Unknown,Transcriptomics,Genomics,Proteomics

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Gene expression rhythms in rainbow darter male (Etheostoma caeruleum) across an annual reproductive cycle

ABSTRACT: We have been studying intersex in male rainbow darter (Etheostoma caeruleum) associated with exposure to sewage effluents. To understand changes in the gene transcriptome associated with intersex it was necessary to have a better understanding of normal annual changes in the transcriptome. The goal of this research is to identify patterns of gene expression associated with the different stages of gonad development during the annual cycle. The studies of molecular pathways involve in ovarian or testis development has been poorly studied. While most studies focus on female ovarian changes, there is a gap in understanding testis development. A customized second generation microarray for rainbow darter (8x15k) was used to identify patterns of gene expression -in terms of mRNA abundance- in male rainbow darter gonads during an annual cycle. Rainbow darter males were collected on field work surveys in May (spawning), August (post-spawning), and October (recrudescence) 2011, and January (developing) and March (pre-spawning) 2012, using a back pack electrofisher from a clean area at the Grand River, ON, Canada.

ORGANISM(S): Etheostoma caeruleum

SUBMITTER: Paulina Bahamonde

PROVIDER: E-GEOD-57865 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:We have constructed a rainbow trout high-density oligonucleotide microarray by using all the available tentative consensus (TC) sequences from the Rainbow Trout Gene Index database (The Computational Biology and Functional Genomics Lab., Dana Farber Cancer Institute and Harvard School of Public Health). The Rainbow Trout Gene Index integrates research data from all available international rainbow trout genomic research projects. The newly designed microarray incorporates 37,394 unique transcript-specific oligonucleotide probes, 60-mer long each. The microarray was printed according to our design by Agilent Technologies using the 4 X 44-design format and contains 1417 Agilent control spots. The performance of the new microarray platform was evaluated by analyzing gene expression associated with the rainbow trout vitellogenesis-induced muscle atrophy. These chips can be ordered from Agilent using design number 016320. This microarray is anticipated to open new avenues of research that will aid in the development of novel strategies to enhance growth efficiency and quality in salmonid species. Keywords: Development of an oligo-array for rainbow trout The performance of the new microarray platform was evaluated by analyzing transcriptome response associated with the rainbow trout vitellogenesis-induced muscle atrophy. Severe muscle deterioration accompanies the physiological responses of the energetic demands of the rainbow trout spawning/vitellogenesis. Atrophying muscle of fertile fish had 11% less extractable muscle (g/bw) and 11% less protein content compared to non-atrophying muscle of sterile fish (p<0.01). The rainbow trout was used to profile changes in gene expression of atrophying muscles. Gene expression levels were determined by comparing the amount of mRNA transcript present in the experimental sample (fertile fish) to the control (sterile fish). RNAs isolated from each experimental fish were run on separate microarrays in independent experiments, with no pooling. A total of 8 fish were used in the microarray experiments (4 replicates x 2 groups). Fluorophors (Cy3 and Cy5) were randomly assigned to RNA from each of the atrophying and nonatrophying muscles to limit the dye effect.

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Gene expression rhythms in rainbow darter male (Etheostoma caeruleum) across an annual reproductive cycle

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