Aztiria Eugenio, Gotti Cecilia, Domenici Luciano
International School for Advanced Studies (ISAS-SISSA), 34014 Trieste, Italy.
J Comp Neurol. 2004 Dec 20;480(4):378-91. doi: 10.1002/cne.20358.
In the present paper we analyzed the expression pattern of the alpha4 and alpha7 nicotinic acetylcholine receptor (nAChR) subunits in the rat visual cortex through postnatal development, to clarify whether their expression is developmentally regulated and whether eventual developmental changes are regulated by visual experience. We found that both alpha4 and alpha7 mRNA levels accumulate from postnatal day 12 (P12) before eye opening, to around P35. The immunohistochemical results indicated that both subunits are expressed throughout all cortical laminae, except layer I. Alpha4 subunit immunohistochemistry revealed significant increments in the number of positive cells in layers V and VI after eye opening. In the case of the alpha7 subunit, the number of immunoreactive cells increased in all cortical layers soon after eye opening, except in layer VI, matching the results found at the transcriptional level. In animals reared in darkness from P9 to P22, the relative amount of the alpha4 mRNA and the number of immunoreactive cells exhibited no changes. 3H-epibatidine binding experiments showed that the number of heteromeric nAChR subunits in dark-reared rats did not change with respect to age-matched controls, thus confirming the immunohistochemical results. The mRNA of the alpha7 subunit remained stable in dark-reared rats, whereas the number and distribution of immunoreactive cells changed. Moreover, the number of 125I alphabungarotoxin-binding nAChRs was significantly increased in dark-reared animals. These results indicate that visual cortex stimulation by visual input is an essential step for alpha7 nAChR normal expression, suggesting a possible role for these receptors in an experience-dependent fashion on the maturation of this cortical area.
在本论文中,我们分析了大鼠视觉皮层中α4和α7烟碱型乙酰胆碱受体(nAChR)亚基在出生后发育过程中的表达模式,以阐明它们的表达是否受发育调控,以及最终的发育变化是否受视觉经验调控。我们发现,α4和α7的mRNA水平在出生后第12天(P12)睁眼之前开始积累,到P35左右达到高峰。免疫组织化学结果表明,除了I层外,这两个亚基在所有皮质层均有表达。α4亚基免疫组织化学显示,睁眼后V层和VI层阳性细胞数量显著增加。对于α7亚基,睁眼后除VI层外,所有皮质层免疫反应性细胞数量均增加,这与转录水平的结果相符。在从P9到P22饲养在黑暗环境中的动物中,α4 mRNA的相对含量和免疫反应性细胞数量没有变化。3H-埃博霉素结合实验表明,黑暗饲养大鼠中异源nAChR亚基的数量与年龄匹配的对照组相比没有变化,从而证实了免疫组织化学结果。α7亚基的mRNA在黑暗饲养大鼠中保持稳定,而免疫反应性细胞的数量和分布发生了变化。此外,黑暗饲养动物中125Iα-银环蛇毒素结合的nAChRs数量显著增加。这些结果表明,视觉输入对视觉皮层的刺激是α7 nAChR正常表达的关键步骤,提示这些受体可能以经验依赖的方式在该皮质区域的成熟过程中发挥作用。