ABSTRACT: Microarray evidence for off target effects and the optimisation of targeted RNAi mediated gene silencing in the cattle tick Rhipicephalus (Boophilus) microplus
Project description:Organophosphate resistant and susceptible tick larvae from laboratory strains of the southern cattle tick, Rhipicephalus (Boophilus) microplus were exposed to low doses of the organophosphate (OP) acaricide, coumaphos. Serial Analysis of Gene Expression (SAGE) was used to analyze differential gene expression in response to OP treatment and to compare the responses in OP-treated and untreated resistant and susceptible tick larvae. ** note: Contact person is Felix D. Guerrero. email: felix.guerrero@ars.usda.gov Keywords: SAGE, acaricide response, organophosphate.
Project description:Organophosphate resistant and susceptible tick larvae from laboratory strains of the southern cattle tick, Rhipicephalus (Boophilus) microplus were exposed to low doses of the organophosphate (OP) acaricide, coumaphos. Serial Analysis of Gene Expression (SAGE) was used to analyze differential gene expression in response to OP treatment and to compare the responses in OP-treated and untreated resistant and susceptible tick larvae. ** note: Contact person is Felix D. Guerrero. email: felix.guerrero@ars.usda.gov Keywords: SAGE, acaricide response, organophosphate. four samples make up this series: Munoz Treated: 6030 tags Munoz Untreated: 7806 tags SanRoman Treated: 7714 tags SanRoman Untreated: 9065 tags
Project description:This trial was undertaken to examine the perhipheral cellular and antibody response of cattle following infestation with the cattle tick, Rhipicephalus microplus. The information from the Affymetrix gene expression data is used to complement other measurements of immune function such as cellular subset composition and antibody response in cattle of high (Brahman) and low (Holstein-Friesian) resistance to the cattle tick. Keywords: Disease state analysis
Project description:This experiment was undertaken to document changes in gene expression in the skin of tick-resistant Brahman (Bos indicus) and tick-susceptible Holstein-Friesian (Bos taurus) cattle prior to, and following, infestation with the cattle tick Rhipicephalus (Boophilus) microplus Keywords: Disease state analysis
Project description:Transcriptional profiling of midgut tissues isolated from Rhipicephalus microplus and Rhipicephalus decoloratus females at day 20 post infestation. This enabled the identification of transcripts that are species-specific or shared between the two tick species tested.
Project description:A R. microplus microarray was used to study differential gene expression in acaricide exposed larvae from an amitraz-resistant strain. The acaricide treatments were: organophosphate (OP), pyrethroid, ivermectin, and amitraz. The microarrays contained over 13,000 probes corresponding to each member of R. microplus gene index ESTs previously described (http://compbio.dfci.harvard.edu/tgi/cgi-bin/tgi/gimain.pl?gudb=b_microplus). Serial analysis of gene expression (SAGE) data from the OP treated R. microplus was used to verify the OP microarray data. The expression profiles of selected transcripts were verified by real time PCR. Among the significantly differentially expressed genes, were a tick legumain, involved in blood digestion, gluthathione S-transferase (GST), a detoxification enzyme involved in pesticide resistance, acyltransferase, several putative salivary sulfotransferases, and a glutamate receptor. **Note: contact person: Felix D. Guerrero email: felix.guerrero@ars.usda.gov Keywords: Rhipicephalus (Boophilus) microplus, acaricide resistance genes, organophosphates OP, microarrays, detoxification enzymes.
Project description:Comparative microarray analysis of Rhipicephalus (Boophilus) microplus expression profiles of larvae pre-attachment and feeding adult female stages on Bos indicus and B. taurus cattle Global analysis of gene expression changes in R. microplus during larval, pre-attachment and early adult stages of its life cycle feeding on Bos indicus and Bos taurus cattle were compared using gene expression microarray analysis. Among the 13 601 R. microplus transcripts from BmiGI Version 2 we identified 297 up and 17 down regulated transcripts were differentially expressed between R. microplus feeding on tick resistant cattle [Bos indicus (Brahman)] compared to R. microplus feeding on tick susceptible cattle [Bos taurus (Holstein-Friesian)]. These include genes encoding enzymes involved in primary metabolism, and genes related to stress, defence, cell wall modification, cellular signaling, receptor and cuticle. Microarrays were validated by qRT-PCR analysis of selected transcripts including the validation of three housekeeping genes. The analysis of all tick stages under survey suggested a coordinated regulation of defence proteins, proteases, and protease inhibitors to achieve successful attachment and survival of R. microplus on different host breeds particularly Bos indicus cattle.
Project description:This trial was undertaken to examine the perhipheral cellular and antibody response of cattle following infestation with the cattle tick, Rhipicephalus microplus. The information from the Affymetrix gene expression data is used to complement other measurements of immune function such as cellular subset composition and antibody response in cattle of high (Brahman) and low (Holstein-Friesian) resistance to the cattle tick. Experiment Overall Design: RNA was extracted from white blood cells during a period of successive, heavy infestations with R. microplus. RNA samples from 3 Holstein-Friesian and 3 Brahman animals were analysed on individual slides.
Project description:Transcriptional profiling of midgut tissues isolated from Rhipicephalus microplus and Rhipicephalus decoloratus females at day 20 post infestation. This enabled the identification of transcripts that are species-specific or shared between the two tick species tested. A direct comparison (Balanced block) was performed: Midgut tissues collected on day 20 (post infestation) from R. microplus was compared to that of R. decoloratus. Biological replicates: 3 ; Technical replicates: 1.
Project description:A R. microplus microarray was used to study differential gene expression in acaricide exposed larvae from an amitraz-resistant strain. The acaricide treatments were: organophosphate (OP), pyrethroid, ivermectin, and amitraz. The microarrays contained over 13,000 probes corresponding to each member of R. microplus gene index ESTs previously described (http://compbio.dfci.harvard.edu/tgi/cgi-bin/tgi/gimain.pl?gudb=b_microplus). Serial analysis of gene expression (SAGE) data from the OP treated R. microplus was used to verify the OP microarray data. The expression profiles of selected transcripts were verified by real time PCR. Among the significantly differentially expressed genes, were a tick legumain, involved in blood digestion, gluthathione S-transferase (GST), a detoxification enzyme involved in pesticide resistance, acyltransferase, several putative salivary sulfotransferases, and a glutamate receptor. **Note: contact person: Felix D. Guerrero email: felix.guerrero@ars.usda.gov Keywords: Rhipicephalus (Boophilus) microplus, acaricide resistance genes, organophosphates OP, microarrays, detoxification enzymes. In the study presented here, 10 hybridizations were preformed. Two technical replicates for each treatment. There were no biological replicates included in this study due to limitations on biomaterials. Expression profiles for 13601 unique ESTs were analyzed.
