Project description:Ecotoxicogenomics in field experiments have yielded valuable mechanistic information for organisms present in polluted environments. The Queen conch (Strombus gigas) is a threatened species and populations are declining due to anthropogenic impact that includes pollution from boating activities. In the British Virgin Islands (BVI), local Queen conch populations have exhibited imposex, a condition in which both male and female gonadal characteristics are present and studies in the BVI suggest that tributyl tin (TBT), a chemical used in boat paint, is correlated to increased incidence imposex. This present study utilized a previously validated 8 x 15K Queen conch microarray to characterize the response of the ovarian transcriptome in conch found in polluted environments with high TBT in the BVIs. There polluted sites, Road Harbour (RH) and Trellis Bay (TB), are harbours with high boating activity while the reference sites, Guana Island (GI) and Anegada (AN), are areas with low boating activity. Microarray analysis revealed that there were 17 transcripts with high homology to known genes that were differentially expressed in the environments with high TBT and these included 6 induced and 11 down-regulated transcripts (p<0.01). These differentially expressed transcripts included phosphoenolpyruvate carboxylase, transposase, and high-affinity phosphate transporter PT1. When considering both RH and TB together in comparison to GI, functional enrichment showed that the biological processes and molecular functions of calcium ion binding, immune response, and negative regulation of cell proliferation were over represented in the polluted sites. Gene set enrichment analysis revealed that transcripts involved in the biological processes of general metabolism, immune, lipid metabolism, and stress were affected in polluted environments. Although difficult to directly link changes at the transcriptomics level to TBT in the harbour, this analysis provides novel insight into pathways impacted in regions that experience heavy boating activity in the BVIs.
Project description:Ecotoxicogenomics in field experiments have yielded valuable mechanistic information for organisms present in polluted environments. The Queen conch (Strombus gigas) is a threatened species and populations are declining due to anthropogenic impact that includes pollution from boating activities. In the British Virgin Islands (BVI), local Queen conch populations have exhibited imposex, a condition in which both male and female gonadal characteristics are present and studies in the BVI suggest that tributyl tin (TBT), a chemical used in boat paint, is correlated to increased incidence imposex. This present study utilized a previously validated 8 x 15K Queen conch microarray to characterize the response of the ovarian transcriptome in conch found in polluted environments with high TBT in the BVIs. There polluted sites, Road Harbour (RH) and Trellis Bay (TB), are harbours with high boating activity while the reference sites, Guana Island (GI) and Anegada (AN), are areas with low boating activity. Microarray analysis revealed that there were 17 transcripts with high homology to known genes that were differentially expressed in the environments with high TBT and these included 6 induced and 11 down-regulated transcripts (p<0.01). These differentially expressed transcripts included phosphoenolpyruvate carboxylase, transposase, and high-affinity phosphate transporter PT1. When considering both RH and TB together in comparison to GI, functional enrichment showed that the biological processes and molecular functions of calcium ion binding, immune response, and negative regulation of cell proliferation were over represented in the polluted sites. Gene set enrichment analysis revealed that transcripts involved in the biological processes of general metabolism, immune, lipid metabolism, and stress were affected in polluted environments. Although difficult to directly link changes at the transcriptomics level to TBT in the harbour, this analysis provides novel insight into pathways impacted in regions that experience heavy boating activity in the BVIs. Adult female queen conchs were collected from field sites in the British Virgin Islands with varying levels of TBT contamination and imposex incidence. Contaminated sites included Rhode Harbour (RH, n=3) and Trellis Bay (TB, n=3), and the low-pollution site was Guana Island (GA, n=5). Ovaries were preserved in RNAlater, transported to the laboratory, and RNA was prepared using a phenol-chloroform extraction and a CsCl gradient. RNA was labeled with Cy3 for a single-dye design, per the Agilent protocol.
Project description:This experiment was designed to identify gene expression differences in young bees raised in colonies with a queen, with queen mandibular pheromone (QMP), or without a queen. The source colony was split into three colonies, allocating roughly equal quantities of adult bees, brood, and pollen and nectar stores. One colony retained the original queen (QR), one was left queenless (QL), and the third was given a strip that contained 10 Qeq (QMP+), a dose shown to mimic a live queen (20). The three colonies were transferred to a different apiary > 2 mi away so they would not return to the site of their natal hive. Prior to the colony split, bees (N ~1500) were collected 0-36 h after eclosion, marked on the dorsal thorax with a paint dot (Testors Paint), and ~500 were placed in each of the three colonies. Two days later, the marked bees were collected (N = 100). Bees were collected into liquid nitrogen, and heads were stored as above.
Project description:Deep sequencing of mRNA from queen and virgin queen of two Ants: Camponotus floridanus and Harpegnathos saltator Total RNA were isolated from virgin queens and queens by pulverizing the entire animal after removal of gut and poison glands in liquid N2 and then dissolving in TRIzol. PolyA+ RNA were prepared for deep sequencing.
Project description:Deep sequencing of smRNA from queen and virgin queen of two Ants: Camponotus floridanus and Harpegnathos saltator Total RNA were isolated from virgin queens and queens by pulverizing the entire animal after removal of gut and poison glands in liquid N2 and then dissolving in TRIzol. Gel-purified 18-30 nts RNAs were used for sequencing.
Project description:Reproduction of queen conchs at nearshore sites in the Florida Keys is known to be impaired (Delgado et al. 2004). A recent microarray study of male queen conchs collected from the Florida Keys during the early part of the reproductive season (February, 2007) indicated that NS conchs show differences in expression of spermatogenesis-related and small GTPase signaling transcripts (Spade et al. 2010). The current study investigates gene expression in the ovary of female queen conchs from the same sampling effort in February, 2007.