Abstract
The rhodents, such as mice, rats, guinea-pigs and rabbits, both males and females, were used, and tissues of various organs as shown in Fig. 1 were examined, namely of squamous epithelium of oral mucosa, tongue, oesophagus, salaivary gand, stomach, intestine, liver, pancreas, trachea, lung, heart, spleen, kidney, testis, epididymis, uterus, FALLOPIan tube, ovary, thyroid gland and adrenal gland. Fresh tissues were cut into 10 to 20μ sections in the cryostat at- 20°C in accordance with COO's modification of LINDERSTRÖM-LANG technique. The fresh frozen sections were dried in room temperature for 20 to 30 minutes and fixed in buffered 10% formalin for 10 minutes and rinsed briefly in distilled water. Both histochemical techniques were utilized which were used by BURSTONE & FOLK and NACHLAS, CRAWFORD & SELIGMAN. According to the BURSTONE-FOLK's azo coupling method (1950) specimens were incubated with hydrochloride of L-leucyl-β-naphthyl amine resp. DL-alanyl-β-naphthylamine as the substrate. When the diazotized o-aminoazotoluene (Garnet GBC) as the dye coupler was employed the specimens gained desirable result, but diazotized p-nitro-o-anisidin (Diazo red B) or p-nitro-p-amino-2, 5-dimethoxy-diphehylamide (Black salt K) were able to be used instead of Garnet GBC. While in these experiment the incubated solution was the mixed substrate of 1% stocke substrate solution, 40ml of distilled water, 10ml of 0.2M phosphate buffer (pH 6.8) and 30mg of Diazo blue B. The specimens were incubated for an hour at room temperature. Furthermore the method of NACHLAS et al. was applied. The sections were incubated at 37°C for an hour, and then rinsed in distilled water and chelated in 0.1M cupper sulfate.
Histochemical demonstration of aminopeptidase in rhodent organs was performed in employing both methods.
1. The histochemical reactions of aminopeptidase in both methods of BURSTONE & FOLK (1956) and NACHLAS, CRAWFORD & SELIGMAN (1957) surveyed the similar localization.
2. The aminopeptidase activity of various tissues of guinea-pig was more intensive than that of mouse, rat and rabbit.
3. High activities of aminopeptidase were observed in the kindney and intestine of all animals as well as in the submaxillary gland and prancreas of mouse and in the submaxillary gland of rabbit.
4. Inhibiting effects of several substances on the enzyme activity are shown in Fig. 2. A complete inhibiting took place in 0.001M Fe (SO4)3, and a moderate one in 0.001M CuSO4. A complete inhibition was also observed in 0.01M Ba (OH)2 and a moderate one in 0.01M CuSO4, Pb (NO3)2 and AL2(SO4)3.
