Biochemistry of methanol poisoning. 4. The effect of methanol and its metabolites on retinal metabolism.
Biochemistry of methanol poisoning. 4. The effect of methanol and its metabolites on retinal metabolism.
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甲醇中毒的生物化学。
DOI：
10.1042/bj0820164
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发表时间：
1962
期刊：
影响因子：
--
通讯作者：
J. Cooper
中科院分区：
文献类型：
--
作者：
M. Kini;J. Cooper
材料和方法Pi，无机磷酸盐。材料.吡啶核苷酸、细胞色素c和酵母己糖激酶购自Sigma Chemical Co.。ATP的钾盐购自Mann Research Laboratories Inc.，纽约。均匀标记的[14 C]-葡萄糖（[U-14 C]葡萄糖）从新英格兰核公司获得，[32 P]磷酸盐从橡树岭国家实验室获得。所用的所有其他化学品均为试剂级纯度。牛的眼睛在动物死亡后立即在屠宰场被移除，放置在碎冰中并运送到实验室。沿锯齿缘沿着作一圆形切口，切开眼球，挤出玻璃体液，在视网膜与视盘的连接被切断后，用钝镊子轻轻提起视网膜。大约300毫克。在扭力天平中称量视网膜组织的重量，并将其悬浮于包含在冷却的瓦尔堡烧瓶中的孵育培养基中。然后将烧瓶连接到压力计上，转移到370 ℃的水浴中，用常规技术进行差示测压，通常持续1小时。从动物死亡到开始孵育的时间间隔不超过3小时。当[14 C]葡萄糖为底物时，加入0-2 ml终止反应。的30%三氯乙酸，并继续振荡30分钟。以确保中心井中的碱完全吸收14 CO2。干重测定表明，100 mg.按上述方法制备的全视网膜湿重相当于12 mg。干重
MATERIALS AND METHODS Abbreviation. Pi, inorganic phosphate. Materials. Pyridine nucleotides, cytochrome c and yeast hexokinase were purchased from Sigma Chemical Co. The potassium salt of ATP was obtained from Mann Research Laboratories Inc., New York. Uniformly labelled [14C]-glucose ([U-14C] glucose) was obtained from New England Nuclear Corp. and [32P] phosphate from the Oak Ridge National Laboratory. All other chemicals used were of reagent-grade purity.Experimental procedures. Ox eyes were removed at the abattoir immediately after the death of the animal, placed in crushed ice andtransported to the laboratory. The eyeball was opened by a circular incision along the ora serrata, the vitreous humour squeezed out and the retina gently lifted by means of a blunt forceps after its attach-ment to the optic disk had been severed. Approximately 300 mg. of retinal tissue was weighed in a torsion balance and suspended in incubation media contained in chilled Warburg flasks. The flasks were then attached to the manometers, transferred to a water bath at 370 and differential manometry was carried out by conventional techniques for a period of usually 1 hr. The time interval from the death of the animal to commencement of the incubation was not more than 3 hr. When [14C] glucose was the substrate, the reaction was terminated by the addition of 0-2 ml. of 30% trichloroacetic acid, and shaking was continued for a period of 30min. to ensure complete absorption of 14CO2 by the alkali in the centre well. Dry-weight determinations indicated that 100 mg. of whole retina, wet weight, prepared as described, was equivalent to 12 mg. dry wt.