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Organogenesis and distribution of the ocular lymphatic vessels in the anterior eye
Yifan Wu, Young Jin Seong, Kin Li, Dongwon Choi, Eunkyung Park, George H. Daghlian, Eunson Jung, Khoa Bui, Luping Zhao, Shrimika Madhavan, Saren Daghlian, Patill Daghlian, Desmond Chin, Il-Taeg Cho, Alex K. Wong, Martin Heur, Sandy Zhang-Nunes, James C. Tan, Masatsugu Ema, Tina T. Wong, Alex S. Huang, Young-Kwon Hong
Yifan Wu, Young Jin Seong, Kin Li, Dongwon Choi, Eunkyung Park, George H. Daghlian, Eunson Jung, Khoa Bui, Luping Zhao, Shrimika Madhavan, Saren Daghlian, Patill Daghlian, Desmond Chin, Il-Taeg Cho, Alex K. Wong, Martin Heur, Sandy Zhang-Nunes, James C. Tan, Masatsugu Ema, Tina T. Wong, Alex S. Huang, Young-Kwon Hong
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Research Article Development Ophthalmology

Organogenesis and distribution of the ocular lymphatic vessels in the anterior eye

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Abstract

Glaucoma surgeries, such as trabeculectomy, are performed to lower intraocular pressure to reduce risk of vision loss. These surgeries create a new passage in the eye that reroutes the aqueous humor outflow to the subconjunctival space, where the fluid is presumably absorbed by the conjunctival lymphatics. Here, we characterized the development and function of the ocular lymphatics using transgenic lymphatic reporter mice and rats. We found that the limbal and conjunctival lymphatic networks are progressively formed from a primary lymphatic vessel that grows from the nasal-side medial canthus region at birth. This primary lymphatic vessel immediately branches out, invades the limbus and conjunctiva, and bidirectionally encircles the cornea. As a result, the distribution of the ocular lymphatics is significantly polarized toward the nasal side, and the limbal lymphatics are directly connected to the conjunctival lymphatics. New lymphatic sprouts are produced mainly from the nasal-side limbal lymphatics, posing the nasal side of the eye as more responsive to fluid drainage and inflammatory stimuli. Consistent with this polarized distribution of the ocular lymphatics, a higher drainage efficiency was observed in the nasal side than the temporal side of the eye when injected with a fluorescent tracer. In contrast, blood vessels are evenly distributed at the anterior surface of the eyes. Also, we found that these distinct vascular distribution patterns were conserved in human eyes. Together, our study demonstrated that the ocular surface lymphatics are more densely present in the nasal side and uncovered the potential clinical benefits in selecting the nasal side as a glaucoma surgery site to improve fluid drainage.

Authors

Yifan Wu, Young Jin Seong, Kin Li, Dongwon Choi, Eunkyung Park, George H. Daghlian, Eunson Jung, Khoa Bui, Luping Zhao, Shrimika Madhavan, Saren Daghlian, Patill Daghlian, Desmond Chin, Il-Taeg Cho, Alex K. Wong, Martin Heur, Sandy Zhang-Nunes, James C. Tan, Masatsugu Ema, Tina T. Wong, Alex S. Huang, Young-Kwon Hong

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Figure 1

Visualization of the ocular lymphatic vessels and Schlemm’s canal.

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Visualization of the ocular lymphatic vessels and Schlemm’s canal.
(A–C)...
(A–C) Lateral view of the ocular lymphatics (arrows) and Schlemm’s canal (arrowheads) in the anterior segment of the eye of a Prox1-EGFP transgenic mouse. The limbus area in panel B is enlarged 2.3 times in panel C, where lymphatic valves (asterisks) are identified by stronger EGFP (Prox1) expression than luminal LECs. (D–F) Lyve1 whole-mount staining (red) showing the limbal lymphatics and Schlemm’s canal in the eye of a Prox1-EGFP mouse. While EGFP (Prox1) is expressed in both Schlemm’s canal (arrowhead) and the limbal lymphatics (arrows), Lyve1 is detected only in the limbal lymphatics, not in Schlemm’s canal. (G–L) Anti-Cd31 staining (green) of the limbal blood and lymphatic vessels and Schlemm’s canal in the eyes of a Prox1-tdTomato reporter mouse. Schlemm’s canal (arrowhead) and the limbal lymphatics (arrow), but not limbal blood vessels (double arrows), are selectively shown by Prox1-tdTomato reporter (red). (M–O) Anti-Vegfr3 staining (green) in the eyes of a Prox1-tdTomato reporter mouse. Vegfr3 is expressed in both the limbal lymphatics (arrow) and Schlemm’s canal (arrowhead). Adult mice (6–10 weeks old, both sexes, n = 4) were used for all panels. One representative image is shown for each panel. Co, cornea. Scale bar: 100 μm.

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