Rapid cold-hardening in larvae of the Antarctic midge Belgica antarctica: cellular cold-sensing and a role for calcium

2008 ◽  
Vol 294 (6) ◽  
pp. R1938-R1946 ◽  
Author(s):  
Nicholas M. Teets ◽  
Michael A. Elnitsky ◽  
Joshua B. Benoit ◽  
Giancarlo Lopez-Martinez ◽  
David L. Denlinger ◽  
...  
Keyword(s):  
Cell Survival ◽  
Freezing Tolerance ◽  
Fat Body ◽  
Malpighian Tubules ◽  
Cold Hardening ◽  
Rapid Cold Hardening ◽  
Belgica Antarctica ◽  
The Antarctic ◽  
Cold Sensing

In many insects, the rapid cold-hardening (RCH) response significantly enhances cold tolerance in minutes to hours. Larvae of the Antarctic midge, Belgica antarctica, exhibit a novel form of RCH, by which they increase their freezing tolerance. In this study, we examined whether cold-sensing and RCH in B. antarctica occur in vitro and whether calcium is required to generate RCH. As demonstrated previously, 1 h at −5°C significantly increased organismal freezing tolerance at both −15°C and −20°C. Likewise, RCH enhanced cell survival of fat body, Malpighian tubules, and midgut tissue of larvae frozen at −20°C. Furthermore, isolated tissues retained the capacity for RCH in vitro, as demonstrated with both a dye exclusion assay and a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT)-based viability assay, thus indicating that cold-sensing and RCH in B. antarctica occur at the cellular level. Interestingly, there was no difference in survival between tissues that were supercooled at −5°C and those frozen at −5°C, suggesting that temperature mediates the RCH response independent of the freezing of body fluids. Finally, we demonstrated that calcium is required for RCH to occur. Removing calcium from the incubating solution slightly decreased cell survival after RCH treatments, while blocking calcium with the intracellular chelator BAPTA-AM significantly reduced survival in the RCH treatments. The calmodulin inhibitor N-(6-aminohexyl)-5-chloro-1-naphthalenesulfonamide hydrochloride (W-7) also significantly reduced cell survival in the RCH treatments, thus supporting a role for calcium in RCH. This is the first report implicating calcium as an important second messenger in the RCH response.

Characterization of drought-induced rapid cold-hardening in the Antarctic midge, Belgica antarctica

Polar Biology ◽  
2019 ◽  
Vol 42 (6) ◽  
pp. 1147-1156 ◽  
Author(s):  
Yuta Kawarasaki ◽  
Nicholas M. Teets ◽  
Benjamin N. Philip ◽  
Leslie J. Potts ◽  
J. D. Gantz ◽  
...  
Keyword(s):  
Cold Hardening ◽  
Rapid Cold Hardening ◽  
Belgica Antarctica ◽  
The Antarctic

Modeling seasonal changes in intracellular freeze-tolerance of fat body cells of the gall fly Eurosta solidaginis (Diptera, Tephritidae)

1997 ◽  
Vol 200 (1) ◽  
pp. 185-192 ◽  
Author(s):  
V Bennett ◽  
R Lee
Keyword(s):  
Cell Survival ◽  
Seasonal Changes ◽  
Cold Hardiness ◽  
Fat Body ◽  
Freeze Tolerance ◽  
Cellular Level ◽  
Eurosta Solidaginis ◽  
Freeze Tolerant

Although seasonal changes in the freeze-tolerance of third-instar larvae of Eurosta solidaginis have been well documented for the whole organism, the nature of this cold-hardiness at the cellular level has not been examined. Seasonal changes in the survival of fat body cells from E. solidaginis larvae were assessed using fluorescent vital dyes after freezing at -10, -25 or -80 °C for 24 h both in vivo and in vitro. Cells frozen in vitro were frozen with glycerol, with sorbitol (both of which enhanced cell survival) or without cryoprotectants. Both cellular and organismal survival were low in August when larvae were not freeze-tolerant, then increased dramatically during September and October before leveling off from November to January. This observation for cells frozen without cryoprotectants indicates that the cells themselves have adapted. The single most important factor influencing cell survival, as determined by logistic regression modeling, was the time of larval collection, which reflects the level of cold-hardiness achieved by field acclimation. Cells frozen in vivo exhibited greater survival than did those frozen in vitro, even with the addition of cryoprotectants. Since no differences were observed between cells frozen with glycerol or sorbitol, the role of the multi-component cryoprotectant system present in E. solidaginis should be investigated.

Stress tolerance in a polyextremophile: the southernmost insect

2015 ◽  
Vol 93 (9) ◽  
pp. 679-686 ◽  
Author(s):  
R.E. Lee ◽  
D.L. Denlinger
Keyword(s):  
Biotic Interactions ◽  
Cross Tolerance ◽  
Atp Production ◽  
Genome Wide ◽  
Rapid Cold Hardening ◽  
Belgica Antarctica ◽  
Genes Encoding ◽  
Shock Proteins ◽  
The Antarctic ◽  
Autophagy Inhibition

Since biotic interactions within the simple terrestrial communities on the Antarctic Peninsula are limited compared with tropical and temperate regions, survival is largely dictated by the numerous abiotic challenges. Our research focuses on adaptations to environmental stresses experienced by the Antarctic midge (Belgica antarctica Jacobs, 1900), the southernmost free-living insect. Midge larvae can survive freezing and anoxia year-round. Not only can frozen larvae undergo rapid cold-hardening (RCH) at temperatures as low as –12 °C, but RCH develops more rapidly in frozen compared with supercooled larvae. Whether larvae overwinter in a frozen state or cryoprotectively dehydrated may depend on hydration levels within their hibernacula. Larvae constitutively up-regulate genes encoding heat shock proteins, as well as the antioxidant enzymes superoxide dismutase and catalase. Larvae accumulate osmoprotectants in response to freezing, desiccation, and exposure to seawater; exposure to one of these osmotic stressors confers cross-tolerance to the others. Molecular responses to dehydration stress include extensive genome-wide changes that include differential expression of aquaporins among tissues, upregulation of pathways associated with autophagy, inhibition of apoptosis, and downregulation of metabolism and ATP production.

scholarly journals Brief exposure to a diverse range of environmental stress enhances stress tolerance in the polyextremophilic Antarctic midge, Belgica antarctica

2020 ◽  
Author(s):  
J. D. Gantz ◽  
B. N. Philip ◽  
N. M. Teets ◽  
Y. Kawarasaki ◽  
L. J. Potts ◽  
...  
Keyword(s):  
Environmental Stress ◽  
Stress Tolerance ◽  
Cold Tolerance ◽  
Abiotic Stresses ◽  
Environmental Cues ◽  
Ambient Conditions ◽  
Diverse Range ◽  
Rapid Cold Hardening ◽  
Belgica Antarctica ◽  
The Antarctic

AbstractInsects use rapid acclimation to enhance their tolerance of abiotic stresses within minutes to hours. These responses are critical adaptations for insects and other small ectotherms to tolerate drastic changes in temperature, hydration, or other factors that can fluctuate precipitously with ambient conditions or as a result of behavior. Rapid cold-hardening, where insects use brief exposure to modest chilling as a cue to enhance their cold tolerance, is the most thoroughly-studied of these responses and relatively little is known about rapid acclimation that is either triggered by or enhances tolerance of other abiotic stresses. Here, we used larvae of the Antarctic midge, Belgica antarctica, a polar extremophile that routinely experiences numerous stresses in nature, to investigate how 2 h exposure to modest environmental stresses affect stress tolerance in insects. Brief pretreatment by various stresses, including hyperosmotic challenge, hypoosmotic challenge, acidity, basicity, and UV irradiation enhanced stress tolerance in B. antarctica larvae relative to untreated controls. These results indicate that numerous environmental cues can trigger rapid acclimation in insects and that these responses can enhance tolerance of multiple stresses.

The warburg effect and tumour cell survival in human GBMs

2007 ◽  
Vol 30 (4) ◽  
pp. 97 ◽  
Author(s):  
A Wolf ◽  
J Mukherjee ◽  
A Guha
Keyword(s):  
Cytochrome C ◽  
Cell Survival ◽  
Expression Profile ◽  
Tumour Cell ◽  
Warburg Effect ◽  
Glycolytic Enzymes ◽  
Cytochrome C Release ◽  
Apoptotic Pathway ◽  
The Warburg Effect

Introduction: GBMs are resistant to apoptosis induced by the hypoxic microenvironment and standard therapies including radiation and chemotherapy. We postulate that the Warburg effect, a preferential glycolytic phenotype of tumor cells even under aerobic conditions, plays a role in these aberrant pro-survival signals. In this study we quantitatively examined the expression profile of hypoxia-related glycolytic genes within pathologically- and MRI-defined “centre” and “periphery” of GBMs. We hypothesize that expression of hypoxia-induced glycolytic genes, particularly hexokinase 2 (HK2), favours cell survival and modulates resistance to tumour cell apoptosis by inhibiting the intrinsic mitochondrial apoptotic pathway. Methods: GBM patients underwent conventional T1-weighted contrast-enhanced MRI and MR spectroscopy studies on a 3.0T GE scanner, prior to stereotactic sampling (formalin and frozen) from regions which were T1-Gad enhancing (“centre”) and T2-positive, T1-Gad negative (“periphery”). Real-time qRT-PCR was performed to quantify regional gene expression of glycolytic genes including HK2. In vitro functional studies were performed in U87 and U373 GBM cell lines grown in normoxic (21% pO2) and hypoxic (< 1%pO2) conditions, transfected with HK2 siRNA followed by measurement of cell proliferation (BrdU), apoptosis (activated caspase 3/7, TUNEL, cytochrome c release) and viability (MTS assay). Results: There exists a differential expression profile of glycolytic enzymes between the hypoxic center and relatively normoxic periphery of GBMs. Under hypoxic conditions, there is increased expression of HK2 at the mitochondrial membrane in GBM cells. In vitro HK2 knockdown led to decreased cell survival and increased apoptosis via the intrinsic mitochondrial pathway, as seen by increased mitochondrial release of cytochrome-C. Conclusions: Increased expression of HK2 in the centre of GBMs promotes cell survival and confers resistance to apoptosis, as confirmed by in vitro studies. In vivo intracranial xenograft studies with injection of HK2-shRNA are currently being performed. HK2 and possibly other glycolytic enzymes may provide a target for enhanced therapeutic responsiveness thereby improving prognosis of patients with GBMs.

scholarly journals Macropinocytosis requires Gal-3 in a subset of patient-derived glioblastoma stem cells

2021 ◽  
Vol 4 (1) ◽  
Author(s):  
Laetitia Seguin ◽  
Soline Odouard ◽  
Francesca Corlazzoli ◽  
Sarah Al Haddad ◽  
Laurine Moindrot ◽  
...  
Keyword(s):  
Stem Cells ◽  
Cell Survival ◽  
Small Gtpases ◽  
Molecular Signature ◽  
Carbohydrate Binding ◽  
Pancreatic Cancers ◽  
Galectin 3 ◽  
Pharmacologic Inhibition

AbstractRecently, we involved the carbohydrate-binding protein Galectin-3 (Gal-3) as a druggable target for KRAS-mutant-addicted lung and pancreatic cancers. Here, using glioblastoma patient-derived stem cells (GSCs), we identify and characterize a subset of Gal-3high glioblastoma (GBM) tumors mainly within the mesenchymal subtype that are addicted to Gal-3-mediated macropinocytosis. Using both genetic and pharmacologic inhibition of Gal-3, we showed a significant decrease of GSC macropinocytosis activity, cell survival and invasion, in vitro and in vivo. Mechanistically, we demonstrate that Gal-3 binds to RAB10, a member of the RAS superfamily of small GTPases, and β1 integrin, which are both required for macropinocytosis activity and cell survival. Finally, by defining a Gal-3/macropinocytosis molecular signature, we could predict sensitivity to this dependency pathway and provide proof-of-principle for innovative therapeutic strategies to exploit this Achilles’ heel for a significant and unique subset of GBM patients.

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