ecdysterone and Starvation

Autophagy and apoptosis, which could be induced by common stimuli, play crucial roles in development and disease. The functional relationship between autophagy and apoptosis is complex, due to the dual effects of autophagy. In the Bombyx Bm-12 cells, 20-hydroxyecdysone (20E) treatment or starvation-induced cell death, with autophagy preceding apoptosis. In response to 20E or starvation, BmATG8 was rapidly cleaved and conjugated with PE to form BmATG8-PE; subsequently, BmATG5 and BmATG6 were cleaved into BmATG5-tN and BmATG6-C, respectively. Reduction of expression of BmAtg5 or BmAtg6 by RNAi decreased the proportion of cells undergoing both autophagy and apoptosis after 20E treatment or starvation. Overexpression of BmAtg5 or BmAtg6 induced autophagy but not apoptosis in the absence of the stimuli, but promoted both autophagy and apoptosis induced by 20E or starvation. Notably, overexpression of cleavage site-deleted BmAtg5 or BmAtg6 increased autophagy but not apoptosis induced by 20E or starvation, whereas overexpression of BmAtg5-tN and BmAtg6-C was able to directly trigger apoptosis or promote the induced apoptosis. In conclusion, being cleaved into BmATG5-tN and BmATG6-C, BmATG5 and BmATG6 mediate apoptosis following autophagy induced by 20E or starvation in Bombyx Bm-12 cells, reflecting that autophagy precedes apoptosis in the midgut during Bombyx metamorphosis.

Topics: Animals; Apoptosis; Autophagy; Bombyx; Cell Line; Ecdysterone; Insect Proteins; Models, Biological; Starvation

Insect midgut secretes a semi-permeable peritrophic membrane (PM), which plays important roles in protecting the midgut and helping with food digestion. The lepidopteran larvae produce type 1 PM, which is degraded when insects develop into the metamorphic stages. However, the underlying mechanism is unclear. In the present study, two peritrophin-like proteins (peritrophin-57 and 37) were identified from the midgut expression sequence tag library and transcriptome of the common cutworm, Spodoptera litura. The temporal and spatial expression patterns and responses to the induction of 20-hydroxyecdysone (20E) and starvation were examined by real-time quantitative polymerase chain reaction according to their common sequence region. The chitin-binding activity was also studied using a competitor, calcofluor. The open reading frames are 1 554 and 1 020 bp, respectively. They shared four highly conserved peritrophin-A domains and were expressed only in the midgut rather than in the other tissues, including fat body, epidermis, Malpighian tube and hemolymph. Their transcriptional expression could only be detected at the larval stages rather than in eggs, prepupae, pupae and adults. The purified protein of peritrophin-37 bound to chitin in a dose-dependent manner. These results indicate that the two proteins are peritrophins, the structural components of PM. In addition, the messenger RNA levels of the two peritrophins were significantly down-regulated by 20E injection, whereas feeding/starvation had no effect on the expression. These findings suggest that the increase of 20E titer may be an important factor which controls the degradation of PM during metamorphosis.

Topics: Animals; Base Sequence; Chitin; Digestive System; Ecdysterone; Expressed Sequence Tags; Gene Library; Insect Proteins; Larva; Metamorphosis, Biological; Molecular Sequence Data; Pupa; Spodoptera; Starvation; Transcriptome

Insect chitinases are hydrolytic enzymes that are required for the degradation of glycosidic bonds of chitin. In this study, we identified and characterized a full-length cDNA of the chitinase gene (BdCht2) in the oriental fruit fly, Bactrocera dorsalis. The cDNA contains an open reading frame (ORF) of 1449 bp that encodes 483 amino acid residues and 126- and 296-bp non-coding regions at the 5'- and 3'-ends, respectively. The BdCht2 genome has four exons and three introns. The predicted molecular mass of the deduced BdCht2 is approximately 54.3 kDa, with an isoelectric point of 5.97. The 977 bp 5' flanking region was identified and the transcription factor binding sites were predicted. Bioinformatic analyses showed that the deduced amino acid sequence of BdCht2 had 34%-66% identity to that of chitinases identified in other insect species. Quantitative real-time PCR (qPCR) analyses indicated that BdCht2 was mainly expressed during the larval-pupal and pupal-adult transitions. The tissue-specific expression showed that the highest expression was in the integument, followed by the fat body and other tissues. Moreover, the expression of BdCht2 was upregulated significantly upon 20-hydroxyecdysone (20E) at different dose injections after 8 h compared to that of the control. Starvation also increased the expression of BdCht2 in the third-instar larvae and was suppressed again by re-feeding the insects. These results suggest that BdCht2 plays an important role in the molting process of B. dorsalis larvae and can be regulated by 20E.

Topics: Amino Acid Sequence; Animals; Base Sequence; Binding Sites; Chitinases; Diptera; DNA, Complementary; Ecdysterone; Larva; Molecular Sequence Data; Phylogeny; Starvation; Tephritidae; Transcription Factors; Transcription, Genetic

Female reproduction includes maturation of oocytes and the synthesis of yolk proteins (vitellogenin, Vg) in the fat body and their deposition into the oocytes. Our recent studies showed that juvenile hormone (JH) regulates Vg synthesis and 20-hydroxyecdysone (20E) regulates oocyte maturation in the red flour beetle (Tribolium castaneum). Here, we report on the role of nutritional signaling on vitellogenesis and oogenesis. Comparison of gene expression between fed and starved beetles by microarray analysis showed the up-regulation of genes involved in energy homeostasis and down-regulation of genes involved in egg production in the starved beetles. The RNA interference (RNAi) aided knock-down in the expression of genes involved in insulin and TOR signaling pathways showed that both these signaling pathways play key roles in Vg synthesis and oocyte maturation. Starvation of female beetles resulted in a block in Vg synthesis but not in the progression of primary oocyte development to the resting stage. Feeding after starvation induced Vg synthesis and the progression of primary oocytes from the resting stage to the mature stage. However, in the beetles where JH or 20E synthesis or action was blocked by RNAi, both Vg synthesis and oocyte maturation were affected suggesting that both these hormones (JH and 20E) and nutritional signaling and their cross-talk regulate vitellogenesis and oogenesis.

Topics: Animals; Base Sequence; Down-Regulation; Ecdysterone; Female; Gene Expression Regulation, Developmental; Insect Proteins; Insulin; Juvenile Hormones; Microarray Analysis; Molecular Sequence Data; Nutritional Physiological Phenomena; Oocytes; Oogenesis; Reproduction; RNA Interference; RNA, Messenger; Signal Transduction; Starvation; TOR Serine-Threonine Kinases; Tribolium; Up-Regulation; Vitellogenesis; Vitellogenins

In animals, appendages develop in proportion to overall body size; when individual size varies, appendages covary proportionally. In insects with complete metamorphosis, adult appendages develop from precursor tissues called imaginal disks that grow after somatic growth has ceased. It is unclear, however, how the growth of these appendages is matched to the already established body size. We studied the pattern of cell division in the tobacco hornworm Manduca sexta and found that both the rate of cell division and the duration of growth of the wing imaginal disks depend on the size of the body in which they develop. Moreover, we found that both of these processes are controlled by the level and duration of secretion of the steroid hormone ecdysone. Thus, proportional growth is under hormonal control and indirectly regulated by the central nervous system.

Topics: Animals; Body Size; Cell Division; Central Nervous System; Ecdysone; Ecdysterone; Larva; Manduca; Pupa; Starvation; Wings, Animal

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.

Topics: Animals; Dopamine; Drosophila; Ecdysterone; Female; Juvenile Hormones; Reproduction; Starvation; Time Factors

The steroid hormone 20-hydroxyecdysone (20E) triggers the major developmental transitions in Drosophila, including molting and metamorphosis, and provides a model system for defining the developmental and molecular mechanisms of steroid signaling. 20E acts via a heterodimer of two nuclear receptors, the ecdysone receptor (EcR) and Ultraspiracle, to directly regulate target gene transcription.. Here we identify the genomic transcriptional response to 20E as well as those genes that are dependent on EcR for their proper regulation. We show that genes regulated by 20E, and dependent on EcR, account for many transcripts that are significantly up- or downregulated at puparium formation. We provide evidence that 20E and EcR participate in the regulation of genes involved in metabolism, stress, and immunity at the onset of metamorphosis. We also present an initial characterization of a 20E primary-response regulatory gene identified in this study, brain tumor (brat), showing that brat mutations lead to defects during metamorphosis and changes in the expression of key 20E-regulated genes.. This study provides a genome-wide basis for understanding how 20E and its receptor control metamorphosis, as well as a foundation for functional genomic analysis of key regulatory genes in the 20E signaling pathway during insect development.

Topics: Animals; Cells, Cultured; DNA-Binding Proteins; Drosophila melanogaster; Drosophila Proteins; Ecdysterone; Gene Expression Regulation, Developmental; Genes, Insect; Genes, Regulator; Genome, Insect; Immunity; Larva; Metamorphosis, Biological; Microarray Analysis; Mutation; Pupa; Receptors, Steroid; Reproducibility of Results; RNA, Messenger; Starvation

Ecdysone, the major steroid hormone of Drosophila melanogaster, is known for its role in development and reproduction. Flies that are heterozygous for mutations of the ecdysone receptor exhibit increases in life-span and resistance to various stresses, with no apparent deficit in fertility or activity. A mutant involved in the biosynthesis of ecdysone displays similar effects, which are suppressed by feeding ecdysone to the flies. These observations demonstrate the importance of the ecdysone hormonal pathway, a new player in regulating longevity.

Topics: Animals; Body Weight; Crosses, Genetic; Drosophila melanogaster; Ecdysone; Ecdysterone; Female; Fertility; Genes, Insect; Ligands; Longevity; Male; Mutation; Oxidative Stress; Phototropism; Receptors, Steroid; Starvation; Temperature

The mechanisms that control the growth rate of internal tissues during postembryonic development are poorly understood. In insects, the growth rate of imaginal disks varies with nutrition and keeps pace with variation in somatic growth. We describe here a mechanism by which the growth of wing imaginal disks is controlled. When wing imaginal disks of the butterfly Precis coenia are removed from the larva and placed in a standard nutrient-rich tissue culture medium they stop growing, suggesting that nutrients alone are not sufficient to support normal growth. Such disks can be made to grow at a normal rate by supplementing the culture medium with an optimal concentration of the steroid hormone 20-hydroxyecdysone and with hemolymph taken from growing larvae. The growth-promoting activity of the hemolymph is caused by a heat-stable factor that can be extracted from the CNS and appears to be identical to the neurohormone bombyxin, a member of the insulin family of proteins. Synthetic bombyxin stimulates growth at concentrations as low as 30 ngml, and specific antibodies to bombyxin completely remove growth-promoting activity from the hemolymph. Bombyxin evidently acts together with 20-hydroxyecdysone to stimulate cell division and growth of wing imaginal disks. It appears that the level of bombyxin in the hemolymph is modulated by the brain in response to variation in nutrition and is part of the mechanism that coordinates the growth of internal organs with overall somatic growth.

Topics: Animals; Butterflies; Ecdysterone; Growth Substances; Hemolymph; Insulin; Larva; Multigene Family; Neuropeptides; Nutritional Physiological Phenomena; Organ Culture Techniques; Organ Size; Starvation; Wings, Animal

A double-species ecologo-genetical model, including Drosophila and yeast, has been used as a new methodological instrument for investigation of the physiological mechanism of recombination. Incubation of Drosophila females in the medium containing yeast of the strain mutant for ergosterol synthesis leads to suppression of temperature-induced crossing over. The mass-spectrum analysis of steroid fraction from Drosophila females has shown that incubation of the yeast medium without ergosterol results in arrest of ecdysterone synthesis. These data are explained by the absence of ecdysterone synthesis precursor in the fly organism. The endocrinal control of crossing over is discussed in the light of hormonal regulation of meiosis.

Topics: Animals; Crossing Over, Genetic; Drosophila; Ecdysterone; Female; Hot Temperature; Mass Spectrometry; Models, Genetic; Recombination, Genetic; Saccharomyces cerevisiae; Starvation; Sterols; Suppression, Genetic

In contrast to previously described transformations of insect epidermal cells, morphological changes in the larval epidermis of Manduca sexta involve large changes in cell volume: a threefold increase at the beginning and a twofold decrease at the end of the last feeding phase. The volume changes are determined, in part, or entirely, by changes in cell height (cell elongation and cell shortening). Initial cell elongation occurs in a region-specific manner, whereas subsequent cell shortening affects all of the epidermis equally. As shown by ligation experiments and hormone treatments in tissue culture, larval changes in cell height, unlike cell elongation and differentiation in prepupae, are not regulated by developmental hormones (juvenile hormones, ecdysteroids). Instead, the maintenance of a normal columnar epithelium in fifth instars depends on continuous growth. Lack of food, especially protein, results in reversible cell shortening at any time during the feeding phase. Intake of water does not mitigate this cellular response; cell shortening is also insensitive to ouabain but inhibited by cold treatments. We propose that during larval growth epidermal cell height is under specific homeostatic control, independent of mechanisms regulating cell width, ploidy levels, or mitotic activity.

Topics: Animals; Body Fluids; Culture Techniques; Diet; Ecdysterone; Epidermal Cells; Epithelial Cells; Homeostasis; Larva; Lepidoptera; Ligation; Morphogenesis; Starvation; Trypsin

The three yolk proteins of Drosophila melanogaster begin to be synthesized at eclosion. Transcription of the genes is regulated by the genes tra, tra-2 and dsx and also by the insect hormones, juvenile hormone and 20-hydroxyecdysone. We show that there is yet another level of control which is dependent upon feeding. Females that are starved from eclosion show a basal level of yolk protein gene transcription, which is rapidly increased when a complete diet is supplied. We show that the effect is not due to incorrect development of the fat body and is unlikely to be solely due to a general effect on protein synthesis. Later in development, cessation of feeding leads to selective inhibition of yolk protein synthesis and hence egg production. The effects of starvation can be partially overcome by 20-hydroxyecdysone, juvenile hormone, casein, amino acid mix or sucrose, but only a complete medium or live yeast brings about total recovery. Using yp1-Adh fusions (fusions of the promoter region of yp1 to the structural gene for Adh), the DNA sequence required for this diet-enhanced transcription has been located within an 890 bp fragment upstream of the yp1 gene. The insect hormones do not operate on this same DNA fragment.

Topics: Animals; Diet; Drosophila melanogaster; Ecdysterone; Egg Proteins; Gene Expression Regulation; Genes; Juvenile Hormones; Protein Biosynthesis; RNA; Starvation; Transcription, Genetic