ecdysterone and Nerve-Degeneration

Steroid hormones mediate a wide variety of developmental and physiological events in insects, yet little is known about the genetics of insect steroid hormone biosynthesis. Here we describe the Drosophila dare gene, which encodes adrenodoxin reductase (AR). In mammals, AR plays a key role in the synthesis of all steroid hormones. Null mutants of dare undergo developmental arrest during the second larval instar or at the second larval molt, and dare mutants of intermediate severity are delayed in pupariation. These defects are rescued to a high degree by feeding mutant larvae the insect steroid hormone 20-hydroxyecdysone. These data, together with the abundant expression of dare in the two principal steroid biosynthetic tissues, the ring gland and the ovary, argue strongly for a role of dare in steroid hormone production. dare is the first Drosophila gene shown to encode a defined component of the steroid hormone biosynthetic cascade and therefore provides a new tool for the analysis of steroid hormone function. We have explored its role in the adult nervous system and found two striking phenotypes not previously described in mutants affected in steroid hormone signaling. First, we show that mild reductions of dare expression cause abnormal behavioral responses to olfactory stimuli, indicating a requirement for dare in sensory behavior. Then we show that dare mutations of intermediate strength result in rapid, widespread degeneration of the adult nervous system.

Topics: Amino Acid Sequence; Animals; Base Sequence; DNA Primers; Drosophila; Ecdysterone; Ferredoxin-NADP Reductase; Gene Expression Regulation, Developmental; Genes, Insect; Humans; Insect Hormones; Molecular Sequence Data; Mutation; Nerve Degeneration; Sequence Homology, Amino Acid; Smell; Steroids

A decline in circulating 20-hydroxyecdysone permits the emergence of the adult Manduca sexta moth; this endocrine signal also triggers the death of approximately half of the neurons in the unfused abdominal ganglia of the moth central nervous system. This programmed death of neurons was markedly reduced by treatment with either actinomycin D (an RNA synthesis inhibitor) or cycloheximide (a protein synthesis inhibitor). Similar results were found after addition of these agents to ventral nerve cord cultures. The effectiveness of these treatments in delaying or blocking neuronal death depended upon their time of administration relative to the normal time of post-emergence death in the particular motoneuron under study: late-dying neurons, for example, could still be saved by these treatments even after early-dying neurons had already initiated degeneration. In both intact moths and cultured ventral nerve cords, the ability of actinomycin D to prevent neuronal death waned at the same time at which replacement of the steroid hormone could no longer block neuronal death. This suggests that the steroid commitment point represents the time at which the genes that mediate cell death are transcribed. Cycloheximide remained effective in delaying or blocking neuronal death until shortly before the onset of degeneration, suggesting that ongoing protein synthesis is essential for the initiation of the degeneration response.

Topics: Animals; Cell Death; Cells, Cultured; Cycloheximide; Dactinomycin; Ecdysterone; Ganglia, Invertebrate; Histocytochemistry; Larva; Moths; Motor Neurons; Nerve Degeneration; Nerve Tissue Proteins; Neurons

Adult emergence at the end of metamorphosis in the moth Manduca sexta is followed by the death of abdominal interneurons and motoneurons. Abdominal ganglia removed from insects before this period of naturally occurring cell death and maintained in vitro showed neuronal death confined to the same cells that normally die in vivo. Addition of physiological levels of the steroid 20-hydroxyecdysone to the culture system prevented the selective death of these motoneurons.

Topics: Animals; Cell Survival; Culture Techniques; Ecdysterone; Ganglia; Interneurons; Lepidoptera; Metamorphosis, Biological; Moths; Motor Neurons; Nerve Degeneration; Neurons

The emergence of the adult Manduca sexta moth is followed by the programmed degeneration of about 50% of the insect's abdominal interneurons and motoneurons. Neurons in implanted ganglia undergo degeneration in concert with neurons of the host, thereby indicating that a hormonal cue is important. The neuronal death follows the normal decline in the levels of the steroid hormones, ecdysteroids, that occurs at the end of metamorphosis. Manipulations that cause a precocious ecdysteroid decline result in an early death of the neurons. Also, prolongation of the ecdysteroid titer extend the life of the neurons. By properly timed steroid application it was possible to dissociate the neuronal degeneration from the behavioral events and muscle degeneration that also occur at emergence. Also the stereotyped sequence of death among identified motoneurons could be stopped at various points in the sequence by the carefully timed application of steroid. It was concluded that the signal for neuronal death was the withdrawal of ecdysteroids at the end of metamorphosis and that the death was probably a direct response of the individual neurons to the hormone withdrawal.

Topics: Animals; Ecdysterone; Ganglia; Interneurons; Kinetics; Lepidoptera; Moths; Motor Neurons; Nerve Degeneration