Metabolic consequences of the extent and disposition of the aqueous intracellular environment

Clegg, James S.

doi:10.1002/jez.1402150308

Cited by 27 publications

(3 citation statements)

References 37 publications

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“…Even now, water remains elusive in terms of its extent of and disposition within the intracellular environment (20). Just as Wilson (21), in 1925, was unaware of the specific details of the hyaloplasm, present researchers are confronted with trying to understand intracellular compartment organization in terms of the nature of soluble enzymes and the physical properties of the aqueous compartments.…”

mentioning

confidence: 99%

Combined proton NMR imaging and spectral analysis of locust embryonic development

Gassner

Lohman²

1987

Proc. Natl. Acad. Sci. U.S.A.

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Images and spectra of desert locust[Schistocerca gregaria (Forskal)] embryos were collected in vivo using a 4.7-tesla proton ('H) NMR imaging spectrometer. The 100 x 100 x 500 ,um inmage resolution and 125-nl localized spectral volumes were obtained within minutes of each other. The dynamics of embryonic development were slow enough that the time delay between imaging and spectral measurements is negligible. Thus, image and spectral data correspond closely and approximate real-time observations of embryonic changes. The above procedure was applied every 12 hr during the entire course of development. This analytical approach demonstrates that imaging and localized spectroscopy can be used to visualize and assess changes in embryonic water and lipid content in concert with the development of a living embryo.NMR studies of developing plants and animals have scientific merit because they offer the opportunity to carry out research on living organisms in a nondestructive way. Developmental geneticists need to understand the sequential and compartmental processes in early embryonic development (1-6).Cryopreservationists need to identify chemical and structural changes that are critical to maintaining the integrity of the organism during freezing and thawing (7). Physiologists need to track relevant nuclei through plant (8) and animal (9) vascular systems. These and many other areas of in vivo research could benefit from the nondestructive methods available using NMR technology.Here we show that proton NMR spectroscopy designed to combine imaging and selected volume spectral analysis offers great promise for research on developmental dynamics. We use this NMR method [image-selected in vivo spectroscopy, ISIS (10)] to follow the course of development of the locust Schistocerca gregaria (Forskal) from a few hours after fertilization to the formation and hatching of the first nymphal stage. This technique demonstrates that the near real-time dynamics of a developmental system can be visualized and chemically analyzed noninvasively. Embryonic movements, the formation of anatomical structures, and the dynamics of water and lipid in selected submicroliter volumes can be evaluated.Locust embryos develop in a centrolecithal ooplasm where the yolk is distributed within a cytoplasmic matrix that is continuous with a peripheral layer of yolk-free cytoplasm. The first embryonic divisions take place in a substrate that consists of yolk elements that differ in kind and size. After several energid divisions, nuclei with their associated cytoplasms migrate to the surface to form the presumptive embryonic germ band and embryonic membranes; nuclei remaining in (or returning to) the interior give rise to vitellophags. In the locust, the germ band forms as a 300-,um disc on the yolk periphery near the posterior dorsal margin. The remaining energids contribute to (i) membranes that envelop the embryo, (ii) yolk bodies, or (iii) lytic cells that digest the yolk. These cytological details and the subsequent features of the locust emb...

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confidence: 99%

Combined proton NMR imaging and spectral analysis of locust embryonic development

Gassner

Lohman²

1987

Proc. Natl. Acad. Sci. U.S.A.

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“…As dehydrative freezing progresses to impinge on restricted water, cell structures can become unstable leading to irreversible injury under some slow cooling conditions [36,45]. A second factor observed at higher cooling rates was related to intracellular ice formation (IIF), with the incidence of injury due to IIF increasing with increasing cooling rate [209].…”

Section: Cryoprotectants: Solutes That Modulate the Liquid Water To Imentioning

confidence: 99%

Cryoprotectants: A review of the actions and applications of cryoprotective solutes that modulate cell recovery from ultra-low temperatures

Elliott

Wang

Fuller³

2017

Cryobiology

415

292

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Cryopreservation has become a central technology in many areas of clinical medicine, biotechnology, and species conservation within both plant and animal biology. Cryoprotective agents (CPAs) invariably play key roles in allowing cells to be processed for storage at deep cryogenic temperatures and to be recovered with high levels of appropriate functionality. As such, these CPA solutes possess a wide range of metabolic and biophysical effects that are both necessary for their modes of action, and potentially complicating for cell biological function. Early successes with cryopreservation were achieved by empirical methodology for choosing and applying CPAs. In recent decades, it has been possible to assemble objective information about CPA modes of action and to optimize their application to living systems, but there still remain significant gaps in our understanding. This review sets out the current status on the biological and chemical knowledge surrounding CPAs, and the conflicting effects of protection versus toxicity resulting from the use of these solutes, which are often required in molar concentrations, far exceeding levels found in normal metabolism. The biophysical properties of CPAs that allow them to facilitate different approaches to cryogenic storage, including vitrification, are highlighted. The topics are discussed with reference to the historical background of applying CPAs, and the relevance of cryoprotective solutes in natural freeze tolerant organisms. Improved cryopreservation success will be an essential step in many future areas such as regenerative medicine, seed banking, or stem cell technology. To achieve this, we will need to further improve our understanding of cryobiology, where better and safer CPAs will be key requirements.

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“…In both cases, microcalorimetric studies have shown that the metabolic rate falls rapidly to no more than a few per cent of the active rate, with an almost complete cessation of carbohydrate metabolism (Hand and Gnaiger, 1988;Glasheen and Hand, 1989). The trigger to hypometabolism in anhydrobiotic embryos seems to be linked to ce11 dehydration and this has led to interesting considerations about the physical state of ce11 water (Clegg, 1981;1984). Recent work has also shown that dehydrated embryos are rich in "compatible" organic solutes, which may help to keep the integrity of macromolecular structures (Glasheen and Hand, 1989).…”

Section: Metabolic Depression: the Brine Shrimp Embryo As A Modelmentioning

confidence: 99%

Crustaceans as experimental animals for metabolic and transport physiology

Truchot

1993

Aquat. Living Resour.

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This paper develops briefly four examples illustrating the use of crustaceans as experimental animals in the field of metabolic and transport physiology. In cells of euryhaline crabs, organic substances such as free amino acids contribute much more than inorganic ions to osmotic adaptation, a feature that is now realized to be of general importance in living organisms. Anhydrobiotic and anaerobically dormant embryos of the brine shrimp Artemia provide very convenient models to study cellular control of metabolic depression. Studies on amphibious crabs have highlighted the constraints imposed upon gas exchange and acid-base balance by the very contrasted properties of oxygen and carbon dioxide in water and in air. Finally, the blood oxygen transport system of decapod crustaceans has been a mode1 of choice to demonstrate the significance of an increased oxygen affinity for adaptation to environmental hypoxia. These exarnples illustrate the role played in the past as well as the great future potential of crustaceans as experimental animal models for general studies in Comparative Physiology.

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Metabolic consequences of the extent and disposition of the aqueous intracellular environment

Cited by 27 publications

References 37 publications

Combined proton NMR imaging and spectral analysis of locust embryonic development

Combined proton NMR imaging and spectral analysis of locust embryonic development

Cryoprotectants: A review of the actions and applications of cryoprotective solutes that modulate cell recovery from ultra-low temperatures

Crustaceans as experimental animals for metabolic and transport physiology

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