Surface localization of 5'-nucleotidase on the mouse lymphocyte.

Abstract

The optimal conditions for the cytochemical localization of 5'-nucleotidase (AMPase) in the mouse lymphocyte have been established. Quantitative monitoring of the effects of fixation and the components of the cytochemical medium showed that the cytochemistry can be performed under conditions that do not lead to loss of AMPase activity, and also under conditions where penetration of the substrate into the cell has occurred. The cytochemical reaction product was seen only on the surface of a proportion of splenic lymphocytes, regardless of the fixative used. Biochemical data confirmed that AMPase is an ectoenzyme and is the only protein in splenic lymphocytes capable of catalysing the hydrolysis of AMP. The activity of 5'-nucleotidase was studied also by harvesting cells either from thymus or spleen of A/ST or Cd-1 mouse strains. The enzymatic activity in splenic lymphocytes was more than six time higher than the activity of intact thymus cells. Cytochemically it was evident that within splenic lymphocytes there was a distinct population of lymphocytes with readily demonstrable AMPase activity, and another with no cytochemically demonstrable AMPase activity. It was concluded that murine lymphocytes vary in their activity of AMPase, and that the enzyme is exclusively confined to the cell surface.

Distribution of 5'-nucleotidase in human lymphoid tissues.

“…Peripheral blood mononUclear cells and tonsillar cells have a value for 5'-nucleotidase similar to B lymphoblasts. Observations by others in human and other mammalian lymphoid tissue support these data (19)(20)(21)(22)(23)(24).…”

Ultrastructural and ultracytochemical examination of the effects of preadministration of Xiao-Chai-Hu-Tang on hepatic disorders induced by D-galactosamine HCL.

“…For light microscopy, frozen sections of 15 p m in thickness were cut with an electrofreezing microtome. They were incubated in a medium of 5'-nucleotidase (24) for 30 min at 37°C, alkaline phosphatase (15) for 60 min at 37°C, and glucose-6-phosphatase (25) for 30 min at room temperature. Then the sections were washed in distilled water, treated with 1 % yellow anmonium sulfide, washed in distilled water, mounted on slide glasses and observed under a light microscope.…”

Ultracytochemical and biochemical investigations of alkaline phosphatase and 5'-nucleotidase activities in carcinogen-induced well-and poorly-differentiated adenocarcinoma of the rat stomach.

“…Histochemical and ultracytochemical procedures Six cases of well-differentiated adenocarcinoma from Group I, 2 of poorly-differentiated carcinoma from Group II and the normal glandular stomach from 2 untreated controls at the age of about 30 weeks were used for histochemical and ultracytochemical studies. Some frozen tissue samples were cut on a cryostat for histochemical ALPase reaction with conventional azodye method (3) and for 5'-Nase reaction with lead-based method (43). For ultracytochemistry, fresh tissues which were cut into longitudinal strips of approximately 0.2 cm width with razor blades were fixed in cold (4°C) 2% paraformaldehyde plus 1% glutarardehyde in 0.1 M cacodylate buffer (pH 7.4) for 1.0 hr.…”