Cold shock damage is due to lipid phase transitions in cell membranes: A demonstration using sperm as a model
TL;DR: The lipid phase behavior was consistent with the temperature range over which cooling was damaging for pig and shrimp sperm, and the with the extent of damage produced in pig and human sperm, the first direct evidence that cold shock results from lipid phase transitions in cell membranes.
Abstract: When cells are cooled to temperatures above the freezing point of water at rates greater than a few degrees per minute, they sustain irreversible injury. Reduction of this "cold shock" damage could increase the survival of animals and plants at low environmental temperatures and improve the cryopreservation of plant and animal cells. Leakage of solutes across membranes, associated with thermotropic phase transitions in membrane lipids, is thought to be responsible, but this hypothesis has not been tested directly. Using Fourier transform infrared spectroscopy (FTIR), we measured the lipid phase transitions in intact, living sperm, the animal cell in which cold shock has been studied most extensively. A shift in the CH2 absorbance peaks indicates the transition from liquid-crystalline to gel phase. The phase transition in sperm membranes occurred at a lower temperature for a marine shrimp than for the pig. In each case, potassium leakage, which is a hallmark of cold shock damage, increased abruptly near the end of the phase transition. Human sperm are quite resistant to cold shock, and an abrupt lipid phase transition was not detected. This phase behavior is typical of membranes containing a high proportion of cholesterol, and human sperm have an unusually high sterol content. High cholesterol levels are known to stabilize membranes during cooling. Overall, the lipid phase behavior was consistent with the temperature range over which cooling was damaging for pig and shrimp sperm, and the with the extent of damage produced in pig and human sperm. This is the first direct evidence that cold shock results from lipid phase transitions in cell membranes.
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TL;DR: Evidence for new approaches for improving the performance of cryopreserved semen is offered and factors affecting the proportion of survivors and functional status of survivors are reviewed.
1,383 citations
Cites background from "Cold shock damage is due to lipid p..."
...However, it is well known that a major phase change Ž .occurs in the vicinity of 5–158C Drobnis et al., 1993 , and this may well be the prime temperature range for temperature dependent injury....
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TL;DR: The possible roles of cryoprotectants and additives are considered in the context of their putative interactions with the sperm plasma membrane and modern approaches to the laboratory assessment of spermatozoa after freeze-thawing are discussed.
648 citations
Cites background from "Cold shock damage is due to lipid p..."
...Ž .lipid phase transition effects was presented by Drobnis et al. 1993 ....
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...Spermatozoa undergo these lipid phase transitions Crowe et .al., 1989; Drobnis et al., 1993; Holt and North, 1984; 1986; Parks and Lynch, 1992 typically within the temperature range 17–368C....
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...Drobnis et al., 1993 ....
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TL;DR: Successful semen cryopreservation enhances the advantages of AI over natural breeding, as long-term storage facilitates semen transport over distances, permits the quarantine of semen, and enables extended use of superior germplasm, even after the sire's death.
Abstract: The genetic improvement of agriculturally important species and disease control are of fundamental importance to the success of a sustainable agri-food industry. In this sense, artificial insemination (AI) is arguably the most important tool contributing to the advancement of modern animal production. Through AI, 1 ejaculate from a genetically superior male can be used to impregnate multiple females to maximize the distribution of agriculturally favorable genes. As well, AI eliminates physical contact between animals, thus limiting the spread of sexually transmitted diseases. Successful semen cryopreservation enhances these advantages of AI over natural breeding. Long-term storage facilitates semen transport over distances, permits the quarantine of semen, and enables extended use of superior germplasm, even after the sire’s death. For agriculturally important animals, semen cryopreservation is an established industry worldwide, particularly for dairy cattle. Genome resource banking to preserve the biodiversity of endangered species or valuable transgenic lines also would benefit from sperm cryopreservation. For many mammals; however, effective semen cryopreservation is not a reality because a large number of sperm are apparently infertile following freezing and thawing. Compared to fresh, 8 times more cryopreserved bovine sperm were required to achieve equivalent fertilization rates in vivo (Shannon and Vishwanath, 1995). Although AI in pigs with fresh, cooled semen is increasingly popular, the fertility of cryopreserved semen remains about half that of fresh semen (Crabo, 1991). Following either transuterine or oviductal insemination of superovulated ewes, conception rates and percentage of fer-
585 citations
Cites background from "Cold shock damage is due to lipid p..."
...The susceptibility of the plasma membrane to undergo lipidphase transitions during cooling is inversely related to the proportion of cholesterol present (Drobnis et al, 1993)....
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TL;DR: Dynamics in adhesive and fusion properties, molecular composition and architecture of the sperm plasma membrane, as well as membrane derived signalling are reviewed.
580 citations
TL;DR: It is argued that species differences in female tract anatomy, subtle differences in sperm transport mechanisms, ability to time inseminations and deliver spermatozoa effectively are powerful determinants of fertility with cryopreserved spermatozosa.
558 citations
References
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TL;DR: It is proposed that, for T c where T c is the transition temperature, perylene is excluded from the hydrocarbon interior of the membranes, whereas, T c this probe may be accommodated in the membrane interior to a large extent.
955 citations
TL;DR: It is suggested that, because the non-lamellar lipids tend to undergo a liquid-crystalline to gel-phase transition at higher temperatures than lamellar-forming lipids, these will tend to phase separate into a gel phase domain rich in these lipids.
399 citations
TL;DR: Cold-induced changes in the organization of sperm plasma membrane components were demonstrated by the use of fast-freezing combined with freeze-fracture electron microscopy and species-dependent differences incold-induced ultrastructural changes were considered to be determined by lipid composition and asymmetry of the plasma membrane, and might be related to differences in cold resistance between species.
222 citations
TL;DR: It is shown here that Fourier transform infrared spectroscopy provides a rapid, convenient method for detecting lipid phase transitions in intact cells and that the transitions can nevertheless be detected in other intact cells, including those of four plant species and sperm of three animals.
179 citations
TL;DR: It is proposed that this type of irreversible change within the sperm plasma membrane may contribute to the loss of motility and fertility suffered by spermatozoa after cooling and freezing and suggested that protective substances such as egg yolk may exert their effects by countering these deleterious changes, rather than by preventing their occurrence.
Abstract: In an effort to investigate the nature of the cellular injury caused when mammalian spermatozoa are cooled prior to cryopreservation, the occurrence of thermal phase transitions amongs the lipid components of the sperm plasma membrane was investigated by the use of freeze-fracture electron microscopy. The mechanisms by which glycerol and egg yolk exert protective effects during semen cooling and freezing were also examined. Ram and blackbuck spermatozoa, maintained at 30 degrees C prior to fixation at this temperature, exhibited randomly distributed intramembranous particles over the acrosomal, postacrosomal, and flagellar regions of the plasma membrane. In contrast, spermatozoa fixed at 5 degrees C after slow cooling to this temperature exhibited particle clustering over the postacrosomal region of the head as well as over the tail. These effects were not influenced by the presence of egg yolk or glycerol during the cooling procedure, although these substances protected the spermatozoa against loss of motility. Particle clustering over the sperm tail, induced by the slow cooling process, was found to be only partially reversible. The extensive areas of particle-free lipid, noted to result from the cooling procedure, were absent if the spermatozoa were rewarmed to 30 degrees C; however, the original distribution of particles was not restored and numerous small particle-free domains persisted. It is proposed that this type of irreversible change within the sperm plasma membrane may contribute to the loss of motility and fertility suffered by spermatozoa after cooling and freezing. Furthermore, it is suggested that protective substances such as egg yolk may exert their effects by countering these deleterious changes, rather than by preventing their occurrence.
135 citations