In Vivo Analysis of Optic Fissure Fusion in Zebrafish: Pioneer Cells, Basal Lamina, Hyaloid Vessels, and How Fissure Fusion is Affected by BMP

• Published in: International journal of molecular sciences 2020-04, Vol.21 (8), p.2760
• Main Authors: Eckert, Priska, Knickmeyer, Max D, Heermann, Stephan
• Format: Article
• Language: English
• Subjects: Animals; Animals, Genetically Modified - metabolism; Basal lamina; Basement Membrane - metabolism; Blindness; Blood Vessels - anatomy & histology; BMP; Bone morphogenetic proteins; Bone Morphogenetic Proteins - genetics; Bone Morphogenetic Proteins - metabolism; Cell fusion; Coloboma - metabolism; Coloboma - pathology; Cytology; Growth factors; hyaloid vessel; Localization; Markers; Mesenchyme; Morphogenesis; Morphology; Optic Disk - abnormalities; optic fissure; pioneer cells; POM; Retina; Retinal Pigment Epithelium - cytology; Retinal Pigment Epithelium - metabolism; Stem cells; Time-Lapse Imaging; Veins; Veins & arteries; Zebrafish; Zebrafish - metabolism; Zebrafish Proteins - genetics; Zebrafish Proteins - metabolism
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms21082760

Colobomata, persistent optic fissures, frequently cause congenital blindness. Here, we focused on optic fissure fusion using in vivo time-lapse imaging in zebrafish. We identified the fusion initiating cells, which we termed "pioneer cells." Based on morphology, localization, and downregulation of the neuroretinal (NR) precursor marker , these cells could be considered as retinal pigment epithelial (RPE) progenitors. Notably, pioneer cells regain expression and integrate into the NR after fusion, indicating that they do not belong to the pool of RPE progenitors, supported by the lack of RPE marker expression in pioneer cells. They establish the first cellular contact between the margins in the proximal fissure region and separate the hyaloid artery and vein. After initiation, the fusion site is progressing distally, increasing the distance between the hyaloid artery and vein. A timed BMP (Bone Morphogenetic Protein) induction, resulting in coloboma, did not alter the morphology of the fissure margins, but it did affect the expression of NR and RPE markers within the margins. In addition, it resulted in a persisting basal lamina and persisting remnants of periocular mesenchyme and hyaloid vasculature within the fissure, supporting the necessity of BMP antagonism within the fissure margins. The hampered fissure fusion had severe effects on the vasculature of the eye.