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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Development genes and evolution 206 (1996), S. 297-304 
    ISSN: 1432-041X
    Keywords: Key words Ascidian embryogenesis ; Polarity of unfertilized egg ; Cell fusion ; Cytoplasmic transfer ; Cytoplasmic determinants
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Cytoplasmic determinants that specify the fate of endoderm, muscle and epidermis cells are known to be localized in specific areas of fertilized eggs of ascidians. The presence of such cytoplasmic determinants in unfertilized eggs was demonstrated in previous studies, but no information has yet been proved about their distribution. To investigate the distribution of cytoplasmic determinants in unfertilized eggs, we devised a method for distinguishing the polarity of unfertilized eggs using vital staining and we performed cytoplasmic-transfer experiments by fusing blastomeres and cytoplasmic fragments from various identified regions of unfertilized eggs. Cytoplasmic fragments, that contained cortical and subcortical material, from five different positions along the animal-vegetal axis were prepared, and they were fused with a4.2 (presumptive-epidermis) or A4.1 (non-epidermis) blastomeres. The ectopic development of endoderm, muscle and epidermis cells that was promoted by the transplanted cytoplasm was assessed by examining the expression of alkaline phosphatase (ALP), myosin and epidermis-specific antigen, respectively. Differentiation of endoderm and muscle was observed at higher frequencies as cytoplasmic fragments closer to the vegetal pole were transplanted. Conversely, formation of epidermis was observed at higher frequencies as cytoplasmic fragments closer to the animal pole were transplanted. The results suggest that, in cortical and subcortical regions of unfertilized ascidian eggs, endoderm and muscle determinants are widely distributed along a gradient, with maximum activity at the vegetal pole, whilst epidermis determinants are also distributed along a gradient but with maximum activity at the animal pole.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-2013
    Keywords: Key words Ruthenium red ; Smooth muscle ; Ca2+-dependent K+ channel ; Ca2+ channel ; Ryanodine ; Urinary bladder
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract  Three major ionic currents, Ca2+-dependent K+ current (I K-Ca), delayed rectifier type K+ current (I kd) and Ca2+ current (I Ca), were activated by depolarization under whole-cell clamp in single smooth muscle cells isolated from guinea-pig urinary bladder. Externally applied ruthenium red (RuR) reduced the amplitude of I K-Ca and I Ca at 0 mV (IC50 values were 4.2 and 5.6 μM, respectively), but did not affect I Kd. Spontaneous transient outward currents (STOCs) and caffeine-induced outward currents (I caf) at –30 mV were reduced by external 10 μM RuR. When 10 μM RuR was added to the pipette solution, I K-Ca during depolarization, STOCs and I caf significantly decreased with time. RuR did not change the unitary current amplitude of the large-conductance Ca2+-dependent K+ (BK) channels, but reduced the open probability of the channel under excised patch-clamp recording mode. RuR reduced the channel activity more effectively from the cytosolic face than from the other. This inhibition decreased when the cytosolic Ca2+ concentration was increased. These results indicate that RuR blocks BK and Ca2+ channels in urinary bladder smooth muscle cells. The decrease in I K-Ca, STOCs and I caf by RuR is attributable to the direct inhibition of BK channel activity, probably in addition to the inhibition of Ca2+ release from storage sites. The direct inhibition of BK channel activity by RuR may be related to the interaction of RuR with the Ca2+-binding sites of the channel protein.
    Type of Medium: Electronic Resource
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