Inducible Slc4a11 Knockout Triggers Corneal Edema Through Perturbation of Corneal Endothelial Pump.
Inducible Slc4a11 Knockout Triggers Corneal Edema Through Perturbation of Corneal Endothelial Pump.
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可诱导的Slc4a11基因敲除通过角膜内皮泵的扰动触发角膜水肿。
DOI：
10.1167/iovs.62.7.28
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发表时间：
2021-06-01
影响因子：
4.4
通讯作者：
Bonanno JA
中科院分区：
文献类型：
--
作者：
Ogando DG;Shyam R;Kim ET;Wang YC;Liu CY;Bonanno JA
常规的Slc 4a 11敲除（KO）在睁眼时显示出显著的角膜水肿，这一事实使导致水肿的初始事件的研究复杂化。诱导型KO将提供检查Slc 4a 11活性丧失后的早期事件的机会。 将Slc 4a 11 Flox（SF）小鼠与表达雌激素受体Cre蛋白酶融合蛋白的小鼠杂交，并喂食他莫昔芬（Tm）两周。通过OCT测量角膜厚度（CT）。在8周终点，测定氧化损伤、紧密连接完整性、基质乳酸浓度、内皮通透性、差异表达的转运蛋白和连接蛋白。单独地，还检查了仅角膜细胞可诱导的Slc 4a 11 KO。在Tm诱导后四周，Slc 4a 11转录物水平为对照的2%。八周后，角膜厚度逐渐增加，比野生型（WT）厚50%，内皮形态显着改变，硝基酪氨酸染色增加，基质乳酸显着升高，乳酸转运蛋白表达和Na-K ATP酶活性降低，ATP升高，紧密连接和粘附连接的表达改变，荧光素渗透性增加。WT和仅角膜细胞Slc 4a 11 KO之间的CT没有发现显著差异。Slc 4a 11诱导型KO显示出与常规KO相似的表型的发展，从而验证了该模型并提供了进一步用于通过使用非侵入性体内生理探针来检查细胞事件序列的工具。
The conventional Slc4a11 knockout (KO) shows significant corneal edema at eye opening, a fact that complicates the study of the initial events leading to edema. An inducible KO would provide opportunities to examine early events following loss of Slc4a11 activity. Slc4a11 Flox (SF) mice were crossed with mice expressing the estrogen receptor Cre Recombinase fusion protein and fed tamoxifen (Tm) for two weeks. Corneal thickness (CT) was measured by OCT. At eight weeks endpoint, oxidative damage, tight junction integrity, stromal lactate concentration, endothelial permeability, differentially expressed transporters, and junction proteins were determined. Separately, a keratocyte only inducible Slc4a11 KO was also examined. At four weeks post-Tm induction Slc4a11 transcript levels were 2% of control. Corneal thickness increased gradually and was 50% greater than Wild Type (WT) after eight weeks with significantly altered endothelial morphology, increased nitrotyrosine staining, significantly higher stromal lactate, decreased expression of lactate transporters and Na-K ATPase activity, higher ATP, altered expression of tight and adherens junctions, and increased fluorescein permeability. No significant differences in CT were found between WT and keratocyte only Slc4a11 KO. The Slc4a11 inducible KO shows development of a similar phenotype as the conventional KO, thereby validating the model and providing a tool for further use in examining the sequence of cellular events by use of noninvasive in vivo physiological probes.
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影响因子：
3.7
作者：
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影响因子：
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影响因子：
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作者：
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通讯作者：
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影响因子：
5.2
作者：
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影响因子：
3.5
作者：
Vilas GL;Loganathan SK;Liu J;Riau AK;Young JD;Mehta JS;Vithana EN;Casey JR
通讯作者：
Casey JR