Cell biology of deep-sea multicellular organisms

Abstract

Establishing tissue cultures derived from deep-sea multicellular organisms has been extremely difficult because of the serious damage they sustain upon decompression and exposure to the high temperature of surface seawater. We developed a novel pressure-stat aquarium system for the study of living deep-sea multicellular organisms under pressure. Using this system, we have succeeded in maintaining a variety of deep-sea multicellular organisms under pressure and atmospheric conditions after gradual, slow decompression. Furthermore, we successfully cultivated and freeze-stocked pectoral fin cells of the deep-sea eel Simenchelys parasiticus collected at a depth of 1,162 m under atmospheric pressure conditions. This review describes novel capture and maintenance devices for deep-sea organisms and cell culture studies of the organisms under atmospheric and pressure conditions.

Diving under a Microscope—A New Simple and Versatile In Vitro Diving Device for Fluorescence and Confocal Microscopy Allowing the Controls of Hydrostatic Pressure, Gas Pressures, and Kinetics of Gas Saturation

“…This system allowed performing trans-illumination through samples and fluorescence microscopy under high hydrostatic pressure. It supported, for example, the analysis of the effect of pressure on calcium homeostasis (Crenshaw & Salmon, 1996), cytoskeleton (Crenshaw et al, 1996), or cell division (Koyama, 2007). Nevertheless, the interest of those approaches remains limited in the field of human diving studies: during human diving, not only hydrostatic pressure but also different gas partial pressure (N 2 , O 2 , etc.)…”

Electrical Retrieval of Living Microorganisms from Cryopreserved Marine Sponges Using a Potential-Controlled Electrode

“…1 . To obtain the sponge-associated microorganisms, we carefully removed any microbial contaminants attached to the surface of the sponge tissue using previously described deep-sea animal cell and tissue culture techniques (Koyama and Aizawa 2000 , 2008 ; Koyama et al 2003 , 2005 ; Koyama 2007 ). The frozen marine sponge tissue was transferred into ice-cold sterile artificial seawater (30 g of NaCl, 0.7 g of KCl, 5.3 g of MgSO 4 ⋅7H 2 O, 10.8 g of MgCl 2 ⋅6H 2 O, 1 g of CaCl 2 ⋅2H 2 O L −1 ) and incubated for 2 h. The thawed or fresh marine sponge tissue was rinsed with fresh sterile artificial seawater cooled to 4 °C.…”

Electrical Collection of Membrane-intact and Dehydrogenase-positive Symbiotic Bacteria from the Deep-sea Bivalve <i>Calyptogena Okutanii</i>

“…13,14 2.3 Collection of symbiont bacteria from deep-sea bivalves using the potential-controlled electrode To obtain intracellular symbiotic bacteria from gill tissues of C. okutanii, we used the previously described deep-sea animal cell culture techniques for preventing the contamination of tissueattached bacteria. 16 The mucus was removed from freshly dissected gill tissues of the deep-sea bivalve with paper towels, and then the tissues were washed with artificial seawater at 4°C and dipped into 70% ethanol for 30 s (Fig. 1A).…”