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Anhydrobiosis: Cellular Adaptation to Extreme Dehydration

John H. Crowe, Lois M. Crowe, John E. Carpenter, Steven Petrelski, Folkert A. Hoekstra, Pedro De Araujo, Anita D. Panek

10.1002/cphy.cp130220

Source: Supplement 30: Handbook of Physiology, Comparative Physiology

Originally published: 1997

Published online: January 2011

Full Article on Wiley Online Library



Abstract

The sections in this article are:

1 Induction of Anhydrobiosis
2 Biochemical Adaptations in Anhydrobiotes
2.1 Accumulation of Sugars by Anhydrobiotes
2.2 Sugars and the Wafer Replacement Hypothesis
2.3 Other Organic Compounds in Anhydrobiotic Plants
3 Sugars Stabilize Dry Proteins
3.1 Stabilization of Dried Proteins
3.2 Evidence for Direct Interaction between Sugars and Dry Proteins
3.3 Freezing Proteins
3.4 Preferential Interaction Mechanism and Cryoprotection of Proteins
4 Are Freezing and Dehydration Equivalent Stress Vectors?
5 Physical Properties of Phospholipids and Consequences of Dehydration
5.1 The Hydration Force
5.2 Effects of Water on the Physical Properties of Phospholipids
5.3 Physiological Consequences of Dehydration
6 Stabilization of Dry Liposomes
6.1 Retention of Trapped Solutes
6.2 Is the Bulk Concentration of Trehalose Important for Preservation?
6.3 Effects of Other Sugars
6.4 Mechanism of Stabilization of Dry Bilayers
6.5 Sugars and Lipid Phase Transitions
6.6 A Corollary to the Phase Transition Model
6.7 Effects of Trehalose on Phase Transitions in Dry DPPC
7 Mechanism of Interactions Between Sugars and Phospholipids
7.1 Evidence for Direct Interaction
7.2 When During the Drying Process Does Direct Interaction Occur?
8 Vitrification: An Alternative to the Water Replacement Hypothesis?
9 Extension of the Phase Transition Hypothesis to Native Membranes
10 Extension of the Phase Transition Hypothesis to Intact Cells
10.1 Effects on Phase Transitions in Intact Cells
10.2 The Phenomenon of Imbibitional Damage
10.3 Escape from Imbibitional Damage
10.4 Effects of Temperature on Leakage
10.5 What Is the Mechanism of Leakage?
10.6 Evidence for Gel‐to‐Liquid Crystalline Phase Transitions During Imbibition
10.7 Imbibition, Lipid Phase Transitions, and Germination
10.8 A Hydration‐Dependent Phase Diagram for Dry Cells
10.9 General Applicability of FTIR for Studies on Anhydrobiotes
10.10 Depression of Tm in Dry Pollen
10.11 Lipid Phase Transitions and Imbibitional Leakage in Dry Yeast
11 Toward a Mechanism for Stabilizing Dry Cells
11.1 Potential Routes for the Introduction of Trehalose into Cells
11.2 Studies on Genetics of Trehalose Synthesis
11.3 Survival of Drying by Mutants
11.4 A Transport System for Trehalose in Yeasts
11.5 Conditions for Expression of the Trehalose Transporter
11.6 Prospectus for Stabilizing Dry Cells
12 Are Additional Adaptations Required in Anhydrobiosis?
12.1 Studies on Nematodes
12.2 Studies on Pollen
12.3 Mechanism of Destabilization of Membranes by Fatty Acids
12.4 Generation of Free Fatty Acids in Dry Bilayers: Oxidation
12.5 Enzymatic Deesterification of Fatty Acids: Lipases
12.6 Is PLA2 Active in Dry Bilayers?
12.7 Inhibition of PLA2
13 Summary of Adaptations to Dehydration: Is Trehalose Sufficient?
Figure 1. Figure 1.

Chemical structures of some molecules accumulated at high concentrations by anhydrobiotic organisms.
Figure 2. Figure 2.

Comparison of abilities of various sugars to stabilize phosphofructokinase during freeze‐drying.

Data from 37
Figure 3. Figure 3.

Infrared spectra of trehalose in the fingerprint region for sugars. Trehalose freeze‐dried alone (A), freeze‐dried in the presence of lysozyme (B) or bovine serum albumin (C), or hydrated (D).

Data from 36
Figure 4. Figure 4.

Amide band region for hydrated lysozyme (dotted line), lysozyme freeze‐dried alone (dashed line), and lysozyme freeze‐dried in the presence of trehalose (solid line).

Data from 36
Figure 5. Figure 5.

A. Effects of trehalose on wave number for the amide II band of freeze‐dried lysozyme. B.Comparison of the recovery of phosphofructokinase activity after freeze‐thawing (triangles) or freeze‐drying and rehydration (circles).

Data from 36
Figure 6. Figure 6.

Left: Second derivative infrared spectra in the amide I region of a hydrated (upper) and a dry (lower) protein, basic fibroblast growth factor (bFGF). Right: Second derivative spectrum of bFGF freeze‐dried in the presence of 200 mg/ml sucrose.

Data from 142
Figure 7. Figure 7.

Leakage of carboxyfluorescein from vesicles of dimyristoylphosphatidylcholine as they are heated through phase transition.

Data from 58
Figure 8. Figure 8.

Retention of trapped solutes by liposomes that had previously been freeze‐dried in the presence of the indicated amounts of trehalose.

Data from 69
Figure 9. Figure 9.

Comparison of the abilities of various sugars to prevent leakage of trapped solutes from liposomes during drying.

Data from 55
Figure 10. Figure 10.

Residual water content in liposomes freeze‐dried in the presence of trehalose. Also shown are data for retention of trapped solutes by the same liposomes.

Data from 56
Figure 11. Figure 11.

Effects of various sugars on fusion between liposomes during drying.

Data from 69
Figure 12. Figure 12.

Effects of trehalose on calorimetric transitions in liposomes. Also shown are data for retention of trapped solutes by the same liposomes.

Data from 69
Figure 13. Figure 13.

Model for mechanism of stabilization of dry liposomes by trehalose.
Figure 14. Figure 14.

Effects of incubating frozen liposomes above and below on retention of trapped solutes. After incubation at indicated temperatures, liposomes were freeze‐dried and rehydrated and retention was measured.
Figure 15. Figure 15.

Retention of trapped solutes by liposomes freeze‐dried with trehalose alone, dextran alone, or mixtures of trehalose and dextran. Amount of trehalose is indicated on abscissa, with amount of dextran written on each curve.
Figure 16. Figure 16.

Typical infrared spectra in CH2 stretching region, illustrating change in frequency with temperature. Spectra are for intact bacteria (Escherichia coli). Similar spectra have been obtained with pure phospholipids, isolated membranes, and a wide variety of intact cells 62.
Figure 17. Figure 17.

Wave number of CH2 stretch in intact Escherichia coli under indicated conditions.
Figure 18. Figure 18.

Relationship between temperature of imbibition and survival of rehydration in pollen grains and vibrational frequency of membrane phospholipids. Open circles, wave numbers for samples containing 0.36 g H2O/g dry weight; closed circles, wave numbers for samples containing 0.06 g H2O/g dry weight; triangles, germination following rehydration at indicated temperatures. In samples containing 0.36 g/g, viability approached 100% when pollen grains were placed in H2O at 4°C.

Data from 60
Figure 19. Figure 19.

Phase diagram for membrane phospholipids in pollen, showing effects of hydration on Tm (open circles) and similar data for germination (closed squares). Data were extracted from plots similar to those shown in Figure. 18.

Data from 101
Figure 20. Figure 20.

Biosynthetic pathway for trehalose.
Figure 21. Figure 21.

Effects of phospholipase A2 on stability of dry liposomes.


Figure 1.

Chemical structures of some molecules accumulated at high concentrations by anhydrobiotic organisms.



Figure 2.

Comparison of abilities of various sugars to stabilize phosphofructokinase during freeze‐drying.

Data from 37


Figure 3.

Infrared spectra of trehalose in the fingerprint region for sugars. Trehalose freeze‐dried alone (A), freeze‐dried in the presence of lysozyme (B) or bovine serum albumin (C), or hydrated (D).

Data from 36


Figure 4.

Amide band region for hydrated lysozyme (dotted line), lysozyme freeze‐dried alone (dashed line), and lysozyme freeze‐dried in the presence of trehalose (solid line).

Data from 36


Figure 5.

A. Effects of trehalose on wave number for the amide II band of freeze‐dried lysozyme. B.Comparison of the recovery of phosphofructokinase activity after freeze‐thawing (triangles) or freeze‐drying and rehydration (circles).

Data from 36


Figure 6.

Left: Second derivative infrared spectra in the amide I region of a hydrated (upper) and a dry (lower) protein, basic fibroblast growth factor (bFGF). Right: Second derivative spectrum of bFGF freeze‐dried in the presence of 200 mg/ml sucrose.

Data from 142


Figure 7.

Leakage of carboxyfluorescein from vesicles of dimyristoylphosphatidylcholine as they are heated through phase transition.

Data from 58


Figure 8.

Retention of trapped solutes by liposomes that had previously been freeze‐dried in the presence of the indicated amounts of trehalose.

Data from 69


Figure 9.

Comparison of the abilities of various sugars to prevent leakage of trapped solutes from liposomes during drying.

Data from 55


Figure 10.

Residual water content in liposomes freeze‐dried in the presence of trehalose. Also shown are data for retention of trapped solutes by the same liposomes.

Data from 56


Figure 11.

Effects of various sugars on fusion between liposomes during drying.

Data from 69


Figure 12.

Effects of trehalose on calorimetric transitions in liposomes. Also shown are data for retention of trapped solutes by the same liposomes.

Data from 69


Figure 13.

Model for mechanism of stabilization of dry liposomes by trehalose.



Figure 14.

Effects of incubating frozen liposomes above and below on retention of trapped solutes. After incubation at indicated temperatures, liposomes were freeze‐dried and rehydrated and retention was measured.



Figure 15.

Retention of trapped solutes by liposomes freeze‐dried with trehalose alone, dextran alone, or mixtures of trehalose and dextran. Amount of trehalose is indicated on abscissa, with amount of dextran written on each curve.



Figure 16.

Typical infrared spectra in CH2 stretching region, illustrating change in frequency with temperature. Spectra are for intact bacteria (Escherichia coli). Similar spectra have been obtained with pure phospholipids, isolated membranes, and a wide variety of intact cells 62.



Figure 17.

Wave number of CH2 stretch in intact Escherichia coli under indicated conditions.



Figure 18.

Relationship between temperature of imbibition and survival of rehydration in pollen grains and vibrational frequency of membrane phospholipids. Open circles, wave numbers for samples containing 0.36 g H2O/g dry weight; closed circles, wave numbers for samples containing 0.06 g H2O/g dry weight; triangles, germination following rehydration at indicated temperatures. In samples containing 0.36 g/g, viability approached 100% when pollen grains were placed in H2O at 4°C.

Data from 60


Figure 19.

Phase diagram for membrane phospholipids in pollen, showing effects of hydration on Tm (open circles) and similar data for germination (closed squares). Data were extracted from plots similar to those shown in Figure. 18.

Data from 101


Figure 20.

Biosynthetic pathway for trehalose.



Figure 21.

Effects of phospholipase A2 on stability of dry liposomes.

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Article Title: Anhydrobiosis: Cellular Adaptation to Extreme Dehydration
 

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John H. Crowe, Lois M. Crowe, John E. Carpenter, Steven Petrelski, Folkert A. Hoekstra, Pedro De Araujo, Anita D. Panek. Anhydrobiosis: Cellular Adaptation to Extreme Dehydration. Compr Physiol 2011, Supplement 30: Handbook of Physiology, Comparative Physiology: 1445-1477. First published in print 1997. doi: 10.1002/cphy.cp130220