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Genes involved in thoracic exoskeleton formation during the pupal-to-adult molt of a social insect model, Apis mellifera

ABSTRACT: The exoskeleton gives to insects its shape, waterproofing and a range of other roles, besides serving as means of locomotion, the motor function being accomplished by the attachment of somatic muscles. Exoskeleton is renewed every time the insect molts under the control of ecdysteroid hormones. The last molt, which in the holometabolous transforms pupae in adults, involves biosynthesis of the definitive exoskeleton by the subjacent epidemis, followed by progressive differentiation. We used whole genome-based oligonucleotide microarray hybridization and RNA extracted from the honey bee thoracic dorsum, at three time-points of the pupal-to-adult molt, as a strategy to screen genes involved in exoskeleton formation. Gene ontology analysis separated the differentially expressed genes (DEGs) in distinct Biological Processes and Molecular Functions dependently on the time-point approached, thus revealing the functional categories required for adult exoskeleton formation. Approximately 546 of a list of 1253 unique DEGs were up-regulated in the thoracic dorsum during adult cuticle formation, including 23 among the 24 identified cuticle protein (CP) genes, and 29 muscle-related genes. Conserved sequence motifs allowed to include the CP genes in the CPR, Tweedle, Apidermin, CPF, CPLCP1 and Analogous-to-Peritrophins families. Two genes, which do not pertain to any known CP families, were abundantly expressed in the epidermis during adult cuticle formation as shown by in situ hybridization, thus strongly suggesting that they are genuine CP genes. The search for cis-regulatory motifs in the 5’untranslated region of the DEGs revealed potential binding sites for known transcript factors. Several of them were shared by CP genes and thoracic muscle genes, thus allowing to infer that they are co-regulated during differentiation of the thoracic exoskeleton. Together, these data add new information on molecular aspects of exoskeleton formation in the context of the ecdysteroid-coordinated pupal-to-adult molt.

ORGANISM(S): Apis mellifera

PROVIDER: GSE43047 | GEO | 2012/12/29

SECONDARY ACCESSION(S): PRJNA184919

REPOSITORIES: GEO

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Project description:The exoskeleton gives to insects its shape, waterproofing and a range of other roles, besides serving as means of locomotion, the motor function being accomplished by the attachment of somatic muscles. Exoskeleton is renewed every time the insect molts under the control of ecdysteroid hormones. The last molt, which in the holometabolous transforms pupae in adults, involves biosynthesis of the definitive exoskeleton by the subjacent epidemis, followed by progressive differentiation. We used whole genome-based oligonucleotide microarray hybridization and RNA extracted from the honey bee thoracic dorsum, at three time-points of the pupal-to-adult molt, as a strategy to screen genes involved in exoskeleton formation. Gene ontology analysis separated the differentially expressed genes (DEGs) in distinct Biological Processes and Molecular Functions dependently on the time-point approached, thus revealing the functional categories required for adult exoskeleton formation. Approximately 546 of a list of 1253 unique DEGs were up-regulated in the thoracic dorsum during adult cuticle formation, including 23 among the 24 identified cuticle protein (CP) genes, and 29 muscle-related genes. Conserved sequence motifs allowed to include the CP genes in the CPR, Tweedle, Apidermin, CPF, CPLCP1 and Analogous-to-Peritrophins families. Two genes, which do not pertain to any known CP families, were abundantly expressed in the epidermis during adult cuticle formation as shown by in situ hybridization, thus strongly suggesting that they are genuine CP genes. The search for cis-regulatory motifs in the 5M-bM-^@M-^Yuntranslated region of the DEGs revealed potential binding sites for known transcript factors. Several of them were shared by CP genes and thoracic muscle genes, thus allowing to infer that they are co-regulated during differentiation of the thoracic exoskeleton. Together, these data add new information on molecular aspects of exoskeleton formation in the context of the ecdysteroid-coordinated pupal-to-adult molt. The microarray experiments compared differential gene expression in the thoracic integuments of newly-ecdysed pupae (Pw phase), pupae-in-apolysis (Pp phase) and pharate-adults (Pbl phase) collected at the same time from a single honey bee colony. Two separate pools of 10 thoracic integuments each were prepared for each developmental phase (Pw, Pp and Pbl). Dye swaps were done for each comparison and two slides were used to evaluate the differential expression.

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Genes involved in thoracic exoskeleton formation during the pupal-to-adult molt of a social insect model, Apis mellifera

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