Microarray-based gene expression profiles of silkworm brains
Ontology highlight
ABSTRACT: Molecular genetic studies of Bombyx mori have led to profound advances in understanding the regulation of development. Bombyx mori brain, as a main endocrine organ, plays important regulatory roles in various biological processes. The microarray technology will allow the genome-wide analysis of gene expression patterns in silkworm brains. We reported microarray-based gene expression profiles in silkworm brains at four stages including V7, P1, P2 and P3. A total of 4,550 genes were transcribed in at least one selected stage. Of these, clustering algorithms separated the expressed genes into stable expressed genes and variable expressed genes. The results of the gene ontology (GO) and kyoto encyclopedia of genes and genomes (KEGG) analysis of stable 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 variable expressed genes. Thirdly, thirty-two neuropeptide hormones genes, nine neuropeptide-like precursor genes, and 117 cuticular protein genes were expressed in selected developmental stages. The present study defined major characteristics of the transcriptional profiles in the brains of Bombyx mori at the specific development stages. Our data will provide abundant information that will be useful in future research. Transcription profiling experiments, 4 developmental stages (samples) were analyzed. Dual-channel experiments, with test samples labeled by Cy5 and common reference samples labeled by Cy3. Common reference sample was used for data normalization. One Biological replicate. No dye-swaps.
Project description:Polycomb group (PcG) proteins are involved in chromatin modifications for maintaining gene repression that play important roles in the regulation of gene expression, tumorigenesis, chromosome X-inactivation, and genomic imprinting in Drosophila melanogaster, mammals, and even plants. PcG proteins act together in three multimeric complexes, Polycomb repressive complex 1 (PRC1), Polycomb repressive complex 2 (PRC2), and Pleiohomeotic repressive complex (PhoRC), to repress transcription of the target genes. Here, we identified Polycomb target genes in Bombyx mori using genome-wide expression screening based on the knockdown of the BmSCE, BmESC, BmPHO, or BmSCM gene, which represent the distinct complexes. As a result, most genes were up-regulated after knocking down these four PcG genes, which indicated a potential epigenetic mechanism on the regulation of these genes expression by the PcG system. The further analysis of our data will provide some important information for the regulation mediated by PcG proteins in Bombyx mori. Transcription profiling experiments, knockdowns of four Polycomb genes (four samples) in silkworm BmN4-SID1 cells, were analyzed. Dual-channel experiments, with test samples labeled by Cy5 and common reference samples labeled by Cy3. The common reference sample, knockdown of the EGFP gene in BmN4-SID1 cells, was used for data normalization. One biological replicate. No dye-swaps.
Project description:The molting of insects is a complex biological progress which requires varies of genes to participate in the event. MicroRNAs are considered to be one of the key roles involved in development in many organisms. Microarray technology was employed to examine the expression profile of multiple genes after extrinsical up-regulation of miR-1 level by injection of miR-1 mimics in silkworm (Bombyx mori.L), which caused altered expression profile of numerous genes, especially those which involved in the processes of cuticle renewing and development. 814 genes were up-regulated and 636 were down-regulated after treated with miR-1 mimics, the rest 2961 detectable genes were considered to have no significant changes in expression level. Transcription profiling experiments, 3 pairs of samples (miR-1 injected and negative control) were analyzed. Dual-channel experiments, with control samples labeled by Cy5 and miR-1 injected samples labeled by Cy3. Fold change was calculated via comparing signal intensity of Cy3 vs Cy5. 3 biological replicates were performed.
Project description:Transcriptional characteristics of genes in the midgut of domestic silkworms after 24 h exposure to phoxim through whole-genome oligonucleotide microarray. Transcription profiling experiments, phoxim-treated midgut (samples) were analyzed. Dual-channel experiments, with test samples labeled by Cy5 and control samples labeled by Cy3. Three Biological replicate. No dye-swaps.
Project description:Insect cuticle plays essential roles in multiple physiological functions. During molting and metamorphosis, tremendous changes occur in silkworm cuticles. Silkworm is a model of Lepidoptera insects; however, little is known about the stage expression profiles of genes in cuticles of silkworm. In the present study, we selected 16 developmental stages, ranging from day 1 of the first instar larvae to day 8 of pupae, to perform microarray-based expression profiles. The data told us that various functions and physiological pathways were activated in the cuticle. Moreover, the expression profiles of cuticular protein genes, as the important components of cuticle, were investigated. The current study provides important insights for the functional study of insect cuticle and the regulation of insect cuticular protein genes. Transcription profiling experiments, 16 developmental stages (samples) were analyzed. Dual-channel experiments, with test samples labeled by Cy5 and common reference samples labeled by Cy3. Common reference sample was used for data normalization. One biological replicate. No dye-swaps.
Project description:Microsporidia have attracted much attention because they infect a variety of species ranging from protists to mammals, including immunocompromised patients with AIDS or cancer. Aside from the study on Nosema ceranae, few works have focused on elucidating the mechanism in host response to microsporidia infection. Nosema bombycis is a pathogen of silkworm pM-CM-)brine that causes great economic losses to the silkworm industry. Detailed understanding of the host (Bombyx mori) response to infection by N. bombycis is helpful for prevention of this disease. The 23 K silkworm genome array was used to investigate host responses (i.e., Bombyx mori) occurring at 2, 4, 6 and 8 d post-infection by Nosema bombycis.We focused on elucidating the mechanism of the host response to microsporidia infection, especially for the investigation of host immune response . The third instar molted silkworm larvae were in oral infected by Nosema bombycis. In order to known the silkworm host response to Nosema bombycis infection at different time points, samples of infected larvae (i.e., the treatment set) and uninfected larvae (i.e., the control set) were collected at 2, 4, 6 and 8 dpi for RNA extraction and array hybridization. The obtained data were usd to investigate on the interplay of the genome-wide expression profile of hosts.
Project description:In the silkworm, Bombyx mori, juvenile hormone (JH) and 20-hydroxyecdysone (20E) levels are high during the final larval molt (4M) but both absent during the feeding stage of 5th instar (5F), while JH level is low and 20E level is high during the prepupal stage (PP). Fat body is the important organs in insect, we want to find out differentially expressed genes which are respectively regulated by the two hormones. Total RNA from 4th molting,5th feeding and prepupa stages Bombyx fat body were used to generate target cDNA, and then hybridized to 48k Bombyx genome Array Genechips, representing about 23000 characterized genes
Project description:Comparative analysis of tobacco leaves transcriptomes unveils carotenoid pathway potentially determined the characteristics of aroma compounds in different environmental regions. Tobacco (Nicotiana tabacum) is a sensitive crop to environmental changes, and a tobacco with unique volatile aroma fractions always formed in specific ecological conditions. In order to investigate the differential expressed genes caused by environmental changes and reveal the formation mechanism of characteristics of tobacco in three different aroma tobacco regions of Guizhou Province, Agilent tobacco microarray was adapted for transcriptome comparison of tobacco leaves in medium aroma tobacco region Kaiyang and light aroma tobacco regions Weining and Tianzhu. Results showed that there was big difference among the gene expression profiles of tobacco leaves in different environmental conditions. A total of 517 differential expressed genes (DEGs) between Weining and Tianzhu were identified, while 733 and 1,005 genes differentially expressed between Longgang and another two tobacco regions Weining and Tianzhu, respectively. Compared with Longgang, up-regulated genes in Weining and Tianzhu were likely involved in secondary metabolism pathways, especially carotenoid pathway, including PHYTOENE SYNTHASE, PHYTOENE DEHYDROGENASE, LYCOPENE ε-CYCLASE, CAROTENOID β-HYDROXYLASE and CAROTENOID CLEAVAGE DIOXYGENASE 1 genes, while most down-regulated genes played important roles in response to temperature and light radiation, such as heat shock proteins. Gene Ontology and MapMan analyses demonstrated that the DEGs among different environmental regions were significantly enriched in light reaction of photosystem II, response of stimulus and secondary metabolism, suggesting they played crucial roles in environmental adaptation and accumulation of aroma compounds in tobacco plants. Through comprehensive transcriptome comparison, we not only identified several stress response genes in tobacco leaves from different environmental regions but also highlighted the importance of carotenoid pathway genes for characteristics of aroma compounds in specific growing regions. Our study primarily laid the foundation for further understanding the molecular mechanism of environmental adaptation of tobacco plants and molecular regulation of aroma substances in tobacco leaves. In order to investigate the differential expressed genes caused by environmental changes and reveal the formation mechanism of characteristics of tobacco in three different aroma tobacco regions of Guizhou Province, Agilent tobacco microarray was adapted for transcriptome comparison of tobacco leaves in medium aroma tobacco region Kaiyang and light aroma tobacco regions Weining and Tianzhu.
Project description:Hpa1Xoo-Mediated Transcriptome in Transgenic Cotton Reveals the Constitutively Expressed Diverse Defense genes in Multiple Signaling Pathways Associated with Hypersensitive Cell Death Like other harpins, harpinXoo enables plants to acquire multiple resistance to pathogen and insect attack. However, the molecular model is not fully understood, especially in transgenic plant harpinXoo for genome-wide constitutive expression. Here, we showed that 530 cDNAs differentially expressed in transgenic hpa1Xoo-harboring cotton (T-34) compared to its receiptor (Z35), through analyzing the transcriptome profile in a customized cotton 12k cDNA microarray, which was enriched in 34 pathways. Among them, 123 genes were identified from T-34 as hypersensitive reaction-mediated defense genes, involved in reactive oxygen species-, salicylic acid-, jasmonates-, ethylene-, auxin-, abscisic acid-, and Ca2+-mediated signaling pathways, and we uncovered various components of defense responses associated with recognition of the pathogen-derived elicitor. Apart from elevated genes for basic defense, harpinXoo activated leucine-rich-repeat plant receptor kinases and mitogen-activated protein kinase cascade to strengthen downstream defense responses, including production of antimicrobial compounds in T-34. Defense genes from T-34 were expressed at a much higher level in response to pathogen infection compared with Z35. Simultaneous up- and down-regulation in differentially-expressed genes, and increased expression of genes encoding energy producing and consuming pathways, suggested that energy balance was maintained in the genetically modified cotton without biotic stress. High-energy demand only occurred following pathogen infection. Using kanamycin-resistance tests, positive plants were successfully screened from transgenic cotton T-34 T6 progenies; all the receiptor Z35 plants gave negative results. We randomly selected three positive plants for polymerase chain reaction (PCR) analysis; the different positive bands (420, 310 and 180 bp) were confirmed as harpinXoo, the 35s promoter, and the NOS terminator, respectively, by sequencing and BLAST searches (www.ncbi.nlm.nih.gov.). For further analysis, the positive results were confirmed by Southern and western blots. Three positive bands were obtained in each T-34 sample located at about 4, 6 and 10 kb in the Southern blot. This pattern suggests that T-34 incorporated the harpinXoo transgene at multiple chromosomal locations. Western blot analysis showed that the harpinXoo protein was constitutively expressed in T-34 transgenic lines, whereas it was barely detectable in Z35 (data not shown). The transgenic plants contained a selectable ‘marker’ gene, neomycin phosphotransferase II (nptII). Previous research showed that expression of marker genes (e.g. nptII, uidA, hph and uidA-gfp) did not contribute to visible phenotypes in transgenic plants. The non-targeted transgenic and non-transgenic plants were equivalent in their global patterns of transcription (El Ouakfaoui and Miki, 2005). Thus, the present study examined the many differences between T-34 and its receiptor Z35 in regard to the whole genome expression of G. hirsutum using microarrays to monitor changes in individual gene expression level effected by harpinXoo. All experiments were conducted using the 12k cotton cDNA microarray, and cotton true leaves and roots at the 4–5-true-leaf-stage. The use of this microarray enabled a quantitative approach to examine changes in transcript level for all genes within the G. hirsutum genome; as a result, inferences can be drawn regarding the molecular effects of transgenic cotton expressing harpinXoo. There were three biological replicates in the experiment for each organ.
Project description:This SuperSeries is composed of the following subset Series: GSE26376: Microarray profiling of monocytic differentiaion reveals miRNA-mRNA intrinsic correlation (miRNA) GSE26377: Microarray profiling of monocytic differentiaion reveals miRNA-mRNA intrinsic correlation (gene expression) Refer to individual Series
Project description:The microarray test aims to find the miRNAs involved in somatic embryogenesis in Arabidopsis. The plant microRNA array V2.0 (CapitalBio Corp., Beijing, China) contained 426 non-reduplicated plant miRNA probes, including 188 in Arabidopsis. Finally, 75 plant miRNAs expressed differentially, 36 increased and 39 decreased included. Two critical period samples of somatic embryogenesis were chosen for the microarray test: the edge of embryonic calli in embryonic callus-inducing medium (ECIM) for 14 days and the secondary somatic embryos protuberances in somatic embryo-inducing medium (SEIM) for 2 days (marked as “14D” and “J2D” accordingly) . Three chip were test in each sample.