scholarly journals Freeze tolerance of Cyphoderris monstrosa (Orthoptera: Prophalangopsidae)

2016 ◽  
Vol 148 (6) ◽  
pp. 668-672 ◽  
Author(s):  
Jantina Toxopeus ◽  
Jacqueline E. Lebenzon ◽  
Alexander H McKinnon ◽  
Brent J. Sinclair
Keyword(s):  
Cold Tolerance ◽  
Freeze Tolerance ◽  
Malpighian Tubules ◽  
Ice Formation ◽  
Freezing And Thawing ◽  
Tolerance Strategy ◽  
Late Instar ◽  
Freeze Tolerant ◽  
Overwintering Survival

AbstractThe great grig, Cyphoderris monstrosa Uhler (Orthoptera: Prophalangopsidae), is a large (20–30 mm, >1 g), nocturnal ensiferan that inhabits montane coniferous forests in northwestern North America. Cyphoderris monstrosa overwinters as a late instar nymphs, but its cold tolerance strategy has not previously been reported. We collected nymphs from near Kamloops, British Columbia, Canada in late spring to determine their cold tolerance strategy. Cyphoderris monstrosa nymphs were active at low temperatures until they froze at −4.6±0.3 °C. The nymphs survived internal ice formation (i.e., are freeze tolerant), had a lethal temperature between −9 °C and −12 °C, and could survive for between five and 10 days at −6 °C. Isolated C. monstrosa gut, Malpighian tubules, and metafemur muscle tissues froze at temperatures similar to whole nymphs, and likely inoculate freezing in vivo. Hemolymph osmolality was 358±51 mOsm, with trehalose and proline comprising ~10% of that total. Glycerol was not detectable in hemolymph from field-fresh nymphs, but accumulated after freezing and thawing. The control of ice formation and presence of hemolymph cryoprotectants may contribute to C. monstrosa freeze tolerance and overwintering survival.

scholarly journals Evidence for non-colligative function of small cryoprotectants in a freeze-tolerant insect

2019 ◽  
Vol 286 (1899) ◽  
pp. 20190050 ◽  
Author(s):  
Jantina Toxopeus ◽  
Vladimír Koštál ◽  
Brent J. Sinclair
Keyword(s):  
Fat Body ◽  
Ex Vivo ◽  
Freeze Tolerance ◽  
Field Cricket ◽  
Ice Formation ◽  
Low Molecular Weight ◽  
Manipulative Experiments ◽  
Gryllus Veletis ◽  
Freeze Tolerant

Freeze tolerance, the ability to survive internal ice formation, facilitates survival of some insects in cold habitats. Low-molecular-weight cryoprotectants such as sugars, polyols and amino acids are hypothesized to facilitate freeze tolerance, but their in vivo function is poorly understood. Here, we use a combination of metabolomics and manipulative experiments in vivo and ex vivo to examine the function of multiple cryoprotectants in the spring field cricket Gryllus veletis . Cold-acclimated G. veletis are freeze-tolerant and accumulate myo -inositol, proline and trehalose in their haemolymph and fat body. Injecting freeze-tolerant crickets with proline and trehalose increases survival of freezing to lower temperatures or for longer times. Similarly, exogenous myo -inositol and trehalose increase ex vivo freezing survival of fat body cells from freeze-tolerant crickets. No cryoprotectant (alone or in combination) is sufficient to confer freeze tolerance on non-acclimated, freeze-intolerant G. veletis . Given that each cryoprotectant differentially impacts survival in the frozen state, we conclude that small cryoprotectants are not interchangeable and likely function non-colligatively in insect freeze tolerance. Our study is the first to experimentally demonstrate the importance of non-colligative cryoprotectant function for insect freeze tolerance both in vivo and ex vivo , with implications for choosing new molecules for cryopreservation.

Freeze-thaw injury in erythrocytes of the freeze-tolerant wood frog, Rana sylvatica

1991 ◽  
Vol 261 (6) ◽  
pp. R1346-R1350 ◽  
Author(s):  
J. P. Costanzo ◽  
R. E. Lee
Keyword(s):  
Cell Injury ◽  
Osmotic Fragility ◽  
Rana Sylvatica ◽  
Freeze Tolerance ◽  
Wood Frog ◽  
In Vitro Tests ◽  
Freezing And Thawing ◽  
Freeze Tolerant ◽  
High Concentrations

Erythrocytes from the freeze-tolerant wood frog (Rana sylvatica) were subjected to in vitro tests of freeze tolerance, cryoprotection, and osmotic fragility. The responses of cells from frogs acclimated to 4 or 15 degrees C were similar. Erythrocytes that were frozen in saline hemolyzed at -4 degrees C or lower. The addition of high concentrations (150 and 1,500 mM) of glucose or glycerol, cryoprotectants produced naturally by freeze-tolerant frogs, significantly reduced cell injury at -8 degrees C, but concentrations of 1.5 or 15 mM were ineffective. Hemolysis was reduced by 94% with 1,500 mM glycerol and by 84% with 1,500 mM glucose; thus glycerol was the more effective cryoprotectant. Mean fragility values for frog erythrocytes incubated in hypertonic and hypotonic saline were 1,938 and 49 mosM, respectively. Survival in freeze tolerance and cryoprotection experiments was comparable for erythrocytes from frogs and humans, suggesting that these cells may respond similarly to freezing-related stresses. However, the breadth of osmotic tolerance, standardized for differences in isotonicity, was greater for frog erythrocytes than for human erythrocytes. Our data suggest that erythrocytes from R. sylvatica are adequately protected by glucose under natural conditions of freezing and thawing.

Freeze tolerance in turtles: visual analysis by microscopy and magnetic resonance imaging

1994 ◽  
Vol 267 (4) ◽  
pp. R1078-R1088 ◽  
Author(s):  
B. Rubinsky ◽  
J. S. Hong ◽  
K. B. Storey
Keyword(s):  
Magnetic Resonance Imaging ◽  
Magnetic Resonance ◽  
Visual Analysis ◽  
Abdominal Cavity ◽  
Free Water ◽  
Freeze Tolerance ◽  
The Body ◽  
Resonance Imaging ◽  
Freezing And Thawing

Two visual techniques were used to analyze the patterns of natural freezing and thawing in freeze-tolerant hatchling painted turtles Chrysemys picta marginata. Directional solidification plus light microscopy of liver, heart, and skeletal muscle slices was used to compare freezing at -4 degrees C (a survivable temperature in vivo) and -20 degrees C (not survivable). At -4 degrees C tissues showed large amounts of ice in expanded extracellular and vascular spaces, occupying 36% (liver) and 61% (muscle) of total tissue volume. Cells at -4 degrees C were shrunken, but intracellular water remained; at -20 degrees C, however, cells showed little evidence of free water. Liver micrographs showed novel spherical shells of water associated with intracellular particles (apparently glycogen granules) suggesting that a noncolligative method of cell water retention was employed. Proton magnetic resonance imaging was used for noninvasive analysis of freezing and thawing in the intact animal. Images showed that freezing propagated in a directional manner through the body with ice formed first in extraorgan spaces (e.g., abdominal cavity, brain ventricles). However, thawing occurred uniformly throughout the body core, and organs melted more rapidly than the extraorgan ice surrounding them.

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.

Fat body cells and calcium phosphate spherules induce ice nucleation in the freeze-tolerant larvae of the gall fly Eurosta solidaginis (Diptera, Tephritidae)

1996 ◽  
Vol 199 (2) ◽  
pp. 465-471 ◽  
Author(s):  
J Mugnano ◽  
R Lee ◽  
R Taylor
Keyword(s):  
Calcium Phosphate ◽  
Fat Body ◽  
Freeze Tolerance ◽  
Ice Nucleation ◽  
Malpighian Tubules ◽  
Whole Body ◽  
Eurosta Solidaginis ◽  
New Class ◽  
Ice Nucleators ◽  
Freeze Tolerant

During the autumn, the third-instar larvae of the gall fly Eurosta solidaginis acquire freeze tolerance and their crystallization temperatures increase into the -8 to -10 °C range. Despite conflicting reports, efficient endogenous ice nucleators have not been identified in this freeze-tolerant insect. We found large crystalloid spheres within the Malpighian tubules of overwintering larvae. Energy-dispersive X-ray microanalysis and infrared spectroscopy indicated that the spherules were a hydrate of tribasic calcium phosphate. To test for ice-nucleating activity, we placed the calcium phosphate spherules in 10 µl of Schneider's insect medium and cooled them in a refrigerated bath. The addition of spherules increased the crystallization temperature of Schneider's medium by approximately 8 C, from -18.4±0.8 °C to -10.1±0.9 °C (mean ± s.e.m., N=20). Ice-nucleating activity (-10.9±0.9 °C) was also demonstrated in fat body cells suspended in 10 µl of Schneider's medium. Both calcium phosphate spherules and fat body cells have ice-nucleating activity sufficiently high to explain whole-body crystallization temperatures. Furthermore, other crystalloid deposits, commonly found in diapausing or overwintering insects, also exhibited significant ice-nucleating activity. These endogenous crystalloid deposits represent a new class of heterogeneous ice nucleators that potentially regulate supercooling and promote freeze tolerance in E. solidaginis and possibly in other overwintering insects.

EFFECT OF PROPYLTHIOURACIL ON THE IODINATION AND MATURATION OF RAT THYROGLOBULIN

1966 ◽  
Vol 53 (2) ◽  
pp. 271-285 ◽  
Author(s):  
Claude Simon ◽  
Marie Roques ◽  
Janine Torresani ◽  
Serge Lissitzky
Keyword(s):  
Single Injection ◽  
Freezing And Thawing ◽  
Isotopic Equilibrium ◽  
Sedimentation Constant

ABSTRACT The effect of propylthiouracil on the maturation of rat thyroglobulin in vivo has been investigated. Newly iodinated thyroglobulin dimer is labile to freezing and thawing. This observation has been used to interpret the findings in the present experiments. From experiments using rats in isotopic equilibrium with 125I, and treated with propylthiouracil or propylthiouracil and tri-iodothyronine and also given a single injection of 131I, the following conclusions were formulated 1) the appearance of iodinated S12 thyroglobulin monomer is due to the dissociation of labile iodinated thyroglobulin dimer and appears more readily if the dimer is poorly iodinated, 2) uniodinated thyroglobulin dimer is the most probable substrate for iodination in vivo, 3) maturation of thyroglobulin dimer (as shown by increasing sedimentation constant from 16—17 to 19) is accompanied by increasing amounts of iodine in the molecule, 4) it is not possible to say at present if iodination and iodothyronine formation is the cause or the consequence of thyroglobulin dimer maturation, 5) propylthiouracil might inhibit thyroglobulin maturation by decreasing iodine organification.

The anterior and posterior ‘stomach’ regions of larval Aedes aegypti midgut: regional specialization of ion transport and stimulation by 5-hydroxytryptamine

1999 ◽  
Vol 202 (3) ◽  
pp. 247-252 ◽  
Author(s):  
T.M. Clark ◽  
A. Koch ◽  
D.F. Moffett
Keyword(s):  
Steady State ◽  
Aedes Aegypti ◽  
Ion Transport ◽  
Malpighian Tubules ◽  
Transport Mechanisms ◽  
Regional Specialization ◽  
Negative Deflection ◽  
Electrical Polarity

The ‘stomach’ region of the larval mosquito midgut is divided into histologically distinct anterior and posterior regions. Anterior stomach perfused symmetrically with saline in vitro had an initial transepithelial potential (TEP) of −66 mV (lumen negative) that decayed within 10–15 min to a steady-state TEP near −10 mV that was maintained for at least 1 h. Lumen-positive TEPs were never observed in the anterior stomach. The initial TEP of the perfused posterior stomach was opposite in polarity, but similar in magnitude, to that of the anterior stomach, measuring +75 mV (lumen positive). This initial TEP of the posterior stomach decayed rapidly at first, then more slowly, eventually reversing the electrical polarity of the epithelium as lumen-negative TEPs were recorded in all preparations within 70 min. Nanomolar concentrations of the biogenic amine 5-hydroxytryptamine (5-HT, serotonin) stimulated both regions, causing a negative deflection of the TEP of the anterior stomach and a positive deflection of the TEP of the posterior stomach. Phorbol 12,13-diacetate also caused a negative deflection of the TEP of the anterior stomach, but had no effect on the TEP of the posterior stomach. These data demonstrate that 5-HT stimulates region-specific ion-transport mechanisms in the stomach of Aedes aegypti and suggest that 5-HT coordinates the actions of the Malpighian tubules and midgut in the maintenance of an appropriate hemolymph composition in vivo.

Anti-diuresis in the blood-feeding insect Rhodnius prolixus Stål: the peptide CAP2b and cyclic GMP inhibit Malpighian tubule fluid secretion.

1997 ◽  
Vol 200 (17) ◽  
pp. 2363-2367 ◽  
Author(s):  
M C Quinlan ◽  
N J Tublitz ◽  
M J O'Donnell
Keyword(s):  
Cyclic Gmp ◽  
Physiological Role ◽  
Malpighian Tubule ◽  
Fluid Secretion ◽  
Blood Feeding ◽  
Rhodnius Prolixus ◽  
Malpighian Tubules ◽  
Tubule Fluid ◽  
Secretion Rates

Rhodnius prolixus eliminates NaCl-rich urine at high rates following its infrequent but massive blood meals. This diuresis involves stimulation of Malpighian tubule fluid secretion by diuretic hormones released in response to distention of the abdomen during feeding. The precipitous decline in urine flow that occurs several hours after feeding has been thought until now to result from a decline in diuretic hormone release. We suggest here that insect cardioacceleratory peptide 2b (CAP2b) and cyclic GMP are part of a novel mechanism of anti-diuresis. Secretion rates of 5-hydroxytryptamine-stimulated Malpighian tubules are reduced by low doses of CAP2b or cyclic GMP. Maximal secretion rates are restored by exposing tubules to 1 mmol l-1 cyclic AMP. Levels of cyclic GMP in isolated tubules increase in response to CAP2b, consistent with a role for cyclic GMP as an intracellular second messenger. Levels of cyclic GMP in tubules also increase as urine output rates decline in vivo, suggesting a physiological role for this nucleotide in the termination of diuresis.

The Symptoms and Histopathology of Poisoning by Maneb in Oncopeltus fasciatus (Dallas),

1965 ◽  
Vol 97 (11) ◽  
pp. 1200-1208 ◽  
Author(s):  
R. D. McMullen
Keyword(s):  
Oxygen Consumption ◽  
Malpighian Tubules ◽  
Oncopeltus Fasciatus ◽  
Secretory Cells ◽  
Cell Nuclei ◽  
Gut Epithelium

AbstractManeb (manganous ethylene bisdithiocarhamate) applied topically to Oncopeltus fasciatus nymphs causes death after 7 to 10 days. The gross symptoms of intoxication, histopathology and effect on oxygen consumption are described. Activities such as feeding and walking are slightly reduced after 24 hours and completely inhibited after 3 to 4 days. The tissues most severely affected by the treatment are the secretory cells of the mid-gut epithelium and the cells of the Malpighian tubules. These at first show extreme vacuolization, reduction of the size of cell nuclei and finally cytolysis. Oxygen consumption in vivo is reduced by the treatment.

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