Cytochemical Localization of Adenosine Triphosphatase in the Mitotic Apparatus of Hela and Sarcoma 180 Tissue Culture Cells

Abstract

ATPase was localized in distinct regions of the mitotic apparatus of HeLa and Sarcoma 180 tissue culture cells. ATPase was demonstrated in the metaphase spindle of HeLa and Sarcoma 180 cells fixed in cold buffered 2 per cent formalin (pH 6.5 to 6.8) containing 2 X l0 -a CaCl2. A high concentration of ATPase was frequently observed at the poles of the spindle. ATPase was also demonstrated in the interzonal region of both cell types during anaphase. The narrowing of the band of ATPase activity localized in the interzonal region during telophase indicates that ATPase activity is associated with the central spindle. In polar views of Sarcoma 180 cells fixed in cold, unbuffered, 2 per cent formalin, ATPasc was frequently localized in granules in the region of the inner circumference of the ring of chromosomes formed at metaphase. ATPase in the mitotic apparatus of HeLa and Sarcoma 180 cells was shown not to be due to non-specific alkaline phosphatase. Mitotic apparatus ATPase in Sarcoma 180 cells was suppressed by an --SH inhibitor.

Ultrastructure and ATPase activity of the centriolar body at the mitotic stage in neoblasts of the planarianDugesia sp.

“…Mazia et al (1961) first studied the ATPase activity of the mitotic apparatus in the isolated spindle of sea urchin eggs. Later their findings were confirmed by others (Miki 1963;Hartmann 1964); Shifrin and Levine 1968). However, previous studies were carried out by light microscopic and biochemical methods, so the exact location of ATPase activity could not be determined.…”

“…Mazia et al (1961) first reported the presence of ATPase in the spindle body of the mitotic apparatus of sea urchin eggs and its presence was later confirmed biochemically (Miki 1962;Weisenberg and Taylor 1968) and morphologically by light microscopy (Miki 1962;Hartmann 1964;Shifrin and Levine 1968) but this only gives poor resolution of the distribution of ATPase activity. Since Gibbons and Rowe (1965) demonstrated that a special protein (dynein), with the properties of ATPase, was attached to the outer doublets of microtubules of cilia, it is possible that ATPase is present in spindle microtubules.…”

Cytoplasmic Microtubules and Radial‐Segmented Nuclei (Rieder Cells)

“…Except for spindle elongation in anaphase, cell movement and cell division are associated with structures that contain ATPase activity, use ATP as energy source and are suppressed by sulfhydryl reagents (Hoffmann-Berling 1959, Mazia et al 1961, Hartmann 1964. Thus, the failure of NEM and PHMB to inhibit the formation of RS nuclei s:ems to be more compatible with the hypothesis, that RS is due to shrinking cytoplasmic microtubules.…”

Cytoplasmic Microtubules and Radial‐Segmented Nuclei (Rieder Cells)

“…The suggestion that the tension of chromosome migration was provided by microtubule shortening due to loss of oriented monomeres from the microtubules (InouC & Sato 1967) seems difficult to reconcile with current theories about the basic mechanism of contractile processes. Furthermore, the contractile capacity of microtubules per se is not generally accepted (Kane & Forer 1965, Bajer 1968), although an actin-like amino acid composition of microtubules (Adelman et al 1968) and an ATPase activity in the mitotic apparatus (Hartmann 1964, Weisen-Bajer 1968, Nicklas 1969, it would probberg & Taylor 1968) have been demon-ably at present be better to conceive them strated. Since the microtubules are thought as being ultrastructures that convert the to be rigid structures associated to cell physical and chemical energy provided, i.e.…”