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Effects of agouti overexpression in the brain and skin transcriptome of zebrafish

ABSTRACT: Twenty transgenic fish overexpressing agouti-signaling protein (ASIP, Ceinos et al., Pigment Cell and Melanoma Research 28:196-209, 2015) and wild type (WT) zebrafish were placed in 6-liter tanks and acclimated during 30 days prior to sampling to minimize physiological differences. Animals were sacrificed and the brain+pituitary and the skin dissected under the stereo microscope, placed in trizol and stored at -80C until RNA extraction. Six biological replicates (3 females and 3 males) of each tissue (skin and brain) of each group (ASIP vs WT) were analyzed in a total of 6 slides containing 4 microarrays in the 4 × 44 k format. Global gene expression analyses were performed to search for both sex-dependent and transgene-dependent effects as well as the interaction between both factors (sex x phenotype) on the brain+pituitary and skin transcriptome. Wild type (Tu) or transgenic zebrafish overexpression agouti-signaling protein (ASIP) were reared in similar conditions. Brain+pituitary and skin were sampled from 6 ASIP (3 males + 3 females) and WT (3 males + 3 females) zebrafish. Samples were individually hybridized in a total of 6 slides containing 4 microarrays in the 4 × 44 k format. Microarrays included 60-mer probes to the unique transcripts from Ensembl and Unigene which were annotated by functional categories of GO and pathways of KEGG with bioinformatics package STARS

ORGANISM(S): Danio rerio

SUBMITTER: Aleksei Krasnov

PROVIDER: E-GEOD-72908 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Effects of agouti overexpression in the brain and skin transcriptome of zebrafish

Project description:Twenty transgenic fish overexpressing agouti-signaling protein (ASIP, Ceinos et al., Pigment Cell and Melanoma Research 28:196-209, 2015) and wild type (WT) zebrafish were placed in 6-liter tanks and acclimated during 30 days prior to sampling to minimize physiological differences. Animals were sacrificed and the brain+pituitary and the skin dissected under the stereo microscope, placed in trizol and stored at -80C until RNA extraction. Six biological replicates (3 females and 3 males) of each tissue (skin and brain) of each group (ASIP vs WT) were analyzed in a total of 6 slides containing 4 microarrays in the 4 × 44 k format. Global gene expression analyses were performed to search for both sex-dependent and transgene-dependent effects as well as the interaction between both factors (sex x phenotype) on the brain+pituitary and skin transcriptome.

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Transcriptional profiles associated with Context dependent behavior in Rock X Sand F1 in Lake Malawi Cichlids

Project description:We sought to evaluate the brain gene expression profiles of male courtship display. To assess male display and courtship behavior, we designed a courtship preference assay. We evaluated social interactions between males and females using a 40 gallon tank design with a ‘rock’ habitat at one end and ‘sand’ at the other, separated by glass bottom. When parental rock species (Petrotilapia nigra (TaxId 526958), Maylandia zebra (TaxId 106582), Labeotropheus feulliborni) are placed in this tank paradigm, males court females over the rocks. Males of sand species (Mchenga conophorus, Aulonocara baenschi (TaxId 143496), Tramitichromis intermedius (TaxId 323801)) court females over sand and construct species appropriate bowers. When single rock x sand F1 males were placed in this set up with F1 females, males invariably courted females over the ‘rock’ habitat, suggesting genetic dominance. When two rock x sand F1 males were allowed to compete for F1 females in this tank paradigm, something interesting happened. One male, typically the larger, courted females over the rock habitat, and the other simultaneously constructed bowers to court females in the sand. We detected no difference in GSI (gonadal somatic index) between F1 males behaving as ‘socially rock’ vs. ‘socially sand.’ This observation of divergent behavior among interacting F1 brothers suggests an interaction between the genome and the social environment in these males.

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Danio rerio

Project description:Effects of agouti overexpression in the brain and skin transcriptome of zebrafish

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Seminal Fluid Proteins of cactophilic Drosophila

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Differentially Expressed Genes in the Skin of Tumor Susceptible K14-Agouti Transgenic Mice

Project description:The mouse agouti gene is normally expressed in the skin and regulates coat color pigmentation. Dominant regulatory mutations in the agouti gene, such as viable-yellow (Avy), cause ubiquitous over-expression of the wild-type gene product in every tissue in the body. As a result of this altered agouti expression, viable-yellow mice have solid yellow hair, they are obese and diabetic, and they have increased susceptibility to spontaneous and chemically induced tumors in a wide variety of tissues (e.g., skin, lung, liver, mammary gland and urinary bladder). Additionally, liver-specific expression of the agouti gene was shown to promote hepatocellular carcinogenesis, even in the absence of obesity and diabetes. Our lab recently extended these findings to show that the agouti gene also promotes skin cancer in the absence of obesity and diabetes. In K14-Agouti transgenic mice, the wild-type agouti gene is over-expressed in the skin under the regulatory control of the keratin 14 (K14) promoter. Over-expression of agouti in the skin results in yellow coat color without the obesity and diabetes. In two-stage skin carcinogenesis experiments the agouti protein was shown to act as a tumor promoter since it promotes the development of skin tumors in the K14-Agouti transgenic mice, but only after the skin was initiated with a single dose of 7, 12-dimethylbenz[a]anthrance (DMBA). These experiments set the stage for determining the mechanism of action of agouti protein in tumor promotion, which remains completely uncharacterized. As a first step in determining the role of agouti in skin cancer, we used cDNA microarray and quantitative real time polymerase chain reaction (QRT-PCR) analysis to identify and validate a set of differentially expressed genes in the skin of K14-Agouti transgenic mice at the promotion stage of carcinogenesis. Keywords: promotion stage of carcinogenesis, biological replicates,

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