Menger N, Seidenbecher C I, Gundelfinger E D, Kreutz M R
Department of Neuroanatomy, MPI for Brain Research, Frankfurt, Germany.
Cell Tissue Res. 1999 Oct;298(1):21-32. doi: 10.1007/s004419900060.
Caldendrin is a novel calcium-binding protein confined to the somatodendritic compartment of neurons. Here we have studied the expression pattern of caldendrin in the rat retina. First we assessed the distribution of caldendrin transcripts in the adult and developing retina by in situ hybridization. In the adult retina, transcripts are expressed mainly in the inner half of the inner nuclear layer (INL) and to a lesser extent in the ganglion cell layer (GCL). During development labeling of the inner part of the cytoblast layer, where amacrine cells reside, is already present at postnatal day 1 (P1). The intensity of hybridization signal in this sublamina of the developing INL increases up to P8, whereas significant labeling in the GCL was first found at P14, coinciding with eye opening. Immunodetection with a polyclonal antibody revealed intensive staining of cells in the inner retina, which are presumably mainly amacrine and significantly fewer bipolar and ganglion cells. All parvalbumin-containing All amacrines were immunopositive for caldendrin. Colocalization with calbindin was found in cone bipolar cells, the majority of AII amacrines, and calbindin-positive cells in the GCL. In the GCL, caldendrin was also colocalized with calretinin-immunopositive cells. Most caldendrin-positive amacrine cells in the adult rat retina were glycinergic and only a few were GABAergic. In retinal flat mounts, it was confirmed that less than 10% of retrogradely labeled retinal ganglion cells (RGC) are caldendrin-positive. Caldendrin immunoreactivity does not colocalize with tyrosine hydroxylase, VIP, substance P and somatostatin immunoreactivity. In summary, caldendrin expression is regulated differentially in retinal cell types during development and is restricted to a subpopulation of amacrine, bipolar, and ganglion cells, suggesting specific functions in the developing and mature retina.
钙调蛋白是一种新型的钙结合蛋白,局限于神经元的树突体部分。在此,我们研究了钙调蛋白在大鼠视网膜中的表达模式。首先,我们通过原位杂交评估了成年和发育中视网膜中钙调蛋白转录本的分布。在成年视网膜中,转录本主要在内核层(INL)的内半部表达,在神经节细胞层(GCL)中表达较少。在发育过程中,成神经细胞层内部(无长突细胞所在部位)在出生后第1天(P1)就已有标记。发育中INL的该亚层中的杂交信号强度在P8时增加,而GCL中的明显标记首先在P14时发现,与睁眼时间一致。用多克隆抗体进行免疫检测显示,视网膜内层细胞有强烈染色,这些细胞可能主要是无长突细胞,双极细胞和神经节细胞明显较少。所有含小白蛋白的无长突细胞对钙调蛋白均呈免疫阳性。在视锥双极细胞、大多数AII无长突细胞以及GCL中的钙结合蛋白阳性细胞中发现了与钙结合蛋白的共定位。在GCL中,钙调蛋白也与钙视网膜蛋白免疫阳性细胞共定位。成年大鼠视网膜中大多数钙调蛋白阳性无长突细胞是甘氨酸能的,只有少数是γ-氨基丁酸能的。在视网膜铺片中,证实逆行标记的视网膜神经节细胞(RGC)中少于 10%是钙调蛋白阳性。钙调蛋白免疫反应性与酪氨酸羟化酶、血管活性肠肽、P物质和生长抑素免疫反应性不共定位。总之,钙调蛋白的表达在发育过程中在视网膜细胞类型中受到不同调节,并且局限于无长突细胞、双极细胞和神经节细胞的一个亚群,表明其在发育中和成熟视网膜中具有特定功能。