Simultaneous visualization of alkaline phosphatase and gamma-glutamyl transpeptidase in kidney sections

An histochemical method is presented to simultaneously localize, for the first time, alkaline phosphatase (ALP) and gamma-glutamyltranspeptidase (gamma-GT) in the kidney. The reaction product of ALP activity appears as a dark brown precipitate of lead sulfide, while a bright red copper chelate of an azo dye (Fast blue BBN salt) final product indicates sites of gamma-GT activity. The amalgamation of Mayahara's (ALP) and Rutenberg's (gamma-GT) techniques resulted in the demonstration of various categories of kidney tubules, according to the staining reaction of the cell brush borders: Black tubules where ALP predominates; Intermediate tubules showing a mixture of brown and red precipitates; Red tubules indicating a prevalence of gamma-GT activity; Negative tubules. A possible relation might exist between the staining characteristics observed and the different proximal tubule segments, thus allowing their distinction. In addition, this technique has the advantage to permit the concomitant study of ALP and gamma-GT distribution on the same tissue section instead of serial sections, so reducing the number of manipulations and observations as well as the amount of tissue required.