Microarray-based gene expression profiles of silkworm brains

<p>Abstract</p> <p>Background</p> <p>Molecular genetic studies of <it>Bombyx mori </it>have led to profound advances in our understanding of the regulation of development. <it>Bombyx mori </it>brain, as a main endocrine organ, plays important regulatory roles in various biological processes. Microarray technology will allow the genome-wide analysis of gene expression patterns in silkworm brains.</p> <p>Results</p> <p>We reported microarray-based gene expression profiles in silkworm brains at four stages including V7, P1, P3 and P5. A total of 4,550 genes were transcribed in at least one selected stage. Of these, clustering algorithms separated the expressed genes into stably expressed genes and variably expressed genes. The results of the gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) analysis of stably expressed genes showed that the ribosomal and oxidative phosphorylation pathways were principal pathways. Secondly, four clusters of genes with significantly different expression patterns were observed in the 1,175 variably expressed genes. Thirdly, thirty-two neuropeptide genes, six neuropeptide-like precursor genes, and 117 cuticular protein genes were expressed in selected developmental stages.</p> <p>Conclusion</p> <p>Major characteristics of the transcriptional profiles in the brains of <it>Bombyx mori </it>at specific development stages were present in this study. Our data provided useful information for future research.</p

A syntenic coding region for vitelline membrane proteins in four lepidopteran insects

The vitelline membrane is the inner layer of the eggshell, but the genomic information available for vitelline membrane proteins (VMPs) in Lepidoptera is limited. In the present study, we identified a syntenic coding region for VMPs in four lepidopteran genomes (Bombyx mori, Manduca sexta, Danaus plexippus and Heliconius melpomene) and four putative VMP coding genes located within it. RT-PCR results showed Bombyx VMP coding genes expressed prior to the early choriogenesis stage in follicles. Alignment analyses revealed that the vitelline membrane domain was shared between Lepidoptera and Diptera. However, the third cysteine residue conserved in dipteran VMPs was absent in those of Lepidoptera. In addition, another conserved region was identified in lepidopteran VMPs

Expression profile of cuticular genes of silkworm, Bombyx mori

<p>Abstract</p> <p>Background</p> <p>Insect cuticle plays essential roles in many physiological functions. During molting and metamorphosis tremendous changes occur in silkworm cuticle where multiple proteins exist and genes encoding them constitute about 1.5% of all <it>Bombyx mori </it>genes.</p> <p>Results</p> <p>In an effort to determine their expression profiles, a microarray-based investigation was carried out using mRNA collected from larvae to pupae. The results showed that a total of 6676 genes involved in various functions and physiological pathways were activated. The vast majority (93%) of cuticular protein genes were expressed in selected stages with varying expression patterns. There was no correlation between expression patterns and the presence of conserved motifs. Twenty-six RR genes distributed in chromosome 22 were co-expressed at the larval and wandering stages. The 2 kb upstream regions of these genes were further analyzed and three putative elements were identified.</p> <p>Conclusions</p> <p>Data from the present study provide, for the first time, a comprehensive expression profile of genes in silkworm epidermal tissues and evidence that putative elements exist to allow massive production of mRNAs from specific cuticular protein genes.</p

Proteomic Analysis of Larval Midgut from the Silkworm (Bombyx mori)

The midgut is the major organ for food digestion, nutrient absorption and also a barrier for foreign substance. The 5th-instar larval stage of silkworm is very important for larval growth, development, and silk production. In the present study, we used 2-DE and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) to analyze the midgut proteins from the 5th-instar larvae as well as the midgut proteins under starvation condition. A total of 96 proteins were identified in this study; and among them, 69 proteins were observed in midgut for the first time. We also found that the silkworm larval midgut responded to starvation by producing a 10 kDa heat shock protein and a diapause hormone precursor

MicroRNAs of Bombyx mori identified by Solexa sequencing

<p>Abstract</p> <p>Background</p> <p>MicroRNA (miRNA) and other small regulatory RNAs contribute to the modulation of a large number of cellular processes. We sequenced three small RNA libraries prepared from the whole body, and the anterior-middle and posterior silk glands of <it>Bombyx mori</it>, with a view to expanding the repertoire of silkworm miRNAs and exploring transcriptional differences in miRNAs between segments of the silk gland.</p> <p>Results</p> <p>With the aid of large-scale Solexa sequencing technology, we validated 257 unique miRNA genes, including 202 novel and 55 previously reported genes, corresponding to 324 loci in the silkworm genome. Over 30 known silkworm miRNAs were further corrected in their sequence constitutes and length. A number of reads originated from the loop regions of the precursors of two previously reported miRNAs (bmo-miR-1920 and miR-1921). Interestingly, the majority of the newly identified miRNAs were silkworm-specific, 23 unique miRNAs were widely conserved from invertebrates to vertebrates, 13 unique miRNAs were limited to invertebrates, and 32 were confined to insects. We identified 24 closely positioned clusters and 45 paralogs of miRNAs in the silkworm genome. However, sequence tags showed that paralogs or clusters were not prerequisites for coordinated transcription and accumulation. The majority of silkworm-specific miRNAs were located in transposable elements, and displayed significant differences in abundance between the anterior-middle and posterior silk gland.</p> <p>Conclusions</p> <p>Conservative analysis revealed that miRNAs can serve as phylogenetic markers and function in evolutionary signaling. The newly identified miRNAs greatly enrich the repertoire of insect miRNAs, and provide insights into miRNA evolution, biogenesis, and expression in insects. The differential expression of miRNAs in the anterior-middle and posterior silk glands supports their involvement as new levels in the regulation of the silkworm silk gland.</p

MicroRNAs show diverse and dynamic expression patterns in multiple tissues of Bombyx mori

<p>Abstract</p> <p>Background</p> <p>MicroRNAs (miRNAs) repress target genes at the post-transcriptional level, and function in the development and cell-lineage pathways of host species. Tissue-specific expression of miRNAs is highly relevant to their physiological roles in the corresponding tissues. However, to date, few miRNAs have been spatially identified in the silkworm.</p> <p>Results</p> <p>We establish for the first time the spatial expression patterns of nearly 100 miRNAs in multiple normal tissues (organs) of <it>Bombyx mori </it>females and males using microarray and Northern-blotting analyses. In all, only 10 miRNAs were universally distributed (including bmo-let-7 and bmo-bantam), while the majority were expressed exclusively or preferentially in specific tissue types (e.g., bmo-miR-275 and bmo-miR-1). Additionally, we examined the developmental patterns of miRNA expression during metamorphosis of the body wall, silk glands, midgut and fat body. In total, 63 miRNAs displayed significant alterations in abundance in at least 1 tissue during the developmental transition from larvae to pupae (e.g., bmo-miR-263b and bmo-miR-124). Expression patterns of five miRNAs were significantly increased during metamorphosis in all four tissues (e.g., bmo-miR-275 and bmo-miR-305), and two miRNA pairs, bmo-miR-10b-3p/5p and bmo-miR-281-3p/5p, showed coordinate expression.</p> <p>Conclusions</p> <p>In this study, we conducted preliminary spatial measurements of several miRNAs in the silkworm. Periods of rapid morphological change were associated with alterations in miRNA expression patterns in the body wall, silk glands, midgut and fat body during metamorphosis. Accordingly, we propose that corresponding ubiquitous or tissue-specific expression of miRNAs supports their critical roles in tissue specification. These results should facilitate future functional analyses.</p

MicroRNA expression profiling during the life cycle of the silkworm (Bombyx mori)

<p>Abstract</p> <p>Background</p> <p>MicroRNAs (miRNAs) are expressed by a wide range of eukaryotic organisms, and function in diverse biological processes. Numerous miRNAs have been identified in <it>Bombyx mori</it>, but the temporal expression profiles of miRNAs corresponding to each stage transition over the entire life cycle of the silkworm remain to be established. To obtain a comprehensive overview of the correlation between miRNA expression and stage transitions, we performed a whole-life test and subsequent stage-by-stage examinations on nearly one hundred miRNAs in the silkworm.</p> <p>Results</p> <p>Our results show that miRNAs display a wide variety of expression profiles over the whole life of the silkworm, including continuous expression from embryo to adult (miR-184), up-regulation over the entire life cycle (let-7 and miR-100), down-regulation over the entire life cycle (miR-124), expression associated with embryogenesis (miR-29 and miR-92), up-regulation from early 3^rdinstar to pupa (miR-275), and complementary pulses in expression between miR-34b and miR-275. Stage-by-stage examinations revealed further expression patterns, such as emergence at specific time-points during embryogenesis and up-regulation of miRNA groups in late embryos (miR-1 and bantam), expression associated with stage transition between instar and molt larval stages (miR-34b), expression associated with silk gland growth and spinning activity (miR-274), continuous high expression from the spinning larval to pupal and adult stages (miR-252 and miR-31a), a coordinate expression trough in day 3 pupae of both sexes (miR-10b and miR-281), up-regulation in pupal metamorphosis of both sexes (miR-29b), and down-regulation in pupal metamorphosis of both sexes (miR-275).</p> <p>Conclusion</p> <p>We present the full-scale expression profiles of miRNAs throughout the life cycle of <it>Bombyx mori</it>. The whole-life expression profile was further investigated via stage-by-stage analysis. Our data provide an important resource for more detailed functional analysis of miRNAs in this animal.</p

Microarray-based gene expression profiles in multiple tissues of the domesticated silkworm, Bombyx mori

Using a genome-wide oligonucleotide microarray, gene expression was surveyed in multiple silkworm tissues on day 3 of the fifth instar, providing a new resource for annotating the silkworm genome

SilkDB: a knowledgebase for silkworm biology and genomics

The Silkworm Knowledgebase (SilkDB) is a web-based repository for the curation, integration and study of silkworm genetic and genomic data. With the recent accomplishment of a ∼6X draft genome sequence of the domestic silkworm (Bombyx mori), SilkDB provides an integrated representation of the large-scale, genome-wide sequence assembly, cDNAs, clusters of expressed sequence tags (ESTs), transposable elements (TEs), mutants, single nucleotide polymorphisms (SNPs) and functional annotations of genes with assignments to InterPro domains and Gene Ontology (GO) terms. SilkDB also hosts a set of ESTs from Bombyx mandarina, a wild progenitor of B.mori, and a collection of genes from other Lepidoptera. Comparative analysis results between the domestic and wild silkworm, between B.mori and other Lepidoptera, and between B.mori and the two sequenced insects, fruitfly and mosquito, are displayed by using B.mori genome sequence as a reference framework. Designed as a basic platform, SilkDB strives to provide a comprehensive knowledgebase about the silkworm and present the silkworm genome and related information in systematic and graphical ways for the convenience of in-depth comparative studies. SilkDB is publicly accessible at http://silkworm.genomics.org.cn