Biomaterial Database

Ultrastructural demonstration of phosphatases by perfusion fixation followed by perfusion incubation of rat liver. 1975

H Glaumann

An alternative to previous methods (tissue chopper, frozen sections) for the ultrastructural demonstration of phosphatases is described. The present approach is based on a short vascular perfusion of rat liver with glutaraldehyde through the inferior caval vein, followed by vascular perfusion incubation with a medium containing the enzyme substrates. The effect of glutaraldehyde on three different types of phosphatases was investigated, namely a lysosomal enzyme (acid phosphatase) a tightly bound microsomal enzyme (G6Pase) and a loosely bound microsomal enzyme (IDPase). It is demonstrated that by perfusion with glutaraldehyde for three minutes good cellular morphology is obtained and that 50-60% of the initial activity of glucose-6-phosphatase, inosine-diphosphatase and acid phosphatase remains. The localization and deposition of G6Pase activity were distinct and observed throughout the endoplasmic reticulum and the nuclear envelope. For acid phosphatase, the reaction product was confined to various types of lysosomes including presumed autophagic vacuoles. No signs of enzyme diffusion were noted. The present approach seems to offer some advantages: it is simple and requires no extra equipment, penetration of the fixative and incubation enzyme medium is good, and finally freeze artifacts are avoided.

UI	MeSH Term	Description	Entries
D007292	Inosine Nucleotides		Inosine Phosphates,Nucleotides, Inosine,Phosphates, Inosine
D008099	Liver	A large lobed glandular organ in the abdomen of vertebrates that is responsible for detoxification, metabolism, synthesis and storage of various substances.	Livers
D008247	Lysosomes	A class of morphologically heterogeneous cytoplasmic particles in animal and plant tissues characterized by their content of hydrolytic enzymes and the structure-linked latency of these enzymes. The intracellular functions of lysosomes depend on their lytic potential. The single unit membrane of the lysosome acts as a barrier between the enzymes enclosed in the lysosome and the external substrate. The activity of the enzymes contained in lysosomes is limited or nil unless the vesicle in which they are enclosed is ruptured or undergoes MEMBRANE FUSION. (From Rieger et al., Glossary of Genetics: Classical and Molecular, 5th ed).	Autolysosome,Autolysosomes,Lysosome
D008297	Male		Males
D008862	Microsomes, Liver	Closed vesicles of fragmented endoplasmic reticulum created when liver cells or tissue are disrupted by homogenization. They may be smooth or rough.	Liver Microsomes,Liver Microsome,Microsome, Liver
D010477	Perfusion	Treatment process involving the injection of fluid into an organ or tissue.	Perfusions
D010744	Phosphoric Monoester Hydrolases	A group of hydrolases which catalyze the hydrolysis of monophosphoric esters with the production of one mole of orthophosphate.	Phosphatase,Phosphatases,Phosphohydrolase,Phosphohydrolases,Phosphomonoesterase,Phosphomonoesterases,Phosphoric Monoester Hydrolase,Hydrolase, Phosphoric Monoester,Hydrolases, Phosphoric Monoester,Monoester Hydrolase, Phosphoric
D002467	Cell Nucleus	Within a eukaryotic cell, a membrane-limited body which contains chromosomes and one or more nucleoli (CELL NUCLEOLUS). The nuclear membrane consists of a double unit-type membrane which is perforated by a number of pores; the outermost membrane is continuous with the ENDOPLASMIC RETICULUM. A cell may contain more than one nucleus. (From Singleton & Sainsbury, Dictionary of Microbiology and Molecular Biology, 2d ed)	Cell Nuclei,Nuclei, Cell,Nucleus, Cell
D004721	Endoplasmic Reticulum	A system of cisternae in the CYTOPLASM of many cells. In places the endoplasmic reticulum is continuous with the plasma membrane (CELL MEMBRANE) or outer membrane of the nuclear envelope. If the outer surfaces of the endoplasmic reticulum membranes are coated with ribosomes, the endoplasmic reticulum is said to be rough-surfaced (ENDOPLASMIC RETICULUM, ROUGH); otherwise it is said to be smooth-surfaced (ENDOPLASMIC RETICULUM, SMOOTH). (King & Stansfield, A Dictionary of Genetics, 4th ed)	Ergastoplasm,Reticulum, Endoplasmic
D005952	Glucose-6-Phosphatase	An enzyme that catalyzes the conversion of D-glucose 6-phosphate and water to D-glucose and orthophosphate. EC 3.1.3.9.	Glucosephosphatase,Glucose 6-Phosphatase,Glucose-6-Phosphate Phosphohydrolase,Glucose 6 Phosphatase