scholarly journals Cytochemical localization of nicotinamide adenine dinucleotide phosphatase(NADPase) and nucleoside diphosphatase activities in the Golgi apparatus of the rat epididymis.

1989 ◽  
Vol 22 (4) ◽  
pp. 431-441
Author(s):  
MASAYA MATSUSHITA ◽  
TAKAYUKI AKAHOSHI ◽  
TAKUMA SAITO
Keyword(s):  
Golgi Apparatus ◽  
Nicotinamide Adenine Dinucleotide ◽  
Nicotinamide Adenine ◽  
Cytochemical Localization ◽  
Rat Epididymis ◽  
Nucleoside Diphosphatase

scholarly journals Coupling of uridine-5'-diphosphate (UDP) formation and nicotinamide adenine dinucleotide (NAD+) reduction for cytochemical localization of glycosyltransferases.

1983 ◽  
Vol 31 (10) ◽  
pp. 1175-1182 ◽  
Author(s):  
G R Matyas ◽  
D J Morré
Keyword(s):  
Golgi Apparatus ◽  
Rat Liver ◽  
Nicotinamide Adenine Dinucleotide ◽  
Nicotinamide Adenine ◽  
Cytochemical Localization ◽  
Permeabilized Cells ◽  
Exogenous Glucose ◽  
Dense Deposits ◽  
Donor And Acceptor ◽  
Reduced Nicotinamide Adenine Dinucleotide

A technique applicable to the cytochemical localization of glycosyltransferases through a series of coupled enzyme reactions is described. Uridine-5'-diphosphate (UDP) formed by glycosyltransferases is first phosphorylated to uridine-5'-triphosphate (UTP) by nucleoside 5'-diphosphate kinase. The UTP plus exogenous glucose-1-phosphate is converted into UDP-glucose by uridine-5'-diphosphoglucose pyrophosphorylase. UDP-glucose is then oxidized by uridine-5'-diphosphoglucose dehydrogenase to form UDP-glucuronic acid and reduced nicotinamide adenine dinucleotide (NADH). The NADH is utilized by membrane-located NADH-ferricyanide oxidoreductases in the presence of a copper salt to form electron-dense deposits of cupric ferrocyanide (Hatchett's brown). Using this technique, galactosyltransferase has been localized in cisternae (including the central midregions of the cisternae) of Golgi apparatus isolated from rat liver. Reactivity is absent from the cis-most cisternae and membrane elements. The reaction is dependent on UDP-galactose and inhibited by ethylene diaminetetraacetic acid and puromycin. the latter is a known inhibitor of galactosyltransferase of rat liver Golgi apparatus. The reaction is adaptable by varying the sugar nucleotide donor and acceptor to any glycosyltransferase utilizing UDP-sugars (except UDP-glucose). Presently it is restricted to isolated membrane fractions and permeabilized cells due to the need for accessibility of reagents and coupling enzymes.

scholarly journals CYTOCHEMICAL LOCALIZATION OF BRAIN NICOTINAMIDE ADENINE DINUCLEOTIDE PHOSPHATE (OXIDIZED)-DEPENDENT DEHYDROGENASES QUALITATIVE AND QUANTITATIVE DISTRIBUTIONS

1974 ◽  
Vol 22 (1) ◽  
pp. 7-19 ◽  
Author(s):  
K. L. SIMS ◽  
F. C. KAUFFMAN ◽  
E. C. JOHNSON ◽  
V. M. PICKEL
Keyword(s):  
Nervous System ◽  
Nicotinamide Adenine Dinucleotide ◽  
Molecular Layer ◽  
Nicotinamide Adenine Dinucleotide Phosphate ◽  
Neuronal Cell ◽  
Nicotinamide Adenine ◽  
Experimental Conditions ◽  
Cytochemical Localization ◽  
Neuronal Cell Bodies ◽  
Neuronal Processes

This study compares the histochemical and microchemical localizations of nicotinamide adenine dinucleotide phosphate (reduced) and nicotinamide adenine dinucleotide (reduced) diaphorases and four nicotinamide adenine dinucleotide phosphate (oxidized)-dependent enzymes (glucose 6-phosphate, 6-phosphogluconate, malate and isocitrate dehydrogenases) in areas of rat metencephalon and spinal cord. For the four nicotinamide adenine dinucleotide phosphate (NADP) enzymes, the pattern of localization following use of a modified tetrazolium procedure was compared with quantitative data obtained by microdissection from the same areas in adjacent sections. Optimal experimental conditions for reaction pH, temperature, substrate, cofactor and divalent cation concentrations were used for both the quantitative analysis following microdissection and the histochemical tetrazolium procedure. Consecutive sections were also examined for isocitrate dehydrogenase (nicotinamide adenine dinucleotide (oxidized)) and nicotinamide adenine dinucleotide (reduced) diaphorase activities in addition to seriatim thionine reference sections. Our results indicate that, within the central nervous system, certain characteristic qualitative differences exist in the distribution of the nicotinamide adenine dinucleotide phosphate (oxidized)- and nicotinamide adenine dinucleotide (oxidized)-dependent dehydrogenase enzymes. Nicotinamide adenine, dinucleotide enzymes are visualized predominantly in neuronal cell bodies or neuropil consisting primarily of neuronal processes; in adjacent sections, NADP enzyme activities are visualized almost exclusively in glial elements with two important exceptions. The first is the cerebellar molecular layer where the results from both micro- and histochemical techniques indicate high levels of the NADP enzymes relative to other dehydrogenases and high activity relative to the levels of these NADP enzymes in other nervous system areas. The second exception occurs in those neuronal groups known to contain high levels of catecholamines; these data are the subject of a companion report.

scholarly journals Ultrastructural localization of nicotinamide adenine dinucleotide phosphatase (NADPase) activity to the intermediate saccules of the Golgi apparatus in rat incisor ameloblasts.

1980 ◽  
Vol 28 (1) ◽  
pp. 16-26 ◽  
Author(s):  
C E Smith
Keyword(s):  
Golgi Apparatus ◽  
Nicotinamide Adenine Dinucleotide ◽  
Secretory Granules ◽  
Nicotinamide Adenine Dinucleotide Phosphate ◽  
Ultrastructural Localization ◽  
Nicotinamide Adenine ◽  
General Phenomenon ◽  
Rat Incisor ◽  
Selective Staining ◽  
Dense Bodies

Cytochemical evidence for the existence of a Golgi-associated phosphatase activity that hydrolyzes nicotinamide adenine dinucleotide phosphate (NADP) at acid pH in rat incisor ameloblasts was obtained by incubating sections from glutaraldehyde-fixed teeth in a medium containing NADP as substrate and lead ions as capture agent. Following incubation for 1 hr at 37 degrees C and pH 5.0, the Golgi saccules situated between those at the cis (immature) and trans (mature) faces of the ameloblast Golgi apparatus were marked by reaction product with the heaviest deposit in the middle saccule. Reaction product was otherwise seen in trace amounts only over some elements of the GERL system as well as a few lysosomal dense bodies and immature secretory granules. Control experiments established that the selective staining of intermediate Golgi saccules at pH 5.0 could only be duplicated by using substrates that resembled the complete NADP molecule, and not just the portion containing the adenosine 2'-monophosphate group. As well, no deposits of reaction product were seen within the Golgi saccules of ameloblasts incubated at pH 5.0 with nictoinamide adenine dinucleotide (NAD) as the substrate or that were incubated at pH 7.2 or pH 9.0 with NADP as the substrate. It was concluded that a specific, acid-NADPase activity is present in the intermediate Golgi saccules of secretory ameloblasts. Preliminary observations on other cells suggest that the localization of NADPase activity to Golgi saccules may constitute a general phenomenon.

scholarly journals Cytochemical localization of NADH oxidase in Candida albicans.

1977 ◽  
Vol 25 (3) ◽  
pp. 193-199 ◽  
Author(s):  
M Borgers ◽  
S De Nollin ◽  
F Thoné ◽  
H Van Belle
Keyword(s):  
Candida Albicans ◽  
Nicotinamide Adenine Dinucleotide ◽  
Oxidase Activity ◽  
Nadh Oxidase ◽  
Lethal Effect ◽  
Nicotinamide Adenine ◽  
Strong Increase ◽  
Cytochemical Localization ◽  
Prolonged Incubation ◽  
Reduced Nicotinamide Adenine Dinucleotide

The application of a recently published technique to localize reduced nicotinamide adenine dinucleotide oxidase activity is described in glutaraldehyde-fixed Candida albicans. The reaction product appears as a finely granular precipitate on the mitochondrial cristae and on the central vacuolar membrane, and, if present, on the vacuolar contents. Fixation should be kept to a minimum and prolonged incubation times up to 2 hr are necessary to show these reactive sites. The reaction appears to be strongly substrate-dependent and not affected by cyanide. Exposure of C. albicans cells to the antimycotic miconazole resulted in a strong increase in reduced nicotinamide, adenine dinucleotide and oxidase activity. The hypothesis is put forward that this enzyme, together with peroxidative and catalatic enzymes, may be implicated in the mechanism by which miconazole exerts its lethal effect on C. albicans.

Effect of Niacin on Mitochondria of Allium Cepa Root Cells

1967 ◽  
Vol 25 ◽  
pp. 78-79
Author(s):  
M. Arif Hayat
Keyword(s):  
Nicotinamide Adenine Dinucleotide ◽  
Hydrogen Transfer ◽  
Nicotinamide Adenine Dinucleotide Phosphate ◽  
Nicotinamide Adenine ◽  
Root Tips ◽  
Root Cells ◽  
Transfer Processes ◽  
Standard Procedures ◽  
A Cell ◽  
Critical Interest

Although it is recognized that niacin (pyridine-3-carboxylic acid), incorporated as the amide in nicotinamide adenine dinucleotide (NAD) or in nicotinamide adenine dinucleotide phosphate (NADP), is a cofactor in hydrogen transfer in numerous enzyme reactions in all organisms studied, virtually no information is available on the effect of this vitamin on a cell at the submicroscopic level. Since mitochondria act as sites for many hydrogen transfer processes, the possible response of mitochondria to niacin treatment is, therefore, of critical interest.Onion bulbs were placed on vials filled with double distilled water in the dark at 25°C. After two days the bulbs and newly developed root system were transferred to vials containing 0.1% niacin. Root tips were collected at ¼, ½, 1, 2, 4, and 8 hr. intervals after treatment. The tissues were fixed in glutaraldehyde-OsO4 as well as in 2% KMnO4 according to standard procedures. In both cases, the tissues were dehydrated in an acetone series and embedded in Reynolds' lead citrate for 3-10 minutes.

Nicotinamide Adenine Dinucleotide and the Metabolism of Ethanol and Acetaldehyde

1967 ◽  
Vol 28 (2) ◽  
pp. 213-224 ◽  
Author(s):  
E. Majchrowicz ◽  
B. L. Bercaw ◽  
W. M. Cole ◽  
D. H. Gregory
Keyword(s):  
Nicotinamide Adenine Dinucleotide ◽  
Nicotinamide Adenine
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