Piezotolerance of the cytoskeletal structure in cultured deep-sea fish cells using DNA transfection and protein introduction techniques

We used DNA transfection and protein introduction techniques to investigate the pressure tolerance of cytoskeletal structures in pectoral fin cells derived from the deep-sea fish Simenchelys parasiticus (habitat depth, 366–2,630 m). The deep-sea fish cells have G418 resistance. The cell number increased until day 6 of cultivation and all cells had died by day 35 when cultured in 35-mm Petri dishes in medium containing G418. Enhanced yellow fluorescent protein-tagged human β-actin (EYFP-actin) was stably expressed by 1 in 100,000 deep-sea fish cells. Because almost none of the EYFP-actin was incorporated into actin filaments of the cells, we replaced the relatively large EYFP tag with a chemical fluorescent compound and succeeded in incorporating fluorescently labeled rabbit actins into the deep-sea fish actin filaments. Most of the filament structure in the cells with rabbit actin inserted underwent depolymerization when subjected to pressure of 100 MPa for 20 min, in contrast to control cells. There were no differences in the tubulin filament structure between control cells and deep-sea fish cells with fluorescein-labeled bovine tubulin inserted after the application of pressure ranging from 40 to 100 MPa for 20 min.     

Comparative study of the spectral sensitivities of mesopelagic crustaceans

The spectral sensitivities of 12 species of mesopelagic crustaceans were studied by means of electrophysiological recordings. Nine of the species are vertical migrators, while 3 are not, and 9 species possess bioluminescent organs, while 3 are not bioluminescent. All species had a single peak of spectral sensitivity with maxima between 470 nm and 500 nm. There was no apparent correlation between sensitivity maxima and daytime depth distribution, migratory behavior, or the presence or absence of bioluminescent organs. With the exception of the hyperiid amphipod Phronima sedentaria, the spectral sensitivities of these mesopelagic crustaceans demonstrate a better match for maximum sensitivity to bioluminescence than to downwelling light. Accepted: 29 June 1999     

Culturability and survival of an extreme thermophile isolated from deep-sea hydrothermal vents

The culturability of a strictly anaerobic, extremely thermophilic archaeon, Thermococcus peptonophilus (optimal growth temperature: 85° C), was studied during survival stages at various temperatures (98, 85, 70, and 4° C). Total cell number (determined by DAPI staining), active cells (rhodamine-stained cells), and culturable cells (using most-probable-number) were counted over time. The number of culturable cells decreased under each condition tested. The total number of cells significantly decreased only at temperatures close to the maximum for growth (98° C); at this temperature, the cells spontaneously lysed. Our results suggested that survival at 4° C in oxygenated waters might be a mechanism for the dispersion of extreme thermophiles in the ocean. In addition, we proved the existence of T. peptonophilus cells in several physiological states: culturable cells, active non-culturable cells, inactive non-culturable cells, and dead cells. Cell death was caused by cellular lysis. Received: 5 February 1996 / Accepted: 16 April 1996