■α-突触核蛋白(aSyn)是帕金森病(PD)等神经退行性疾病的关键参与者,路易体痴呆症,或多系统萎缩。Syn是表达在整个大脑,并且还可以在各种外周组织中检测到。事实上,PD的初始症状是非运动的,包括自主神经功能障碍,提示外周可能在疾病的早期发展中起重要作用。aSyn在非中央组织中的表达水平相对较低,这给其在不同组织中的检测和定量带来了挑战。
■我们的目标是通过毛细管电泳(CE)免疫印迹评估中央和外周小鼠组织中aSyn检测的灵敏度,以传统的SDS-PAGE免疫印迹为现行标准。
■提取来自野生型小鼠的中枢和非中枢来源的组织,包括中脑,内耳,食道/胃。aSyn检测通过免疫印迹使用简单的Western大小为基础的CE和SDS-PAGE进行评估。
■CE免疫印迹显示在中枢和外周组织中一致检测到aSyn。通过SDS-PAGE,免疫印迹显示出与aSyn对应的可靠信号,特别是在膜固定之后。
■我们的结果表明,使用CESimpleWestern免疫印迹系统可以可靠地检测中枢和外周组织中的aSyn。当旨在将CE与SDS-PAGE进行正式比较时,这些观察结果可以用作初步数据集,以及使用该技术进一步表征aSyn。
UNASSIGNED: Alpha-synuclein (aSyn) is a key player in neurodegenerative diseases such as Parkinson\'s disease (PD), dementia with Lewy bodies, or multiple system atrophy. aSyn is expressed throughout the brain, and can also be detected in various peripheral tissues. In fact, initial symptoms of PD are non-motoric and include autonomic dysfunction, suggesting that the periphery might play an important role in early development of the disease. aSyn is expressed at relatively low levels in non-central tissues, which brings challenges for its detection and quantification in different tissues.
UNASSIGNED: Our goal was to assess the sensitivity of aSyn detection in central and peripheral mouse tissues through capillary electrophoresis (CE)
immunoblot, considering the traditional SDS-PAGE
immunoblot as the current standard.
UNASSIGNED: Tissues from central and non-central origin from wild type mice were extracted, and included midbrain, inner ear, and esophagus/stomach. aSyn detection was assessed through immunoblotting using Simple Western size-based CE and SDS-PAGE.
UNASSIGNED: CE immunoblots show a consistent detection of aSyn in central and peripheral tissues. Through SDS-PAGE, immunoblots revealed a reliable signal corresponding to aSyn, particularly following membrane fixation.
UNASSIGNED: Our results suggest a reliable detection of aSyn in central and peripheral tissues using the CE Simple Western
immunoblot system. These observations can serve as preliminary datasets when aiming to formally compare CE with SDS-PAGE, as well as for further characterization of aSyn using this technique.