Stress-induced accumulation of glycerol in the flesh fly, Sarcophaga bullata: Evidence indicating anti-desiccant and cryoprotectant functions of this polyol and a role for the brain in coordinating the response
Nondiapausing larvae of the flesh fly, Sarcophaga bullata, responded to several forms of short-term environmental stress (low temperature, anoxia and desiccation) by accumulating glycerol. Elevation of this polyol, regardless of the type of stress that induced accumulation, conferred cold resistance: larvae with high glycerol levels were 3-4 times more tolerant of a 2h exposure to -10 degrees C than unstressed larvae. Protection against low temperature injury, as well as dehydration, was also attained by injection of exogenous glycerol into third instar larvae. This artificially induced cold hardiness was only temporary: when glycerol-injected larvae were exposed to -10 degrees C immediately after injection, survival was high, but none survived if they were injected and then held at 25 degrees C for 2 days before the -10 degrees C exposure. Larvae ligated behind the brain immediately after low temperature exposure failed to accumulate glycerol, but glycerol did accumulate in larvae ligated 6-24h after cold treatment, thus implying a critical role for the brain in initiating glycerol production. Interestingly, a much shorter exposure (2h) to low temperature was sufficient to reduce the maximum rate of water loss. Collectively, these observations suggest that multiple pathways may be exploited in response to stress: one pathway is most likely associated with rapid cold hardening (RCH) which generates immediate protection, and a second pathway remains activated for a longer period to enhance the initial protection afforded by glycerol.
"Although the alterations in these sugars are small, they may have beneficial effects on membrane function (Overgaard et al., 2014). Moreover, Yoder et al. (2006) demonstrated that an increase in the glycerol levels significantly improved chill tolerance, as may also happen for G. coquereliana. Together, our results demonstrate that the tropical cockroach G. coquereliana responds to cold stress by a shift in biochemical pathways , which results in the synthesis of compounds that protect the insect from or desensitize it to temperature changes. "
[Show abstract][Hide abstract] ABSTRACT: Insects cope with thermal stressors using mechanisms such as rapid cold hardening and acclimation. These mechanisms have been studied in temperate insects, but little is known about their use by tropical insects in response to cold stress. Here, we investigated whether cold stress (1 × 8 h and 3 × 8 h at 4 °C) triggers a metabolic response in the Madagascar cockroach Gromphadorhina coquereliana. We examined the effects of cold on the levels of selected metabolites in the fat body tissue of G. coquereliana. After cold exposure, we found that the quantity of total protein increased significantly in the insect fat body, whereas glycogen decreased slightly. Using antibodies, we observed upregulation of AQP-like proteins and changes in the HSP70 levels in the fat body of G. coquereliana when exposed to cold. We also examined the content and nature of the free sugars in the G. coquereliana hemolymph and discovered an increase in the levels of polyols and glucose in response to cold stress. These results suggest an important role of the fat body tissue of tropical insects upon cold exposure.
Comparative Biochemistry and Physiology - Part A Molecular & Integrative Physiology 01/2015; 183. DOI:10.1016/j.cbpa.2015.01.007 · 1.97 Impact Factor
"Glycerol and sorbitol, as the most common cryoprotectant molecules, lower the supercooling points and protect against protein denaturation 69. Glycerol kinase (CL3214.Contig1) is a rate-limiting enzyme in glycerol utilization 70, increasingly expressed during diapause to promote glycerol biosynthesis which was stress-induced and related to high cold resistance (Figure 8) 71. In a specific way, sorbitol dehydrogenase (CL3305.Contig1), related to diapause termination in eggs of B. mori 72, was upregulated in LD (Figure 8). "
[Show abstract][Hide abstract] ABSTRACT: Bactrocera minax is a major citrus pest distributed in China, Bhutan and India. The long pupal diapause duration of this fly is a major bottleneck for artificial rearing and underlying mechanisms remain unknown. Genetic information on B. minax transcriptome and gene expression profiles are needed to understand its pupal diapause. High-throughput RNA-seq technology was used to characterize the B. minax transcriptome and to identify differentially expressed genes during pupal diapause development. A total number of 52,519,948 reads were generated and assembled into 47,217 unigenes. 26,843 unigenes matched to proteins in the NCBI database using the BLAST search. Four digital gene expression (DGE) libraries were constructed for pupae at early diapause, late diapause, post-diapause and diapause terminated developmental status. 4,355 unigenes showing the differences expressed across four libraries revealed major shifts in cellular functions of cell proliferation, protein processing and export, metabolism and stress response in pupal diapause. When diapause was terminated by 20-hydroxyecdysone (20E), many genes involved in ribosome and metabolism were differentially expressed which may mediate diapause transition. The gene sets involved in protein and energy metabolisms varied throughout early-, late- and post-diapause. A total of 15 genes were selected to verify the DGE results through quantitative real-time PCR (qRT-PCR); qRT-PCR expression levels strongly correlated with the DGE data. The results provided the extensive sequence resources available for B. minax and increased our knowledge on its pupal diapause development and they shed new light on the possible mechanisms involved in pupal diapause in this species.
International journal of biological sciences 09/2014; 10(9):1051-63. DOI:10.7150/ijbs.9438 · 4.51 Impact Factor
"Cold sensing may be centrally coordinated via brain control to initiate glycerol production (Yoder et al., 2006). Alternatively, calcium influx in all tissues exposed to low temperature may autonomously induce RCH (Teets et al., 2013). "
[Show abstract][Hide abstract] ABSTRACT: Insects in temperate zones survive low temperatures by migrating or tolerating the cold. The diamondback moth, Plutella xylostella, is a serious insect pest on cabbage and other cruciferous crops worldwide. We showed that P. xylostella became cold-tolerant by expressing rapid cold hardiness (RCH) in response to a brief exposure to moderately low temperature (4°C) for 7 h along with glycerol accumulation in hemolymph. Glycerol played a crucial role in the cold-hardening process because exogenously supplying glycerol significantly increased the cold tolerance of P. xylostella larvae without cold acclimation. To determine the genetic factor(s) responsible for RCH and the increase of glycerol, four glycerol kinases (GKs), and glycerol-3-phosphate dehydrogenase (PxGPDH) were predicted from the whole P. xylostella genome and analyzed for their function associated with glycerol biosynthesis. All predicted genes were expressed, but differed in their expression during different developmental stages and in different tissues. Expression of the predicted genes was individually suppressed by RNA interference (RNAi) using double-stranded RNAs specific to target genes. RNAi of PxGPDH expression significantly suppressed RCH and glycerol accumulation. Only PxGK1 among the four GKs was responsible for RCH and glycerol accumulation. Furthermore, PxGK1 expression was significantly enhanced during RCH. These results indicate that a specific GK, the terminal enzyme to produce glycerol, is specifically inducible during RCH to accumulate the main cryoprotectant.
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