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Cell fusion-mediated improvement in transfection competence for repair-deficient mutant of mouse T cell line (1988)

Shiomi, Tadahiro ; Hieda-Shiomi, Naoko ; Sato, Koki ; [et al.]

Springer

Somatic cell and molecular genetics 14 (1988), S. 195-203

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ISSN: 1572-9931

Source: Springer Online Journal Archives 1860-2000

Topics: Biology , Medicine

Notes: Abstract A multiple mutagen-sensitive mutant (XUM1) of mouse T-cell lymphoma line, L5178Y, is hypersensitive to ionizing radiation, ultraviolet (UV) light, and cross-linking agents (such as mitomycin C). The frequency of transfection for XUML cells after exposure to calcium phosphate-coprecipitated pSV2neo DNA was more than 104-fold less effective than that for Ltk−aprt− (LTA) cells. Other transfection methods (DEAE-dextran and polybrene-DMSO) were not effective for L5178Y and XUM1 cells. The transfection-proficient trait of LTA cells was demonstrated to be genetically dominant by examining the transfection frequency in hybrid clones constructed between XUM1 and LTA cells. To circumvent the problem with XUM1, the LTA genes necessary for transformation processes were introduced into XUM1 cells by constructing hybrids between XUM1 and LTA cells irradiated with X-rays which causes directional chromosome elimination for hybrid cells. Four of 194 hybrid clones tested were transfection-proficient and hypersensitive to UV (XL102, XL107, XL215, and XL216). All four clones were not hypersensitive to X-rays or mitomycin C. The frequencies of transfection for XL102 and XL216 were nearly the same level as that for LTA cells. The efficiency of transfection for XL107 and XL215 was 10 to 100-fold lower than that for LTA cells.

Type of Medium: Electronic Resource

URL: http://dx.doi.org/10.1007/BF01534404

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Implication that potassium flux and increase in intracellular calcium are necessary for the initiation of sperm motility in salmonid fishes (1994)

Tanimoto, Satomi ; Nakazawa, Tohru ; Kudo, Yoshihisa ; [et al.]

New York, NY [u.a.] : Wiley-Blackwell

Molecular Reproduction and Development 39 (1994), S. 409-414

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ISSN: 1040-452X

Keywords: K+ channel blockers ; K+ electrode ; Fura-2 ; Life and Medical Sciences ; Cell & Developmental Biology

Source: Wiley InterScience Backfile Collection 1832-2000

Topics: Biology

Notes: Flux of K+ and changes in intracellular Ca2+ in the sperm of salmonid fishes were measured with spectrophotometry, ion electrode, microscopic fluorometry, and radioisotope accumulation. Release of K+ occurred at the initiation of sperm motility which is induced by decrease in external K+ and the K+ efflux and sperm motility were inhibited by K+ channel blockers. Intracellular Ca2+ increased within a short period in K+- free condition, and the accumulation of 45Ca in sperm cells was higher in motile sperm than that in immotile sperm. The efflux of K+ and the increase in intracellular Ca2+ were suppressed when external K+ concentration increased, i.e., sperm remained immotile. These results suggest that efflux of K+ through K+ channel and subseqent increase in intracellular Ca2+ are prerequisite for the initiation of sperm motility. © 1994 Wiley-Liss, Inc.

Additional Material: 7 Ill.