Localization of pigment epithelium derived factor (PEDF) in developing and adult human ocular tissues.
Localization of pigment epithelium derived factor (PEDF) in developing and adult human ocular tissues.
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发表时间：
2001-06
期刊：
影响因子：
2.2
通讯作者：
P. Karakousis;Sinoj K. John;K. Behling;E. Surace;Julie E. Smith;A. Hendrickson;W. Tang;J. Bennett-J.-Benn
中科院分区：
文献类型：
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作者：
P. Karakousis;Sinoj K. John;K. Behling;E. Surace;Julie E. Smith;A. Hendrickson;W. Tang;J. Bennett-J.-Benn
目的探讨色素上皮衍生因子（PEDF）在发育和成人眼部组织中的定位。方法利用PEDF 327-343氨基酸的多克隆抗体（pAb）或PEDF c -末端155氨基酸的单克隆抗体（mAb），通过免疫荧光法定位PEDF在胎儿和成人眼睛中的位置。抗体的特异性经Western blotting证实。原位杂交法在成人视网膜中定位PEDF mRNA。结果在发育中的视网膜（7.4 ~ 21.5胎周，Fwks）， pAb抗pedf标记的视网膜色素上皮（RPE）颗粒、发育中的视锥细胞、部分神经母细胞和神经节细胞层（GCL）中的许多细胞。在成人视网膜中，pAb抗pedf标记杆状细胞和视锥细胞的细胞质和细胞核，而不是视锥细胞。内皮细胞、睫状体细胞、脉络膜细胞、角膜上皮和内皮细胞、睫状体细胞也呈pAb - pedf阳性。免疫肽对pAb抗pedf的预吸附阻断了视网膜和除光感受器外的其他组织的特异性标记。与pAb抗PEDF的免疫定位一致，原位杂交在成人视网膜的RPE、光感受器、内核层细胞和神经节细胞中发现PEDF mRNA。在18周及以上的发育视网膜和成人视网膜中，mAb抗pedf标记了光感受器间基质（IPM）。使用pAb抗pedf的视网膜、角膜和睫状体/虹膜的Western印迹产生了约46 kDa的条带。使用mAb抗pedf，视网膜产生约46 kDa的条带；角膜和睫状体/虹膜在约46 kDa处有多条带状。PEDF最初被报道为RPE细胞的产物，存在于发育和成人眼睛的光感受器和视网膜内细胞类型中。光感受器和RPE可将PEDF分泌到IPM中。
PURPOSE To localize pigment epithelium-derived factor (PEDF) in developing and adult human ocular tissues. METHODS PEDF was localized in fetal and adult eyes by immunofluorescence with a polyclonal antibody (pAb) against amino acids 327-343 of PEDF, or a monoclonal antibody (mAb) against the C-terminal 155 amino acids of PEDF. Specificity of the antibodies was documented by Western blotting. PEDF mRNA was localized in adult retina by in situ hybridization. RESULTS In developing retinas (7.4 to 21.5 fetal weeks, Fwks), pAb anti-PEDF labeled retinal pigment epithelium (RPE) granules, developing cones, some neuroblasts and many cells in the ganglion cell layer (GCL). In adult retinas, pAb anti-PEDF labeled rod and cone cytoplasm and nuclei of rods but not cones. Cells in the INL and GCL, choroid, corneal epithelium and endothelium, and ciliary body were also pAb PEDF-positive. Preadsorption of pAb anti-PEDF with the immunizing peptide blocked specific labeling in retina and other tissues, except for photoreceptor outer segments. In agreement with the immunolocalization with pAb anti-PEDF, in situ hybridization revealed PEDF mRNA in the RPE, photoreceptors, inner nuclear layer cells and ganglion cells in adult retina. In developing retinas 18 Fwks and older, and in adult retinas, mAb anti-PEDF labeled the interphotoreceptor matrix (IPM). Western blots of retina, cornea, and ciliary body/iris with pAb anti-PEDF produced several bands at about 46 kDa. With mAb anti-PEDF, retina produced one band at about 46 kDa; cornea and ciliary body/iris had several bands at about 46 kDa. CONCLUSIONS PEDF, originally reported as a product of RPE cells, is present in photoreceptors and inner retinal cell types in developing and adult human eyes. Photoreceptors and RPE may secrete PEDF into the IPM.