Greferath Ursula, Vessey Kirstan A, Jobling Andrew I, Mills Samuel A, Bui Bang V, He Zheng, Nag Nupur, Ohtsu Hiroshi, Fletcher Erica L
Department of Anatomy and Neuroscience, The University of Melbourne, Parkville, Victoria, Australia.
Department of Optometry and Vision Sciences, The University of Melbourne, Parkville, Victoria, Australia.
PLoS One. 2014 Dec 29;9(12):e116025. doi: 10.1371/journal.pone.0116025. eCollection 2014.
The role of histamine in the retina is not well understood, despite it regulating a number of functions within the brain, including sleep, feeding, energy balance, and anxiety. In this study we characterized the structure and function of the retina in mice that lacked expression of the rate limiting enzyme in the formation of histamine, histidine decarboxylase (Hdc-/- mouse). Using laser capture microdissection, Hdc mRNA expression was assessed in the inner and outer nuclear layers of adult C57Bl6J wildtype (WT) and Hdc(-/-)-retinae. In adult WT and Hdc(-/-)-mice, retinal fundi were imaged, retinal structure was assessed using immunocytochemistry and function was probed by electroretinography. Blood flow velocity was assessed by quantifying temporal changes in the dynamic fluorescein angiography in arterioles and venules. In WT retinae, Hdc gene expression was detected in the outer nuclear layer, but not the inner nuclear layer, while the lack of Hdc expression was confirmed in the Hdc-/- retina. Preliminary examination of the fundus and retinal structure of the widely used Hdc-/- mouse strain revealed discrete lesions across the retina that corresponded to areas of photoreceptor abnormality reminiscent of the rd8 (Crb1) mutation. This was confirmed after genotyping and the strain designated Hdcrd8/rd8. In order to determine the effect of the lack of Hdc-alone on the retina, Hdc-/- mice free of the Crb1 mutation were bred. Retinal fundi appeared normal in these animals and there was no difference in retinal structure, macrogliosis, nor any change in microglial characteristics in Hdc-/- compared to wildtype retinae. In addition, retinal function and retinal blood flow dynamics showed no alterations in the Hdc-/- retina. Overall, these results suggest that histamine plays little role in modulating retinal structure and function.
尽管组胺在大脑中调节多种功能,包括睡眠、进食、能量平衡和焦虑,但它在视网膜中的作用尚未得到充分了解。在本研究中,我们对缺乏组胺形成限速酶(组氨酸脱羧酶,Hdc-/-小鼠)表达的小鼠视网膜的结构和功能进行了表征。使用激光捕获显微切割技术,评估了成年C57Bl6J野生型(WT)和Hdc(-/-)视网膜内核层和外核层中Hdc mRNA的表达。对成年WT和Hdc(-/-)小鼠的眼底进行成像,使用免疫细胞化学评估视网膜结构,并通过视网膜电图检测功能。通过量化小动脉和小静脉动态荧光血管造影中的时间变化来评估血流速度。在WT视网膜中,在外核层检测到Hdc基因表达,但在内核层未检测到,而在Hdc-/-视网膜中证实缺乏Hdc表达。对广泛使用的Hdc-/-小鼠品系的眼底和视网膜结构进行初步检查,发现整个视网膜存在离散病变,这些病变对应于类似于rd8(Crb1)突变的光感受器异常区域。基因分型后得到证实,并将该品系命名为Hdcrd8/rd8。为了确定单纯缺乏Hdc对视网膜的影响,培育了不含Crb1突变的Hdc-/-小鼠。这些动物的眼底看起来正常,与野生型视网膜相比,Hdc-/-小鼠的视网膜结构、大胶质细胞增生或小胶质细胞特征均无差异。此外,Hdc-/-视网膜的视网膜功能和视网膜血流动力学也没有改变。总体而言,这些结果表明组胺在调节视网膜结构和功能方面作用不大。