de Winter Fred, Cui Qi, Symons Natalie, Verhaagen Joost, Harvey Alan R
Graduate School for Neurosciences Amsterdam, Netherlands Institute for Brain Research, Amsterdam, The Netherlands.
Invest Ophthalmol Vis Sci. 2004 Dec;45(12):4554-62. doi: 10.1167/iovs.04-0173.
Semaphorins comprise a family of molecules that influence the growth and guidance of neuronal processes. Class-3 semaphorins are secreted proteins, and their effects are mediated by neuropilin (NP) and plexin (Plx) receptors. There is considerable information on mechanisms that influence axonal guidance and plasticity in the mammalian visual system, but a role for semaphorins has received less scrutiny. The purpose of the current study was to survey class-3 semaphorin and cognate receptor expression in young and adult rat retinas.
The mRNA expression of five class-3 semaphorins (3A, 3B, 3C, 3E, and 3F) and receptor subtypes NP-1 and -2 and plexins A1 and A2 was determined, by using riboprobes and in situ hybridization on cryosections of newborn (postnatal day [P]0), juvenile (P14), and adult rat retinas. Retinal ganglion cells (RGCs) were identified by retrograde labeling after injections of a fluorescent tracer (Fluorogold, FG) into the superior colliculus. Hybridized sections were also immunostained to identify specific retinal cell classes.
mRNA expression for all five members of the class-3 semaphorin family was seen in adult FG-labeled RGCs. Qualitatively, expression was highest for semaphorins 3B and 3C, and lowest for 3A. Levels of mRNA expression in RGCs were lower in newborn retinas but were adult-like by P14. Expression by different cell types in the inner nuclear layer was also seen, especially at P14. Expression of NP-2 and PlxA2 mRNAs was evident in developing inner nuclear and ganglion cell layers at birth. Expression increased postnatally and was maintained into adulthood. NP-1 and PlxA1 expression was also present, but at comparatively lower levels.
The presence of class-3 semaphorins and their receptors in neonatal and adult rat retina suggests a potential role for these proteins in retinal development and in the maturation, stabilization, and plasticity of mammalian primary visual pathways.
信号素是一类影响神经元突起生长和导向的分子家族。3类信号素是分泌蛋白,其作用由神经纤毛蛋白(NP)和丛状蛋白(Plx)受体介导。关于影响哺乳动物视觉系统轴突导向和可塑性的机制已有大量信息,但信号素的作用受到的关注较少。本研究的目的是调查3类信号素及其同源受体在幼年和成年大鼠视网膜中的表达情况。
采用核糖探针和原位杂交技术,对新生(出生后第[P]0天)、幼年(P14)和成年大鼠视网膜的冰冻切片进行检测,以确定5种3类信号素(3A、3B、3C、3E和3F)以及受体亚型NP-1、NP-2和丛状蛋白A1、A2的mRNA表达。通过向上丘注射荧光示踪剂(荧光金,FG)进行逆行标记,以鉴定视网膜神经节细胞(RGC)。杂交切片也进行免疫染色,以识别特定的视网膜细胞类型。
在成年FG标记的RGC中可见3类信号素家族所有5个成员的mRNA表达。定性来看,信号素3B和3C的表达最高,3A的表达最低。新生视网膜中RGC的mRNA表达水平较低,但到P14时已接近成年水平。在内核层不同细胞类型中也可见表达,尤其是在P14时。出生时,NP-2和丛状蛋白A2 mRNA在发育中的内核层和神经节细胞层有明显表达。出生后表达增加,并持续到成年。NP-1和丛状蛋白A1也有表达,但水平相对较低。
3类信号素及其受体在新生和成年大鼠视网膜中的存在表明,这些蛋白在视网膜发育以及哺乳动物初级视觉通路的成熟、稳定和可塑性方面可能发挥作用。