Franzen Anderson J, Cunha Marcel M L, Batista Evander J O, Seabra Sergio H, De Souza Wanderley, Rozental Sonia
Laboratório de Biologia Celular de Fungos, Instituto de Biofísica Carlos Chagas Filho (IBCCF), Centro de Ciências da Saúde (CCS), Universidade Federal do Rio de Janeiro (UFRJ), Bloco G, Ilha do Fundão, Rio de Janeiro, RJ, 21949-900, Brazil.
Microsc Res Tech. 2006 Sep;69(9):729-37. doi: 10.1002/jemt.20344.
The influence of tricyclazole (5-methyl-1,2,4-triazol[3,4]benzothiazole), a specific DHN-melanin inhibitor, on the cell walls and intracellular structures of Fonsecaea pedrosoi conidia and sclerotic cells was analyzed by transmission electron microscopy (TEM), deep-etching, and field emission scanning electron microscopy. The treatment of the fungus with 16 microg mL(-1) of tricyclazole (TC) did not significantly affect fungal viability, but electron microscopy observations showed several important morphological differences between TC-treated and non-TC treated cells. Control sclerotic cells presented patched granules, with an average diameter of 47 nm, on the cell surface, which were absent in TC-treated cells. Also, TC-treated sclerotic cells showed an undulated relief. TC treatment leads to an accumulation of electron lucent vacuoles in the fungal cytoplasm of both conidia and sclerotic cells, and treated conidia observed by deep etching showed a relevant thickening of the fungal cell wall. Together, these observations support the previous data of our group that F. pedrosoi synthesizes melanin in intracellular organelles. In addition, we suggest that melanin is not only an extracellular constituent but could also be dispersing all over the cell walls and could have an effective role in cross-linking different cell wall compounds that help maintain the regular shape of the cell wall.
通过透射电子显微镜(TEM)、深度蚀刻和场发射扫描电子显微镜,分析了特定的二羟基萘(DHN)黑色素抑制剂三环唑(5-甲基-1,2,4-三唑并[3,4]苯并噻唑)对佩德罗分支孢分生孢子和硬化细胞的细胞壁及细胞内结构的影响。用16μg/mL的三环唑(TC)处理该真菌,对真菌活力没有显著影响,但电子显微镜观察显示,经TC处理的细胞与未处理的细胞之间存在一些重要的形态学差异。对照硬化细胞在细胞表面呈现平均直径为47nm的斑块状颗粒,而经TC处理的细胞中不存在这些颗粒。此外,经TC处理的硬化细胞呈现出起伏的表面。TC处理导致分生孢子和硬化细胞的真菌细胞质中出现电子透明空泡的积累,通过深度蚀刻观察到,经处理的分生孢子的真菌细胞壁有明显增厚。总之,这些观察结果支持了我们小组之前的数据,即佩德罗分支孢在细胞内细胞器中合成黑色素。此外,我们认为黑色素不仅是一种细胞外成分,还可能分散在整个细胞壁上,并在交联不同细胞壁化合物方面发挥有效作用,有助于维持细胞壁的正常形状。