Assembly and Annotation of the Starfish Egg Transcriptome: The Search for the Elusive Molecular Controllers of Fertilization

Abstract

Fertilization is the process of sperm‐egg interaction leading to a permanent block to polyspermy and re‐initiation of the cell cycle. Release of Ca2+ from the endoplasmic reticulum is the integral molecular mechanism responsible for activation of fertilization; however, the molecular players involved in the initiation and regulation of the Ca2+ release are not well‐established in any organism. Research in the fertilization field has been hampered due to difficulty in studying the sperm‐egg cell system. Eggs and sperm of the starfish Patiria miniata present an attractive model to study the molecular aspects of fertilization due to the ease of obtaining millions of viable sperm and eggs, as well as, their amenability to in vitro fertilization and biochemical assays. However, the lack of specific antibodies for the identification of proteins in the starfish model system has delayed discovery. To address this deficiency, we produced the first P. miniata egg transcriptome by de novo RNA sequencing, which will allow us to predict which proteins could be present and function in controlling the fertilization signaling pathway. The goal of this project is to annotate the starfish egg transcriptome and categorize the transcripts into known protein families. We present evidence for the presence of all major signaling pathways and confirm biologically‐relevant RNAs via RT‐PCR against freshly isolated starfish egg RNA. This work will be used to establish a potential proteome for the identification of proteins that may function in the fertilization signaling pathway.Support or Funding InformationNational Institutes of Health R15This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.