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猫视觉皮层中与发育和可塑性相关的蛋白质表达模式变化:一种荧光二维差异凝胶电泳方法。

Development and plasticity-related changes in protein expression patterns in cat visual cortex: a fluorescent two-dimensional difference gel electrophoresis approach.

作者信息

Van den Bergh Gert, Clerens Stefan, Firestein Bonnie L, Burnat Kalina, Arckens Lutgarde

机构信息

Laboratory of Neuroplasticity and Neuroproteomics, Katholieke Universiteit Leuven, Leuven, Belgium.

出版信息

Proteomics. 2006 Jul;6(13):3821-32. doi: 10.1002/pmic.200500570.

Abstract

During early postnatal brain development, changes in visual input can lead to specific alteration of function and connectivity in mammalian visual cortex. In cat, this so-called critical period exhibits maximal sensory-driven adaptations around postnatal day 30 (P30), and ceases toward adulthood. We examined the molecular framework that directs age- and experience-dependent plasticity in cat visual cortex, by comparing protein expression profiles at eye opening (postnatal day 10 (P10), when experience-dependent plasticity starts), the peak of the critical period (P30), and in adulthood. Using 2-D DIGE, we performed comparisons of P10-P30 and P30-adult brain protein samples. Sixty protein spots showed statistically significant intensity changes in at least one comparison. Fifty-one spots were identified using quadrupole-TOF MS/MS or LC-MS/MS, containing 37 different proteins. The progressive increase or decrease in protein expression levels could be correlated to age-dependent postnatal brain development. Four spots containing transferrin, 14-3-3 alpha/beta and cypin, showed maximal protein expression levels at P30, thereby showing a positive correlation to critical period plasticity. Western analysis indeed revealed a clear effect of visual deprivation on cypin expression in cat visual cortex. Our results therefore demonstrate the power of 2-D DIGE as a tool toward understanding the molecular basis of nervous system development and plasticity.

摘要

在出生后早期大脑发育过程中,视觉输入的变化可导致哺乳动物视觉皮层功能和连接性的特定改变。在猫中,这个所谓的关键期在出生后第30天(P30)左右表现出最大的感觉驱动适应性,并在成年期停止。我们通过比较睁眼时(出生后第10天(P10),此时依赖经验的可塑性开始)、关键期峰值(P30)和成年期的蛋白质表达谱,研究了指导猫视觉皮层年龄和经验依赖性可塑性的分子框架。使用二维差异凝胶电泳(2-D DIGE),我们对P10 - P30和P30 - 成年大脑蛋白质样本进行了比较。60个蛋白质斑点在至少一次比较中显示出统计学上显著的强度变化。使用四极杆 - 飞行时间串联质谱(quadrupole - TOF MS/MS)或液相色谱 - 质谱联用(LC - MS/MS)鉴定了51个斑点,包含37种不同的蛋白质。蛋白质表达水平的逐渐增加或减少可能与出生后年龄依赖性大脑发育相关。含有转铁蛋白、14 - 3 - 3α/β和cypin的四个斑点在P30时显示出最大蛋白质表达水平,从而与关键期可塑性呈正相关。蛋白质免疫印迹分析确实揭示了视觉剥夺对猫视觉皮层中cypin表达的明显影响。因此,我们的结果证明了二维差异凝胶电泳(2-D DIGE)作为一种工具在理解神经系统发育和可塑性分子基础方面的强大作用。

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