Document Detail


Histochemical demonstration of phospholipase B (lysolecithinase) activity in rat tissues.
MedLine Citation:
PMID:  17121389     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
A method has been developed for the histochemical demonstration of phospholipase B (lysolecithinase) of rat tissues. The enzyme attacks lysolecithin with liberation of 1 mole of glycerylphosphorylcholine and 1 mole of fatty acid. The recommended procedure involves use of 6-10 micro frozen sections, fixed in cold calcium-formol and incubated at 37 degrees C in Tris buffered medium at pH 6.6 containing 2.2 X 10(-3) M lysolecithin and 1% cobalt acetate. The fatty acid liberated by enzymatic hydrolysis is trapped as a cobalt precipitate and is then converted to a black-brown precipitate by treatment with dilute ammonium sulfide in cold isotonic saline. Equivalent amounts of fatty acid and glycerylphosphorylcholine are recovered by extraction and analysis of the incubated sections and of the incubation medium, thus proving that lysolecithin hydrolysis occurs under the proposed reaction conditions. Staining is reduced by treating the sections with copper ions, mercury compounds, alcohols, acetone and by heating at 60 degrees C prior to incubation with substrate. Lowering of the pH of the incubation medium has similar effect. These findings are interpreted as evidence of the enzymatic nature of the reaction. Cells exhibiting a positive staining are found in the lamina propria of the intestinal villi and crypts, in the red pulp of the spleen and in the interstitial tissue of lung, liver and thymus. Similar elements are present in bone marrow smears and in leukocyte preparations obtained by peritoneal lavage. The morphologic and staining characteristics of these cells correspond to those of the eosinophilic leukocytes. Physical and chemical agents (x-irradiation, corticosteroids) which sharply decrease the number of eosinophils also reduce the number of cells shown histochemically to hydrolyze lysolecithin. A correspondent diminution of phospholipase B activity of homogenates of the same tissues can be shown in vitro. Differences in tissue distribution and chemical properties distinguish the phospholipase B from less specific esterases and lipases.
Authors:
A Ottolenghi; J P Pickett; W B Greene
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society     Volume:  14     ISSN:  0022-1554     ISO Abbreviation:  J. Histochem. Cytochem.     Publication Date:  1966 Dec 
Date Detail:
Created Date:  2006-11-23     Completed Date:  2006-12-22     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9815334     Medline TA:  J Histochem Cytochem     Country:  United States    
Other Details:
Languages:  eng     Pagination:  907-14     Citation Subset:  IM    
Affiliation:
Department of Physiology and Pharmacology, Duke University Medical Center, Durham, North Carolina, USA.
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MeSH Terms
Descriptor/Qualifier:
Acetone / pharmacology
Alcohols / pharmacology
Animals
Bone Marrow / drug effects,  enzymology*,  ultrastructure
Copper / pharmacology
Enzyme Activation
Fatty Acids / metabolism
Glycerylphosphorylcholine / metabolism
Hydrogen-Ion Concentration
Hydrolysis
Immunohistochemistry
Intestines / drug effects,  enzymology*,  ultrastructure
Leukocytes / drug effects,  enzymology,  ultrastructure
Lung / drug effects,  enzymology*,  ultrastructure
Lysophosphatidylcholines / metabolism
Lysophospholipase / antagonists & inhibitors,  metabolism*
Male
Mercury Compounds / pharmacology
Rats
Rats, Inbred Strains
Spleen / drug effects,  enzymology*,  ultrastructure
Sulfides / pharmacology
Temperature
Thymus Gland / drug effects,  enzymology*,  ultrastructure
Chemical
Reg. No./Substance:
0/Alcohols; 0/Fatty Acids; 0/Lysophosphatidylcholines; 0/Mercury Compounds; 0/Sulfides; 12135-76-1/ammonium sulfide; 563-24-6/Glycerylphosphorylcholine; 67-64-1/Acetone; 7440-50-8/Copper; EC 3.1.1.5/Lysophospholipase

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


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