Summary
A method has been developed for electron microscopic histochemical demonstration of phospholipase B, (lecithinase B, E C 3.1.1.5, lysolecithin acyl hydrolase), which hydrolyzes α- and β-positions of phospholipids in mouse liver, kidney and adrenal tissues. Tissues either fixed in cold 1% paraformaldehyde or unfixed were cut into 40 μm frozen sections and were incubated at 37° C in a medium at pH 6.6 or 4.5 containing 2 μM lysolecithin and 0.25 mM CaCl2 for 20 min. The fatty acids liberated by enzymatic hydrolysis were trapped as calcium precipitate and were converted to lead precipitate by treatment with lead nitrate. The reaction products were observed by electron microscopy to be localized on the end of the smooth endoplasmic reticulum at pH 6.6 and in lysosomes and lipid droplets at pH 4.5. It is concluded that the products showed the localization of phospholipase B activity.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Commission on Enzymes of the International Union of Biochemistry (1961) Report of the commision on enzymes of the international union of biochemistry. Pergamon Press, London
Epstein B, Shapiro B (1959) Lecithinase and lysolecithinase of intestinal mucosa. Biochem J 71:615–619
Gomeri C (1945) The microtechnical demonstration of lipase activity. Proc Soc Exp Biol Med 58:362–363
Marples EA, Thompson RHS (1960) The distribution of phospholipase B in mammalian tissues. Biochem J 74:123–127
Morii S, Takiguchi N, Sugihara H, Murakami H (1972) Enzymic digestive techniques for simple lipid esters and phospholipids. Takeuchi T, Ogawa K, Fujita S (eds) Proc 4th Intern Cong Histochem Cytochem. Nakanishi Printing, Kyoto, pp 253–254
Murata F, Yokota S, Nagata T (1968) Electron microscopic demonstration of lipase in the pancreatic acinar cells of mice. Histochemie 13:215–222
Nagata T, Ohno S (1975) Electron microscopic demonstration of phosphatide acyl hydrolase. Acta Anat Nippon 50:61–61
Ottolenghi A, Pickett JP, Green WB (1967) Histochemical demonstration of phospholipase B (lysolecithinase) activity in rat tissues. J Histochem Cytochem 14:907–914
Reynolds ES (1963) The use of lead citrate at high pH as an electron opaque stain in electron microscopy. J Cell Biol 17:208–212
Takeuchi T (1956) Histochemical demonstration of lecithase A. Acta Pathol Jpn 6:13–18
Yamada K (1969) Studies on the incorporation of14C- or3H-labelled fatty acids,3H-labelled lyso-phosphatidylcholine and14C-labelled glycerol into phospolipids of rat liver slices. Sapporo Med J 35:78–90
Watson ML (1958) Staining of tissue sections for electron microscopy with heavy metals. J Biophys Biochem Cytol 4:475–478
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Nagata, T., Iwadare, N. Electron microscopic demonstration of phospholipase B activity in the liver and the kidney of the mouse. Histochemistry 80, 149–152 (1984). https://doi.org/10.1007/BF00679989
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00679989