Deparment of Neurosurgery, University of Michigan, Ann Arbor, MI, USA ; Department of Neurology, The 1 Hospital of Jilin University, Changchun, Jilin, China.
Transl Stroke Res. 2013 Feb;4(1):130-4. doi: 10.1007/s12975-012-0232-3. Epub 2012 Dec 12.
Quantification of acute brain injury in basal ganglia is essential for mechanistic and therapeutic studies in experimental intracerebral hemorrhage (ICH). Using conventional counting of degenerating cells based on morphological or immunohistochemical criteria, it is hard to define the boundary of the whole lesion area. Dopamine- and cAMP-regulated phosphoprotein, Mr 32 kDa (DARPP-32) is a cytosolic protein highly enriched in medium-sized spiny neurons of the striatum. We developed new methods for quantifying lesion area by detecting the difference of the DARPP-32 negative area and the hematoma clot, and by measuring DARPP-32 protein level for semi-qualification in rat model of ICH. We found that DARPP-32 negative area around hematoma was present at day-1, peaked at day-3, and decreased at day-14 after ICH, a time course paralleled by DARPP-32 Western blots. The DARPP-32 negative area matched well with the necrotic area determined using propidium iodide. Treatment with an iron chelator, deferoxamine, attenuated the ICH-induced reduction in DARPP-32 protein levels. These results suggest that DARPP-32 is a simple and quantifiable indicator of ICH-induced neuronal death in basal ganglia.
基底节急性脑损伤的量化对于实验性脑出血（ICH）的机制和治疗研究至关重要。使用基于形态学或免疫组织化学标准的常规退化细胞计数方法，很难定义整个病变区域的边界。多巴胺和 cAMP 调节的磷蛋白，Mr32kDa（DARPP-32）是一种富含纹状体中型棘突神经元的胞浆蛋白。我们开发了新的方法来通过检测 DARPP-32 阴性区域和血肿凝块之间的差异，以及通过测量 DARPP-32 蛋白水平进行半定量，来量化 ICH 大鼠模型中的病变区域。我们发现血肿周围的 DARPP-32 阴性区域在 ICH 后第 1 天出现，在第 3 天达到峰值，并在第 14 天减少，这一时间过程与 DARPP-32 Western blot 相吻合。DARPP-32 阴性区域与使用碘化丙啶确定的坏死区域非常吻合。铁螯合剂去铁胺的治疗减轻了 ICH 诱导的 DARPP-32 蛋白水平降低。这些结果表明，DARPP-32 是基底节中 ICH 诱导的神经元死亡的简单和可量化的指标。
