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Effect of feeding and females on male salivary glands

ABSTRACT: A differential display experiment was done to compare genes expressed in salivary glands from unfed males, males fed in the presence of females and males fed in the absence of females. Possible differential bands were excised, cloned and sequenced. A macroarray study was done on these clones as probes. RNA from the salivary glands of ticks from the three conditions was labelled and hybridized. Two sets of primers were used in the differential display, and these same two were used in the labelling, so there are two platforms, corresponding to what primer pair was used. Two independent RNA isolations were done, Experiment 1 used one, 2 and 3 the other. Experiment 2 used exposure to X-ray film, while 1 and 3 used a phosphorimager plate. Two exposures of the X-ray film were found to give acceptable results, 1 and 5 min. These were averaged in the analysis. Keywords = salivary_gland Keywords = feeding Keywords = male Keywords = female Keywords: other

ORGANISM(S): Dermacentor andersoni

PROVIDER: GSE1320 | GEO | 2004/04/21

SECONDARY ACCESSION(S): PRJNA89893

REPOSITORIES: GEO

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Project description:Although the salivary transcriptome of adult mosquitoes has been thoroughly described in several recent papers, very little information exists regarding the biological role of larval salivary glands in the Culicidae. We used whole-transcriptome AffymetrixÂ® chips to compare the transcriptional profiles of Anopheles gambiae larval (L4) salivary glands and whole larvae. We identified a total of 277 transcripts as being significantly enriched in the salivary glands. Based on available annotation for the known or predicted protein sequences encoded by these transcripts, 41 were identified as corresponding to secreted proteins, 233 as corresponding to non-secreted (housekeeping) proteins, and 3 as encoding proteins of unknown function. Based on functional annotation of the putatively secreted gene products, we propose that larval salivary secretions have roles in nutrient digestion, detoxification, immunity, and mouthpart lubrication. Interestingly, several components of the larval saliva (e.g. apyrase and serine proteases) have also been reported to exist in adult female saliva, where they are though to help regulate a vertebrate hostâs immune response to bloodfeeding. In conclusion, our results suggest that the salivary glands are important components of both the digestive and immune systems of larval mosquitoes, and that their study might provide clues about the evolution of adaptations to bloodfeeding observed in adults. Experiment Overall Design: We attempted to identify transcripts that are both present and significantly enriched in the salivary glands (SG) of the 4th instar Anopheles gambiae larva. Transcripts were considered as present in the SG if the built-in Affymetrix detection call scored them as such in all three biological replicates of the experiment. Expression values (i.e. signal intensities) of transcripts from the salivary glands were compared to those of a matching group of whole 4th instar larvae using a t-test. Transcripts showing an expression value two-fold or higher (p < 0.01; false discovery rate=1%) in the SG as compared to whole larvae were considered to be significantly enriched.

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Effect of feeding and females on male salivary glands

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