from gross appearance of islets. © 1993 Wiley-Liss, Inc.
Additional Material:
5 Ill.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1002/ar.1092370409
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