Activities of key enzymes of purine degradation and re-utilization in human trophoblastic cells.
Using density gradient centrifugation, human trophoblastic cells were enriched from mixed cell populations of enzymatically dispersed first- and third-trimester placentae. Over 95 per cent of the cells recovered were of epithelial (i.e., trophoblastic) origin, as evidenced by their cytokeratin intermediate filament positivity and vimentin negativity, examined using indirect immunofluorescence, and also by their high content of human chorionic gonadotrophin. The activities of key enzymes involved in purine degradation and re-utilization (5'-nucleotidase; AMP-deaminase; hypoxanthine phosphoribosyltransferase (HPRT); xanthine dehydrogenase/oxidase) as well as the total activity of alkaline phosphatase were measured in the trophoblastic cells. A six-fold increase in the trophoblastic alkaline phosphatase activity was noted between the first and third trimester. A 40 per cent decrease was noted in the activity of 5'-nucleotidase, which, on the basis of kinetic properties, appears to have a dominant role in the dephosphorylation of placental nucleoside-5'-monophosphates. The trophoblastic activities of AMP-deaminase, HPRT, and xanthine dehydrogenase/oxidase did not change as a function of the gestational age. In view of the relative activities of the latter two enzymes, hypoxanthine formed in the trophoblast appears more likely to be re-utilized than degraded to uric acid.
Vettenranta
K
K
Children's Hospital, University of Helsinki, Finland.
Raivio
K O
KO
D015720:5'-Nucleotidase; D000243:Adenosine Deaminase / Q000378:metabolism*; D000469:Alkaline Phosphatase / Q000378:metabolism; D002469:Cell Separation; D005260:Female; D005455:Fluorescent Antibody Technique; D006801:Humans; D007041:Hypoxanthine Phosphoribosyltransferase / Q000378:metabolism*; D007658:Ketone Oxidoreductases / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D009700:Nucleoside Deaminases / Q000378:metabolism*; D009708:Nucleotidases / Q000378:metabolism*; D011247:Pregnancy; D011261:Pregnancy Trimester, First; D011263:Pregnancy Trimester, Third; D011687:Purines / Q000378:metabolism; D014327:Trophoblasts / Q000166:cytology / Q000201:enzymology*; D014968:Xanthine Dehydrogenase / Q000378:metabolism*
D011687:Purines; D014968:Xanthine Dehydrogenase; D007658:Ketone Oxidoreductases; D007041:Hypoxanthine Phosphoribosyltransferase; D009708:Nucleotidases; D000469:Alkaline Phosphatase; D015720:5'-Nucleotidase; D009700:Nucleoside Deaminases; D000243:Adenosine Deaminase