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.

Stress-reactivity and juvenile hormone degradation in Drosophila melanogaster strains having stress-related mutations.

Juvenile hormone (JH) degradation was studied under normal and stress conditions in young and matured females of Drosophila melanogaster strains having mutations in different genes involved in responses to stress It was shown that (1) the impairment in heat shock response elicits an alteration in stress-reactivity of the JH system; (2) the impairment JH reception causes a decrease of JH-hydrolysing activity and of stress-reactivity in young females, while in mature ones stress reactivity is completely absent; (3) the absence of octopamine results in higher JH-hydrolysis level under normal conditions and altered JH stress-reactivity; (4) the higher dopamine content elicits a dramatic decrease of JH degradation under normal conditions and of JH stress-reactivity. Thus, the impairments in any component of the Drosophila stress reaction result in changes in the reponse of JH degradation system to stress. The role of JH in the development of the insect stress reaction is discussed.

Stress-reactivity of a Drosophila melanogaster strain with impaired juvenile hormone action.

Met(27) is a null allele of the Methoprene-tolerant gene of D. melanogaster that shows resistance to the toxic effects of both juvenile hormone (JH) and a JH analog, methoprene. The mechanism of resistance appears to be altered JH reception. We measured fertility, JH-hydrolyzing activity, and dopamine (DA) levels in Met(27) and Met(+) flies under normal (25 degrees C) and heat-stress (38 degrees C) conditions. We show that under normal conditions Met(27) females have JH-hydrolyzing activity and fertility lower than Met(+), but DA content did not differ between the two strains. At 38 degrees C Met(27) flies show no impairment in JH-hydrolyzing activity in response to stress, but they do show lower DA levels and impaired reproduction. The results with Met(27) are consistent with the previous hypothesis that the alteration in fertility that follows heat stress in D. melanogaster could result from alteration in the JH endocrine system.

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.

A comparative analysis of juvenile hormone metabolyzing enzymes in two species of Drosophila during development.

The course of changes in the activities of enzymes degrading juvenile hormone (JH), epoxyde hydrolase (JHEH) and JH-esterase (JHE) was studied in two lines of Drosophila virilis (101 and 147) and in two lines of D. melanogaster (Canton-S and 921283). It was established for D. virilis that changes in the JH titre during pupal-adult development is determined by the activity level of JHE rather then JHEH, while in D. melanogaster developmental changes in JH titre are related to changes in the activity level of both JHE and JHEH. In adults of D. virilis, the high level of JH-hydrolysing activity is determined by JHE and in those of D. melanogaster by JHEH. Differences in the course of changes in the JHE activity level between adults of lines 101 and 147 of D. virilis were found, and also in the JHEH activity level between adults of lines Canton S and 921283 of D. melanogaster. It was shown that attainment of a definite JHE activity level in females of lines 101 and 147 agrees well with the onset of oviposition of fertilized eggs. The possible role of JHE in reproduction of D. virilis is discussed.

The genetics of esterases in Drosophila. VIII. The gene regulating the activity of JH-esterase in D. virilis.

The content of JH-esterase was assayed by radial immunodiffusion in Drosophila virilis pupae under normal conditions and under the effects of extreme factors. It was found that JH-esterase content is the same (not different from the control) in pupae showing a high activity of the enzyme and in those not showing it. These data are evidence for a gene controlling JH-esterase activity. It was also shown that a regulatory factor converts inactive into active JH-esterase when homogenates of pupae, with active and inactive forms, were mixed and incubated together. It was demonstrated that the source of the activating factor is the larval brain. Sublines 147-R and 147-I were produced by introducing the second chromosome pair of stocks 103 and 101, which are heat resistant, into the genome of individuals of stock 147, which is heat sensitive. Sublines 160-III, 160-IV, 160-V, and 160-VI were produced by introducing the third, fourth, fifth, and sixth chromosome pairs of stock 147 into the genome of stock 160S, which is heat-resistant. The results of analysis of JH-esterase activity and the viability of individuals of these sublines at high temperatures indicated that the gene regulating the activity of JH-esterase is located in the sixth chromosome of D. virilis.

Genetics of esterases in Drosophila. VII. Genetic control of the activity level of juvenile hormone (JH)-esterase and heat resistance in D. virilis at high temperatures.

The heat-resistant subline 147S was obtained in Drosophila virilis by selecting for viability individuals of heat-sensitive stock 147. It was shown that in the heat-treated 147S pupae the activity of juvenile hormone (JH)-esterase is decreased and, consequently, the titer of juvenile hormone is increased compared with those in the control pupae. These changes are consistent with those observed earlier for resistant stock 101. Heat-resistant stocks 101 and 147S were crossed with heat-sensitive stock 147, whose heat-treated larvae show earlier activation and higher activity of JH-esterase than control larvae. The viability and electrophoretic esterase patterns were analyzed in the F1 and F2 hybrids at different temperatures. It was found that the F1 hybrid is resistant to the effect of high temperature and its activity level of JH-esterase is lower compared with controls. In the F2 hybrid, there was a 3:1 segregation of viability and a 1:2:1 segregation of the activity level of JH-esterase at high temperatures. It is concluded that the activity level of JH-esterase and heat resistance in D. virilis are monogenically controlled at high temperatures.

Genetics of esterases in Drosophila. V. Characteristics of the "pupal" esterase in D. virilis.

From analysis of the properties of the "pupal" esterase (p-esterase) in Drosophila virilis, it is concluded that it is heat stable, its electrophoretic detection depends on culture density, its expression is stage specific, and it is not a variant of esterase 2. It was also demonstrated that p-esterase, like esterase 6, is activated by injections of the juvenile hormone into larvae. Heat treatment of heat-resistant D. virilis stocks led to decreased activities of the juvenile hormone dependent esterases but did not affect those of the heat-sensitive stocks. It is suggested that heat resistance in D. virilis is related to some functional features of the system of modifier genes controlling the phenotypic expression of esterases.