Studies on Rapidly Frozen Suspensions of Yeast Cells by Differential Thermal Analysis and Conductometry

Mazur, Péter

doi:10.1016/s0006-3495(63)86824-1

Cited by 52 publications

(16 citation statements)

References 10 publications

(11 reference statements)

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“…These results indicate that eutectic crystallization of the suspending medium may influence the nucleation temperature under certain conditions, but cannot be considered to be the singular cause of intracellular nucleation. Thus, although nucleation of yeast cells (13) or sea urchin eggs (Strongyocentrotus nudus) (2) is not associated with eutectic crystallization under the conditions imposed, nucleation of eggs of another species of sea urchin (Hemicentrotus pulcherrimus) is associated with the eutectic crystallization (3).…”

Section: Discussionmentioning

confidence: 96%

Effect of Cold Acclimation on Intracellular Ice Formation in Isolated Protoplasts

Dowgert¹,

Steponkus²

1983

Plant Physiol.

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When cooled at rapid rates to temperatures between -10 and -30°C, the incidence of intracellular ice formation was less in protoplasts enzymically isolated from cold acclimated leaves of rye (Secale cereale L. cv Puma) than that observed in protoplasts isolated from nonacclimated leaves. The extent of supercooling of the intracellular solution at any given temperature increased in both nonacclimated and acclimated protoplasts as the rate of cooling increased. There was no unique relationship between the extent of supercooling and the incidence of intracellular ice formation in either nonacclimated or acclimated protoplasts. In both nonacclimated and acclimated protoplasts, the extent of intracellular supercooling was snimlar under conditions that resulted in the greatest difference in the incidence of intracellular ice formation-cooling to -15 or -20°C at rates of 10 or 16'C/mitute. Further, the hydraulic conductivity determined during freeze-induced dehydration at -5°C was similar for both nonacclimated and acclimated protoplasts. A major distinction between nonacclimated and acclimated protoplasts was the temperature at which nucleation occurred. In nonacclimated protoplasts, nucleation occurred over a relatively narrow temperature range with a median nucleation temperature of -15°C, whereas in acclimated protoplasts, nucleation occurred over a broader temperature range with a median nucleation temperature of -42°C. We conclude that the decreased incidence of intracellular ice formation in acclimated protoplasts is attributable to an increase in the stability of the plasma membrane which precludes nucleation of the supercooled intracellular solution and is not attributable to an increase in hydraulic conductivity of the plasma membrane which purportedly precludes supercooling of the intracellular solution.It is commonly observed that the incidence of intracellular ice formation is a function of the cooling rate. As early as 1886, observed that rapid cooling resulted in intracellular ice formation whereas during slow cooling ice crystals were confined to extracellular spaces. Similar rates; and the higher the permeability to water, the lower the probability of intracellular ice formation. Further, because the deplasmolysis time of cells in cold hardy tissues was less than that for cells in nonhardy tissues, they inferred that water permeability increased following cold acclimation and that the incidence of intracellular ice formation would be less in acclimated tissues. In 1938, Siminovitch and Scarth (23) experimentally demonstrated that intracellular ice formation occurs less frequently in hardy tissues than nonhardy tissues when cooled at comparable rates. They too attributed this difference to the purported increase in water permeability of the plasma membrane following cold acclimation as proposed by Levitt and Scarth (10). Although this concept has been widely accepted and frequently cited, direct experimental verification of an increase in water permeability following cold acclimation has been...

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Section: Discussionmentioning

confidence: 96%

Effect of Cold Acclimation on Intracellular Ice Formation in Isolated Protoplasts

Dowgert¹,

Steponkus²

1983

Plant Physiol.

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“…Despite the advances that have been made, cryopreservation still induces considerable damage to sperm as a result of the extreme temperature and osmolality changes to which cells are exposed [1–3]. With rapidly decreasing temperatures, extracellular solutes begin to freeze and the cell is exposed to an increasingly hyperosmolal environment, which turns extremely hypo-osmotic during thawing [4,5]. This fluctuation in osmolality causes substantial membrane stretching, which can disrupt the actin cytoskeleton and decrease the lateral packing of the lipids, leaving the cell membrane extremely vulnerable to oxidative damage [3,6,7].…”

Section: Introductionmentioning

confidence: 99%

Antioxidant treatment in the absence of exogenous lipids and proteins protects rhesus macaque sperm from cryopreservation-induced cell membrane damage

McCarthy

Meyers

2011

Theriogenology

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Osmotic stress caused oxidative stress in rhesus macaque sperm, which was alleviated by antioxidant supplementation. The objective of the present study was to demonstrate that cryopreservation of rhesus macaque sperm also induces (reactive oxygen species) ROS production, and to determine whether ROS have an important role in cryopreservation-induced membrane. Additionally, we evaluated the antioxidant capacity of TEST buffer (with 20% egg yolk and 13% skim milk) and supplementation with antioxidants, superoxide dismutase (SOD), catalase (CAT), and α-tocopherol. There was a substantial level of ROS production in both the presence (15% increase in superoxide, P < 0.01; 14% increase in hydrogen peroxide, P < 0.01) and absence of egg yolk (EY) and skim milk (SM; 33% increase in superoxide, P < 0.001; 48% increase in hydrogen peroxide, P < 0.001). Superoxide dismutase provided little membrane protection against ROS, but increased post-thaw total and progressive motility by 10% (P < 0.01) and 15% (P < 0.05), respectively. Supplementation with CAT and α-tocopherol in the presence of EY and SM decreased H 2 O 2 by 55% (P < 0.01) and 49% (P < 0.001), whereas supplementation with CAT and α-tocopherol in the absence of EY and SM reduced the level of lipid peroxidation by 61% (P < 0.05) and 28% (P < 0.01). In conclusion, this is apparently the first report that cryopreservation of rhesus macaque sperm induced a significant increase in ROS and that antioxidant supplementation can significantly decrease the extent of ROS-induced membrane damage.

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“…Although many advances have been made over the past 30 years to enhance the efficiency of NHP sperm cryopreservation, a substantial amount of sublethal and lethal damage still results from exposure to the extreme temperature and osmolality effects that cryopreservation introduces [1,3,4]. As the cell is exposed to plunging temperatures, extracellular ice crystallization begins, which results in concentration of the surrounding solutes in the unfrozen aqueous channels between ice crystals [3,5,6]. In response to the developing hyperosmolal environment, cells lose water and shrink in volume until the intracellular and extracellular solute concentrations equilibrate [6].…”

Section: Introductionmentioning

confidence: 99%

Osmotic Stress Induces Oxidative Cell Damage to Rhesus Macaque Spermatozoa1

McCarthy

Baumber

Kass

et al. 2010

Biology of Reproduction

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Cryopreservation introduces extreme temperature and osmolality changes that impart lethal and sublethal effects on spermatozoa survival. Additionally, evidence indicates that the osmotic stress induced by cryopreservation causes oxidative stress to spermatozoa as well. Our objective was to determine the effect of reactive oxygen species (ROS) on rhesus macaque (Macaca mulatta) sperm function and to determine whether osmotic stress elicits the production of ROS. In the first experiment, the xanthine-xanthine oxidase (X-XO) system was used to generate the ROS superoxide anion (O(2)(-.)) and hydrogen peroxide (H(2)O(2)) in the presence or absence of the ROS scavengers superoxide dismutase and catalase, respectively. In the second experiment, osmotic stress was introduced by incubation of spermatozoa in a series of anisosmotic media ranging from 100 to 1000 mOsmol/kg in the presence or absence of the antioxidant alpha-tocopherol. Treatment with the X-XO system resulted in a significant increase in the generation of O(2)(-.) and H(2)O(2) that was detectable using flow cytometry. The ROS generated by the X-XO system was dose dependent, and as the concentration of ROS increased, motility decreased and lipid peroxidation increased while no affect was observed on viability. Incubation of spermatozoa in anisosmotic media also resulted in an increase in O(2)(-.) generation and lipid peroxidation that was significantly decreased in the presence of the powerful antioxidant alpha-tocopherol. These results clearly indicate that osmotic stress causes oxidative stress in rhesus macaque spermatozoa, which strongly supports the hypothesis that cryopreservation-induced osmotic stress may lead to oxidative cell damage.

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Studies on Rapidly Frozen Suspensions of Yeast Cells by Differential Thermal Analysis and Conductometry

Cited by 52 publications

References 10 publications

Effect of Cold Acclimation on Intracellular Ice Formation in Isolated Protoplasts

Effect of Cold Acclimation on Intracellular Ice Formation in Isolated Protoplasts

Antioxidant treatment in the absence of exogenous lipids and proteins protects rhesus macaque sperm from cryopreservation-induced cell membrane damage

Osmotic Stress Induces Oxidative Cell Damage to Rhesus Macaque Spermatozoa1

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