Harvesting and separation of two populations of lysosomes from porcine endometrium.
The lysosomes present in homogenates of porcine endometrium epithelium equilibrate in two density regions of Percoll gradients. Patterns with varying proportions between high and low density peaks are observed, when aliquots of a tissue sample are processed with different all-glass Potter-Elvejhem homogenizers. The described constant-tolerance shearing device (CTSD), in contrast, provides homogenate fractions with higher latencies and steady distribution patterns. They are characteristic for each of the six lysosomal markers and the six other structure-bound enzymes measured in gradient fractions of the particulate matter harvested between 600g and 17,000g. The 17,000g sediments of CTSD homogenates contain more than 40% of the total lysosomal enzymatic activities. Recoveries from Percoll gradients are between 93 and 101%. Enrichments in the high density region range from 35-fold (beta-glucosidase) to 82-fold (acid ribonuclease). Both lysosomal populations exhibit latencies between 89 and 94%. Our results indicate that light lysosomes can be artificially generated by inappropriate homogenization, which should be considered in experiments on the formation and maturation of lysosomes.
Adamski
J
J
Max-Planck-Institut für experimentelle Endokrinologie, Hannover, Bundesrepublik Deutschland.
Sierralta
W D
WD
Jungblut
P W
PW
D000818:Animals; D002458:Cell Fractionation; D002499:Centrifugation, Density Gradient; D004717:Endometrium / Q000032:analysis / Q000201:enzymology / Q000648:ultrastructure*; D005260:Female; D008247:Lysosomes / Q000032:analysis* / Q000201:enzymology; D008854:Microscopy, Electron; D013552:Swine