Expression of CHOP and CReP in Zebrafish Embryos

Poster Number

50

Lead Author Major

Pre-Pharmacy and Pre Denistr

Format

Poster Presentation

Faculty Mentor Name

Doug Weiser

Faculty Mentor Department

Biological Sciences

Abstract/Artist Statement

The endoplasmic reticulum, or ER, plays an important role in protein synthesis and protein folding. The accumulation of unfolded proteins induces ER stress, which can lead to apoptosis. In response, a pathway called UPR, or unfolded protein response, activates to free the ER of excess protein. UPR involves the phosphorylation of eIF-2alpha, which inhibits protein synthesis. Phosphorylated eIF-2alpha promotes CHOP, a gene important in reducing ER stress. After ER stress, two other genes, GADD34 and CReP, are involved in the eIF- 2alpha dephosphorylation to stop UPR. In humans, GADD34 is up-regulated after inducing stress while CReP remains at a constant low baseline level. While both genes have the same function, they have different expressions. This research desires to discover the exact role of CReP in ER stress pathways. To do so, CReP was expressed. RNA probes made from CReP were inserted into two types of zebrafish embryos. One was treated with thapsigargin to induce ER stress while the other remained untreated. Comparing the two embryos revealed where CReP was stimulated when the organism underwent ER stress. The same procedure was done with CHOP, the positive control. Zebrafish was chosen as the experimental organism because of its quick development period and clear embryo, allowing easy observation of the probes. In addition, they would survive the removal of CReP and GADD34. In previous experiments, mice that had both genes removed died too quickly to make any observations while the removal of just one gene did not seem to have an effect.

Location

DeRosa University Center, Ballroom

Start Date

20-4-2013 1:00 PM

End Date

20-4-2013 3:00 PM

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Apr 20th, 1:00 PM Apr 20th, 3:00 PM

Expression of CHOP and CReP in Zebrafish Embryos

DeRosa University Center, Ballroom

The endoplasmic reticulum, or ER, plays an important role in protein synthesis and protein folding. The accumulation of unfolded proteins induces ER stress, which can lead to apoptosis. In response, a pathway called UPR, or unfolded protein response, activates to free the ER of excess protein. UPR involves the phosphorylation of eIF-2alpha, which inhibits protein synthesis. Phosphorylated eIF-2alpha promotes CHOP, a gene important in reducing ER stress. After ER stress, two other genes, GADD34 and CReP, are involved in the eIF- 2alpha dephosphorylation to stop UPR. In humans, GADD34 is up-regulated after inducing stress while CReP remains at a constant low baseline level. While both genes have the same function, they have different expressions. This research desires to discover the exact role of CReP in ER stress pathways. To do so, CReP was expressed. RNA probes made from CReP were inserted into two types of zebrafish embryos. One was treated with thapsigargin to induce ER stress while the other remained untreated. Comparing the two embryos revealed where CReP was stimulated when the organism underwent ER stress. The same procedure was done with CHOP, the positive control. Zebrafish was chosen as the experimental organism because of its quick development period and clear embryo, allowing easy observation of the probes. In addition, they would survive the removal of CReP and GADD34. In previous experiments, mice that had both genes removed died too quickly to make any observations while the removal of just one gene did not seem to have an effect.