20-hydroxyecdysone and juvenile hormone influence tyrosine hydroxylase activity in Drosophila females under normal and heat stress conditions.

The effects of exogenous 20-hydroxyecdysone (20E) and the juvenile hormone (JH) on the activity of the tyrosine hydroxylase (TH), the first and rate-limiting DA biosynthetic enzyme, has been studied in young females of wild type D. virilis and D. melanogaster under normal conditions and under heat stress (38 degrees C). Both 20E feeding of the flies and JH application led to a substantial rise in TH activity. A rise in JH and 20E levels was found not to prevent the response of TH to heat stress, but to change the intensity of its response to the stress exposure. Putative mechanisms of regulation of DA level by 20E and JH in Drosophila females are discussed.

Effects of juvenile hormone and 20-hydroxyecdysone on alkaline phosphatase activity in Drosophila under normal and heat stress conditions.

The effect of 20-hydroxyecdysone (20E) and the juvenile hormone (JH) on the activity of the alkaline phosphatase (ALP) has been studied in young females of wild-type Drosophila virilis and Drosophila melanogaster under normal conditions and under heat stress (38 degrees C). Both 20E feeding of the flies and JH application led to a substantial rise in ALP activity. ALP activity was also measured in young females of a JH-deficient strain of D. melanogaster, apterous(56f). A decrease in the enzyme activity was observed in the mutant females as compared to wild type. A rise in JH and 20E levels was found not to prevent the response of ALP to heat stress, but to change its stress-reactivity. Mechanisms of regulation of dopamine (DA) level by gonadotropins in Drosophila are discussed.

[Ecdysone 20-monooxygenase activity in Drosophila virilis strains varying in ecdysteroid response to heat stress].

The effect of various duration of heat stress (38 degrees C) on the activity of ecdysone 20-monooxygenase converting ecdysone into 20-hydroxyecdysone has been studied in D. virilis of wild type and mutant strain females, which differ by the mode of heat stress response of ecdysone and 20-hydroxyecdysone. We are the first to show that heat stress induces activity of ecdysone 20-monooxygenase in Drosophila females and enzyme activity correlates with the level of 20-hydroxyecdysone.

[The mechanism of the effect of apterous56f mutation on the reproductive function of Drosophila melanogaster].

The effects of L-dihydroxyphenylalanine (L-DOPA) and 20-hydroxyecdysone (20E) were studied with respect to the content of dopamine (DA), intensity of the juvenile hormone (JH) degradation, and fecundity of the wildtype flies (Canton S) and JH-deficient apterous56f mutants (in young females, carrying this mutation, the levels of DA and 20E production were strongly increased). Fly feeding with L-DOPA proved to increase the level of DA in a dose-dependent manner and reduce JH degradation in 2-day-old females of both strains. Feeding with 20E produced the same effect. Treating the wild-type flies with 2.5 mg L-DOPA caused a 24-h delay in beginning of oviposition and reduction in fecundity throughout the experiment. An L-DOPA dose of 1 mg caused no such changes. An experimental increase in 20E titer led to reduced fecundity of the wild-type flies, though no delay in oviposition was observed. In mutant flies, an increase in DA and 20E levels accelerated beginning of oviposition and increased fecundity of young females, though the latter parameter was reduced in mature individuals. Thus, an increase in endogenous DA and 20E characteristic of young apterous56f females is assumed to be a compensatory response that leads to a higher JH titer and induction of vitellogenesis.

Effects of dopamine on juvenile hormone metabolism and fitness in Drosophila virilis.

The effects of dopamine (DA) on juvenile hormone (JH) metabolism and fitness (estimated as fecundity and viability levels under heat stress (38 degrees C)) in Drosophila virilis have been studied. An increase of DA level obtained by feeding with DA reduced fitness of wild-type (wt) flies under stress, and decreased JH degradation in young wt females while increasing it in sexually mature wt females. A decrease in DA levels resulted from 3-iodo-tyrosine treatment and caused a decrease in JH degradation in sexually mature wt and heat sensitive (hs) mutant females (DA level in hs females is twice as high in wt females). A dramatic decrease in viability under stress and fecundity under normal conditions in wt, but not hs, females was observed. 3-iodo-tyrosine treatment also reduced the number of oocytes at stages 8-14, delayed oocyte transition to stage 10 and resulted in the accumulation of mature eggs in wt females. It delayed maturation of wt, but not hs, males as well, but did not affect their fertility. This advances our understanding of the regulation of JH metabolism by DA in Drosophila and suggests a crucial role for the basal DA level in fitness.

Juvenile hormone, 20-hydroxyecdysone and dopamine interaction in Drosophila virilis reproduction under normal and nutritional stress conditions.

To elucidate the role of the juvenile hormone (JH) in the control of Drosophila reproduction under stress, JH degradation, dopamine (DA) content and reproduction were studied upon 20E treatment in Drosophila virilis females of wild type (wt) and a mutant, with increased 20E level and decreased fertility, under normal and nutritional stress conditions. 20E treatment of wt flies for 7 days results in an increase of DA content in young females, but a decrease in mature females, a decrease of JH degradation in both young and mature females, an 1-day delay in onset of oviposition and a decrease of fecundity to the level typical of mutant flies. One day of 20E treatment in 7-day-old fed and starved flies results in a small decrease of JH degradation in the fed females and a great decrease in the starved ones. Fecundity decreases in the fed flies to the levels of the starved untreated flies in both wt and mutant strains. An oviposition arrest is observed in the treated and the untreated starved, but not in the treated fed, females of both strains. The data obtained suggest ecdysone control of JH metabolism mediated via DA.

Role of DOPA decarboxylase and N-acetyl transferase in regulation of dopamine content in Drosophila virilis under normal and heat stress conditions.

Activity levels of DOPA decarboxylase (DDC), an enzyme responsible for dopamine (DA) synthesis, and of N-acetyl transferase (NAT), an enzyme responsible for its degradation, were studied under normal and stress conditions in two lines of Drosophila virilis which are in contrast concerning their DA content under normal conditions and its alterations under stress. Interlinear differences in DDC activity were detected, and genetic analysis was carried out. It was found out that neither DDC nor NAT activity in D. virilis changed under stress. Based on the results of this study and data obtained earlier by the authors, a mechanism of DA content control under normal and stress conditions was proposed.

The role of juvenile hormone in the control of reproductive function in Drosophila virilis under nutritional stress.

To investigate the role of juvenile hormone (JH) in the control of Drosophila reproduction under stress, JH degradation and reproduction were studied under nutritional stress and JH treatment in Drosophila virilis females of wild type (wt) and a heat stress (hs) mutant: this mutant does not respond to heat stress by alterations in JH metabolism and has decreased JH level and fertility under normal conditions. One day of starvation results in a decrease of JH degradation, a delay in oocyte maturation, degradation of early vitellogenic egg chambers, accumulation of mature oocytes and a 24 h oviposition arrest in both wt and hs females. A fertility decrease was observed in both wt and hs females 24 h following the end of starvation. JH treatment leads to a decrease of JH degradation and an arrest of oviposition for 24 h in fed females. JH treatment prior to starvation seems to protect some oocytes from resorption: in JH-treated wt females, fertility increases rapidly following the end of starvation. The dynamics of JH degradation and fertility are similar following starvation and JH treatment. The role of JH in the accumulation of mature oocytes and the delay of oviposition under stress are discussed.

[Alteration in the content of biogenic amines in two stocks of Drosophila virilis and their hybrids in ontogenesis and under heat stress]. / Izmenenie soderzhaniia biogennykh aminov u dvukh linii Drosophila virilis i ikh gibridov v ontogeneze i pri teplovom stresse.

The levels of dopamine and serotonin in two stocks of D. virilis and their F1 hybrids throughout ontogenesis and under heat stress (for 60 min at 32 degrees C) were examined. Sharp increase in dopamine levels was reported under pupariation and with emergence of adults; alterations in serotonin levels were not so obvious. The concentration of dopamine in adults was shown to be considerably higher (more than twice) in the stock 147 than that in the stock 101. Significant increase of dopamine quantity in both stocks was observed under heat stress (serotonin remaining unchanged), while the level of response in the stock 101 was twice as high as in adults of the stock 147. The response typical of the stock 147 was prevailing in the hybrids F1 (147 x 101).

[Genetic analysis of differences in dopamine metabolism in two Drosophila virilis lines under normal conditions and under heat stress]. / Geneticheskii analiz razlichii v metabolizme dofamina u dvykh linii Drosophila virilis v norme i pri teplovom stresse.

The results of genetic analysis of the differences in dopamine (DA) content between two lines of Drosophila virilis in response to heat stress are presented. The gene(s) controlling the differences in DA content under normal conditions is located on the X-chromosome, and the gene(s) controlling the differences in responsiveness of DA system to stress was found to be autosomal. By means of polyacrylamide gel electrophoresis was also examined phenoloxidase activity in these lines under normal conditions and short-term heat stress (60 min, 38 degrees C). Of the five phenol oxidase fractions identified, three were major, A1, A2 and A3 components, and two appeared occasionally. It seems likely that A1 is monophenol oxidase and A2 is diphenol oxidase. It was shown that neither mono, nor diphenol oxidase activities change under heat stress. It was found that D. virilis lines 147 and 101 differ in diphenol oxidase activity under normal conditions. Genetic analysis of these differences revealed that they are controlled by a single gene (or a group of closely linked genes). The gene controlling diphenol oxidase activity in D. virilis is located on the X-chromosome.

[Genetic analysis of differences in metabolism of juvenile hormone in lines of Drosophila virilis resistant and sensitive to heat stress]. / Geneticheskii analiz razlichii v metabolizme iuvenil'nogo gormona u ustrichivoi i chuvstvitel'noi k teplovomu stressu linii Drosophila virilis.

The metabolism of juvenile hormone (JH) in imagos of two Drosophila species (D. virilis and D. melanogaster) was studied in the normal state and after short-term heat stress. Females of both species respond to stress by a decrease in the activity of JH esterase. This decrease is realized through a decrease in the amount of protein. JH metabolism in males does not change after heat stress. Genetic analysis of differences in the activity of DPF-resistant JH esterase in stress-sensitive and stress-resistant strains of D. virilis was performed. The differences are controlled by one autosomal gene.

[Tyrosine hydroxylase activity in Drosophila virilis lines with contrasting stress response and in their F1 hybrids]. / Aktivnost' tirozingidroksilazy u kontrastnykh po reaktsii na stress linii Drosophila virilis i ikh F1-gibridov.

Activity of tyrosine hydroxylase (TH) was examined in adult flies from stress lines of Drosophila virilis resistant and nonresistant to heat and from their hybrids under heat stress and normal conditions. The resistant line was shown to have higher enzyme activity and dominate for this character. Gene(s) controlling the level of TH activity under normal conditions were not sex-linked. It was shown than 30-min heat stress caused a decrease in TH activity; with 120-min stress, the activity increased. A possible mechanism of these changes is discussed. In the nonresistant line, the TH activity does not change under heat stress; the line dominates at this trait. Gene(s) controlling the level of the enzyme activity under stress are not sex-linked.

[Interaction of hormones in controlling reproductive function of Drosophila females under stressful conditions is genetically determined]. / Vzaimodeistvie gormonov v kontrole reproduktivnoi funktsii samok Drosophila v usloviiakh stressa geneticheski determinirovano.

The effect of heat stress (38 degrees C) on the content of octopamine (OA) and 20-hydroxyecdysone (20HE) was studied under normal and stressful conditions in adult flies of Drosophila virilis lines contrasting in the level of the juvenile hormone (JH). The wild-type flies (line 101) exhibited a pronounced sex dimorphism for the content of both OA and 20HE, which was substantially lower in this line than in flies of the mutant line 147. The level of both hormones increased in flies of line 101 exposed to heat stress, whereas it remained unchanged in flies of line 147 under the same conditions. The effect of heat stress on the level of JH metabolism and fertility was also studied in D. melanogaster wild-type lines and lines carrying mutations in genes responsible for OA and DA syntheses. In octopamineless females of the T beta hnM18 line and in females of the Ste line characterized by a doubled content of DA, JH degradation differed from normal: it was increased in both young and mature T beta hnM18 females, while decreased in young and increased in mature Ste flies. Fertility was substantially lower in the Ste than in the wild-type line. Flies of all of the D. melanogaster lines produced a stress response; however, in mutant lines, both fertility and stress reactivity of the systems controlling JH metabolism differed significantly from that of the wild-type lines. The role of JH, 20HE, OA, and DA interaction in regulation of Drosophila reproduction under stressful conditions is discussed.

Role of ecdysone 20-monooxygenase in regulation of 20-hydroxyecdysone levels by juvenile hormone and biogenic amines in Drosophila.

The effects of increased levels of dopamine (feeding flies with dopamine precursor, L: -dihydroxyphenylalanine) and octopamine (feeding flies with octopamine) on ecdysone 20-monooxygenase activity in young (2 days old) wild type females (the strain wt) of Drosophila virilis have been studied. L: -dihydroxyphenylalanine and octopamine feeding increases ecdysone 20-monooxygenase activity by a factor of 1.6 and 1.7, respectively. Ecdysone 20-monooxygenase activity in the young (1 day old) octopamineless females of the strain Tbetah ( nM18 ), in females of the strain P845 (precursor of Tbetah ( nM18 ) strain) and in wild type females (Canton S) of Drosophila melanogaster have been measured. The absence of octopamine leads to a considerable decrease in the enzyme activity. We have also studied the effects of juvenile hormone application on ecdysone 20-monooxygenase activity in 2-day-old wt females of D. virilis and demonstrated that an increase in juvenile hormone titre leads to an increase in the enzyme activity. We discuss the supposition that ecdysone 20-monooxygenase occupies a key position in the regulation of 20-hydroxyecdysone titre under the conditions that lead to changes in juvenile hormone titre and biogenic amine levels.

Role of arylalkylamine N-acetyltransferase in regulation of biogenic amines levels by gonadotropins in Drosophila.

The effect of 20-hydroxyecdysone (20E) and the juvenile hormone (JH) on the activity of the arylalkylamine N-acetyltransferase (AANAT) was studied in young females of wild-type D. virilis and D. melanogaster. 20E feeding of the flies led to a decrease in AANAT activity in both species when dopamine (DA) was used as substrate, but did not affect the enzyme activity when octopamine (OA) was used as substrate. JH application increased AANAT activity with DA as substrate in both species, but did not change it with OA as substrate. AANAT activity was also measured in young females of a JH-deficient strain of D. melanogaster, apterous ( 56f ). A decrease in the enzyme activity was observed in the mutant females as compared to wild-type. Mechanisms of regulation of DA level by gonadotropins in Drosophila are discussed.

Effects of octopamine on reproduction, juvenile hormone metabolism, dopamine, and 20-hydroxyecdysone contents in Drosophila.

The effect of an experimentally increased octopamine content (feeding flies with OA) on the levels of juvenile hormone (JH) degradation, dopamine (DA), and 20-hydroxyecdysone (20E) contents, oogenesis, and fecundity of wild type Drosophila flies has been studied. OA feeding of the flies was found to (1) cause a considerable decrease in JH degradation in females, but not males, of D. melanogaster and D. virilis; (2) have no effect on DA content in D. melanogaster and D. virilis; (3) increase 20E contents in D. virilis females; (4) decrease to a large extent the number of vitellogenic (stages 8-10) and mature (stage 14) oocytes in D. virilis; and (5) decrease the fecundity of D. melanogaster and D. virilis. A possible mechanism of action of OA as a neurohormone on the reproductive function of Drosophila is discussed.

Dopamine and octopamine regulate 20-hydroxyecdysone level in vivo in Drosophila.

The effects of increased level of dopamine (DA) (feeding flies with DA precursor, L-dihydroxyphenylalanine, L-DOPA) on the level of 20-hydroxyecdysone (20E) and on juvenile hormone (JH) metabolism in young (2-day-old) wild type females (the strain wt) of Drosophila virilis have been studied. Feeding the flies with L-DOPA increased DA content by a factor of 2.5, and led to a considerable increase in 20E level and a decrease of JH degradation (an increase in JH level). We have also measured the levels of 20E in the young (1-day-old) octopamineless females of the strain Tbetah(nM18) and in wild type females, Canton S, of D. melanogaster. The absence of OA led to a considerable decrease in 20E level (earlier it was shown that in the Tbetah(nM18) females, JH degradation was sharply increased). We have studied the effects of JH application on 20E level in 2-day-old wt females of D. virilis and demonstrated that an increase in JH titre results in a steep increase of 20E level. The supposition that biogenic amines act as intermediary between JH and 20E in the control of Drosophila reproduction is discussed.

Stress response in Drosophila melanogaster strain inactive with decreased tyramine and octopamine contents.

Juvenile hormone hydrolysis, tyrosine decarboxylase activity, dopamine content and fitness (viability and fertility) were studied under normal and stress conditions in the adults of Drosophila melanogaster inactive strain carrying a mutation that decreases tyrosine decarboxylase activity and results in the lower contents of tyramine and octopamine. A sexual dimorphism of tyrosine decarboxylase activity, dopamine level and survival under heat stress in inactive flies was found. inactive adults showed higher dopamine levels and lower survival levels under heat stress as compared to wild type (Canton S) adults. Juvenile hormone degradation is decreased in young and increased in mature inactive females as compared to wild type. Fertility of the inactive strain did not differ from that of wild type strain under normal conditions, but after heat exposure the dynamics of its restoration was different. inactive females were found to develop the stress reaction, with juvenile hormone degradation, tyrosine decarboxylase activity, dopamine content and fertility levels used as the reaction indicators.

The effect of mutations altering biogenic amine metabolism in Drosophila on viability and the response to environmental stresses.

Dopamine (DA) content, tyrosine decarboxylase (TDC) activity and survival were studied under normal and environmental stress conditions in the ste and e strains carrying ebony mutation increasing DA level and the octopamineless strain Tbetah(nM18) of Drosophila melanogaster. Wild-type strains Canton S and Oregon R, and strain p845 from which Tbetah(nM18) strain was derived were used as controls. Sexual dimorphism of TDC activity, DA content, and survival in flies of all D. melanogaster strains under study was found. Tbetah(nM18) mutation sharply reduced TDC activity in females, while ebony had no such effect. DA content and survival under heat stress in Tbetah(nM18) flies did not differ from those in the wild type. ste and e flies had drastically increased DA content under normal conditions, dramatically decreased survival under heat stress, but increased survival under starvation. DA content and survival under heat stress were also studied in the reciprocal hybrids (males) F(1) of the cross D. virilis strains 101 (wild type) and 147 with X-linked mutation, which significantly increases DA content. 147x101 males had a considerably higher DA content and lower survival than 101x147 ones. Individuals of all D. melanogaster strains under study developed the stress reaction, as judged by changes in TDC activity and DA levels. The role of biogenic amines in the stress reaction development and adaptation to environmental stresses in Drosophila is discussed. Arch. Insect Biochem. Physiol. 55:55-67, 2004.