Grillo Stephanie L, Stella Salvatore L
Department of Neural and Behavioral Sciences, Penn State College of Medicine, Hershey, Pennsylvania, USA.
Neuroreport. 2018 Jan 17;29(2):118-122. doi: 10.1097/WNR.0000000000000918.
Retinal ganglion cells (RGCs) that express the photopigment melanopsin (mRGCs) are photosensitive and initiate the non-image-forming pathway, where the majority of their axons terminate in the suprachiasmatic nucleus (SCN). RGCs only make up approximately half of the cells in the ganglion cell layer of the retina; therefore, it is important to be able to distinguish them from other cell types. The transgenic Thy-1 YFP mouse line 16 (Thy-1 YFP-16) expresses yellow-fluorescent protein (YFP) in projection neurons, including RGCs. Our objective was to determine whether mRGCs are labeled with YFP in Thy-1 YFP-16 transgenic mice. Paraformaldehyde-fixed retinal wholemounts and frozen vertical sections were prepared from Thy-1 YFP-16 mice and fluorescently labeled with rabbit anti-melanopsin and guinea-pig anti-RNA binding protein with multiple splicing to identify mRGCs and total RGCs, respectively. Thy-1 YFP-16 mouse brains were sectioned coronally and imaged to view RGC axonal projections to the SCN. Confocal images of retinal preparations show that the majority (∼89%) of mRGCs are not YFP-positive in Thy-1 YFP-16 mice, where ∼11% expressed a weak fluorescent signal. In addition, there are almost no YFP-positive axons present in the SCN of coronal brain sections. We conclude that the majority of mRGC somas and axons are not labeled with YFP in the transgenic Thy-1 YFP-16 mouse line; therefore, this mouse model may not suitable for research involving mRGC visual pathways.
表达光色素黑视蛋白的视网膜神经节细胞(mRGCs)具有感光性,并启动非成像通路,其大部分轴突终止于视交叉上核(SCN)。视网膜神经节细胞仅占视网膜神经节细胞层细胞的大约一半;因此,能够将它们与其他细胞类型区分开来很重要。转基因Thy-1黄色荧光蛋白小鼠品系16(Thy-1 YFP-16)在包括视网膜神经节细胞在内的投射神经元中表达黄色荧光蛋白(YFP)。我们的目的是确定在Thy-1 YFP-16转基因小鼠中mRGCs是否用YFP标记。从Thy-1 YFP-16小鼠制备多聚甲醛固定的视网膜全层标本和冷冻垂直切片,分别用兔抗黑视蛋白和豚鼠抗多种剪接RNA结合蛋白进行荧光标记,以鉴定mRGCs和总视网膜神经节细胞。将Thy-1 YFP-16小鼠脑进行冠状切片并成像,以观察视网膜神经节细胞轴突向视交叉上核的投射。视网膜标本的共聚焦图像显示,在Thy-1 YFP-16小鼠中,大多数(约89%)的mRGCs不是YFP阳性,约11%表达微弱的荧光信号。此外,在冠状脑切片中的视交叉上核中几乎没有YFP阳性轴突。我们得出结论,在转基因Thy-1 YFP-16小鼠品系中,大多数mRGCs的胞体和轴突没有用YFP标记;因此,这种小鼠模型可能不适合用于涉及mRGC视觉通路的研究。