Comparative analysis ofCorazonin‐encoding genes (Crz's) inDrosophilaspecies and functional insights intoCrz‐expressing neurons

Abstract

To gain insight into regulatory mechanisms of tissue-specific Corazonin (Crz) gene expression and its functions in Drosophila, we cloned the Crz genes from four Drosophila species (D. melanogaster, D. simulans, D. erecta, and D. virilis) and performed comparative analyses of Crz gene sequences and expression patterns using in situ hybridization and immunohistochemistry. Although Crz gene sequences showed a great deal of diversity, its expression patterns in the CNS were highly conserved in the Drosophila species examined here. In D. melanogaster larva, Crz expression was found in four pairs of neurons per cerebral lobe and in eight pairs of bilateral neurons in the ventral nerve cord; in adult, the number of Crz-producing neurons increased to 6-8 in the pars lateralis of each brain lobe, whereas neurons in the ventral nerve cord were no longer detectable. Crz transcripts were also found in the optic lobes; however, these mRNAs do not seem to be translated. Such adult-like Crz expression patterns were established within 48 hours after pupation. Somata of Crz-neurons in the pars lateralis are located in the vicinity of terminals emanating from PDF-containing pacemaking neurons, indicating a functional connection between the two peptidergic nervous systems. A subset of Crz neurons coexpressed the period clock gene; however, normal Crz transcription was unaffected by central clockworks. Two pairs of ectopic Crz cells were detected in the adult brains of behaviorally arrhythmic Clock(Jrk) or cycle(02) mutants, suggesting that CLOCK and CYCLE proteins negatively regulate Crz transcription in a cell-specific manner.

Neurotoxic protein expression reveals connections between the circadian clock and mating behavior in Drosophila

“…2 A Lower). The exceptions were the neuropeptide mRNAs corazonin and Pdf; both genes are rhythm-related (30,31).…”

Programmed cell death mechanisms of identifiable peptidergic neurons inDrosophila melanogaster

“…To construct P-element containing Crz promoter fused to the gal4-coding sequence (hereafter referred to as Crz-gal4), a genomic region upstream of the Crz gene (-1155 to +78 relative to the transcription start site +1) (Choi et al, 2005) was amplified by PCR, and subcloned into pBluescript at the SmaI site, from which an XbaI-EcoRI fragment was inserted into pPTGAL vector (Sharma et al, 2002). For rpr-gal4 construct, previously defined 1.3 kb rpr upstream sequence (Jiang et al, 2000) was inserted into pPTGAL at BglII/EcoRI sites.…”

“…Whole-mount Crz immunohistochemistry was performed as described previously (Choi et al, 2005). The anti-Crz was previously referred to as anti-CAP, which was raised against Crz-Associated Peptide within the precursor (Choi et al, 2005).…”

“…The anti-Crz was previously referred to as anti-CAP, which was raised against Crz-Associated Peptide within the precursor (Choi et al, 2005). To co-localize EcR-immunoreactivity (IRy) in Crz neurons, rabbit anti-Crz and mouse monoclonal anti-EcR-A (15G1a) or anti-EcR-B1 (AD4.4) were simultaneously applied to the CNSs.…”

Mapeamento histológico das áreas protocerebrais e do complexo retrocerebral de operárias de Melipona quadrifasciata anthidioides (Hymenoptera, Apidae, Meliponini)

“…Esta marcação foi descrita em drosófila (Choi et al, 2005). Os dados aqui apresentados não são, porém, suficientes para afirmar que estas células não estejam presentes em M. quadrifasciata.…”