Project description:The tobacco hornworm, Manduca sexta, is a lepidopteran model species widely used to study insect biochemical processes. While some of its larval hemolymph proteins are well understood, a detailed proteomic analysis was unavailable until 2016, revealing features such as correlation with transcriptome data, formation of immune complexes, and constitution of an immune signaling system. Yet, it is unclear how these may change in other developmental stages. In this paper, we report the proteomes of cell-free hemolymph from prepupae, pupae on days 4 and 13, and young adults. Of the 1,824 proteins identified, 907 have a signal peptide and 215 are related to immunity. Drastic changes in abundance of the storage proteins, for instance, reflect physiological disparities among prepupae, pupae, and adults. Considerably more proteins lacking signal peptide are present in the late pupae, suggesting that plasma acts as a temporary reservoir for intracellular components released from remodeling tissues during metamorphosis. In summary, the proteins and their levels revealed in this study are expected to stimulate focused explorations of humoral immunity in wandering larvae, pupae, and adults of M. sexta and shed light upon functional and comparative genomic research in other holometabolous insects.
Project description:Transcriptional profiling of larval epidermis at specific markings were examined using 11 developmental stages. Gene expression level was compared between mimetic white, cryptic thorax, and cryptic abdomen vs. mimetic black, cryptic eyespot, and cryptic V-shaped markings in all stages. For initial screening of marking-specific genes, 6 developmental stages for mimetic pattern, and 5 developmental stages for cryptic pattern was used. Mimetic white, cryptic thorax, and cryptic abdomen were independently labelled with Cyanine 3-CTP (Cy3), and mimetic black, cryptic eyespot, and cryptic V-shaped markings were independently labelled with Cyanine 5-CTP (Cy5) in all stages.
Project description:Expression profiling of honey bee brains exposed to brood pheromone. Exposure was performed in colonies and young (5 days-old) and old bees (15 days-old) were analyzed .
Project description:Independent Lewis rat data used to confirm the finding of the main experiment "Protection against Mammary Tumorigenesis in Multiple Rat Strains (experiment E-TABM-197: https://www.ebi.ac.uk/arrayexpress/experiments/E-TABM-197).
Project description:Microarray analysis of parity induced gene expression changes in the mammary glands of four strains of rats to identify a common gene signature associated with protection against methylnitrosourea induced mammary tumorigenesis.
Project description:We examined patterns of gene expression in two independent colonies of both M and S molecular forms of Anopheles gambiae at each of three developmental stages of interest: late larvae, sugar-fed virgin females, and gravid females. For each colony, replicates were derived from independent RNA samples extracted from different cohorts to ensure that trends were reproducible. In addition, each replicate was derived from larvae (adults) drawn from three pans (cages) to minimize the contribution of any individual pan to variation between samples. Data were obtained from a total of five biological replicates per mosquito colony.
Project description:Lateral root initiation was used as a model system to study the mechanisms behind auxin-induced cell division. Genome-wide transcriptional changes were monitored during the early steps of lateral root initiation. Inclusion of the dominant auxin signaling mutant solitary root1 (slr1) identified genes involved in lateral root initiation that act downstream of the AUX/IAA signaling pathway. Interestingly, key components of the cell cycle machinery were strongly defective in slr1, suggesting a direct link between AUX/IAA signaling and core cell cycle regulation. However, induction of the cell cycle in the mutant background by overexpression of the D-type cyclin (CYCD3;1) was able to trigger complete rounds of cell division in the pericycle that did not result in lateral root formation. Therefore, lateral root initiation can only take place when cell cycle activation is accompanied by cell fate respecification of pericycle cells. The microarray data also yielded evidence for the existence of both negative and positive feedback mechanisms that regulate auxin homeostasis and signal transduction in the pericycle, thereby fine-tuning the process of lateral root initiation. Experiment Overall Design: Seedlings of both wild type (Col-0) and the lateral root defective mutant (slr-1) were germinated on MS medium supplemented with 10μM NPA (=auxin transport inhibitor). Three days after germination, such seedlings were transferred to MS supplemented with 10μM NAA for 0h, 2h and 6h respectively. The segment between root meristem and root-hypocotyl junction was harvested from about 1500 seedling per time point. All treatments were repeated biologically. 5.8 μg total RNA was used for the preparation of biotinylated cRNA. Labeled RNA was hybridised to ATH1 Affymetrix chips. The resulting data was MAS5.0 normalised.
Project description:We studied the molecular mechanisms underlying the impact of pollen nutrients on honey bee (Apis mellifera) health and how those nutrients improve resistance to parasites. Using digital gene expression, we determined the changes in gene expression induced by pollen intake in worker bees parasitized or not by the mites Varroa destructor, known for suppressing immunity and decreasing lifespan of bees. bees with or without verroa, and fed or not fed pollen
Project description:Ticks are obligate blood feeding ectoparasites that transmit a wide variety of pathogenic organisms to their vertebrate hosts. The tick Amblyomma sculptum is vector of Rickettsia rickettsii, the causative agent of Rock Mountain spotted fever, the most lethal rickettsiosis that affects humans. It is known that the transmission of pathogens by ticks is mainly associated with the physiology of the feeding process. Pathogens that are acquired with the blood meal must first colonize the tick gut and later the salivary glands (SG). Then, to be transmitted during a subsequent blood feeding, pathogens must reach the saliva. Tick saliva contains a complex mixture of bioactive molecules with anti-clotting, anti-platelet aggregation, vasodilatory, anti-inflammatory, and immunomodulatory properties to counteract both the host hemostasis and defense mechanisms, which besides facilitating tick feeding, may also benefits survival and establishment of pathogens in the host. In the current study, we compared the sialotranscriptome of unfed A. sculptum ticks and fed for 72 hours on rabbits using RNA-seq. The total of reads obtained were mapped in 9,560 coding sequences (CDSs) distributed in six major functional classes. Genes encoding secreted proteins, including lipocalins, mucins, protease inhibitors, glycine rich, metalloprotease, and 8.9 kDa superfamily were mostly upregulated by blood feeding. Selected genes were analyzed by RT-qPCR and all of them presented the same transcriptional profile regulation observed in RNA-seq, corroborating the transcriptional findings of this study. Finally, we mapped 116 proteins secreted in tick saliva by mass spectrometry-based proteomic analysis. Identified proteins should be functionally characterized and might be potential targets to develop vaccines for tick control and/or blocking of R. rickettsii transmission as well as pharmacological bioproducts with anti-hemostatic, anti-inflammatory and anti-bacterial activities.