Aldehyde Oxidase Activity Research Articles

Correlation between adult transformation and aldehyde oxidase staining pattern in wing discs ofApterous genotypes inDrosophila melanogaster.

The aldehyde oxidase staining pattern in wing discs ofDrosophila melanogaster bearing the genotypesap blt /ap blt andap blt andap blt /ap 73n showns changes from the wild-type pattern. Extensive areas of the presumptive dorsal posterior wing blade, which are normally unstained, have enzyme activity in these mutants. In wings of these genotypes, dorsal posterior structures are replaced by dorsal anterior wing structures. A strong correlation has been found between the frequencies of various staining patterns in the discs and the extent of transformation in the cuticular structures of the wing, which is consistent with the idea that aldehyde oxidase activity can be used as an indicator in the wing disc of this transformation. Unlike the homoeotic mutationengrailed, apterous has not been interpreted as a selector gene yet the work reported here shows thatapterous alleles can cause changes resembling those of theengrailed phenotype both in aldehyde oxidase staining behaviour and in the cuticular transformation.

Enzyme distribution patterns in the imaginal wing disc ofDrosophila melanogaster and other diptera : A subdivision of compartments into territories.

Analysis of the development of the aldehyde oxidase (AO) pattern in the wing pouch ofD. melanogaster showed that the extension of areas with AO activity occurs in steps. This indicates that the activation of this enzyme is regulated in groups of cells. It is proposed to use the term 'territory' for such a cell group. In the wing pouches ofD. melanogaster, D. simulans andMusca, corresponding parts of the disc become AO positive at comparable developmental stages. This indicates that AO becomes active in individual territories in a specific sequence.Borderlines of the distribution pattern of different enzymes in the wing pouch ofDrosophila and other dipteran species are in agreement with those found for the development of the AO pattern or are complementary to them. This indicates the existence of a common set of territories in the wing pouches of all higher diptera. Borderlines of patterns, as caused by different genetic constitution, are also in accord with this set of territories. The borderlines of some territories coincide with the compartmental A/P or D/V boundary. The results support the idea that both the location of compartmental boundaries and that of borderlines of enzyme territories are determined by a single mechanism.The distribution and the shape of the territories in the wing pouch is best explained by the reaction-diffusion model proposed by Meinhardt (1980), which involves three different gradients.

The effects of molybate, tungstate and lxd on aldehyde oxidase and xanthine dehydrogenase in Drosophila melanogaster.

The effects of dietary sodium molybdate and sodium tungstate on eye color and aldehyde oxidase and xanthine dehydrogenase activities have been determined in Drosophila melanogaster. Dietary sodium tungstate administration has been used as a screening procedure to identify two new lxd alleles. Tungstate administration results in increased frequencies of "brown-eyed" flies in lxd stocks and a coordinate decrease in AO and XDH activities in all genotypes tested. The two new lxd alleles affect AO and XDH in a qualitatively but not quantitatively similar fashion to the original lxd allele. AO and XDH activity and AO-CRM levels appear much more sensitive to mutational perturbations of this gene-enzyme than do XDH-CRM levels in the genotypes tested.

ALDEHYDE OXIDASE DISTRIBUTION IN PICTURE-WINGED HAWAIIAN DROSOPHILA: EVOLUTIONARY TRENDS.

Dipteran imaginal discs, especially those of Drosophila, are a unique system for the study of genetic control in determination and differentiation (Gehring, 1978). Although differentiation of adult structure is limited to metamorphosis, experiments utilizing clonal analysis have shown that groups of cells within imaginal discs become more and more restricted in their determination through the larval period. Morphologically, cells of the discs appear to be similar but Crick and Lawrence (1975) have hypothesized that they may differ from one another by enzyme profile which could be visualized histochemically. Major steps toward this visualization have been taken for the enzyme aldehyde oxidase (AO). A great deal is known of the biochemistry and genetic control of this enzyme in Drosophila. Much of this work is reviewed by Dickinson and Sullivan (1975). Dickinson (1971) was the first to histochemically localize AO in Drosophila melanogaster larvae and adults and find tissue-specific patterns of expression. Patterns of AO acitivity in Drosophila imaginal discs were first observed by Janning (1972, 1973) who also showed, using cell markers in gynandromorphs, that aldehyde oxidase is cell autonomous. Distribution of aldehyde oxidase in larval tissues and imaginal discs was surveyed by Kuhn and Cunningham (1978). These investigators were also able to show that in homoeotic mutants disc specific staining patterns are altered reflecting the change in determination (Kuhn and Cunningham, 1977a, 1977b). It is quite clear from these and other studies that tissue specificity of enzyme activity is a result of genetic regulation. The importance of considering regulatory as well as structural gene changes in evolution was emphasized by King and Wilson (1975). Sprey (1977) surveyed aldehyde oxidase distribution in imaginal discs of several dipterans including D. melanogaster, its sibling, D. simulans, and D. hydei. There is probably no better group of Drosophila in which to examine evolutionary trends in developmental regulation than the Hawaiian species. Precise relationships for 101 species of the picturewinged group have been determined based on banding sequences in the salivary gland polytene chromosomes (Carson et al., 1970; Carson and Kaneshiro, 1976). For these species a great deal is also known about their ecology, behavior, morphology and genetic variability. Dickinson (1979, 1980) has begun to examine these species for tissue-specific gene expression and has demonstrated cis-acting regulatory elements controlling expression of ADH and AO. We have chosen to survey ten species of picture-winged Hawaiian Drosophila, representing several branches of the phylogeny, examining them for larval tissuespecific expression of aldehyde oxidase and patterns of expression within the imaginal discs. This paper describes the AO activity patterns observed in relation to the questions: are patterns of expression species specific and do closely related species manifest similar distribution patterns?

Influence of homoeotic genes on the aldehyde oxidase pattern in imaginal discs of Drosophila melanogaster

AbstractIn a study of the regulation of enzyme patterns in imaginal discs the aldehyde oxidase pattern was determined for some homoeotic mutations of D. melanogaster. Earlier indications that suggested that this pattern follows the determinitive state of compartments within imaginal discs were confirmed by the aldehyde oxidase (AO) pattern of both the wing and haltere discs from en1; bx3, en1; pbx, and en1; bx3 pbx larvae and the antennal discs from Antp73b and ssa larvae.We additionally analyzed whether AO activity depended on the determinative state of an entire compartment or was expressed autonomously in clones. Homozygous engrailed clones were induced by mitotic recombination. From the AO clones found in normally negative areas of the posterior compartment it was concluded that enzyme activity depended upon the determinative state of the cells and was not a function of the compartment as a whole.The results are described with reference to a scheme in which compartmental and subcompartmental selector genes are thought to determine a binary code on which AO patterns depend.

Metabolism of 4-hydroxycyclophosphamide/aldophosphamide in vitro

The metabolism of 4-hydroxycyclophosphamide/aldophosphamide (HCP/AP), the primary metabolitc of cylophosphamide, was studied in vitro. Rabbit muscle glyceraldehyde-3-phosphate dehydrogenase and bovine-liver catalase did not metabolize HCP/AP. Neither did a rat hepatic 1000 g soluble fraction when conditions were made optimal for phosphamidase-catalyzed metabolism. Purified horse liver alcohol dehydrogenase converted HCP/AP to a metabolite tentatively identified as alcophosphamide. This metabolite was also found when HCP/AP was incubated with murine hepatic 105,000 g soluble fractions under conditions optimal for the expression of alcohol dehydrogenase activity. Murine hepatic 105,000 g soluble and solubilized 105,000g particulate fractions catalyzed the oxidation of HCP/AP to carboxyphosphamide when incubation conditions were made optima] for the expression of NAD-linked aldehyde dehydrogenase activity. Acrolein, disulfiram and butyraldehyde inhibited carboxyphosphamide formation in this system whereas pyridoxal, menadione and allopurinol did not. Oxidation of HCP/AP to carboxyphosphamide by murine hepatic 105,000 g soluble or solubilized 105,000 g particulate fractions was not observed in the absence of NAD or when incubation conditions were made optimal for the expression of aldehyde oxidase activity. NAD-linked aldehyde dehydrogenase activity was found in all of the normal and neoplastic tissues examined when butyraldehyde or benzaldehyde was used as the substrate. Aldehyde oxidase activity was found in some of the normal tissues but not in the neoplasms. NAD.linked aldehyde dehydrogenase activity greatly exceeded aldehyde oxidase activity in all tissues. These findings suggest that carboxyphosphamide formation from HCP/AP is predominantly catalyzed by NAD-linked aldehyde dehydrogenases.

THE CONTROL OF ALDEHYDE OXIDASE AND XANTHINE DEHYDROGENASE ACTIVITIES BY THE CINNAMON GENE IN DROSOPHILA MELANOGASTER

The isolation and characterization of 16 alleles of the cinnamon (cin, 1-0.0) locus in Drosophila melanogaster are described. The effects of cin on viability and the maternal effect of cin+ on eye color have been separated from each other as well as from the deficiency for aldehyde oxidase (AO) and xanthine dehydrogenase (XDH) activities. These 16 alleles have been assigned to four complementation groups based on analysis of AO and XDH activities in all heteroallelic female combinations. Zygotic complementation for lethality and eye color has been characterized and allows the ordering of cin alleles in a consistent pattern for the ability to produce viable zygotes and/or complement for the eye color phene. Several complementing cin combinations were analyzed for heat stability of AO. In all cases, AO from allelic heterozygotes was more heat labile than wild-type AO. One cin allele, cin13, produces heat labile AO in combination with cin+ from Oregon-R, hence exhibiting a "dominant" heat stability phenotype.

Notizen: A New Mutant Affecting Aldehyde Oxidase in Drosophila melanogaster

A new locus, Aldox-2, which affects the activity and heat stability of aldehyde oxidase in D. melanogaster is described. The Aldox-2 locus is localized to map position 86 on chromosome 2, between c and px. Aldehyde oxidase activity in aldox-2 homozygotes is approximately 25--30% that of the Oregon-R wild-type control strain. The enzyme from the mutant stock is much more heat labile than is the enzyme from the wild-type strain. Both the activity and heat phenotypes are completely recessive.

The development and function of the female germ line in Drosophila melanogaster: A cell lineage study

X-ray-induced mitotic recombination was used to follow the development and function of the female germ line in Drosophila melanogaster. Clones marked by maternal effect mutations which alter the morphology of the egg [ fs(1)K10] or the phenotype of the resulting progeny ( maroonlike) were produced in trans-heterozygotes irradiated during embryonic, larval, or pupal development or as 5-day-old adults. Judging from the size of clones induced at the blastoderm stage, only five to ten of the pole cells observed on the surface of the embryo contribute to the germ line. Most of the K10 clones induced during embryonic and larval development were associated with mal twin spots, indicating that both daughters of the irradiated germ cell remained in the germ line and gave rise to eggs in the adult. During larval life the number of cells increases logarithmically and reaches a maximum of 110 at 24 hr after pupation. The same value was obtained for 5-day-old adults. In contrast to the mosaic females produced as embryos and larvae, mosaics obtained after pupal and adult irradiations were of two types, those laying only one K10 egg and those laying several K10 eggs distributed over the lifespan of the adult. This result indicates that the stem cell divisions characteristic of the adult period have begun shortly after pupation. About 9 to 11 days are required for an irradiated stem cell to produce its first clonal K10 egg, and two-thirds of this time is spent in the germarium. Each ovariole possesses on the average two to three functioning stem cells. This multiplicity of stem cells was confirmed by the recovery of mosaic ovarioles when mal heterozygotes irradiated as adults or late larvae were stained for aldehyde oxidase activity.

Genetic control of aldehyde oxidase activity and cross-reacting-material in Drosophila melanogaster.

Four different genes are known to affect aldehyde oxidase activity (AO) in Drosophila melanogaster. Mutants at each of these loci eliminate AO activity and simultaneously eliminate detectable AO-crossing reacting material (AO-CRM) even though only one is the structural gene for AO (Aldoxn). The other three genes (cin1, lxd and mal) coordinately "control" the levels of activity of AO and two related enzymes, xanthine dehydrogenase (XDH) and pyridoxal oxidase (PO). Contrary to their effects on AO-CRM, neither of these three mutants eliminate XDH-CRM. A model of interaction of these enzymes and genes controlling their activities is discussed.

The effects of lao on aldehyde, oxidase activity and cross-reacting-material in Drosophila melanogaster.

In Drosophila melanogaster aldehyde oxidase occurs in at least two forms that can be separated electrophoretically. The mutant allele lao (low aldehyde oxidase activity) causes a deficiency of the major form of this enzyme. Immunoelectrophoretic analyses suggest that lao homozygotes produce aldehyde oxidase cross-reacting-material in nearly wild-type levels. Although aldehyde oxidase from the mutant stock is heat labile. properties such as Km and pH optima are not different from the normal enzyme.

Aldehyde oxidase distribution in Drosophila melanogaster mature imaginal discs, histoblasts and rings of imaginal cells

AbstractHistochemical staining characteristics for the enzyme aldehyde oxidase are reported for Drosophila melanogaster imaginal discs, histoblasts, and rings of imaginal cells. Aldehyde oxidase activity was observed in all discs except for the eye disc, was found in all imaginal cell groups except in the imaginal cells of the midgut and was found in the histoblasts. Differential aldehyde oxidase staining patterns were observed for the dorsal prothoracic disc, labial disc, antennal disc, for all leg discs (ventral pro‐, meso‐, and metathoracic discs), for the halere disc (dorsal metathoracic disc), for the wing disc (dorsal mesothoracic disc), and for the genital disc. Patterns observed in the various discs are quite repeatable. Enzyme distribution in the mature wing disc may provide an opportunity to visualize developmental compartments.

Genetic variation of some aldehyde‐oxidizing enzymes in the mouse

1. Twenty-six strains of mice were surveyed by starch gel electrophoresis for genetic variation of four liver enzymes; aldehyde dehydrogenase, aldehyde oxidase, xanthine oxidase and formaldehyde dehydrogenase. 2. A variant of aldehyde dehydrogenase was found in strains ICFW, IS/Cam, NZB, NZW, Simpson and Schneider. A variant of aldehyde oxidase was found in CE. A possible variant of xanthine oxidase was found in SF/Cam. 3. The gene determining the electrophoretic variant of aldehyde oxidase is either the same as, or very closely linked to, the Aox gene which determines aldehyde oxidase activity.

Iron deficiency in the rat: biochemical studies of brain metabolism.

Studies were performed to determine the effects of iron deficiency on brain metabolism in rats. Concentrations of cytochrome pigments, oxidative phosphorylation, and catalase and monoamine oxidase activities in brain tissue were unaffected by iron deficiency. However, activities of aldehyde oxidase, a key enzyme in the pathway of serotonin degradation, were significantly reduced, and concentrations of serotonin and total 5-hydroxyindole compounds were elevated in brain tissue of iron-deficient animals. Aldehyde oxidase activities and concentrations of 5-hydroxyindole compounds in brain tissues returned to approximately normal values one week after treatment of iron deficient animals with iron dextran.

Germ-line dependence of the maroon-like maternal effect in Drosophila

We have used pole cell transplantations to construct germ-line mosaics for maroon-like (mal), a maternal effect mutation in Drosophila. Such mosaics allow one to determine the cell type in which a gene is active. We find that the maroon-like maternal effect is (1) autonomous to the germ line and (2) dose sensitive in germ-line mosaics. Aldehyde oxidase activity is used as a histological probe to investigate the tissue and temporal distribution of mal + activity in the developing ovary. The adult ovary shows mal + activity in the germ line at all discernible stages of oogenesis but no activity is observed in the mesodermally derived follicle cells. Differential mal + activity is observed even in the ovary of the third-instar larvae.

Aldehyde oxidase distribution in the imaginal discs of some diptera

In the imaginal discs ofMusca domestica, Drosophila melanogaster, D. simulans, D. hydei, andZaprionus spec. the enzyme aldehyde oxidase (AO) appeared in a clear-cut pattern. In the leg and eye-antennal discs of these species this pattern shows a high degree of conformity, while that of the wing and haltere discs is species-specific.No aldehyde oxidase activity was detected in the imaginal discs ofCalliphora erythrocephala, Phormia regina orLucilia cuprina, but the discs of these species are characterized by grossly similar patterns of 5'-nucleotidase. Since the other species studied lack this enzyme, the two enzymes may perform similar functions in the morphogenesis of the discs.The coincidence of the sharp boundary of the AO pattern in the leg and wing discs ofD. melanogaster with the boundary between the anterior and posterior disc compartments gives a strong indication for the existence of analogous compartments in other discs showing a similar sharply bounded AO pattern. Compartmentalization may be considered a general phenomenon which occurs in discs of all segments and is not restricted toD. melanogaster. From the changes in the AO pattern during disc development it can be deduced that the localisation of this enzyme is regulated by supracellular determination involving positional information.

The effects of cinnamon on xanthine dehydrogenase, aldehyde oxidase, and pyridoxal oxidase activity during development in Drosophila melanogaster

The cinnamon (cin) eye color mutant of Drosophila melanogaster was characterized to determine biochemical correlations with another mutant, maroon-like. As with maroon-like, cinnamon flies lack three enzymatic activities: xanthine dehydrogenase, aldehyde oxidase, and pyridoxal oxidase. Xanthine dehydrogenase (XDH) is subject to a maternal effect in both mutants; i.e., mutant progeny of heterozygous mothers have XDH activity, resulting in wildtype eye color. However, the maternal effect is stronger in cinnamon than in maroon-like. Whereas maternally affected cinnamon show a large increase in XDH activity during larval stages, and XDH activity is still detectable after eclosion, the magnitude of increase in XDH activity is less in maroon-like, and activity is no longer detectable in second-day pupae and all later stages. The large increase in XDH activity in maternally affected cinnamon suggests that there is de novo synthesis of enzymatically active XDH during development in the absence of the cin + gene. Cinnamon is also unique in that maternally affected flies retain isoxanthopterin (IXP), the product of XDH activity. These flies appear to be deficient in some aspect of either pteridine metabolism or excretion.

Aldehyde oxidase activity in the tumorous-head strain of Drosophila melanogaster

Gross aldehyde oxidase activity from the egg-stage through 10-day-old adults and distribution of the enzyme in eye-antennal imaginal discs in third instar larvae were determined for the tumorous-head strain of Drosophila melanogaster. Aldehyde oxidase activity of several laboratory strains was measured for comparative purposes. Aldehyde oxidase activity was 100% higher during embryogenesis in tuh(ASU) eggs than in Oregon-R-C eggs. A second period of elevated aldehyde oxidase activity was observed during metamorphosis where tuh(ASU) pupae averaged 65% more enzyme activity than Oregon-R-C. Therefore, during determination and differentiation of the eye-antennal imaginal disc, the tuh(ASU) strain possesses a high aldehyde oxidase activity. Wild-type Drosophila melanogaster antennal imaginal discs are aldehyde oxidase positive, whereas attached eye imaginal discs are apparently aldehyde oxidase negative. A sample of eye-antennal imaginal discs from tuh(ASU) third instar larvae revealed that either one or both eye discs of 64% of the larvae were aldehyde oxidase positive. Aldehyde oxidase activity may be correlated with the homoeotic transformation in parts of the eye disc.

New pathway of conversion of pyridoxal to 4-pyridoxic acid.

The only enzyme demonstrated so far to catalyze the formation of 4-pyridoxic acid, the final excretory product of vitamin B6, was aldehyde oxidase. In this paper we have presented an evidence that another enzyme, NAD+-dependent aldehyde dehydrogenase, is capable of catalyzing this reaction. Rat mutants with high, low and no aldehyde oxidase activity excrete the same amount of isotope after a single injection of 3H-pyridoxol in a 12-day experiment; 4-pyridoxic acid is also present in the urine of animals without aldehyde oxidase activity. There is no correlation between the proportion of the excreted acid and the amount of pyridoxal excreted, after the injection of a single dose of the aldehyde form. The Km values for pyridoxal and NAD+, at pH 9.6, have been found to be 75 and 260 mumol/l, respectively. NAD+-dependent pyridoxal dehydrogenase was partially purified from the rat tissues, and the following specific enzyme activities (nmol-min-1-mg-1 protein) were found: red blood cell 71.5 +/- 3.0, intestine 64.9 +/- 9.0, muscle 61.4 +/- 1.6, brain 39.5 +/- 6.0, liver 34.4 +/- 3.3, kidney 21.1 +/- 5.6, heart 18.8 +/- 0.9, and lung 6.5 +/- 2.0. This is believed to be the first report demonstrating pyridoxal oxidation, catalyzed by an NAD+-dependent aldehyde dehydrogenase.

Azo- and nitro-reductases of the cestode Moniezia expansa. Substrate specificity, reaction products and the effects of flavins and other compounds.

1. The effects of various inhibitors and activators on the azo- and nitro-reductases of Moniezia expansa have been studied. Both reductions were partially inhibited by FAD, FMN, riboflavin, allopurinol, dicoumarol, 5-nitro-2-furaldehyde, azide and cyanide at 1 mM. Both reactions were stimulated by hypoxanthine. Menadione, nitrofurantoin, SKF 525-A (2-diethylaminoethyl 2,2-diphenylvalerate) and fluoride were without effect. 2. Xanthine- and aldehyde-oxidase activities were not detected in the enzyme preparation. 3. The substrate specificity of the azo- and nitro-reductases were determined. Azobenzene, 4-dimethylamino-azobenzene and 1,2-dimethyl-4-(4-carboxyphenylazo)-5-hydroxybenzene, nitrobenzene, 4-nitrohippuric acid and the isomers of nitrophenol, nitroanisole, nitrobenzoic acid, nitrobenzaldehyde and nitrobenzyl alcohol were reduced. Nitrobenzaldehyde isomers were not reduced to the alcohols and the coumaric acids were not reduced to the phenylpropionic acids. 4. The products of azo- and nitro-reduction were the corresponding amines; hydroxylamino- and hydrazo-compounds were not detected. 5. The pH optima and cofactor requirements were the same for both azo- and nitro-reduction. Neither reaction was inhibited by oxygen.