Kaser-Eichberger Alexandra, Schrödl Falk, Trost Andrea, Strohmaier Clemens, Bogner Barbara, Runge Christian, Motloch Karolina, Bruckner Daniela, Laimer Martin, Schlereth Simona L, Heindl Ludwig M, Reitsamer Herbert A
University Clinic of Ophthalmology and Optometry Research Program for Experimental Ophthalmology and Glaucoma Research, SALK, Salzburg, Austria.
University Clinic of Ophthalmology and Optometry Research Program for Experimental Ophthalmology and Glaucoma Research, SALK, Salzburg, Austria 2Department of Anatomy, Paracelsus Medical University, Salzburg, Austria.
Invest Ophthalmol Vis Sci. 2015 Jul;56(8):4943-53. doi: 10.1167/iovs.15-16573.
Reports of lymphatics in the anterior human uvea are contradictory. This might be caused due to a certain topography, which has not been considered yet. Therefore, here we systematically analyze iris and adjacent ciliary body with immunohistochemistry by combining various lymphatic markers.
Human iris and ciliary body were obtained from cornea donors and prepared for cryosectioning. Cross sections of tissue blocks at 12/3/6/9 o'clock position and at corresponding intersections (1:30/4:30/7:30/10:30) were processed for immunohistochemistry of LYVE-1, PDPN, PROX1, FOXC2, VEGFR3, and CCL21, and when necessary, these lymphatic markers were combined with CD31, α-smooth muscle-actin, CD68, and 4',6-diamidino-2 phenylindole dihydrochloride (DAPI). Double, triple, and quadruple marker combinations were documented using confocal microscopy.
Numerous podoplanin+ cells were mainly located at the anterior border of the iris while LYVE-1+ cells were distributed throughout the nonpigmented part. Both cell populations were PROX1/FOXC2/CCL21/VEGFR3-. Blood vessels, iris smooth muscles, and individual cells were VEGFR3+. While PDPN+ cells were rarely detected posteriorly of the iris root, many LYVE-1+ cells were present within the ciliary body muscle and villi. Within the muscle, occasionally PDPN+ vessel-like structures were detectable, but these were never colocalized with LYVE-1. Similar vessel-like structures were VEGFR3+/PROX1-/CCL21-, but CD31+. Further, ciliary muscle fibers and ciliary epithelium were immunoreactive for VEGFR3/CCL21, but were LYVE-1/PDPN-. A certain topography of structures at the various uvea-positions investigated was not obvious. The majority of LYVE-1+ cells displayed immunoreactivity for CD68.
Lymphatic vessels colocalizing for at least two lymphatic markers were not detectable. Therefore, if present, putative lymphatic channels of the anterior uvea might display a different marker panel than generally presumed.
关于人类眼前葡萄膜中淋巴管的报道相互矛盾。这可能是由于某种尚未被考虑到的组织结构所致。因此，我们在此通过结合多种淋巴管标志物，运用免疫组织化学方法对虹膜和相邻睫状体进行系统分析。
从角膜捐献者获取人类虹膜和睫状体，并制备用于冷冻切片。在12/3/6/9点钟位置以及相应交叉点（1:30/4:30/7:30/10:30）对组织块进行横断面切片，用于LYVE-1、PDPN、PROX1、FOXC2、VEGFR3和CCL21的免疫组织化学检测，必要时，将这些淋巴管标志物与CD31、α-平滑肌肌动蛋白、CD68以及4',6-二脒基-2-苯基吲哚二盐酸盐（DAPI）联合使用。使用共聚焦显微镜记录双标记、三标记和四标记组合情况。
大量血小板内皮细胞黏附分子（PDPN）阳性细胞主要位于虹膜前缘，而淋巴管内皮透明质酸受体1（LYVE-1）阳性细胞分布于整个无色素部分。这两种细胞群体均为PROX1/FOXC2/CCL21/VEGFR3阴性。血管、虹膜平滑肌和单个细胞为VEGFR3阳性。虽然在虹膜根部后方很少检测到PDPN阳性细胞，但在睫状体肌和绒毛中有许多LYVE-1阳性细胞。在肌肉内，偶尔可检测到PDPN阳性的血管样结构，但这些结构从未与LYVE-1共定位。类似的血管样结构为VEGFR3阳性/PROX1阴性/CCL21阴性，但CD31阳性。此外，睫状肌纤维和睫状体上皮对VEGFR3/CCL21呈免疫反应，但对LYVE-1/PDPN呈阴性。在所研究的不同葡萄膜位置，结构的特定组织结构并不明显。大多数LYVE-1阳性细胞对CD68呈免疫反应。
未检测到至少两种淋巴管标志物共定位的淋巴管。因此，如果存在，眼前葡萄膜的假定淋巴管可能显示出与通常推测不同的标志物组合。
