Document Detail


In vitro methods for studying human placental amino acid transport placental villous fragments.
MedLine Citation:
PMID:  16511986     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
A method is described for measuring amino acid uptake by the syncytiotrophoblast of the human placenta in vitro using taurine as an example. Small fragments of placental villous tissue (2-3 mm3) are tied to a comb that enables them to be moved in concert between a series of incubation and wash buffers. The fragments are incubated in control or Na(+)-free Tyrode's buffer containing 3H taurine for timed intervals. Following incubation, the tissue is washed, lysed in water to release the accumulated isotope, and finally denatured in NaOH to determine protein. The water lysate is counted for radioactivity and the Na(+)-dependent component of 3H taurine uptake calculated as the difference between uptake in control and Na(+)-free conditions. Because the taurine transport protein (system beta) is Na(+)-dependent, the Na(+)-dependent component of 3H taurine uptake gives a measure of system beta activity in the microvillous membrane of the syncytiotrophoblast. The method described here in relation to taurine can be applied to other Na(+)-dependent amino acid transport systems. An advantage of the fragment model is that syncytiotrophoblast metabolism and signaling are retained. A disadvantage is that fragments contain many cell types and carrier-mediated amino acid uptake may be into several compartments including syncytiotrophoblast.
Authors:
Susan L Greenwood; Colin P Sibley
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Publication Detail:
Type:  In Vitro; Journal Article    
Journal Detail:
Title:  Methods in molecular medicine     Volume:  122     ISSN:  1543-1894     ISO Abbreviation:  Methods Mol. Med.     Publication Date:  2006  
Date Detail:
Created Date:  2006-03-03     Completed Date:  2006-04-25     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  101123138     Medline TA:  Methods Mol Med     Country:  United States    
Other Details:
Languages:  eng     Pagination:  253-64     Citation Subset:  IM    
Affiliation:
Academic Unit of Child Health, Human Development and Reproductive Healthcare Academic Group, University of Manchester, St. Mary's Hospital, United Kingdom.
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MeSH Terms
Descriptor/Qualifier:
Amino Acid Transport Systems
Amino Acids / metabolism*
Biological Assay / methods*
Chorionic Villi / metabolism
Female
Humans
Placenta / metabolism*
Pregnancy
Tissue and Organ Harvesting / methods*
Chemical
Reg. No./Substance:
0/Amino Acid Transport Systems; 0/Amino Acids

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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