X-Message-Number: 4720 Date: Fri, 4 Aug 1995 11:08:53 +0200 (MET DST) From: Eugen Leitl <> Subject: nature's ways A short (3 pages of 14, to be continued) excerpt from "Biochemical Adaptation" (Peter W. Hochachka, George N. Somero, Princeton University Press 1984, pp. 436-449). Apart from raising scholary interest concerning boundaries and rates of evolutionary adaption, this fragment elucidates how biological systems adapt to existance in contigous cryohabitats and/or shows some of the mechanisms enabling them to persist through pronounced cold seasons. Especially shotgun cryoprotection strategies utilized by some (ant)arctic organisms which involve simultaneous use of noncolligative macromolecular peptide and glycopeptide as well as the more conventional polyhydroxyl alcohol (polyol) cryoprotectants with at the same time markedly increased dehydration resistance have earned a closer look, I think. The human ability to induce high perfusion end concentrations at high perfusion rates yet hitherto pronounced disability to tackle what is labeled as cryoprotectant toxicity (Fahy et al., 1990) and nature's way of doing things appear remarkably complementary. Perhaps a synthesis might achieve what each isolated technique alone cannot accomplish. -- Eugene -----------------------------cut here--------------------------- Contents: List of Figures List of Tables Preface List of Abbreviations Biochemical Adaptation: Basic Mechanisms and Strategies Design of Cellular Metabolism Adaptation of Enzymes to Metabolic Functions Exercise Adaptations Limiting Oxygen Avalability Metabolic Adaptation to Diving Off-Switches in Metabolism: From Anhydrobiosis to Hibernation Mammalian Developmental Adaptations Respiratory Proteins Water-Solute Adaptations: The Evolution and Regulation of Biological Solutions Temperature Adaption Adaptations to the Deep Sea References Index A fragment from the Temperature Adaptation chapter: Freezing Resistance and Freezing Tolerance. Much as organisms must closely regulate the physical states of their lipid-based systems, for many cold climate species regulating the physical state of the extra- and intracellular fluids assumes critical importance during much or all of the year. With rare exceptions, intracellular ice formation is lethal to cells. While it is true that cryo- preservation methodologies do alloww long-term storage of frozen cells and tissues (Ashwood-Smith and Farrant 1980), the conditions nec- essary to achieve this feat, e.g., the addition of high concentrations of cryoprotectant substances like dimethylsulfoxide, are not accessible to animals in nature. Most organisms are likely to die even if ice for- mation is confined to the extracellular spaces, although there are striking examples of species that not only tolerate freezing of the extra- cellular fluids, but even nucleate ice formation. In the majority of species that experience the threat of freezing, however, mechanisms are employed that prevent ice formation in all body compartments. The unusual macromolecular entities, biological "antifreeze" com- pounds, which retard ice formation and confer freezing resistance to a varied suite of fishes and invertebrates, serve as the primary focus of this final section of the chapter on thermal relationships. The choice of strategy of coping with ambient temperatures that are below the body fluid freezing points of organisms not adapted to resist or tolerate freezing depends on several factors. One important con- sideration relates to the ability of the organism to remain active at these low temperatures. If an ecological factor such as lack of food dictates that dormancy is desirable during the cold season, then the extracellular body fluids of the animal may be modified to facilitate the formation of ice at temperatures close to the freezing point of the blood or hemolymph. In such cases supercooling is largely avoided. These types of animals are termed "freeze-tolerant" because they can with- stand ice formation in the extra-, but not the intracellular fluids. It may seem maladaptive to produce ice-nucleating agents to trigger ice forma- tion at relatively high temperatures, a feat common in freeze-tolerant species (see Zachariassen, 1980) for at first glance there appears to be little basis for concluding that ice formation in the extracellular fluids which is followed by withdrawal of water from the intracellular fluids, could be beneficial to the organisms. It has long been thought that dehydration of the intracellular space, with concomitant increases in inorganic ion concentrations and distortion of intracellular structures, is a major cause of low temperature-induced lethality. However, the point of using nucleating agents to foster ice formation at relatively high freezing temperatures is that, by avoiding supercooling, ice forma- tion will occur in the extracellular spaces at temperatures well above those at which spontaneous ice formation can occur intracellularly. Po- tential damage from dehydration my still exist albeit freeze-tolerant animals have marked capacities to withstand dehydration (Kanwisher, 1955; Murphy and Pierce, 1975). Ice formation in the intracellular space is prevented, however, and this achievement may be most critical for a freeze-tolerant organism. The potential damage caused by ice crystal growth within the cell appears much greater than damage due to dehydration. A second major strategy for dealing with low body temperatures is found in freeze-resistant species, organisms that employ biochemical mechanisms to prevent ice formation in both the extra- and intracellu- lar fluids. In many cases the species that utilize this strategy remain active at potentially freezing temperatures. Sustained activity and the presence of ice-containing extracellular fluids and dehydrated intra- cellular spaces are conditions that seem incompatible. Polar fishes offer especially good examples of freezing resistance, and the peptide and glycopepide antifreezes in the body fluids of these species allow them to remain active in the presence of ice at seawater temperatures of -1.86 deg C (DeVries, 1980, 1982). While many fishes do erect biochemical defences against freezing, it is appropriate to note that many species employ seasonal migrations which lead to removal of the threat of freezing. For example, the long horn sculpin, Myoxocephalus octodecemspinosus, migrates out of near- shore waters in the winter where ice formation occurs, and seeks a deeper, ice-free habitat during those cold months. This behaviour eli- minates the danger of freezing via the seeding of the body fluids (freez- ing points are more than 1 deg C above the freezing point of seawater for fishes lacking antifreezes). However, migration into deeper water would appear to present this sculpin with a reduced food supply as well as exposure to another set of predators. Thus, behavioral avoidance of freezing may carry the costs of existance in a suboptimal habitat for at least part of the year. The development of antifreze molecules has allowed organisms to select habitats on criteria other than than the presence or absence of temperatures that are lethal to freeze-susceptible species. Examples of Freeze-Tolerant Organisms. Prior to reviewing the biochemistry of antifreeze molecules, we shall consider briefly a variety of organisms that tolerate, and often induce, ice formation in their extracellular fluids. These organisms typically are terrestrial species that are dormant in winter. Numerous examples of insects are known that contain ice-nucleating agents that effectively prevent supercooling of the hemolymph (Duman, 1980; Zachariassen, 1980). As mentioned above, this relatively high-temperature freezing prevents more deletorious low-temperature freezing of the cytosol. A somewhat similiar adaptive strategy has recently been reported in the plant, Lobelia telekii, which is native to an Afro-alpine environment where temperatures near -10 deg C may occur at night throughout the year (Krog et al., 1979). This plant, unlike an Arctic beetle, cannot go dormant for weeks to months at a time, but instead must find ways to achieve tolerance for freezing at night while remaining metabolically active and growing during warmer periods of the day. The key factor of this plant's diurnal thermal regulatory strategy is a large, fluid-filled compartment in the inflorscence of the plant. This cavity is filled with a viscous fluid that serves at least two thermally related functions. First, the aqueous fluid has a large heat capacity and thermal inertia, so some of the changes in ambient temperature will be buffered by this fluid. Second, this fluid behaves like the blood of freeze-tolerant insects in that it contains an ice-nucleating agent that triggers freezing near 0 deg C. This ice formation can be beneficial in two ways. As in the case of freeze-tolerant animals, the prevention of supercooling acts as a means for reducing the dangers of intracellular ice formation which could occur if the body temperature of the plant fell well below 0 deg C. In addi- tion, at higher subzero temperatures the heat of fusion released during ice formation can actually warm the plant. Krog et al. (1979) found that as the ambient temperature decreased from about 12 deg C during daytime highs to nearly -8 deg C during the late night, the temperature of the central part of the plant did not fall below 0 deg C. Ice was gener- ated at a sufficient rate to buffer the central temperature. These authors calculated that approximately 2 percent of the central cavity froze under these conditions, suggesting that a significant "heat reservoir" remained should more extreme temperature conditions be experienced. It seems highly unlikely that similiar heat-generating functions of ice-nucleating agents occur in animal body fluids. The relative fluid volumes of L. telekii and animals are grossly different, and the diffi- culsties of controlling heat flow in small animals are obvious. Moreover, -------------------------------------------------------------------------- Hang on, the next stuff is a lot more interesting (but my fingers bleed from typing...). Look at the thing with a proportional font to somewhat reduce layout raggedness. -- Eugene Rate This Message: http://www.cryonet.org/cgi-bin/rate.cgi?msg=4720