Quantitative correlation between secretion and cellular content of catecholamines and dopamine-beta-hydroxylase in cultures of adrenal medulla cells.
Quantitative correlation between secretion and cellular content of catecholamines and dopamine-beta-hydroxylase in cultures of adrenal medulla cells.
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肾上腺髓质细胞培养物中儿茶酚胺和多巴胺-β-羟化酶的分泌和细胞含量之间的定量相关性。
DOI：
10.1016/0006-2952(81)90526-8
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发表时间：
1981
影响因子：
5.8
通讯作者：
Kirshner,N
中科院分区：
文献类型：
--
作者：
Ledbetter,FH;Kirshner,N
方法：如前所述[14]制备分离的牛肾上腺髓质细胞，并以每孔1.2 × 106个细胞的密度在6孔（35 mm）簇皿（Costar 3506）中培养。在使用前，使细胞保持在培养物中至少3天。在室温下用10 μ M尼古丁或50 μ M藜芦碱刺激细胞15分钟，并用20 μ M离子霉素刺激1.5和30分钟。使用一式两份或一式三份威尔斯孔进行所有实验。在温育期结束时，移除培养基并保存用于测定DBH和CA（141.在这些研究中，总CA含量测定为肾上腺素的当量。使用10 μ M对羟基苯甲酸汞以消除内源性抑制剂，一式两份进行所有DBH测定。通过加入1 ml蒸馏水并冷冻裂解细胞。解冻后，加入1 ml 0.3M NaCl，将细胞刮入全玻璃Potter-Elvehjem匀浆器中。匀浆后，将细胞进行两次冻融循环，然后以100，000 g离心1小时。倾析并保存上清液部分，用0.15M NaCl洗涤沉淀两次。将洗涤液与初始裂解物合并，并将沉淀物重悬用于CA和DBH测定。在第二次洗涤后，颗粒中存在少于1%的总CA含量。如Bergmeyer等人所述测定培养基和裂解物中的乳酸脱氢酶活性。[15]第10段。蔗糖培养基由9.75%（w/v）蔗糖、2.2mM CaCl 2、5 mM 4-（2-羟乙基）-1-哌嗪-乙磺酸（HEPES）和10 mM葡萄糖组成，用KOH调节至pH 7.4。Locke溶液的组成和化学品来源已在之前报告/14]。离子霉素是一种新的抗生素，其被表征为钙离子载体（161），由Squibb Institute for Medical Research，Princeton，NJ提供。
MethodsIsolated bovine adrenal medulla cells were prepared as previously described [14] and cultured in 6 well (35 mm) cluster dishes (Costar 3506) at a density of 1.2 X lo6 cells per well. The cells were allowed to remain in culture at least 3 days before use. Cells were stimulated at room temperature for 15 min with 10 PM nicotine or 50 yM veratridine and for 1.5 and 30 min with 20 pM Ionomycin. All experiments were carried out using either duplicate or triplicate wells. At the end of the incubation period, the media were removed and saved for assay of DBH and CA (141. In these studies, the total CA content was determined as equivalents of adrenaline. All DBH assays were carried out in duplicate using 10 PM p-hydroxymercuribenzoate to inactivate endogenous inhibitors. The cells were lysed by adding 1 ml of distilled water and freezing. After thawing, I ml of 0.3 M NaCl was added and the cells were scraped into an all glass Potter-Elvehjem homogenizer. After homogenization, the cells were subjected to two cycles of freezing and thawing and then centrifuged at 100,000 g for 1 hr. The supernatant fraction was decanted and saved, and the pellet was washed twice with 0.15 M NaCI. The washes were combined with the initial lysate and the pellet was resuspended for CA and DBH assay. Less than 1 per cent of the total CA content was present in the pellet after the second wash. Lacticdehydrogenase activity in the media and lysates was determined as described by Bergmeyer et al.[15]. The sucrose medium was composed of 9.75%(w/v) sucrose, 2.2 mM CaClz, 5 mM 4-(2-hydroxyethyl)-l-pipercazine-ethanesulfonic acid (HEPES) and 10 mM glucose adjusted to pH 7.4 with KOH. The composition of Locke’s solution and sources of chemical were reported previously/14]. Ionomycin is a new antibiotic which has been characterized as a calcium ionophore (161 and was supplied by Squibb Institute for Medical Research, Princeton, NJ.
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肾上腺髓质释放儿茶酚胺和多巴胺-β-氧化酶
DOI：
--
发表时间：
1968
期刊：
影响因子：
--
作者：
O. H. Viveros;L. Arqueros;N. Kirshner
通讯作者：
N. Kirshner
DOI：
10.1042/bj1180303
发表时间：
1970
期刊：
The Biochemical journal
影响因子：
--
作者：
H. Winkler;H. Hörtnagl;A. D. Smith
通讯作者：
A. D. Smith
狗淋巴中的多巴胺-β-羟化酶-交感神经激活的作用
影响因子：
6.1
作者：
S. Ngai;W. Dairman;M. Marchelle;S. Spector
通讯作者：
S. Spector
“低钠尿素”介质促进灌注兔心脏钾离子释放的多巴胺β-羟化酶的冲洗
影响因子：
3.3
作者：
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H. Ritzel
关于猫外周交感神经释放后多巴胺-β-羟化酶的命运。
DOI：
10.1111/j.1748-1716.1974.tb05783.x
发表时间：
1974
期刊：
Acta physiologica Scandinavica
影响因子：
--
作者：
S. Ross;H. Eriksson;W. Hellström
通讯作者：
W. Hellström