Sanders Esmond J, Parker Eve, Harvey Steve
Department of Physiology, University of Alberta, Edmonton, Alberta, Canada.
Mol Vis. 2009;15:920-6. Epub 2009 May 4.
Locally synthesized growth hormone (GH) may act as a survival factor in several tissues. Experimental studies with chick retinal ganglion cells (RGCs) suggest that GH, synthesized within the developing retina, may have autocrine or paracrine roles in the regulation of the waves of cell death characteristic of RGC differentiation. There is also evidence that GH may have a similar neuroprotective function in the rat retina, however, there is no information concerning such a role in the human retina. In this paper we extended our earlier work by determining whether the local expression of retinal GH correlates with RGC apoptosis in human retinas.
In the absence of experimental approaches to survival factor function in the human retina, we have used a correlative immunocytochemical technique to determine how the expression of GH relates to cell death in RGCs. We used sections of human retinas, taken postmortem, that we double-labeled for GH and apoptotic cell death using terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL).
We found that approximately 35% of cells in the ganglion cell layer (GCL) were both GH positive and GH receptor (GHR) positive, and that GH colocalized with GHR in these cells. However, none of the apoptotic cells in the GCL were GH immunoreactive. Labeling of sections with the RGC marker, synuclein, indicated that at least 95% of the cells in the GCL were RGCs, leading us to conclude that the majority of the cells that we have examined in the GCL are RGCs.
The results are consistent with the earlier proposal, based on in vivo and in vitro experimental chick embryo studies, that GH promotes survival in adult human RGCs.
局部合成的生长激素(GH)可能在多种组织中充当存活因子。对鸡视网膜神经节细胞(RGCs)的实验研究表明,发育中的视网膜内合成的GH可能在调节RGC分化所特有的细胞死亡波中具有自分泌或旁分泌作用。也有证据表明GH在大鼠视网膜中可能具有类似的神经保护功能,然而,关于其在人类视网膜中的这种作用尚无相关信息。在本文中,我们通过确定视网膜GH的局部表达是否与人类视网膜中的RGC凋亡相关,扩展了我们早期的工作。
由于缺乏针对人类视网膜中存活因子功能的实验方法,我们使用了一种相关免疫细胞化学技术来确定GH的表达与RGC细胞死亡之间的关系。我们使用了死后获取的人类视网膜切片,通过末端脱氧核苷酸转移酶dUTP缺口末端标记法(TUNEL)对其进行GH和凋亡细胞死亡的双重标记。
我们发现神经节细胞层(GCL)中约35%的细胞同时为GH阳性和生长激素受体(GHR)阳性,并且在这些细胞中GH与GHR共定位。然而,GCL中的凋亡细胞均无GH免疫反应性。用RGC标志物突触核蛋白对切片进行标记表明,GCL中至少95%的细胞为RGCs,这使我们得出结论,我们在GCL中检测的大多数细胞是RGCs。
这些结果与基于体内和体外鸡胚实验研究的早期提议一致,即GH促进成人人类RGCs的存活。