Detecting freeze injury and seasonal cold-hardening of cells and tissues in the gall fly larvae, Eurosta solidaginis (Diptera: Tephritidae) using fluorescent vital dyes

This study identified a hierarchy in levels of cold tolerance for diverse tissues from larvae of Eurosta solidaginis. Following freezing at −80 °C, larval survival and the viability of specific tissues were assessed using membrane-permeant DNA stain (SYBY-14) and propidium iodide. Integumentary musc...

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Bibliographic Details
Published in:Journal of insect physiology 2003-11, Vol.49 (11), p.999-1004
Main Authors: Yi, Shu-Xia, Lee, Richard E
Format: Article
Language:English
Subjects:
Acclimatization - physiology
Animals
Cell Survival - physiology
Cold Temperature
Cold-hardening
Cryoprotection
Eurosta solidaginis
Fluorescent Dyes
Freeze tolerance
Freezing
Hemolymph - metabolism
Larva - cytology
Larva - physiology
Microscopy, Fluorescence
Organ Specificity
Seasons
Tephritidae
Tephritidae - physiology
Vital fluorescent dyes
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Summary:This study identified a hierarchy in levels of cold tolerance for diverse tissues from larvae of Eurosta solidaginis. Following freezing at −80 °C, larval survival and the viability of specific tissues were assessed using membrane-permeant DNA stain (SYBY-14) and propidium iodide. Integumentary muscle, hemocytes, tracheae, and the crystal-containing portion of the Malpighian tubules were most susceptible to freezing injury. A second group consisting of fat body, salivary glands, and the proximal region of the Malpighian tubules were intermediate in their susceptibility, while the foregut, midgut, and hindgut were the most resistant to freezing injury. Seasonal increases in larval cold tolerance were closely matched by changes in the cold tolerance of individual tissues. Compared to larvae collected in September, the survival rates for each of the six tissues tested from October-collected larvae increased by 20–30%. The survival rate in all tissues was notably higher than that of whole animals, indicating that larval death could not be explained by the mortality in any of the tissues we tested. This method will be useful for assessing the nature of chilling/freezing injury, the role cryoprotectants, and cellular changes promoting cold tolerance.
ISSN:0022-1910
1879-1611
DOI:10.1016/S0022-1910(03)00168-9