Department of Biology and Salamander Genome Project
This presentation is the property of its rightful owner.
Sponsored Links
1 / 1

Department of Biology and Salamander Genome Project PowerPoint PPT Presentation


  • 74 Views
  • Uploaded on
  • Presentation posted in: General

Department of Biology and Salamander Genome Project. Department of Biology and Amphibian Growth Project. SOME OF OUR MICROARRAY RESULTS. GREEN IS “OFF GENES”; RED IS “ON” GENES.

Download Presentation

Department of Biology and Salamander Genome Project

An Image/Link below is provided (as is) to download presentation

Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author.While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server.


- - - - - - - - - - - - - - - - - - - - - - - - - - E N D - - - - - - - - - - - - - - - - - - - - - - - - - -

Presentation Transcript


Department of biology and salamander genome project

Department of Biology and Salamander Genome Project

Department of Biology and Amphibian Growth Project

SOME OF OUR MICROARRAY RESULTS

GREEN IS “OFF GENES”; RED IS “ON” GENES.

Genes listed here are those that were significant at P < 0.001, exhibited at least a two-fold change in expression when compared to controls, and also have a match to the human genome. There are many other significant patterns of gene expression that do not meet these three criteria and are subject to further analysis.

HOW A MICROARRAY WORKS

theoretically…

…and an example of ours (based on the animal on the above cover!)

28 days after TH immersion

epithelial membrane protein 1 is upregulated in this metamorphosing salamander

PCR verification of microarray

We used PCR methods as a secondary verification of our microarray results. We chose to examine keratin gene expression; these genes were an important part of the identified genes presented above and should be ideal for field diagnostics of endocrine-disrupted metamorphic development (see below). Red arrows indicate significant up-regulated metamorphic genes (“turned on genes”) and green arrows indicated down-regulated metamorphic genes (“turned off genes”).

There are approximately 4800 genes for examination on this GeneChip (or “microarray”).

AND OUR EXPERIMENT

IMMERSION IN T4

CONTROL

0 days

2 days

12 days

This is a “housekeeping” gene, i.e, it is expressed at the same levels all the time and is used as a control and baseline to evaluate up- and down-regulation of the other PCR products.

28 days

21 animals (3 per treatment), thus 21 GeneChips used for analysis.

This work is funded by

This work succeeds because of the continued work of the members of the Amphibian Growth Project: Drew Henry, Janel Richter, Ken Cabarle, Claude Ouedraego, Jeremiah Johnson, Heather Modrow, Judd Entzel and (especially) Karen Pocha-Melby.

Microarray analysis identifies keratin loci as sensitive biomarkers for thyroid hormone disruption in salamanders (Ambystoma)

Robert B. Page, James R. Monaghan, Amy K. Samuels, Jeramiah J. Smith, and Christopher K. Beachy and S. Randal Voss

THE GOAL

To begin characterization of gene expression during salamander metamorphosis

Hypothesis: no variation in gene expression during metamorphosis

For comparison to endocrine disrupted metamorphosis

To identify loci that will be useful in field diagnostics of normal and endocrine disrupted metamorphosis

To correlate patterns of gene expression with morphological, developmental, and life history malformations during endocrine disruption

THE PLAN

Basically…..

Grow axolotls (Ambystoma mexicanum)

Skin

Liver

brain

Collect mRNA

cDNA

MICROARRAY

COMPARING THE STANDARD Gene Expression Profile (GEP) WITH AN ENDOCRINE DISRUPTED GEP

The establishment of the gene expression profile during normal induction of metamorphosis now allows for a comparison to endocrine-disrupted metamorphosis. This project is ongoing; please visit this experiment at the AGP in Moore 232 and 214.

For our microarrays, we use Affymetrix GeneChips to quantify gene expression for approximately 4800 genes, many of which are homologous to human genes. This allows for a comparison to the human genome, and establishes salamander as an excellent model system to understand the role of xenobiotics in human health issues.

no TH

50 nM TH

5 nM TH

Atrazine dosage

0 days

0 g/L (ppb)

2 days

1 g/L (ppb)

12 days

5 g/L (ppb)

28 days


  • Login