Конференции Карельского научного центра РАН, XIII Международный Нематологический симпозиум «Нематоды и другие Ecdysozoa в изменяющейся среде обитания»

Размер шрифта: 
Ultrastructure of unhatched cyst nematodes Globodera rostochiensis and Heterodera schachtii revealed by self-pressurised rapid freezing
Myriam Claeys, N. D. Handayani, P. Lestari, V. V. Yushin, A. Dikin, J. Helders, W. Bert

Изменена: 2019-05-09


Ultrastructural analysis of nematode eggs poses a considerable problem due to the impermeability of the egg-shell. In high pressure freezing, a physical cryo-fixation method, a much faster rate of fixation is obtained and the eggshell does not form a barrier resulting in superior preservation of fine structure. Self-Pressurised Rapid Freezing (SPRF), an alternative low-cost cryo fixation method, was evaluated based on an ultrastructural study of the egg shell and the cuticle of the second stage juveniles of Globodera rostochiensis and Heterodera schachtii in their unhatched native state. The obtained results are also compared with conventional (chemical) fixation. This study clearly demonstrates that SPRF fixation results in a very well-preserved ultrastructure of the entire, intact egg, from the eggshell to the cell details of developing embryos and juveniles. Therefore, SPRF fixation is forwarded as a very accurate, relatively easy-to-use, low-cost and fast technique to study the ultrastructure of unhatched eggs of nematodes. The method may be helpful in a variety of nematology studies, including embryology, functional morphology based on immunolocalization of cell proteins, characterization of vector viruses and symbiotic bacteria, nematicide targets and efficiency. (Support: UGhent TEM-Expertice center, life sciences; RFBR 17-04-00719).