BIOMAT Database Logo BIOMAT Database Logo

Ultrastructural localization of activity of phosphatases by low temperature incubation of unfixed cryostat sections. 1996

J Y Song, and W Tigchelaar, and J P Schellens, and J Van Marle, and C J Van Noorden, and W M Frederiks
Academic Medical Centre, University of Amsterdam, Amsterdam, The Netherlands.

In the present study, we demonstrate the activity of several phosphatases ultrastructurally in long-term (up to 24 months) cold-stored (-80 degrees C) rat tissues. Phosphatase activity was histochemically studied with the use of unfixed cryostat sections in combination with low temperature (4 degrees C) incubation conditions in order to prevent inactivation of enzyme activity and to limit the loss of ultrastructure. 5'-Nucleotidase activity was observed at plasma membranes, mainly at bile canalicular membranes of hepatocytes in liver. Thiamine pyrophosphatase activity was detected not only in trans side cisternae but also in medial and cis side cisternae of Golgi complexes in the parotid gland. Glucose-6-phosphatase activity was localized in endoplasmic reticulum as well as at the outer membrane of the nuclear envelope. Acid phosphatase reaction product was found in lysosomes. Furthermore, the localization patterns of 5'-nucleotidase and thiamine pyrophosphatase activity were compared with those obtained after different fixation procedures such as immediate chemical fixation of tissues or fixation of tissues after freezing and thawing. The results showed similar localization patterns of these enzymes after the different pretreatments. However, with respect to the ultrastructural morphology, some damage was observed in unfixed material after incubation. It can be concluded that the procedure described here enables ultrastructural localization of activity of phosphatases in long-term cold-stored tissues. This procedure will be useful for a retrospective study on archival material when histochemical parameters are needed.

UI MeSH Term Description Entries
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
D008854 Microscopy, Electron Microscopy using an electron beam, instead of light, to visualize the sample, thereby allowing much greater magnification. The interactions of ELECTRONS with specimens are used to provide information about the fine structure of that specimen. In TRANSMISSION ELECTRON MICROSCOPY the reactions of the electrons that are transmitted through the specimen are imaged. In SCANNING ELECTRON MICROSCOPY an electron beam falls at a non-normal angle on the specimen and the image is derived from the reactions occurring above the plane of the specimen. Electron Microscopy
D009685 Nuclear Envelope The membrane system of the CELL NUCLEUS that surrounds the nucleoplasm. It consists of two concentric membranes separated by the perinuclear space. The structures of the envelope where it opens to the cytoplasm are called the nuclear pores (NUCLEAR PORE). Nuclear Membrane,Envelope, Nuclear,Envelopes, Nuclear,Membrane, Nuclear,Membranes, Nuclear,Nuclear Envelopes,Nuclear Membranes
D010306 Parotid Gland The largest of the three pairs of SALIVARY GLANDS. They lie on the sides of the FACE immediately below and in front of the EAR. Gland, Parotid,Glands, Parotid,Parotid Glands
D005615 Freezing Liquids transforming into solids by the removal of heat. Melting
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
D000135 Acid Phosphatase An enzyme that catalyzes the conversion of an orthophosphoric monoester and water to an alcohol and orthophosphate. EC 3.1.3.2. Acid beta-Glycerophosphatase,Acid beta Glycerophosphatase
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia

Related Publications

J Y Song, and W Tigchelaar, and J P Schellens, and J Van Marle, and C J Van Noorden, and W M Frederiks
Localization and cytophotometric analysis of cathepsin B activity in unfixed and undecalcified cryostat sections of whole rat knee joints.
May 1989, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society,
J Y Song, and W Tigchelaar, and J P Schellens, and J Van Marle, and C J Van Noorden, and W M Frederiks
Preparation of unfixed undemineralized bone sections: the Bright bone cryostat.
January 1972, Journal of clinical pathology,
J Y Song, and W Tigchelaar, and J P Schellens, and J Van Marle, and C J Van Noorden, and W M Frederiks
A differential stain for neuronal nucleoli in unfixed cryostat sections.
July 1978, Stain technology,
J Y Song, and W Tigchelaar, and J P Schellens, and J Van Marle, and C J Van Noorden, and W M Frederiks
Improved localization of thiamine pyrophosphatase activity in mounted cryostat sections using gel fixation and gel incubation media.
July 1978, Histochemistry,
J Y Song, and W Tigchelaar, and J P Schellens, and J Van Marle, and C J Van Noorden, and W M Frederiks
Use of epoxy resin slides for handling unfixed cryostat sections intended for histochemistry at the ultrastructural level.
August 1983, The Histochemical journal,
J Y Song, and W Tigchelaar, and J P Schellens, and J Van Marle, and C J Van Noorden, and W M Frederiks
Localization of uric acid oxidase activity in core and matrix of peroxisomes as detected in unfixed cryostat sections of rat liver.
February 1994, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society,
J Y Song, and W Tigchelaar, and J P Schellens, and J Van Marle, and C J Van Noorden, and W M Frederiks
The ultra-structural localization of enzyme activity in unfixed sections.
July 1975, Histochemistry,
J Y Song, and W Tigchelaar, and J P Schellens, and J Van Marle, and C J Van Noorden, and W M Frederiks
Carbonic anhydrase localization in mounted cryostat sections.
March 1980, Stain technology,
J Y Song, and W Tigchelaar, and J P Schellens, and J Van Marle, and C J Van Noorden, and W M Frederiks
A gelatin embedding technique as an aid in the preparation of unfixed cryostat sections.
September 1978, The Histochemical journal,
J Y Song, and W Tigchelaar, and J P Schellens, and J Van Marle, and C J Van Noorden, and W M Frederiks
The use of unfixed cryostat sections for electron microscopic study of D-amino acid oxidase activity in rat liver.
December 1992, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society,
Copied contents to your clipboard!