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成年期进行Y形阻断或幼猫期发生斜视的猫,在关键期前后,其外侧膝状体和视皮层中Cat-301免疫反应性降低。

Pre- and post-critical period induced reduction of Cat-301 immunoreactivity in the lateral geniculate nucleus and visual cortex of cats Y-blocked as adults or made strabismic as kittens.

作者信息

Yin Zheng Qin, Crewther Sheila G, Wang Chun, Crewther David P

机构信息

Southwest Eye Hospital/Southwest Hospital, Chongqing, China.

出版信息

Mol Vis. 2006 Aug 7;12:858-66.

Abstract

PURPOSE

To test the post-critical period stability of perineuronal nets by comparing the expression of antigens on aggrecan (a chondroitin sulfate proteoglycan (CSPG) recognized by the monoclonal antibody Cat-301) in the lateral geniculate nucleus (LGN) and striate cortex (A17) of adult Y-blocked cats and cats made strabismic and amblyopic as kittens. The comparison tested the idea that pre- and post-critical period loss of synchronous activity would differentially affect the perineuronal net of Y-type neurons in LGN and A17.

METHODS

Seven adult cats, two normal, three convergent strabismic amblyopic, and two monocularly Y-blocked cats, were used in this study. The strabismic amblyopic cats had been made monocularly esotropic (by tenotomy) at 14 days of age. The Y-block was created acutely by a pressure cuff placed on the optic nerve behind the left eye in adult cats. The efficacy of both procedures was tested electrophysiologically. Frontal frozen sections were incubated with the Cat-301 antibody and the labeling visualized using a DAB kit. The sections were counterstained with cresyl violet. In each section, Cat-301-stained cells with well-defined nucleoli were counted under a 20x objective with a computer-based quantitative microscope image analysis system.

RESULTS

The percentage of positively labeled cells was reduced in LGN laminae that received input from the deviated eye in amblyopic cats and from the pressure-blocked eye in Y-blocked cats compared with normal cats. Surprisingly, the non-blocked laminae of the Y-blocked cats also showed a significant reduction in positively labeled neurons when compared to normals or to strabismic cats. In the visual cortex of both hemispheres of strabismic and Y-blocked cats, the density of immunopositive neurons was significantly reduced compared with normal. The effect was most pronounced in layers IV-VI for Y-blocked cats and in layer IV for strabismic amblyopic cats.

CONCLUSIONS

The results demonstrate that surface expression of aggrecan in adult cat LGN and A17 of adult cat is reduced both by chronic developmental loss of synchronous input from the two eyes and by acute changes in synchronous input in adulthood. Thus both pre- and post-critical plasticity in the expression of epitopes of aggrecan can be demonstrated. The uniform distribution of Cat-301 labeling tangentially within cortical layers of strabismic amblyopic cats indicates that the reduction in immunoreactivity observed with strabismus induced early in life is not simply eye-specific. Indeed, comparison of the immunopositivity of Y-blocked and strabismic animals, both in LGN and cortex, suggests that even after the critical period is ended, the physical removal of monocular Y-type afferent activity and weakening of binocular feedback connections between cortex and thalamus can alter the stability of the perineuronal nets surrounding the affected neurons.

摘要

目的

通过比较成年Y阻断猫以及幼年时形成斜视和弱视的猫的外侧膝状体核(LGN)和纹状皮质(A17)中聚集蛋白聚糖(一种被单克隆抗体Cat-301识别的硫酸软骨素蛋白聚糖(CSPG))上抗原的表达,来测试神经元周围网在关键期后的稳定性。该比较检验了以下观点:关键期前后同步活动的丧失会对LGN和A17中Y型神经元的神经元周围网产生不同影响。

方法

本研究使用了7只成年猫,其中2只正常,3只会聚性斜视弱视猫,2只单眼Y阻断猫。斜视弱视猫在14日龄时单眼内斜视(通过腱切断术)。成年猫通过在左眼后方的视神经上放置压力袖带急性造成Y阻断。两种手术的效果均通过电生理学进行测试。额部冰冻切片用Cat-301抗体孵育,并用DAB试剂盒使标记可视化。切片用甲酚紫复染。在每个切片中,使用基于计算机的定量显微镜图像分析系统,在20倍物镜下对具有清晰核仁的Cat-301染色细胞进行计数。

结果

与正常猫相比,弱视猫中接受来自偏斜眼输入的LGN层以及Y阻断猫中接受来自压力阻断眼输入的LGN层中,阳性标记细胞的百分比降低。令人惊讶的是,与正常猫或斜视猫相比,Y阻断猫的未阻断层中阳性标记神经元也显著减少。在斜视猫和Y阻断猫的双侧视觉皮质中,免疫阳性神经元的密度与正常猫相比显著降低。对于Y阻断猫,这种影响在IV - VI层最为明显;对于斜视弱视猫,在IV层最为明显。

结论

结果表明,成年猫LGN和A17中聚集蛋白聚糖的表面表达,会因双眼同步输入的慢性发育性丧失以及成年期同步输入的急性变化而降低。因此,可以证明聚集蛋白聚糖表位表达在关键期前后均具有可塑性。斜视弱视猫皮质层内Cat-301标记的切向均匀分布表明,生命早期斜视引起的免疫反应性降低并非简单的眼特异性。实际上,比较Y阻断动物和斜视动物在LGN和皮质中的免疫阳性情况表明,即使在关键期结束后,单眼Y型传入活动的物理去除以及皮质与丘脑之间双眼反馈连接的减弱,也会改变受影响神经元周围神经元周围网的稳定性。

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