Detection of Endoplasmic Reticulum Stress in Zebrafish.
Poster Number
6B
Format
Poster Presentation
Faculty Mentor Name
Doug Weiser
Faculty Mentor Department
Biological Sciences
Abstract/Artist Statement
The unfolded protein response (UPR) is a complex cellular response to the accumulation of unfolded or misfolded proteins in the Endoplasmic Reticulum (ER). The UPR has three primary branches, the ATF6 pathway, the PERK pathway and the IRE1 pathway. Our research focuses on two proteins called GADD34 and CReP that bind to Protein Phosphatase 1 and dephosphorylate eukaryotic initiation factor 2 alpha, which is the substrate of PERK. Thus, GADD34 and CReP act as inhibitors of the PERK pathway. For our research project, we need a marker of cellular UPR signaling. To this end we cloned the zebrafish BIP gene. BIP is a cellular chaperone that increases in expression after ER stress, thus can be used to measure overall ER stress on the cell. We isolated a fragment of the gene using PCR and topo-cloned the gene. We then subcloned the fragment into another plasmid called pCS2+. We then synthesized a ribonucleotide probe antisense to zebrafish BIP mRNA and a control probe sense to the BIP mRNA. This reagent will allow us to conduct in situ hybridization on zebrafish embryos to measure where and when ER stress is occurring.
Location
Information Commons, William Knox Holt Memorial Library and Learning Center
Start Date
29-4-2023 10:00 AM
End Date
29-4-2023 1:00 PM
Detection of Endoplasmic Reticulum Stress in Zebrafish.
Information Commons, William Knox Holt Memorial Library and Learning Center
The unfolded protein response (UPR) is a complex cellular response to the accumulation of unfolded or misfolded proteins in the Endoplasmic Reticulum (ER). The UPR has three primary branches, the ATF6 pathway, the PERK pathway and the IRE1 pathway. Our research focuses on two proteins called GADD34 and CReP that bind to Protein Phosphatase 1 and dephosphorylate eukaryotic initiation factor 2 alpha, which is the substrate of PERK. Thus, GADD34 and CReP act as inhibitors of the PERK pathway. For our research project, we need a marker of cellular UPR signaling. To this end we cloned the zebrafish BIP gene. BIP is a cellular chaperone that increases in expression after ER stress, thus can be used to measure overall ER stress on the cell. We isolated a fragment of the gene using PCR and topo-cloned the gene. We then subcloned the fragment into another plasmid called pCS2+. We then synthesized a ribonucleotide probe antisense to zebrafish BIP mRNA and a control probe sense to the BIP mRNA. This reagent will allow us to conduct in situ hybridization on zebrafish embryos to measure where and when ER stress is occurring.