Tiago C G, Brites A D, Kawauchi G Y
Centro de Biologia Marinha, Universidade de São Paulo, São Sebastião, Brazil.
J Microsc. 2005 Jun;218(Pt 3):240-6. doi: 10.1111/j.1365-2818.2005.01486.x.
The various shapes, size and fine details of ossicles are of paramount importance as systematic characters in the Echinodermata. The ossicles in this group are endodermal and a method to free them from the dermis is essential for their study. To date, a small quantity of hypochlorite solution (usually household liquid bleach) has been used to dissolve a small piece of tissue and, consequently, free these ossicles. A new method, using a proteolytic enzyme instead of hypochlorite solution to dissolve holothurian tissues, is proposed. This method was tested on fragments of body tissues from three species in three different orders of holothurians: Holothuria grisea Selenka, 1867 (Aspidochirotida), Duasmodactyla seguroensis (Deichmann, 1930) (Dendrochirotida) and Synaptula secreta Ancona Lopez (1957) (Apodida). Observations of material prepared using the hypochlorite and the enzyme methods were made using scanning electron microscopy (SEM). The SEM images show the striking differences in the surface of the ossicles obtained by each method, and reveal the advantages of the enzyme method.
在棘皮动物门中，小骨的各种形状、大小和精细细节作为分类特征至关重要。该类群中的小骨是内胚层的，因此一种将它们从真皮中分离出来的方法对于其研究至关重要。迄今为止，少量的次氯酸盐溶液（通常是家用液体漂白剂）被用于溶解一小块组织，从而分离出这些小骨。本文提出了一种新方法，即使用蛋白水解酶而非次氯酸盐溶液来溶解海参组织。该方法在三种不同目海参的三个物种的身体组织片段上进行了测试：1867年的灰海参（Aspidochirotida目）、1930年的杜氏双指参（Dendrochirotida目）以及1957年的隐秘管足参（Apodida目）。使用扫描电子显微镜（SEM）对采用次氯酸盐和酶法制备的材料进行了观察。SEM图像显示了通过每种方法获得的小骨表面的显著差异，并揭示了酶法的优势。
