腹水腫瘍の細胞化学的研究 第1編 脱水素酵素系の細胞化学的研究

Using the Ehrlich ascites tumor cells, the activities of the diphosphopyridine nucleotide diaphorase (DPNase), triphosphopyridine nucleotide diaphorase (TPNase), succinic dehydrogenase (SD) and α-gylcerophosphate dehydrogenase (α-GD) were cytochemically examined, and all of the cytochemical specimens were observed with the phase contrast microscope and formazanhaematoxylin counter staining. Several variations of their activities being caused by the differences of the cytochemical methods were also studied. The results were summarized as follows. 1. In the smear method, in which the ascites is smeared and dried on the object glass, thereafter reacted with the modified Wattenberg's reagents, all cells reveal relatively intense reaction but the differences of the staining intensity of each cell are scarcely noticed. 2. In the suspension method, in which ascites tumor cells are suspended and reacted with the modified Wattenberg's reagents, all cells reveal relatively weak reaction, but the marked differences of the staining intensity of each cell are observed. 3. In order to clarify the relationship between the permeability of the cell membrane and the intensity of the cytochemical reaction, the cells are treated with the benzalkonium chloride solution. This method is called the permeability increasing suspension method, which reveals a intermediate intensity of the staining between the smear method and the suspension method. Each cell shows a definite difference of the staining. 4. No significant difference is observed between the staining intensity of the suspension method and that of the ascites suspension method in which the ascites supernatant is used as the cytochemical reaction medium instead of the inorganic salts solution. 5. The cells treated with the suspension, permeability increasing suspension and ascites suspension method are able to be successively transplanted into the same strain mouse. Therefore, it may be reasonably assumed that in these three methods the cells are cytochemically reacted in the living condition. However, it is impossible to transplant the cells treated with the smear method. 6. By the observation of the staining patterns of these four methods, it may be assumed that these dehydrogenases in the cells are not always activated and the intensities of their enzymatic activities change according to their life cycle. (a) The DPNase activity remarkably increases in the end of the interphase and abruptly decreases to the lowest level at the metaphase. Then the activity gradually increases, keeping low level throughout the first half of the interphase. This enzyme is localized in the granulations of the cytoplasm and also a little in the cytoplasms of the cells with the high activity. (b) In general, the TPNase activity is slightly lower than that of the DPNase, and shows the same mode of the activity and localization as the latter. (c) The SD activity decreases to the lowest level at the metaphase and remains almost constantly low in the other phases, localizing in the mitochondria. (d) The α-GD activity is generally lower than that of the SD and nearly negative, but shows the same changes of the activity and localization as the latter. 7. When the enzymatic activity of Ehrlich ascites tumor cells is compared with that of the normal ascites cells, the DPNase and TPNase activity are remarkably high, while the SD and α-GD activity are low.