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KMID : 0378019670100050070
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New Medical Journal
1967 Volume.10 No. 5 p.70 ~ p.80
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Studies on the Glucose Oxidation, Glycogen Incorporation and Iodine
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Abstract
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It was pointed out that !acetylcholine increased the uptakes of oxygen and iodine by pancrease, brain and thyroid gland when glucose was available as a substrate. The author, in the previous paper, also reported that acetylcholine increased the ability of the salivary gland slices to concentrate iodine.
In view of the recent works by Pastan and Serif that acetylcholine and atropine exert some influences on glucose and iodine metabolisms of the thyroid as well as salivary glands, the present study was aimed to study further the effects of acetylcholine and atropine on the glucose oxidation, glycogen incorporation and iodine uptakes of the salivary gland slices. - Furthermore, the effect of acetylcholine on the iodine metabolism in the parotid gland slice in regards to the differential iodine fractions on the paperchromatography, and on the glucose metabolism in regards to the localization of the radiocarbon particles on the microautoradiography were studied in some more details.
The animals used were the male rats weighing 160 to 180 g and the rabbits weighing 2 kg. After sacrifice the salivary glands were quickly excised and were placed into the Petri dish containing cold Krebs-Ringer bicarbonate. The slices were. gently blotted, weighed, divided into aliquots of 180"to 200 mg, and were placed into the main chamber, of 25 ml. Erlenmeyer flask. The incubation medium in each flask contained 1 mg per ml. of carrier glucose with approximately 0.5 ,uc of radioactivity in the form of glucose-1-04, O. lac of carrier free radioiodine and Krebs-Ringer bicarbonate buffer at pH 7.4 to give a total volume of 2.5 ml.
Acetylcholine chloride, atropine sulfate, eserine salicylate and thiourea were then added as indicated in experimental groups. The gas mixture of 95 % oxygen and 5 % carbon dioxide was flushed in bicarbonate buffer for one minute. The closed flask were immediately incubated at 37C in a Warburg´s shaking water bath, and the reaction was stopped by addition of ION sulfuric acid and carbon dioxide (C1402) was trapped in 10 % KH containing solution. Radioactive carbon dioxide was measured as was described by Field et al.
The incorporation of. glycogen was measured with the sample treated in Van-Slyke Folk apparatus. The paperchromatographic analysis of iodine fraction and the paperchromatographic autoradiography was done as was escnbcd by Taurog et al.. and Serif et al "For the `microautoradiography.of parotid gland `the tissue bl´ s
z4 ere placed dace down on single large sheet of E.~istman´ no screen, -ray film, ; with a brass~ weight on each to ais
insure the good´ contact. The slice sections `were Ness than´´0. 25 mm in thickness. After 20 days´ exposure the ;film was developed for 5 minutes in D-11 at 9.8 F in´a:tray with continuous shakinb. The results were summarized as follows
1. In submaxill
arygland, acetylcholine showed the stimulatory effect on the glucose oxidation, while atropine
´ ´depressed the C1402 production from glucose.
2. In parotid gland, the glucose oxidation was stimulated not only by acetylcholine but also by atropine. ´ 3. Insubmaxillary glared, acetylcholine increased the iodine uptakes, while atropine only negligible or no effects at all.
4. In parotid gland, acetylcholine also increased the iodine uptakes, while atropine decreased.
y 5. In rabbits´ parotid glands, the glycogen incorporation was increased either by acetylcholine or by atropine.
~r ! 6. Paperchromatographic autoradiography ´of hydrolysed salivary gland slices revealed the significant increase "
of diiodotyrosine` and `triiodotyrosine fractcions in the presence of acetylcholine.
7. The increased glycogen´incorporation or uptakes of radioactive carbon particles were observed,´ chiefly in \ the acinous cells and basement membrane of the striated duct of the parotid gland.
8. The mechanisms´ of the effects´ of acetylcholine and atropine on the glucose oxidation, glycogen incorporation and iodine uptakes of the saliva gland were briefly discussed.
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