PUBLICATION

Visualization of the Superior Ocular Sulcus during Danio rerio Embryogenesis

Authors
Yoon, K.H., Widen, S.A., Wilson, M.M., Hocking, J.C., Waskiewicz, A.J.
ID
ZDB-PUB-190416-13
Date
2019
Source
Journal of visualized experiments : JoVE   (145): (Journal)
Registered Authors
Hocking, Jennifer, Waskiewicz, Andrew
Keywords
none
MeSH Terms
  • Animals
  • Embryo, Nonmammalian/cytology
  • Embryo, Nonmammalian/physiology
  • Embryonic Development*
  • Eye/embryology*
  • Fluorescent Antibody Technique
  • Image Processing, Computer-Assisted
  • Iris/embryology*
  • Iris/physiology
  • Lens, Crystalline/embryology*
  • Lens, Crystalline/physiology
  • Mice
  • Organogenesis*
  • Retina/embryology*
  • Retina/physiology
  • Zebrafish/embryology*
PubMed
30985739 Full text @ J. Vis. Exp.
Abstract
Congenital ocular coloboma is a genetic disorder that is typically observed as a cleft in the inferior aspect of the eye resulting from incomplete choroid fissure closure. Recently, the identification of individuals with coloboma in the superior aspect of the iris, retina, and lens led to the discovery of a novel structure, referred to as the superior fissure or superior ocular sulcus (SOS), that is transiently present on the dorsal aspect of the optic cup during vertebrate eye development. Although this structure is conserved across mice, chick, fish, and newt, our current understanding of the SOS is limited. In order to elucidate factors that contribute to its formation and closure, it is imperative to be able to observe it and identify abnormalities, such as delay in the closure of the SOS. Here, we set out to create a standardized series of protocols that can be used to efficiently visualize the SOS by combining widely available microscopy techniques with common molecular biology techniques such as immunofluorescent staining and mRNA overexpression. While this set of protocols focuses on the ability to observe SOS closure delay, it is adaptable to the experimenter's needs and can be easily modified. Overall, we hope to create an approachable method through which our understanding of the SOS can be advanced to expand the current knowledge of vertebrate eye development.
Genes / Markers
Figures
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping