Marciano Paolo G, Brettschneider Julia, Manduchi Elisabetta, Davis Jason E, Eastman Scott, Raghupathi Ramesh, Saatman Kathryn E, Speed Terence P, Stoeckert Christian J, Eberwine James H, McIntosh Tracy K
Departments of Neuroscience, Center for Bioinformatics, University of Pennsylvania, Philadelphia, Pennsylvania 19104, USA.
J Neurosci. 2004 Mar 24;24(12):2866-76. doi: 10.1523/JNEUROSCI.5051-03.2004.
In an effort to understand the complexity of genomic responses within selectively vulnerable regions after experimental brain injury, we examined whether single apoptotic neurons from both the CA3 and dentate differed from those in an uninjured brain. The mRNA from individual active caspase 3(+)/terminal deoxynucleotidyl transferase-mediated biotinylated UTP nick end labeling [TUNEL(-)] and active caspase 3(+)/TUNEL(+) pyramidal and granule neurons in brain-injured mice were amplified and compared with those from nonlabeled neurons in uninjured brains. Gene analysis revealed that overall expression of mRNAs increased with activation of caspase 3 and decreased to below uninjured levels with TUNEL reactivity. Cell type specificity of the apoptotic response was observed with both regionally distinct expression of mRNAs and differences in those mRNAs that were maximally regulated. Immunohistochemical analysis for two of the most highly differentially expressed genes (prion and Sos2) demonstrated a correlation between the observed differential gene expression after traumatic brain injury and corresponding protein translation.
为了了解实验性脑损伤后选择性易损区域内基因组反应的复杂性,我们研究了来自CA3区和齿状回的单个凋亡神经元是否与未受伤大脑中的神经元不同。对脑损伤小鼠中单个活性半胱天冬酶3(+)/末端脱氧核苷酸转移酶介导的生物素化UTP缺口末端标记[TUNEL(-)]以及活性半胱天冬酶3(+)/TUNEL(+)的锥体神经元和颗粒神经元的mRNA进行扩增,并与未受伤大脑中未标记神经元的mRNA进行比较。基因分析表明,mRNA的总体表达随着半胱天冬酶3的激活而增加,并随着TUNEL反应性降低至未受伤水平以下。通过mRNA的区域特异性表达以及最大调控的mRNA差异,观察到凋亡反应的细胞类型特异性。对两个差异表达最高的基因(朊病毒和Sos2)进行免疫组织化学分析,结果表明创伤性脑损伤后观察到的差异基因表达与相应的蛋白质翻译之间存在相关性。