Characterizing the Apoptotic Role of PIG11 During the Embryonic Development of Zebrafish
Location
Alumni Hall South
Department
Biology
Abstract
Apoptosis is a complex, vital, and highly regulated process. This form of programmed cell death ensures proper sculpting of embryonic tissues, and it maintains homeostasis through natural cell turnover. Thus, unregulated apoptosis can lead to all manner of malformations, cancers, cognitive disorders, and/or other life-long disabilities. Our study focuses on a potential key regulator of apoptosis, called p53-induced gene 11 (PIG11). Its overexpression has been noted in multiple types of cancer cells undergoing apoptosis after chemotherapeutic treatment. Interestingly, transcriptional profiling of the zebrafish paralogs, pig11a and pig11b, indicate they are notably expressed up to 5dpf. Although little is known about its role in this period of development, whole mount in situ hybridization (WISH) revealed that Pig11a is spatially restricted to the nervous system beginning at somitogenesis. This suggests it could have an important role in proper neural development. To test the functional conservation of PIG11 and establish zebrafish as a model for discerning its molecular mechanisms, we are developing an inducible mosaic overexpression line using the Tol2kit and Gateway multisite recombination. We’ve engineered a vector containing a heat-shock inducible promoter (Hsp70), our genes of interest (PIG11, Pig11a, or Pig11b), and a bicistronic, nuclear localized GFP reporter (IRES:nlsGFP). A combination of immunostaining and live imaging to track the survival of GFP+ nuclei will allow us to characterize their function in a tissue-specific manner. Based on the paralogs highly conserved amino acid sequences with human PIG11, we predict that the overexpression of these genes will promote abnormal apoptosis during embryonic development in zebrafish.
Faculty Sponsor
Jorge Cantu, Northeastern Illinois University
Characterizing the Apoptotic Role of PIG11 During the Embryonic Development of Zebrafish
Alumni Hall South
Apoptosis is a complex, vital, and highly regulated process. This form of programmed cell death ensures proper sculpting of embryonic tissues, and it maintains homeostasis through natural cell turnover. Thus, unregulated apoptosis can lead to all manner of malformations, cancers, cognitive disorders, and/or other life-long disabilities. Our study focuses on a potential key regulator of apoptosis, called p53-induced gene 11 (PIG11). Its overexpression has been noted in multiple types of cancer cells undergoing apoptosis after chemotherapeutic treatment. Interestingly, transcriptional profiling of the zebrafish paralogs, pig11a and pig11b, indicate they are notably expressed up to 5dpf. Although little is known about its role in this period of development, whole mount in situ hybridization (WISH) revealed that Pig11a is spatially restricted to the nervous system beginning at somitogenesis. This suggests it could have an important role in proper neural development. To test the functional conservation of PIG11 and establish zebrafish as a model for discerning its molecular mechanisms, we are developing an inducible mosaic overexpression line using the Tol2kit and Gateway multisite recombination. We’ve engineered a vector containing a heat-shock inducible promoter (Hsp70), our genes of interest (PIG11, Pig11a, or Pig11b), and a bicistronic, nuclear localized GFP reporter (IRES:nlsGFP). A combination of immunostaining and live imaging to track the survival of GFP+ nuclei will allow us to characterize their function in a tissue-specific manner. Based on the paralogs highly conserved amino acid sequences with human PIG11, we predict that the overexpression of these genes will promote abnormal apoptosis during embryonic development in zebrafish.