Biogenic amines regulate the reproductive function in Drosophila as neurohormones

We studied the influence of experimental increase in the octopamine and dopamine content on the level of juvenile hormone degradation, oogenesis, and fertility in wild type D. virilis flies. Feeding of flies on octopamine led to a significantly decreased level of juvenile hormone degradation (increased titer) in young and sexually mature females, rather than in males, markedly decreased the number of vitellogenic (stages 8-10) and mature (stage 14) oocytes), and sharply reduced fertility. Feeding of flies on dopamine decreased the juvenile hormone degradation (increased titer) in young wild type females and increased it (lowered the juvenile hormone titer) in sexually mature females, as well as decreased the fertility of wild type females to a level characteristic for D. virilis line with a mutation doubling the endogenous dopamine level. A possible mechanism of the influence of these amines on the reproductive function in Drosophila as neurohormones is discussed and a conclusion is drawn that the reduced fertility of females at an increased level of amines appears to be related to an increased level of ecdysteroids, which is caused by an increased, as a result of decreased degradation, juvenile hormone titer.     

Juvenile Hormone Controls Oviposition and Fertility in Drosophila virilis during Starvation

Degradation of juvenile hormone and reproductive function during starvation and experimental increase of the juvenile hormone titer were studied in wild type and mutant D. virilis females incapable to respond to heat stress by changes in juvenile hormone metabolism and fertility. After 24-hour starvation, the females of both lines were characterized by a decreased level of juvenile hormone degradation, 24-hour delay of oviposition, increased oviposition within 3 h after the termination of starvation, and decreased fertility within three days. Application of exogenous juvenile hormone also led to a decreased level of its degradation and 24-hour arrest of oviposition. Experimental increase of the juvenile hormone titer before the beginning of starvation led to a sharply increase fertility (number of laid eggs and number of progenies) within the first 24 h after the termination of starvation. The dynamics of juvenile hormone degradation and of fertility were similar after starvation and upon application of the exogenous hormone. The role of juvenile hormone in the control of egg maturation and laying under stress conditions has been 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 Effects of Stress-Related Hormones on the Stress Resistance in <Emphasis Type="Italic">Drosophila melanogaster</Emphasis> Carrying Mutation in the <Emphasis Type="Italic">Dilp6</Emphasis> Gene

The heat stress resistance of Drosophila melanogaster females carrying a hypomorphic mutation of the DILP6 insulin-like protein gene (dilp6 ⁴¹ ) under a change in the level of stress-related hormones (juvenile hormone and octopamine) is studied. It is revealed that the dilp6 ⁴¹ mutation decreases the heat stress resistance of mature D. melanogaster females. An experimental decrease in the level of juvenile hormone is shown to restore the stress resistance of mutant females to the level of stress resistance observed in wild type Canton S females. These data suggest that the effects of the dilp6 ⁴¹ mutation on the stress resistance of females are mediated by an increased level of juvenile hormone. An experimental increase in the octopamine level that causes an increase in juvenile hormone level supports this hypothesis: the resistance to heat stress decreases in females of both lines and this decrease is more significant in mutant females than in the control line. Thus, it is established for the first time that the effect of the hypomorphic dilp6 gene mutation on the heat stress resistance of D. melanogaster females is mediated by juvenile hormone.     

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 Tβh ^nM18 , in females of the strain P845 (precursor of Tβh ^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.     

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 °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.     

Gene <Emphasis Type="Italic">dilp6</Emphasis> regulates octopamine metabolism in <Emphasis Type="Italic">Drosophila melanogaster</Emphasis>

The effect of strong hypomorphic mutation of the insulin-like protein gene (dilp6) on metabolism of octopamine (one of the main biogenic amines in insects) was studied in Drosophila melanogaster males and females. The activity of tyrosine decarboxylase (the key enzyme of octopamine synthesis) and the activity of octopamine-dependent N-acetyltransferase (the enzyme of its degradation) were measured. It was demonstrated that the activity of both studied enzymes is decreased under normal conditions in the dilp6⁴¹ mutants (as we previously demonstrated, this is correlated with an increased level of octopamine). It was also found that hypomorphic mutation of the dilp6 gene decreases the intensity of tyrosine decarboxylase response to heat stress. Thus, it was demonstrated for the first time that insulin-like DILP6 protein in drosophila influences the level of octopamine (regulating the activity of the enzyme degrading octopamine).     

The <Emphasis Type="Italic">ecdysoneless</Emphasis><Superscript>1</Superscript> Gene Regulates Metabolism of the Juvenile Hormone and Dopamine in <Emphasis Type="Italic">Drosophila melanogaster</Emphasis>

The dopamine (DA) content and the level of juvenile hormone (JH) degradation were studied in females of the wild-type Canton S strain and the ecdysoneless ¹ (ecd ¹) mutant, which does not produce ecdysone at a restrictive temperature (29°C). Exposure at the restrictive temperature considerably increased the JH-hydrolyzing activity and the DA content in five-day ecd ¹ females compared with flies of both strains growing at 19°C and Canton S females exposed at 29°C. In one-day ecd ¹ females, the level of JH degradation also increased at the restrictive temperature, but the DA content was low. The effect of ecdysone deficiency on the stress reaction in Drosophila melanogaster females was studied using changes in DA content and JH degradation as the reaction indicators. The ecd ¹ mutation did not prevent the initiation of the stress reaction in females exposed at the restrictive temperature, but changed its intensity (stress reactivity). The interaction of 20-hydroxyecdysone with JH and DA in regulating Drosophila reproduction under normal conditions and in stress is discussed.     

Juvenile hormone controls egg laying and fertility in Drosophila virilis during starvation

Degradation of juvenile hormone and reproductive function during starvation and experimental increase of the juvenile hormone titer were studied in wild type and mutant D. virilis females incapable to respond to heat stress by changes in juvenile hormone metabolism and fertility. After 24-hour starvation, the females of both lines were characterized by a decreased level of juvenile hormone degradation, 24-hour delay of egg laying, increased egg laying within 3 h after the termination of starvation, and decreased fertility within three days. Application of exogenous juvenile hormone also led to a decreased level of its degradation and 24-hour arrest of egg laying. Experimental increase of the juvenile hormone titer before the beginning of starvation led to a sharply increased fertility (number of laid eggs and number of progenies) within the first 24 h after the termination of starvation. The dynamics of juvenile hormone degradation and of fertility were similar after starvation and upon application of the exogenous hormone. The role of juvenile hormone in the control of egg maturation and laying under stress conditions has been discussed.     

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 arylalkylamine <Emphasis Type="Italic">N</Emphasis>-acetyltransferase in regulation of biogenic amines levels by gonadotropins in <Emphasis Type="Italic">Drosophila</Emphasis>

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.     

Effect of Gonadotropic Hormones on Stress Resistance of <Emphasis Type="Italic">Drosophila melanogaster</Emphasis> Females Infected with Different <Emphasis Type="Italic">Wolbachia pipientis</Emphasis> Genotypes

The effect of gonadotropic hormones (juvenile (JH) and 20-hydroxyecdysone (20E)) on heat stress resistance was for the first time studied in wild type D. melanogaster line females infected with different genotypes of the Wolbachia pipientis alpha-proteobacterium. It was found that an experimental increase in JH level induces a decrease in the heat stress resistance, while an increase in 20E level induces its increase in sixday females both uninfected with the Wolbachia and infected with different bacterium strains (wMelCS, wMelPop, and wMel). However, the intensity of response differs: a decrease in the survival with an increase in JH level and its increase with an increase in 20E level are more pronounced in females infected with pathogenic wMelPop strain and less pronounced in females infected with the wMelCS genotype than in uninfected females and females infected with the wMel genotype. Data obtained suggest that the wMelCS genotype induces a decrease and wMelPop induces an increase in the level of stress hormone (dopamine), since previously we demonstrated that an increase in the JH level in mature females increases the dopamine level, an increase in the 20E level decreases it, and an increase/decrease in the dopamine level, in turn, leads to a decrease/increase of the Drosophila female resistance to heat stress.     

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.     

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.     

Interaction of hormones in controlling reproductive function of Drosophila females under stressful conditions is genetically determined

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.     

Regulation of Reproductive Function in DrosophilaFemales Due to Hormonal Interaction Under Stress Is Genetically Determined

The effect of heat stress (38°C) on the content of octopamine (OA) and 20-hydroxyecdysone (20HE) was studied under normal and stressful conditions in adult flies of Drosophila virilislines 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. melanogasterwild-type lines and lines carrying mutations in genes responsible for OA and DA syntheses. In octopamineless females of the Th ^nM18line and in females of the Steline characterized by a doubled content of DA, JH degradation differed from normal: it was increased in both young and mature Th ^nM18females, while decreased in young and increased in mature Steflies. Fertility was substantially lower in the Stethan in the wild-type line. Flies of all of the D. melanogasterlines 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 Drosophilareproduction under stressful conditions is discussed.     

The mechanism of the effect of apterous 56f 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 wild-type flies (Canton S) and JH-deficient apterous ^56f 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 apterous ^56f females is assumed to be a compensatory response that leads to a higher JH titer in order to induce vitellogenesis.     

Alterations in biogenic amines levels associated with age-related muscular tissue impairment in Drosophila melanogaster

While holding on youth may be a universal wish, aging is a natural process associated with physical and physiological impairment in living organisms. Drosophila provides useful insights into aging-related events. Hence, this study was conducted to investigate the age-related changes in muscle function and architecture in relation to the biogenic amine titers. To achieve this aim, visceral and skeletal muscles performance was tested in newly-eclosed, sexually mature and old adult flies using climbing and gut motility assays. In addition, age-related ultrastructural alterations of muscular tissue were observed using transmission electron microscopy (TEM). The titer of selected biogenic amines was measured using high-performance liquid chromatography (HPLC). The results demonstrated that old flies were dramatically slower in upward movement than either newly-eclosed or sexually mature flies. Similarly, gut contraction rate was significantly lower in old flies than the sexually mature, although it was markedly higher than that in the newly-eclosed flies. In TEM examination, there were several ultrastructural changes in the midgut epithelium, legs and thorax muscles of old flies. Regarding biogenic amine titers, the old flies had significantly lower concentrations of octopamine, dopamine and serotonin than the sexually mature. We concluded that aging has adverse effects on muscular system function and ultrastructure, synchronized with biogenic amine titers changes. Our results highlighted the need for more researches on therapeutics that may balance the levels of age-related alterations in biogenic amines.