Experimental Physiology
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Quarterly Journal of Experimental Physiology 71.2 pp 249-265
© The Physiological Society 1986
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ISOLATION OF DUCTS FROM THE PANCREAS OF COPPER-DEFICIENT RATS

S. Arkle 1, C. M. Lee 2, M. J. Cullen 2, and B. E. Argent 3

1 School of Pharmacy, Portsmouth Polytechnic, Park Building, King Henry 1 Street, Portsmouth PO1 2DZ
2 Departments of Physiological Sciences and Clinical Biochemistry, University Medical School, Framlington Place, Newcastle Upon Tyne NE2 4HH
3 Muscular Dystrophy Research Laboratories, Newcastle General Hospital, Westgate Road, Newcastle Upon Tyne NE4 6BE

A technique is described, involving tissue dissociation and micro-dissection, for the isolation of interlobular ducts from the pancreas of copper-deficient rats. The average length and outside diameter of the isolated ducts were 589·0±18·6 and 78·1±1·6 µm (mean ± S.E.M., n = 425) respectively. Between twenty and fifty ducts could be obtained from each pancreas. Frequently, the smaller intralobular ducts, which had outside diameters of between 15 and 25 µm, were observed as branches of the interlobular ducts. Light and electron microscopy showed that the isolated ducts were structurally intact, and that the epithelial cells possessed all the typical ultrastructural features of duct cells within the gland of copper-replete rats. The isolated ducts consumed oxygen at a rate of 2·27±0·55 ml O2/min.100 g wet weight duct epithelium (n = 6). The concentrations of ATP, ADP and AMP in the ducts were 3·78±0·81, 0·68±0·19 and 0·41±0·13 mmol/l duct epithelium (n = 8) respectively. These data give values for ATP:ADP and ATP:AMP ratios of 5·6:1 and 9·2:1 respectively, and an energy charge of 0·85±0·01 (n = 8) suggesting that the epithelial cells are healthy and in a stable metabolic state. In the presence of the phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine (0·67 mM), the basal concentration of cyclic AMP in the isolated ducts was 17·4±0·7 µmol/1 duct epithelium (n = 3). Secretin (0·1 nM-1 µM) caused a dose-related increase in cyclic AMP content up to a maximum of 376·0±85·3 µmol/l duct epithelium (n = 4). This indicates that the epithelial cells possess secretin receptors, and that these receptors can be functionally linked to adenylate cyclase.

Submitted on August 26, 1985




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