Stevens P, Robinson R L, Van Dyke K, Stitxel R
Pharmacology. 1975;13(1):40-55. doi: 10.1159/000136883.
The perfused bovine adrenal gland was able to take up and phosphorylate adenosine-8-3 H. Stimulation of the gland with carbachol resulted in a simultanious release of catecholamines, adenosine-8-3 H and labelled adenine nucleotides including a small quantity of ATP-3 H. A direct demonstration of the presence of ATP-3 H in isolated chromaffin granules was obrained by modified sucrose density gradient centrifugation. Glands were also perfused with a Ca++-Mg++-free-EDTA medium to inhibit adenosine triphosphatases. Using this medium, significant amounts of ATP-3 H and endogenous ATP were found in the effluent after drug stimulation by either Ba++ of aminophylline. Under Ca++-Mg++-free-EDTA conditions both Ba++ and aminophylline, but not carbachol, stimulated the release of catecholamines, ATP, ATP-3 H with no concomitant increase in total radioactivity. Using normal medium, Ba++ caused the simultaneous release of catecholamines and radioactivity while aminophylline only induced the release of catecholamines. Specific activity measurements of drug-released ATP indicated that the newly synthesized ATP-3 H was being released preferentially during spontaneous activity while the endogenous ATP was preferentially released after either Ba++ or aminophylline-induced secretion.
灌注的牛肾上腺能够摄取并磷酸化腺苷 - 8 -³H。用卡巴胆碱刺激肾上腺会导致儿茶酚胺、腺苷 - 8 -³H和标记的腺嘌呤核苷酸同时释放，其中包括少量的ATP -³H。通过改良的蔗糖密度梯度离心法直接证明了分离的嗜铬颗粒中存在ATP -³H。还用无Ca²⁺ - Mg²⁺ - EDTA培养基灌注肾上腺以抑制腺苷三磷酸酶。使用这种培养基，在钡离子或氨茶碱药物刺激后的流出物中发现了大量的ATP -³H和内源性ATP。在无Ca²⁺ - Mg²⁺ - EDTA条件下，钡离子和氨茶碱都能刺激儿茶酚胺、ATP、ATP -³H的释放，但卡巴胆碱不能，且总放射性没有相应增加。使用正常培养基时，钡离子会导致儿茶酚胺和放射性同时释放，而氨茶碱仅诱导儿茶酚胺的释放。对药物释放的ATP的比活性测量表明，新合成的ATP -³H在自发活动期间优先释放，而内源性ATP在钡离子或氨茶碱诱导的分泌后优先释放。
