Subconjunctival Lymphatics Respond to VEGFC and Anti-Metabolites in Rabbit and Mouse Eyes.
Subconjunctival Lymphatics Respond to VEGFC and Anti-Metabolites in Rabbit and Mouse Eyes.
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DOI：
10.1167/iovs.63.10.16
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发表时间：
2022-09-01
影响因子：
4.4
通讯作者：
中科院分区：
文献类型：
--
作者：
表征兔眼和小鼠眼的结膜下炎症并对其产生影响。家兔接受结膜下注射台盼蓝或可固定荧光葡聚糖。量化了滤过泡相关的流出途径。在天然兔结膜中和注射可固定荧光葡聚糖后，对血管特异性标志物（血小板[podoplanin和LYVE-1]和血管[CD 31]）进行免疫荧光。将血管内皮细胞生长因子-C（VEGFC）结膜下注射于家兔。采用RT-PCR和Western blot检测上述标志物的mRNA和蛋白质。或者，小鼠研究使用Prox 1-tdTomato转基因报告小鼠。结膜下注射条件包括：不注射、平衡盐溶液（BSS）、VEGFC、5-氟尿嘧啶（5 FU）和两种浓度的丝裂霉素C（MMC）。进行了具有不同随访时间和注射次数的两种小鼠注射方案（短和长）。将小鼠眼睛摘除，建立扁平标本，并评估结膜下分支和长度。兔眼表现出清晰的滤过泡相关结膜下流出途径，与血管不同，无鼻/颞偏好。血管特异性标记物的免疫荧光显示兔结膜中存在血管和毛细血管，这些血管与滤过泡相关的结膜下流出途径重叠。结膜下VEGFC增加淋巴管（P = 0.004-0.04）而不是血管（P = 0.77-0.84）mRNA或蛋白。Prox 1-tdTomato转基因报告小鼠表现出天然荧光染色。结膜下VEGFC增加了小鼠淋巴分支和长度（P ≤ 0.001-0.004），而抗代谢物（P ≤ 0.001-0.043）则与长方案相反。在使用不同方法研究的两种动物模型中，结膜下抗炎药对VEGFC和抗代谢药均无反应。这些结果可能对滤过泡形成性青光眼手术或眼部药物递送具有重要意义。
To characterize and pharmacologically influence subconjunctival lymphatics in rabbit and mouse eyes. Rabbits received subconjunctival injections of trypan blue or fixable fluorescent dextrans. Bleb-related outflow pathways were quantified. Immunofluorescence for vessel-specific markers (lymphatics [podoplanin and LYVE-1] and blood vessels [CD31]) were performed in native rabbit conjunctiva and after fixable fluorescent dextran injection. Vascular endothelial cell growth factor-C (VEGFC) was injected subconjunctivally in rabbits. mRNA and protein were assessed for the above markers using RT-PCR and Western blot. Alternatively, mouse studies used Prox1-tdTomato transgenic reporter mice. Subconjunctival injection conditions included: no injection, balanced salt solution (BSS), VEGFC, 5-fluorouracil (5FU) and two concentrations of mitomycin-C (MMC). Two mouse injection protocols (short and long) with different follow-up times and number of injections were performed. Mouse eyes were enucleated, flat mounts created, and subconjunctival branching and length assessed. Rabbit eyes demonstrated clear bleb-related subconjunctival outflow pathways that were distinct from blood vessels and were without nasal/temporal predilection. Immunofluorescence against vessel-specific markers showed lymphatics and blood vessels in rabbit conjunctiva, and these lymphatics overlapped with bleb-related subconjunctival outflow pathways. Subconjunctival VEGFC increased lymphatic (P = 0.004–0.04) but not blood vessel (P = 0.77–0.84) mRNA or protein in rabbits. Prox1-tdTomato transgenic reporter mice demonstrated natively fluorescent lymphatics. Subconjunctival VEGFC increased murine lymphatic branching and length (P ≤ 0.001–0.004) while antimetabolites (P ≤ 0.001–0.043) did the opposite for the long protocol. Subconjunctival lymphatics are pharmacologically responsive to both VEGFC and antimetabolites in two animal models studied using different methodologies. These results may be important for bleb-forming glaucoma surgeries or ocular drug delivery.
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影响因子：
5.1
作者：
Narayanaswamy, Arun;Thakur, Sahil;Nongpiur, Monisha E.;Schmetterer, Leopold;Hong, Young-Kwon;Huang, Alex S.;Wong, Tina T.
通讯作者：
Wong, Tina T.
影响因子：
2
作者：
Bouhenni, Rachida A.;Al Jadaan, Ibrahim;Edward, Deepak P.
通讯作者：
Edward, Deepak P.
影响因子：
3.4
作者：
Akiyama, Goichi;Saraswathy, Sindhu;Huang, Alex S.
通讯作者：
Huang, Alex S.
影响因子：
3
作者：
Huang AS;Saraswathy S;Dastiridou A;Begian A;Legaspi H;Mohindroo C;Tan JC;Francis BA;Caprioli J;Hinton DR;Weinreb RN
通讯作者：
Weinreb RN
影响因子：
2.7
作者：
Oh, SJ;Jeltsch, MM;Wilting, J
通讯作者：
Wilting, J