Rhythm defects caused by newly engineered null mutations in Drosophila's cryptochrome gene.
Much of the knowledge about cryptochrome function in Drosophila stems from analyzing the cryb mutant. Several features of this variant's light responsiveness imply either that CRYb retains circadian-photoreceptive capacities or that additional CRY-independent light-input routes subserve these processes. Potentially to resolve these issues, we generated cry knock-out mutants (cry0's) by gene replacement. They behaved in an anomalously rhythmic manner in constant light (LL). However, cry0 flies frequently exhibited two separate circadian components in LL, not observed in most previous cryb analyses. Temperature-dependent circadian phenotypes exhibited by cry(0) flies suggest that CRY is involved in core pacemaking. Further locomotor experiments combined cry0 with an externally blinding mutation (norpAP24), which caused the most severe decrements of circadian photoreception observed so far. cryb cultures were shown previously to exhibit either aperiodic or rhythmic eclosion in separate studies. We found cry0 to eclose in a solidly periodic manner in light:dark cycles or constant darkness. Furthermore, both cry0 and cryb eclosed rhythmically in LL. These findings indicate that the novel cry0 type causes more profound defects than does the cryb mutation, implying that CRYb retains residual activity. Because some norpAP24 cry0 individuals can resynchronize to novel photic regimes, an as-yet undetermined light-input route exists in Drosophila.
Dolezelova
Eva
E
Department of Biology, Brandeis University, Waltham, Massachusetts 02454, USA.
Dolezel
David
D
Hall
Jeffrey C
JC
D000818:Animals; D030801:Animals, Genetically Modified; D001921:Brain / Q000502:physiology; D002940:Circadian Rhythm; D056931:Cryptochromes; D003624:Darkness; D029721:Drosophila Proteins / Q000235:genetics*; D004331:Drosophila melanogaster / Q000235:genetics*; D005136:Eye Proteins / Q000235:genetics*; D005260:Female; D008027:Light; D008124:Locomotion; D008156:Luciferases / Q000378:metabolism; D008297:Male; D009154:Mutation / Q000235:genetics*; D043562:Receptors, G-Protein-Coupled / Q000235:genetics*; D011995:Recombination, Genetic
D056931:Cryptochromes; D029721:Drosophila Proteins; D005136:Eye Proteins; D043562:Receptors, G-Protein-Coupled; C119828:cry protein, Drosophila; D008156:Luciferases