Robinson Susan W, Nugent Marie L, Dinsdale David, Steinert Joern R
MRC Toxicology Unit, Hodgkin Building, Lancaster Road, Leicester LE1 9HN, UK.
MRC Toxicology Unit, Hodgkin Building, Lancaster Road, Leicester LE1 9HN, UK Department of Genetics, University of Leicester, University Road, Leicester LE1 7RH, UK.
Hum Mol Genet. 2014 Sep 1;23(17):4581-96. doi: 10.1093/hmg/ddu171. Epub 2014 Apr 9.
The cellular prion protein (PrP(C)) has been implicated in several neurodegenerative diseases as a result of protein misfolding. In humans, prion disease occurs typically with a sporadic origin where uncharacterized mechanisms induce spontaneous PrP(C) misfolding leading to neurotoxic PrP-scrapie formation (PrP(SC)). The consequences of misfolded PrP(C) signalling are well characterized but little is known about the physiological roles of PrP(C) and its involvement in disease. Here we investigated wild-type PrP(C) signalling in synaptic function as well as the effects of a disease-relevant mutation within PrP(C) (proline-to-leucine mutation at codon 101). Expression of wild-type PrP(C) at the Drosophila neuromuscular junction leads to enhanced synaptic responses as detected in larger miniature synaptic currents which are caused by enlarged presynaptic vesicles. The expression of the mutated PrP(C) leads to reduction of both parameters compared with wild-type PrP(C). Wild-type PrP(C) enhances synaptic release probability and quantal content but reduces the size of the ready-releasable vesicle pool. Partially, these changes are not detectable following expression of the mutant PrP(C). A behavioural test revealed that expression of either protein caused an increase in locomotor activities consistent with enhanced synaptic release and stronger muscle contractions. Both proteins were sensitive to proteinase digestion. These data uncover new functions of wild-type PrP(C) at the synapse with a disease-relevant mutation in PrP(C) leading to diminished functional phenotypes. Thus, our data present essential new information possibly related to prion pathogenesis in which a functional synaptic role of PrP(C) is compromised due to its advanced conversion into PrP(SC) thereby creating a lack-of-function scenario.
细胞朊蛋白(PrP(C))因蛋白质错误折叠而与多种神经退行性疾病有关。在人类中,朊病毒疾病通常以散发性起源出现,其中未明确的机制诱导PrP(C)自发错误折叠,导致神经毒性PrP-瘙痒病形成(PrP(SC))。错误折叠的PrP(C)信号传导的后果已得到充分表征,但关于PrP(C)的生理作用及其在疾病中的参与情况知之甚少。在这里,我们研究了野生型PrP(C)在突触功能中的信号传导以及PrP(C)内与疾病相关的突变(密码子101处脯氨酸到亮氨酸的突变)的影响。在果蝇神经肌肉接头处野生型PrP(C)的表达导致突触反应增强,这在由增大的突触前囊泡引起的更大的微小突触电流中得以检测到。与野生型PrP(C)相比,突变型PrP(C)的表达导致这两个参数都降低。野生型PrP(C)增强突触释放概率和量子含量,但减小了即时可释放囊泡池的大小。部分这些变化在突变型PrP(C)表达后无法检测到。一项行为测试表明,两种蛋白质的表达都导致运动活动增加,这与突触释放增强和肌肉收缩增强一致。两种蛋白质都对蛋白酶消化敏感。这些数据揭示了野生型PrP(C)在突触处的新功能,PrP(C)中的疾病相关突变导致功能表型减弱。因此,我们的数据提供了可能与朊病毒发病机制相关的重要新信息,其中PrP(C)的功能性突触作用由于其向PrP(SC)的进一步转化而受损,从而导致功能缺失的情况。