Title of Presentation or Performance

Investigating the Development of Blood Vessels in Avian Eyes

Major

Biology

Type of Submission

Poster

Type of Submission (Archival)

Event

Area of Study or Work

Biology

Expected Graduation Date

2023

Location

CNS Atrium, Easel 11

Start Date

4-9-2022 8:30 AM

End Date

4-9-2022 9:45 AM

Abstract

Scleral ossicles are intramembranous bones that form in a concentric ring surrounding the cornea within the eyes of many bird species and are believed to offer structural support to the eye during flight. During eye development, formation of scleral ossicles is dependent on molecular signals secreted from transient thickenings in the overlying conjunctival tissue known as conjunctival papillae. While much is known concerning how conjunctival papillae induce scleral ossicles, recent reports have suggested a separate, less-understood function for conjunctival papillae in regulating the formation and patterning of scleral blood vessels. To better understand the developmental relationship between conjunctival papillae and scleral blood vessels, we sought to observe scleral vasculogenesis in the presence and absence of conjunctival papillae. To achieve the latter, developing embryos were treated ectopically through small windows in the eggshell with the glutamine analogue DON (6-diazo-oxo-L-norleucine), which our previous studies has shown to fully disrupt conjunctival papillae induction. Of note, visualization of developing scleral blood vessels proved experimentally challenging and we outline herein three different strategies we carried out to observe scleral blood vessels and we report the varying degrees of success we had. Our approaches included injecting fluorescent highlighter ink through a major blood vessel and into the bloodstream, using a transgenic quail model that enables fluorescent detection of endothelial cells lining the vasculature and utilizing a specific monoclonal antibody that recognizes a protein on the surface of quail blood vessels. The antibody approach proved successful and enabled us to document scleral blood vessel formation throughout varying stages of development in normal, control embryos that possess conjunctival papillae and DON-treated embryos lacking conjunctival papillae. Our results show DON significantly disrupts scleral blood vessel development to an extent far-exceeding what we would anticipate the conjunctival papillae doing. This suggests DON disrupts papillae induction and vasculogenesis in distinct ways.

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Apr 9th, 8:30 AM Apr 9th, 9:45 AM

Investigating the Development of Blood Vessels in Avian Eyes

CNS Atrium, Easel 11

Scleral ossicles are intramembranous bones that form in a concentric ring surrounding the cornea within the eyes of many bird species and are believed to offer structural support to the eye during flight. During eye development, formation of scleral ossicles is dependent on molecular signals secreted from transient thickenings in the overlying conjunctival tissue known as conjunctival papillae. While much is known concerning how conjunctival papillae induce scleral ossicles, recent reports have suggested a separate, less-understood function for conjunctival papillae in regulating the formation and patterning of scleral blood vessels. To better understand the developmental relationship between conjunctival papillae and scleral blood vessels, we sought to observe scleral vasculogenesis in the presence and absence of conjunctival papillae. To achieve the latter, developing embryos were treated ectopically through small windows in the eggshell with the glutamine analogue DON (6-diazo-oxo-L-norleucine), which our previous studies has shown to fully disrupt conjunctival papillae induction. Of note, visualization of developing scleral blood vessels proved experimentally challenging and we outline herein three different strategies we carried out to observe scleral blood vessels and we report the varying degrees of success we had. Our approaches included injecting fluorescent highlighter ink through a major blood vessel and into the bloodstream, using a transgenic quail model that enables fluorescent detection of endothelial cells lining the vasculature and utilizing a specific monoclonal antibody that recognizes a protein on the surface of quail blood vessels. The antibody approach proved successful and enabled us to document scleral blood vessel formation throughout varying stages of development in normal, control embryos that possess conjunctival papillae and DON-treated embryos lacking conjunctival papillae. Our results show DON significantly disrupts scleral blood vessel development to an extent far-exceeding what we would anticipate the conjunctival papillae doing. This suggests DON disrupts papillae induction and vasculogenesis in distinct ways.