Ultrastructural localization of nicotinamide adenine dinucleotide phosphatase (NADPase), thiamine pyrophosphatase (TPPase), and cytidine monophosphatase (CMPase) in the Golgi apparatus of early spermatids of the rat

1981 ◽  
Vol 201 (4) ◽  
pp. 613-622 ◽  
Author(s):  
Y. Clermont ◽  
M. Lalli ◽  
A. Rambourg
Keyword(s):  
Golgi Apparatus ◽  
Nicotinamide Adenine Dinucleotide ◽  
Ultrastructural Localization ◽  
Nicotinamide Adenine ◽  
Thiamine Pyrophosphatase

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 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 The use of ferricyanide for the electron microscopic demonstration of dehydrogenases in human steroidogenic cells.

1976 ◽  
Vol 24 (11) ◽  
pp. 1194-1203 ◽  
Author(s):  
L Benkoël ◽  
A Chamlian ◽  
E Barrat ◽  
P Laffargue
Keyword(s):  
Nicotinamide Adenine Dinucleotide ◽  
Smooth Endoplasmic Reticulum ◽  
Ultrastructural Localization ◽  
Nicotinamide Adenine ◽  
Reduced Form ◽  
Electron Microscopic ◽  
Ferricyanide Reductase ◽  
Steroidogenic Cells ◽  
Electron Carriers ◽  
Electron Microscopic Demonstration

The ultrastructural localization of 3 beta hydroxysteroid ferricyanide reductase, glucose-6-phosphate ferricyanide reductase and nicotinamide adenine dinucleotide and reduced form-ferricyanide reductase was investigated in some human steroidogenic tissues (corpus luteum of pregnancy, fetal adrenal gland and testis, adult testis and placenta) using ferricyanide as an electron acceptor. Copper ferrocyanide deposits were readily observed in the mitochondria, in the smooth endoplasmic reticulum profiles and in the cytoplasm. The sites of the various dehydrogenase activities could be visualized by using appropriate incubating media. The precise localization of various reactions in different electron transfer chains was determined by using different ferricyanide concentrations and intermediate electron-carriers such as menadione or exogenous nicotinamide adenine dinucleotide and reduced form-diaphorase. The use of respiratory chain inhibitors such as rotenone or antimycine A confirmed these data.

scholarly journals Ultrastructural localization of nicotinamide adenine dinucleotide phosphatase (NADPase) activity within columnar, goblet, and Paneth cells of rat small intestine.

1984 ◽  
Vol 32 (9) ◽  
pp. 989-997 ◽  
Author(s):  
S M Parsons ◽  
C E Smith
Keyword(s):  
Small Intestine ◽  
Epithelial Cells ◽  
Nicotinamide Adenine Dinucleotide ◽  
Ultrastructural Localization ◽  
Paneth Cells ◽  
Nicotinamide Adenine ◽  
Intestinal Epithelial ◽  
Rat Small Intestine ◽  
Golgi Stack ◽  
Cis And Trans

The distribution of nicotinamide adenine dinucleotide phosphatase (NADPase) activity was examined in epithelial cells of rat small intestine. Segments of ileum were fixed with glutaraldehyde and tissue chopper sections were incubated for up to 4 hr at pH 5.0 in cytochemical media prepared with NADP as substrate. NADPase activity was found primarily within the Golgi saccules of columnar, goblet, and Paneth cells. Columnar and goblet cells showed most of the NADPase activity within the saccules which were intermediate between the cis and trans faces of the Golgi stack. Paneth cells, however, showed the heaviest staining within saccules between the intermediate and innermost saccule at the trans aspect of the Golgi stack. Both columnar cells and Paneth cells also contained spotty, and sometimes heavy, deposits of reaction product within an occasional focal area of the GERL system and within an occasional lysosome. Control experiments indicated that the Golgi-associated NADPase activity was enhanced if cells were pretreated for about 12 hr with EGTA prior to incubation. No similar enhancement was apparent if the tissues were pretreated with DMSO. Furthermore, NADPase activity within the Golgi saccules could be inhibited completely by incubating intestinal epithelial cells with NADP in the presence of sodium fluoride or L(+)-tartrate.

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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