Abstract
Histochemical demonstration and colorimetric determination of β-glucuronidase activities have been reported by SELIGMAN, TSOU, RUTENBURG and COHEN (1954) using 6-bromo-2-naphthyl-β-D-glucuronide as the substrate, and COHEN, TSOU, RUTENBURG and SELIGMAN distributed histochemically β-D-galactosidase by employing of 6-bromo-2-naphthyl-β-D-galactopyranoside as the substrate, and also a colorimetric determination of β-glucosidase was introduced by COHEN, RUTIENBURG, TSOU, WOODBUTY and SELIGMAN (1952) in ultilizing as the substrate 6-bromo-2-naphthyl-β-D-glucopyranoside. In these post incubation coupling azo-dye methods for three kinds of β-glycosidase was employed tetrazotized diorthoanisidin as the dye coupler. On the other hand, the histochemical and biochemical determination of β-glucuronidase have been established by FRIDENWALD and BACKER (1948), and recently, 8-hydroxyquinolin glucosiduronic acid as the biological synthetic substrate by FISHMAN and BAKER (1956). The present report notes the localization of β-glucuronidase, β-galactosidase and β-glucosidase activities in rhoden organs, in accordance with post coupling method by SELIGMAN and his co-workers.
The experimental animals such as normal mouse, rat, guinea-pig and rabbit were used for the histochemical study. Fresh sections of about 15μ thick ness were cut by sliding microtome in the -20°C cryostat according to the COON's modification of the LINDERSTRÖM-LANG technique and fresh preparats were dried in room temperature for 30 minutes, fixed briefly in cold 10% neutral formalin for the minutes, rinsed by distilled water, and incubated at 37°C for six hours in each substrate solution. After incubation, the sections were rinsed in tap water and transferred to 4°C solution of 0.02M phosphate buffer at pH 7.5per 1mg of diazo blue B for 5 minutes. The portions which showed the enzymatic activities of β-glycosidase were stained rapidly in color of blue or bluish purple. The results of the hitochemical demonstration of these enzymes in Table 1 may be summarized as follows.
1. The localization of β-glucuronidase, β-galactosidase andβ-glucosidase activities in certain tissues and organs in mouse, rat, guinea-pig and rabbit are almost the same effects except the digestive tubes.
2. In the epithelium of digestive organs such as stomach, duodenum, small intestinum and colon relative intense reactions of β-glucuronidase and β-galactosidase are observed, whereas the activity of β-galactosidase is slight.
3. No organ of the respiratory system shows a intense reaction of these enzymes.
4. In the cortex of the kidney of the rodents were observed a similar localization of stricking enzymatic activities. The reaction of the enzymes is most intense in the cortex of the kidney, while in the outer zone of the medulla much weaker.
5. In the genital organs, there are remarkable difference between the very slight or negative reactions of glycosidase in mouse and guinea-pig, and the stricking and significant reactions of the enzyme in rat and rabbit. The localization and distribution of this enzyme were noted as equal in the uterus, FALLOPIan tube and ovary on the rat and rabbit.
