C. Lantz Ingersoll Project Description
Formation of skeletal structural elements during vertebrate embryogenesis is largely a conserved evolutionary process that employs numerous conserved genes and pathways. One critical gene in this process is the highly conserved vertebrate Type II Collagen alpha 1 (col2α1). It produces multiple splice isoforms that have different functions and expression patterns during skeletogenesis. Two of the primary isoforms are of great interest; variant 1, the primary embryonic isoform, and variant 2, a post embryonic isoform. While the more commonly known post-embryonic isoform has been well studied over the last 30 years, identification of a unique role for the evolutionarily conserved embryonic isoform has remained elusive, even though some evidence has suggested its importance during early development. The objective of this study is to identify whether the highly conserved embryonic isoform of col2α1 which contains a conserved von Willibrand Factor Type-C (vWFC) domain may serve a role in addition to its known structural function during development.
To investigate these potential non-structural function(s) of the conserved col2α1 during embryogenesis, we utilized the model organism Danio rerio, the zebrafish. Other laboratories have identified in other proteins containing conserved vWFC domain(s) as having a role in morphogen gradient regulation, specifically in dorsoventral patterning of developing vertebrate embryos. To investigate this possibility, we executed a three-phased plan to knockout, overexpress, and knockdown the conserved embryonic isoform that contains the conserved vWFC domain to determine its requirement during organogenesis. Our results demonstrate that the knockdown of the embryonic isoform of col2α1 results in abnormalities in formation of craniofacial cartilage and/or tail/trunk structures beginning at the time that col2α1 becomes active during embryogenesis and results in some embryos with similar phenotypes and issues potentially related to dysregulation of TGF-β morphogen gradients.
I would like to thank my thesis advisor, Dr. Rodney M. Dale for his mentorship and unending confidence in me. He has been both a passionate researcher and advisor who constantly pushed forward and motivated me to become a better researcher. I would be remiss to not also thank all of the undergraduate researchers of the Dale Laboratory who dedicated their time and effort to this project’s success; Daniel Brissette, Conrad Stasieluk, Joseph Frazzetta, Miguel Campos, Sophia Lam, Samantha Swank, Dhruti Bangaley, Hind Mothana, and Mayar Azar. I am very grateful and would also like to thank the other members of my committee, Dr. F. Bryan Pickett and Dr. Terry C. Grande, for their advice and support throughout the course of my project.