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碱性成纤维细胞生长因子在正常大鼠和RCS大鼠成熟及发育视网膜中的免疫组织化学定位

Immunohistochemical localization of basic fibroblast growth factor in mature and developing retinas of normal and RCS rats.

作者信息

Connolly S E, Hjelmeland L M, LaVail M M

机构信息

Department of Ophthalmology, University of California School of Medicine, San Francisco 94143-0730.

出版信息

Curr Eye Res. 1992 Oct;11(10):1005-17. doi: 10.3109/02713689209033499.

Abstract

Basic fibroblast growth factor (bFGF) delays photoreceptor degeneration when injected intraocularly in Royal College of Surgeons (RCS) rats with inherited retinal dystrophy. In the present study, we have determined the localization of endogenous bFGF in retinas of normal and RCS rats during the normal developmental period (postnatal days 0-20) and the period of photoreceptor degeneration in RCS rats (days 20-90). bFGF was localized immunohistochemically by indirect immunoperoxidase using two different polyclonal antibodies and one monoclonal antibody against bFGF. bFGF was present in retinas as early as birth, and remained through adult age. Controls using either PBS, non-immune IgG or antibody preabsorbed with bFGF peptide were devoid of label. In normal rats between the ages of birth and postnatal day (P) 4, bFGF was found in developing ganglion cells, superficial blood vessels, some of the innermost cells in the neuroblastic layer, developing horizontal cells, and retinal pigment epithelial (RPE) cells. Between P0 and P4, the intensity of staining increased significantly in horizontal cells. From P6-P10, some cells in the inner nuclear layer remained positive, but horizontal cell staining became less intense in the central retina. The superficial vessels, ganglion cells and RPE cells also remained positive for bFGF. At P20-25, when the retina was essentially mature, bFGF was found in RPE cells, most cells of the ganglion cell layer, and many cells of the inner nuclear layer, but horizontal cells and blood vessels showed a lower concentration of bFGF than they did at younger ages. At P45 and older, blood vessels no longer showed bFGF immunoreactivity. The staining pattern in RCS rats was indistinguishable from that for normal rats at all ages examined. These results show that bFGF is present in the developing and adult rat retina in some neural cells, in addition to vessels and RPE cells. The transient elevated expression of bFGF immunoreactivity in developing horizontal cells and blood vessels suggests a possible role for this growth factor in retinal development. In addition, if RCS retinas possess any difference in bFGF localization or concentration compared to normal retinas, it must be too small to detect by immunohistochemical means, or at least with the reagents used.

摘要

碱性成纤维细胞生长因子(bFGF)经眼内注射到患有遗传性视网膜营养不良的皇家外科学院(RCS)大鼠体内时,可延缓光感受器变性。在本研究中,我们确定了正常发育时期(出生后0 - 20天)以及RCS大鼠光感受器变性时期(20 - 90天)正常和RCS大鼠视网膜内源性bFGF的定位。使用两种不同的抗bFGF多克隆抗体和一种单克隆抗体,通过间接免疫过氧化物酶法对bFGF进行免疫组织化学定位。bFGF在出生时就存在于视网膜中,并持续至成年期。使用PBS、非免疫IgG或用bFGF肽预吸收的抗体作为对照时均未出现标记。在出生至出生后第(P)4天的正常大鼠中,在发育中的神经节细胞、浅层血管、神经母细胞层最内层的一些细胞、发育中的水平细胞以及视网膜色素上皮(RPE)细胞中发现了bFGF。在P0至P4之间,水平细胞中的染色强度显著增加。从P6 - P10,内核层的一些细胞仍为阳性,但中央视网膜中水平细胞的染色变弱。浅层血管、神经节细胞和RPE细胞对bFGF也仍为阳性。在P20 - 25,当视网膜基本成熟时,在RPE细胞、神经节细胞层的大多数细胞以及内核层的许多细胞中发现了bFGF,但水平细胞和血管中bFGF的浓度低于较年轻时。在P45及更大年龄时,血管不再显示bFGF免疫反应性。在所检查的所有年龄段,RCS大鼠的染色模式与正常大鼠无法区分。这些结果表明,除了血管和RPE细胞外,bFGF在发育中和成年大鼠视网膜的一些神经细胞中也存在。bFGF免疫反应性在发育中的水平细胞和血管中短暂升高,提示该生长因子在视网膜发育中可能发挥作用。此外,如果RCS视网膜在bFGF定位或浓度方面与正常视网膜存在任何差异,那么这种差异必定太小,无法通过免疫组织化学方法检测到,或者至少使用所使用的试剂无法检测到。

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