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Insights into the dynamics of hind leg development in honey bee (Apis mellifera L.) queen and worker larvae - A morphology/differential gene expression analysis.

Santos CG, Hartfelder K - Genet. Mol. Biol. (2015)

Bottom Line: In parallel, we generated subtractive cDNA libraries for hind leg discs of queen and worker larvae by means of a Representational Difference Analysis (RDA).From the total of 135 unique sequences we selected 19 for RT-qPCR analysis, where six of these were confirmed as differing significantly in their expression between the two castes in the larval spinning stage.The development of complex structures such as the bees' hind legs, requires diverse patterning mechanisms and signaling modules, as indicated by the set of differentially expressed genes related with cell adhesion and signaling pathways.

View Article: PubMed Central - PubMed

Affiliation: Departamento de Biologia Celular e Molecular e Bioagentes Patogênicos, Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP, Brazil.

ABSTRACT
Phenotypic plasticity is a hallmark of the caste systems of social insects, expressed in their life history and morphological traits. These are best studied in bees. In their co-evolution with angiosperm plants, the females of corbiculate bees have acquired a specialized structure on their hind legs for collecting pollen. In the highly eusocial bees (Apini and Meliponini), this structure is however only present in workers and absent in queens. By means of histological sections and cell proliferation analysis we followed the developmental dynamics of the hind legs of queens and workers in the fourth and fifth larval instars. In parallel, we generated subtractive cDNA libraries for hind leg discs of queen and worker larvae by means of a Representational Difference Analysis (RDA). From the total of 135 unique sequences we selected 19 for RT-qPCR analysis, where six of these were confirmed as differing significantly in their expression between the two castes in the larval spinning stage. The development of complex structures such as the bees' hind legs, requires diverse patterning mechanisms and signaling modules, as indicated by the set of differentially expressed genes related with cell adhesion and signaling pathways.

No MeSH data available.


Related in: MedlinePlus

Characteristics of the queen and worker RDA libraries prepared from queen and worker larval hind leg discs. The assembled sequences were grouped into (i) genes similar (E ≤ e−5)to Drosophila melanogaster, (ii) genes similar to predicted or exclusive transcripts for A. mellifera and other bees represented in the Transcriptome Shotgun Assembly (TSA) database, (iii) genes similar to stem-loop microRNAs (miRBase), (iv) genes with intronic location, (v) genes with a putative UTR location, and (vi) sequences representing unpredicted genes but mapping to the honey bee genome sequence.
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f03: Characteristics of the queen and worker RDA libraries prepared from queen and worker larval hind leg discs. The assembled sequences were grouped into (i) genes similar (E ≤ e−5)to Drosophila melanogaster, (ii) genes similar to predicted or exclusive transcripts for A. mellifera and other bees represented in the Transcriptome Shotgun Assembly (TSA) database, (iii) genes similar to stem-loop microRNAs (miRBase), (iv) genes with intronic location, (v) genes with a putative UTR location, and (vi) sequences representing unpredicted genes but mapping to the honey bee genome sequence.

Mentions: For validation of origin, Blastn and Blastx analyses were done on all library sequences against the nucleotide and the consensual protein databases of predicted A. mellifera genes (Amel Official Gene Set v3.2). Sequences without BLAST matches to predicted honey bee genes were mapped to the assembled honey bee genome (Amel 4.5). Thus, all sequences could be grouped into six categories: (i) sequences with similarity in the nr database, represented herein as similar to Drosophila, since this is the best annotated insect genome; (ii) sequences corresponding to predicted honey bee genes, including those from the bee Transcriptome Shotgun Assembly (TSA) database; (iii) sequences with similarity to stem loop microRNAs; (iv) sequences mapping in intronic regions of predicted coding sequences in the honey bee genome; (v) sequences mapping close to predicted coding sequences in the honey bee genome, thus indicating possible location in an untranslated region (5′ or 3′ UTR); and (vi) sequences without information in public databases, including predicted honey bee genes, but mapping to the honey bee genome. Figure 3 shows the relative representation of sequences for the two libraries within these six categories.


Insights into the dynamics of hind leg development in honey bee (Apis mellifera L.) queen and worker larvae - A morphology/differential gene expression analysis.

Santos CG, Hartfelder K - Genet. Mol. Biol. (2015)

Characteristics of the queen and worker RDA libraries prepared from queen and worker larval hind leg discs. The assembled sequences were grouped into (i) genes similar (E ≤ e−5)to Drosophila melanogaster, (ii) genes similar to predicted or exclusive transcripts for A. mellifera and other bees represented in the Transcriptome Shotgun Assembly (TSA) database, (iii) genes similar to stem-loop microRNAs (miRBase), (iv) genes with intronic location, (v) genes with a putative UTR location, and (vi) sequences representing unpredicted genes but mapping to the honey bee genome sequence.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4612609&req=5

f03: Characteristics of the queen and worker RDA libraries prepared from queen and worker larval hind leg discs. The assembled sequences were grouped into (i) genes similar (E ≤ e−5)to Drosophila melanogaster, (ii) genes similar to predicted or exclusive transcripts for A. mellifera and other bees represented in the Transcriptome Shotgun Assembly (TSA) database, (iii) genes similar to stem-loop microRNAs (miRBase), (iv) genes with intronic location, (v) genes with a putative UTR location, and (vi) sequences representing unpredicted genes but mapping to the honey bee genome sequence.
Mentions: For validation of origin, Blastn and Blastx analyses were done on all library sequences against the nucleotide and the consensual protein databases of predicted A. mellifera genes (Amel Official Gene Set v3.2). Sequences without BLAST matches to predicted honey bee genes were mapped to the assembled honey bee genome (Amel 4.5). Thus, all sequences could be grouped into six categories: (i) sequences with similarity in the nr database, represented herein as similar to Drosophila, since this is the best annotated insect genome; (ii) sequences corresponding to predicted honey bee genes, including those from the bee Transcriptome Shotgun Assembly (TSA) database; (iii) sequences with similarity to stem loop microRNAs; (iv) sequences mapping in intronic regions of predicted coding sequences in the honey bee genome; (v) sequences mapping close to predicted coding sequences in the honey bee genome, thus indicating possible location in an untranslated region (5′ or 3′ UTR); and (vi) sequences without information in public databases, including predicted honey bee genes, but mapping to the honey bee genome. Figure 3 shows the relative representation of sequences for the two libraries within these six categories.

Bottom Line: In parallel, we generated subtractive cDNA libraries for hind leg discs of queen and worker larvae by means of a Representational Difference Analysis (RDA).From the total of 135 unique sequences we selected 19 for RT-qPCR analysis, where six of these were confirmed as differing significantly in their expression between the two castes in the larval spinning stage.The development of complex structures such as the bees' hind legs, requires diverse patterning mechanisms and signaling modules, as indicated by the set of differentially expressed genes related with cell adhesion and signaling pathways.

View Article: PubMed Central - PubMed

Affiliation: Departamento de Biologia Celular e Molecular e Bioagentes Patogênicos, Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP, Brazil.

ABSTRACT
Phenotypic plasticity is a hallmark of the caste systems of social insects, expressed in their life history and morphological traits. These are best studied in bees. In their co-evolution with angiosperm plants, the females of corbiculate bees have acquired a specialized structure on their hind legs for collecting pollen. In the highly eusocial bees (Apini and Meliponini), this structure is however only present in workers and absent in queens. By means of histological sections and cell proliferation analysis we followed the developmental dynamics of the hind legs of queens and workers in the fourth and fifth larval instars. In parallel, we generated subtractive cDNA libraries for hind leg discs of queen and worker larvae by means of a Representational Difference Analysis (RDA). From the total of 135 unique sequences we selected 19 for RT-qPCR analysis, where six of these were confirmed as differing significantly in their expression between the two castes in the larval spinning stage. The development of complex structures such as the bees' hind legs, requires diverse patterning mechanisms and signaling modules, as indicated by the set of differentially expressed genes related with cell adhesion and signaling pathways.

No MeSH data available.


Related in: MedlinePlus