Suehiro Nakanishi

Intralobular distribution of rat liver aldehyde dehydrogenase and alcohol dehydrogenase

1. The activity of liver microsomal high Km-ALDH and mitochondrial low Km-ALDH, which may be primarily responsible for the oxidation of acetaldehyde after ethanol administration was found to be predominantly distributed in the centrilobular area. 2. The activities of other ALDH isozymes in mitochondrial and soluble fractions were evenly distributed in periportal and perivenous regions. 3. The activity of ADH which is involved in production of acetaldehyde was predominantly located in the periportal area. 4. From these results it seems unlikely that a concentration of acetaldehyde after ethanol ingestion is higher in perivenous hepatocytes than in periportal ones. Additional data would be needed to understand fully the mechanism by which ethanol induces predominantly centrilobular liver injury.

Prolongation of the action potential and reduction of the delayed outward K+ current by halothane in single frog atrial cells

Halothane has been shown to produce significant changes in heart rate and strength of contraction. We have studied the mechanism(s) of these effects by recording action potentials and transmembrane ionic currents in single cardiac cells from bullfrog atrium. Our results show that the action potential was prolonged and its plateau was depressed; and that there was a significant inhibition of a potassium current, Ik, and a Ca2+ current, Isi, following halothane (2%) application.

Subcellular aldehyde dehydrogenase activity and acetaldehyde oxidation by isolated intact mitochondria of rat brain and liver after acetaldehyde treatment

The effect of treatment of rats with acetaldehyde on the subcellular NAD+-aldehyde dehydrogenase (EC 1.2.1.3, ALDH) activities and acetaldehyde oxidation by isolated intact mitochondria of the liver and the brain was studied. Inhalation of acetaldehyde caused a significant decrease in the liver mitochondrial low Km-ALDH activity, while brain mitochondrial ALDH activity remained unchanged. Acetaldehyde oxidation by isolated intact liver mitochondria decreased significantly but that by brain mitochondria remained unchanged after acetaldehyde inhalation. These findings raise the possibility that the brain enzyme may be exposed to lower concentration of acetaldehyde than the liver enzyme.

Activity and electrophoretic profiles of brain aldehyde dehydrogenases in mice genetically selected for their ethanol preference.

The activities and electrophoretic profiles of NAD+-aldehyde dehydrogenases (EC 1.2.1.3, ALDHs) in the different brain regions from the mice of inbred C57BL/6J strain (alcohol preferring) and DBA/2 strain (alcohol avoiding) were studied. Kinetic studies on ALDH of the different brain regions revealed the existence of at least two ALDHs in both strains. The ALDH activity in the cerebellum, which was measured spectrophotometrically with 0.1 and 5 mM acetaldehyde as the substrates, was the highest of the three regions and it was about 2-fold higher than that in the cortex in both strains. The ALDH activities of mice of DBA/2 strain were higher than those of mice of C57BL/6J strain in all the regions. Only in the cortex the F1 hybrids (intermediate alcohol preferring) between C57BL/6J and DBA/2 strains had intermediate ALDH activity between their parental strains. Electrophoretic analysis of cortex ALDH revealed that strain differences were observed in the isozyme pattern between pH 7.2 and 7.8 with either acetaldehyde or propionaldehyde as substrate. These findings suggest that the ALDH activity in the cerebral cortex could be related to alcohol preference.

Genetic control of responsiveness of rat liver supernatant aldehyde dehydrogenase to phenobarbital and 3-methylcholanthrene

The responsiveness of the hepatic supernatant NAD+-dependent aldehyde dehydrogenase with a high Km value (high Km-AldDH) to phenobarbital (PB) and 3-methylcholanthrene (3-MC) treatment was studied in male rats of three strains; Wistar, Long-Evans, and Sprague-Dawley.A remarkable strain difference in the response of the enzyme to PB or 3-MC was observed. In rats of the Wistar strain the enzyme activity remained unchanged (“non-responsive”) in all rats after treatment with PB while it increased (“responsive”) 5- to 19-fold in all rats after treatment with 3-MC. The enzyme activity increased 8- to 20-fold and 2- to 8-fold respectively after treatment with PB and 3-MC in all rats of the Long-Evans strain. In rats of the Sprague-Dawley strain the enzyme activity remained unchanged in half of all the rats treated with PB or 3-MC and increased 2- to 7-fold over the basal level in half of the treated rats. The non-responsive rats to PB were all responsive to 3-MC treatment while the responsive rats to PB were responsive in 65% and non-responsive in 35% to 3-MC treatment.

Acetaldehyde level in the blood and liver aldehyde dehydrogenase activities in trichloroethylene-treated rats.

The liver NAD+-dependent aldehyde dehydrogenase (AldDH) activity and the acetaldehyde level in the blood during ethanol metabolism after trichloroethylene (trichlene) exposure were studied in rats. Trichlene inhalation caused large elevations in acetaldehyde levels during ethanol metabolism and caused decreases in the activity of the AldDH with a low Km value in mitochondrial and soluble fractions of liver cells. No significant effects were found in the activity of the high Km-enzyme in mitochondrial, soluble and microsomal fractions. Time course of inhibition of the mitochondrial low Km-enzyme and that of elevations in acetaldehyde levels during ethanol metabolism after trichlene exposure were similar. These findings suggest that acetaldehyde formed from ethanol in vivo is oxidized primarily by the mitochondrial low Km-enzyme.

Circadian rhythms in the activities of brain and liver aldehyde dehydrogenase isozymes in mice.

Circadian variations in the activities of aldehyde dehydrogenase (ALDH) isozymes in the subcellular fractions of the brain and liver were investigated in male and female mice of C57BL/6J strain. The rhythms in high Km-ALDH activities of brain and liver mitochondrial fractions which existed in ordinary light-dark cycle were not observed in animals maintained in the continuous darkness for two weeks. The rhythms in high Km-ALDH activities of hepatic soluble and microsomal fractions existed in both ordinary cycle and total darkness but the rhythmic phases were different. In the low Km-ALDH activity of hepatic mitochondrial fraction, the circadian rhythm was similar in two lighting conditions. There was sex difference in the existence of the circadian rhythm. It seems that the ALDH activity of mice is influenced by light-dark cycle and sex hormones.

Effect of chronic administration of acetaldehyde by inhalation on (Na+ + K+)-activated adenosine triphosphatase activity of rat brain membranes

The effect of chronic acetaldehyde inhalation on (Na+ + K+)-ATPase (EC 3.6.1.3) activities of subcellular fractions of the rat cerebral cortex was studied. Chronic administration of acetaldehyde by inhalation for 4-21 weeks caused significant increases in the enzyme activities of both the synaptosomal plasma membrane (SPM) fraction and the microsomal (MC) fraction. This indicates the change in neural membrane functions of the brain after acetaldehyde treatment. Mg2+-ATPase activities of both subcellular fractions remained unchanged after acetaldehyde treatment.

Effects of pargyline and diethyldithiocarbamate in vivo treatment on aldehyde dehydrogenase activities of submitochondrial fractions

Abstract1) Aldehyde dehydrogenase with a Km for acetaldehyde in the micromolar range (referred to as low Km-AldDH) is located in the matrix and that with a Km in the millimolar range (referred to as high Km-AldDH) is located in both the matrix and the outer membrane of mitochondria. 2) Pargyline in vivo treatment (100 mg/kg i.p.) caused a complete inhibition of the low Km-AldDH activity of the matrix fraction but caused only a 10% inhibition of the high Km-AldDH activities of the matrix and the outer membrane fractions. Diethyldithiocarbamate (DDC) treatment (700 mg/kg i.p.) caused less than 80% inhibition of the low Km-AldDH activity of the matrix fraction and caused a 20% inhibition of the high Km-AldDH activity of the matrix fraction. DDC treatment had no effect on the enzyme activity of the outer membrane fraction. The difference in the degree of inhibition of the low Km-AldDH activity of the matrix fraction by pargyline and DDC treatment may cause different blood acetaldehyde levels during ethanol oxidation.