INFLUENCE OF TESTICULAR HYALURONIDASE ON THE RESISTANCE TO FLOW THROUGH THE ANGLE OF THE ANTERIOR CHAMBER
INFLUENCE OF TESTICULAR HYALURONIDASE ON THE RESISTANCE TO FLOW THROUGH THE ANGLE OF THE ANTERIOR CHAMBER
复制标题
DOI：
10.1111/j.1748-1716.1954.tb01092.x
复制
发表时间：
1954-01-01
期刊：
影响因子：
--
通讯作者：
SCOTCHBROOK, S
中科院分区：
文献类型：
--
作者：
BARANY, EH;SCOTCHBROOK, S
方法：在宰杀后15分钟内摘除牛的眼睛，将其放入碎冰中包装。它们总是被用在同一块粘土上。实验前，通常将它们放入室温(20-22℃)的生理盐水中15分钟。在某些情况下，它们直接在冷状态下使用。在一些实验中，结膜被切除到角膜缘，而在另一些实验中，结膜被留在原位。为了输液，眼睛被放在一个小盘子上。用一根中等大小的针和清洗干净的乳胶管将含有待测溶液的滴管连接到前房。另一针放入前房，后一针用试液冲洗，通过第二针抽出3-5nil，同时向通向第一针的管内注入相应数量的相同溶液。避免了不适当的压力上升，尽管很快发现短期的压力上升，即使是导致角膜混浊的压力上升，对实验的后期过程也没有影响。冲洗花费了大约1分钟的时间，在前房造成了相当大的颠簸，用第二根针进行了反穿刺术，并开始从滴管中输液。时间从第一根针插入的那一刻开始计算。溶液由普通2ml直管滴入0.02ml，与前房针水平平放。流体柱上的压力是由一个保持在大约23CNI的H，@的压缩空气来保持的，但每双眼睛总是相同的。所有进入眼睛的溶液都是每天用从整个玻璃蒸馏器中重新蒸馏的新鲜水配制的。基本溶液由9份0.0%的氯化钠和1份0.15M钠巴比妥缓冲液PII 7组成。在此基础上加入冷冻干燥的睾丸透明质酸酶(来自哈尔辛堡AB Leo的透明质酸酶)或与酶制剂中该稳定剂的量相对应的一定量的甲基明胶。所使用的透明质酸酶已由Malm-Grex(1953)进行了密切的研究。超速离心法和电泳法均一，似乎是一种约11,000分子量的多肽。该制剂不具有蛋白水解性。其活性为每毫克约500个减粘单位(VRU)，比粗制睾丸提取物纯化3*103倍。
Methods.The eyes of cattle were removed within 15 minutes after killing, nisrkcd and packed in crushed ice. They were always used on the same clay. &fore the experiment they were usually put for 15 minutes into saline kept at room temperature (20-22’C). In some instances they were used directly in the cold state instead. In some experiments the conjunctiva was removed up to the limbus, in others it was left in situ. For the infusion the eye was placed on a small dish. A burette containing the solution to be tested was connected to the anterior chamber by a medium-sized needle and well-washed latex tubing. Another needle was put into the anterior chamber and the latter washed with test solution by withdrawing 3--5 nil through the second needle while at the same time injecting a corresponding amount of the same solution into the tubing leading to the first needle. Undue pressure rises were avoiclcd, although it soon was found that a short pressure rise, even one leading to clouding of the cornea, was without influence on the latter course of the experiment. After the washing, which took about 1 minute of time, and which caused considerable turbulence in the anterior chamber, a counter-puncture was made with the second needle and infusion from the burette was started. Time was counted from the moment of insertion of the first needle. The solutions were delivered from ordinary 2 ml straight burettes graduated in 0.02 ml and lying horizontally in the same level as the needle in the anterior chamber. Pressure was excerted on the fluid column by compressed air from a nianostat kept at a pressure of about 23 cni of H,@, but always the same in each pair of eyes. All solutions to go into the eyes were made up every day with freshly redistilled water from a whole-glass still. The basic solution consisted of 9 parts of 0.0 percent NaCl and 1 part of 0.15 M sodiumbarbital buffer, pII 7. To this was added either a lyophilized testicular hyaluroniclase (Hyalas from AB Leo, Halsingborg) or an amount of methyl gelatin corxsponding to the amount of this stabilizer present in the enzyme preparation.The hyaluronidase used has been subjected to close study by MALM-GREX (1953). It is homogeneous on ultracentrifugation and electrophoresis and seems to be a polypeptide of about 11,000 molecular weight. The preparation is free of proteolytic activity. Its activity is given as about 500 viscosity reducing units (VRU) per mg, representing a purification of 3* 103 over the crude testis extract.