Identification of resilin in the leg of cockroach, Periplaneta americana:: confirmation by a simple method using pH dependence of UV fluorescence
Identification of resilin in the leg of cockroach, Periplaneta americana:: confirmation by a simple method using pH dependence of UV fluorescence
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DOI：
10.1016/s1467-8039(00)00014-1
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发表时间：
2000-01-01
影响因子：
2
通讯作者：
Zill, S
中科院分区：
文献类型：
--
作者：
Neff, D;Frazier, SF;Zill, S
我们检查了蟑螂腿的跗节（脚）和胫骨节，以确定含有弹性蛋白节肢素的结构。使用紫外线照射下 420 nm 荧光发射的常规标准以及甲苯胺蓝对整个组织进行组织学染色来测试节肢动物的存在。我们还开发了一种简单的方法，通过随周围介质 pH 值的变化而发生的荧光变化来确认节肢动物的识别。使用仅通过光的常用激发滤光片。 >330 nm，我们发现在中性 pH 值下存在发射，但在低 pH 值下被消除。当介质恢复到较高 pH 值时，它可逆地重新出现。这种效应可归因于在酸性介质中发生的节肢动物氨基酸吸收最大值发生的已知变化（从 330 nm 到 285 nm）。通过检查翅膀铰链的韧带证实了这种识别方法的准确性，此前已证明许多昆虫的翅膀铰链中含有节肢韧带。利用这些技术，我们已经确定了节肢韧带与胫跗关节处的韧带以及腿部第四和第五跗节之间的关节的关系。这些韧带的解剖学排列表明，它们可以通过充当腿部肌肉活动的弹性拮抗剂来帮助在行走过程中产生腿部运动。我们设计的识别方法可以很容易地应用于帮助在其他动物中定位节肢弹性蛋白。 (C) 2000 Elsevier Science Ltd. 保留所有权利。
We have examined the tarsus (foot) and tibial segments of the cockroach leg to identify structures that contain the elastic protein resilin. The presence of resilin was tested using the conventional criteria of fluorescent emission at 420 nm under UV illumination and histological staining of wholemount tissues by toluidine blue. We have also developed a simple method of confirming identification of of resilin through changes in its fluorescence that occur with alteration of pH of the surrounding medium. Using a commonly available excitation filter that only passes light at. >330 nm, we found that the emission was present at neutral pH but was eliminated at low pH. It then reversibly reappeared when medium of higher pH was restored. This effect is attributable to a known shift in the absorption maximum of amino acids of resilin that occurs in acidic media (from 330 to 285 nm). The accuracy of this method of identification was confirmed by examination of ligaments of the wing hinge, which has previously been shown to contain resilin in a number of insects. Using there techniques, we have identified resilin in association with ligaments at the tibio-tarsal joint and in the articulation between the fourth and fifth tarsal segments of the leg. The anatomical arrangement of these ligaments suggests that they could aid in the generation of leg movements during walking by functioning as elastic antagonists to the actions of leg muscles. The method of identification we have devised could readily be applied to aid in the localization of resilin in other animals. (C) 2000 Elsevier Science Ltd. All rights reserved.