Martínez-Moreno Carlos, Andres Alexis, Giterman Daniel, Karpinski Edward, Harvey Steve
Department of Physiology, University of Alberta, Edmonton T6G 2H7, Canada.
Department of Physiology, University of Alberta, Edmonton T6G 2H7, Canada.
Gen Comp Endocrinol. 2014 Jan 1;195:183-9. doi: 10.1016/j.ygcen.2013.10.016. Epub 2013 Nov 13.
Retinal ganglion cells (RGCs) have been shown to be sites of growth hormone (GH) production and GH action in the embryonic (embryo day 7, ED7) chick neural retina. Primary RGC cell cultures were previously used to determine autocrine or paracrine actions of GH in the retina, but the antibody used in their immunopanning (anti-Thy-1) is no longer available. We have therefore characterized an immortalized neural retina (QNR/D) cell line derived from ED7 embryonic quail as a replacement experimental model. These cells express the GH gene and have GH receptor (GHR)-immunoreactivity. They are also immunoreactive for RGC markers (islet-1, calretinin, RA4) and neural fibers (neurofilament, GAP 43, vimentin) and they express the genes for Thy-1, neurotrophin 3 (NTF3), neuritin 1 (NRN1) and brn3 (POU4F). These cells are also electrically active and therefore resemble the RGCs in the neural retina. They are also similarly responsive to exogenous GH, which induces overexpression of the neurotrophin 3 and insulin-like growth factor (IGF) 1 genes and stimulates cell survival, as in the chick embryo neural retina. QNR/D cells are therefore a useful experimental model to assess the actions of GH in retinal function.
视网膜神经节细胞(RGCs)已被证明是胚胎期(胚胎第7天,ED7)鸡神经视网膜中生长激素(GH)产生和GH发挥作用的部位。先前曾使用原代RGC细胞培养来确定GH在视网膜中的自分泌或旁分泌作用,但用于其免疫淘选的抗体(抗Thy-1)已不再可用。因此,我们鉴定了一种源自ED7胚胎鹌鹑的永生化神经视网膜(QNR/D)细胞系,作为替代实验模型。这些细胞表达GH基因并具有GH受体(GHR)免疫反应性。它们对RGC标志物(胰岛-1、钙视网膜蛋白、RA4)和神经纤维(神经丝、GAP 43、波形蛋白)也具有免疫反应性,并且它们表达Thy-1、神经营养因子3(NTF3)、神经突蛋白1(NRN1)和brn3(POU4F)的基因。这些细胞也具有电活性,因此类似于神经视网膜中的RGCs。它们对外源性GH的反应也相似,外源性GH会诱导神经营养因子3和胰岛素样生长因子(IGF)1基因的过表达,并刺激细胞存活,就像在鸡胚神经视网膜中一样。因此,QNR/D细胞是评估GH在视网膜功能中作用的有用实验模型。