Transcriptome, histological, and physiological responses of Amur sleeper (Perccottus glenii) during cold stress, freezing, and recovery

Abstract

Freeze tolerance is a survival strategy employed by some ectotherms living in extremely cold environments. Some fish in extremely cold areas can recover from their frozen state, but they also have to endure cold stress. Amur sleeper (Perccottus glenii) can recover from a completely frozen state. To explore the response of freeze-resistant fish to low temperatures, we analyzed histological alterations, and antioxidant and carbohydrate-lipid metabolizing enzymes of P. glenii under low temperatures. So far, sensory genes regulating P. glenii during cold stress, freezing, and recovery have not been identified. Ultrastructure results indicated that glycogen content and mitochondrial ridge decreased during cold stress and freezing, whereas the number of endoplasmic reticulum increased during recovery. Plasma glucose and glycerol levels of the three treatment groups significantly increased. Lactate dehydrogenase and pyruvate kinase levels significantly increased during cold stress and freezing, and hexokinase levels significantly increased during cold stress. In total, 30,560 unigenes were found (average length 1724 bp, N50 2843 bp). In addition, 7370 differentially expressed genes (DEGs; including 2938 upregulated genes and 4432 downregulated genes) were identified. KEGG analysis revealed that the DEGs were enriched in carbohydrate and lipid metabolism, lipid synthesis, immune system, and anti-apoptosis. Genes involved in glycolysis and phospholipid metabolism were significantly upregulated during cold stress; genes related to circadian rhythm, oxidative phosphorylation, and lipid synthesis were significantly upregulated during freezing; and genes involved in the immune system and anti-apoptosis were significantly upregulated during recovery. Our results attempt to offer new insights into the physiological mechanisms of complex adaptation in P. glenii and provide useful information for future studies on the mechanism underlying freezing/recovery in animals.