Dynamics of platelet-activating-factor release and uptake in a human neutrophil suspension.
The present study has examined the dynamics of platelet-activating-factor (PAF) synthesis, release and uptake in order to understand better the trafficking of PAF between cells and medium. Initial experiments indicated that the amount of PAF found on the outside of the cell remained constant well after the synthesis from a precursor had apparently stopped, and in spite of a continued capacity of the cell to take up and catabolize PAF. These results suggested that PAF produced and stored within the cell is eventually released to the outside of the cell at a rate proportional to that of cellular uptake. In order to estimate the amount of PAF released from the cell, the processes of PAF release and uptake were modelled using simple mathematical functions. It was found that, under the experimental conditions used in this study, the uptake of PAF could be expressed as an exponential function tending to a non-zero baseline. Utilizing this rate constant for the uptake of PAF with the amount of PAF outside the cell, the amount of total PAF released from the cell was estimated. Data from the model suggested PAF was released in amounts 10-fold higher than could actually be measured over 30 min. In fact, the model predicted more PAF could be released from the cell than is synthesized, suggesting that a portion of the PAF which is released is taken up and then released again to the outside of the cell. The potential for PAF and/or its intermediates to be recycled was verified by demonstrating that a large proportion of exogenously provided 1-alkyl-2-lyso-sn-glycero-3-phosphocholine is taken up by the neutrophil, converted into PAF and then released again by the cell. These results suggest that PAF trafficking between the cell and medium is complex and involves many processes, which include synthesis, release, uptake, catabolism and recycling.
Cluzel
M
M
Johns Hopkins Asthma and Allergy Center, Johns Hopkins University School of Medicine, Baltimore, MD 21224.
Rohde
C A
CA
MacGlashan
D W
DW
Undem
B J
BJ
Chilton
F H
FH
D000001:Calcimycin / Q000494:pharmacology; D006801:Humans; D007700:Kinetics; D008962:Models, Theoretical; D009206:Myocardium / Q000737:chemistry; D009504:Neutrophils / Q000378:metabolism*; D010972:Platelet Activating Factor / Q000378:metabolism*
D010972:Platelet Activating Factor; D000001:Calcimycin