PrP genotype and lamb birth weight in a scrapie-free environment: Is there an association?

The National Scrapie Plan (NSP) for Great Britain includes selective breeding programmes. These aim to reduce the frequencies of the ovine prion protein (PrP) genotypes at high risk of developing clinical disease (principally those containing the VRQ allele) within the national flock, and select for the low-risk ARR-containing genotypes. The aim of this work was to use available data from embryo transfer programmes in guaranteed scrapie-free environments to investigate whether there was a difference in lamb birth weight associated with the lamb's PrP genotype, which might affect survival. The first analysis utilised data from 355 Cheviot lambs born between 2001 and 2004. The analysis was then repeated using a larger data set of 737 Cheviot, Dorset and Suffolk lambs born between 1999 and 2003. The most important determinants of mean lamb birth weight were litter size, sex and recipient ewe breed, plus lamb breed in the three breed analysis. In both analyses the effects of all these variables are consistent and of the same order of magnitude. In the first analysis there was evidence for an apparent increase in mean lamb birth weight for the ARR-containing genotypes of 0.3 kg (95% C.I. 0.1–0.5 kg). Whereas, in the second data set there was evidence for a decrease in mean lamb birth weight for the VRQ-containing genotypes of − 0.6 kg (95% C.I. − 0.8 to − 0.4 kg). This had been masked in the first analysis by the categorisation of these genotypes with the ARQ/ARQ genotype. Within these flocks, in a scrapie-free environment, neither finding provides support for concerns that selection for low risk ARR-containing genotypes would be detrimental to lamb birth weight, which is considered to be the most important determinant of lamb survival. Extension of this work into field flocks, where scrapie-status cannot be easily assured, will require the collection of sufficient data to adjust for known direct effects as well as any additional potential confounding effects.     

实时荧光定量PCR检测PrP基因在金黄地鼠脑组织的动态表达

金黄地鼠是研究动物传染性海绵状脑病的理想模型动物之一,其脑组织内朊蛋白基因动态表达数据的测定对探讨该类疾病的发生、发展和分子致病机理具有重要意义。我们利用实时荧光定量RT-PCR技术,对不同年龄金黄地鼠大脑、小脑、丘脑和脑干PrP基因的表达进行了定量。结果发现,脑的四个检测部位都呈现高的表达量,但是同一年龄段不同组织每纳克总RNA中朊蛋白基因的表达量和每毫克组织中朊蛋白基因的表达量有显著的差别,不同组织在不同年龄出现表达高峰。本研究的结果对于探讨朊蛋白的基本功能和脑组织在传染性海绵状脑病病理发生中的作用,提供了基础数据。     

Fate of prions in soil: Degradation of recombinant prion in aqueous extracts from soil and casts of two earthworm species

Prions represent the active agent in transmissible spongiform encephalopathy (TSE) diseases and can remain infective to mammals even after prolonged periods in soil. The influence of mesofauna on prion dispersal and degradation in soil, however, remains unknown. In this study the effect of earthworms on the retention/dissemination of TSEs in soil was evaluated using a model recombinant prion protein (recPrP) and aqueous extracts from soil and fresh casts of two earthworm species, Lumbricus terrestris and Aporrectodea caliginosa. Our results showed that earthworm gut-derived enzymes did not enhance the degradation of recPrP in comparison to soil, even though non-prion related proteolytic activity was higher in fresh worm excrements than in soil samples. Complete degradation of recPrP occurred in the aqueous extracts from all samples within up to 6 days at +15 °C. The proteolytic enzymes responsible for degrading recPrP were inhibited by aprotinin and leupeptin and studies in pure cultures suggested these were most probably of soil microbial origin.     

PrPc重组蛋白脑内接种金黄地鼠对mRNA表达的影响

为明确将牛PrPc重组蛋白接种金黄地鼠脑内是否引起异常临床表现、脑组织病理学变化及对其mRNA表达产生影响,为进一步研究PrPc蛋白与异构体PrPsc 的结构转换机制提供基础数据,将纯化的牛PrPc重组蛋白磷酸盐缓冲液制剂进行地鼠颅内接种,约3 μL/只,对照接种磷酸盐缓冲液（PBS）;102 d后取出脑组织：一部分福尔马林固定后进行常规H.E.染色观察, 另一部分提取脑组织总RNA,进行实时荧光定量RT-PCR检测.结果表明：处理未见临床异常表现;大脑、小脑、脑干组织病理学检测未见海绵状空泡变性和淀粉样斑;大脑PrPc mRNA表达水平无显著性差异.上述结果表明,用PrPc重组蛋白接种,在检测时间102 d内未引起金黄地鼠行为和脑组织的异常改变.     

Interactions of prion proteins with soil

Prions, are proteinaceous particles recognized as the agents of a class of neurodegenerative disorders, called transmissible spongiform encephalopathies (TSE), or prion diseases. Epidemiological data suggest that TSE-contaminated environments may serve as source of infectivity, but there is no information about adsorption of prions onto soil. We carried out experiments by mixing, healthy, or scrapie-infected hamster brains homogenates with three types of soil suspended in different buffers: (i) two saline buffers, i.e., phosphate buffer solution (PBS) and CaCl₂ solution; (ii) a mix of nondenaturing detergents, i.e., Triton X-100 and sodium deoxycholate (DOC) solution; (iii) a non-ionic detergent, i.e., lauryl maltoside; (iv) two anionic detergents, i.e., Sarkosyl or sodium dodecyl sulphate (SDS); and (v) a chaotropic agent, i.e., urea. The unbound prion proteins were detected in the supernatants (after centrifugation of soil suspension) by Western blotting. Results clearly demonstrate that both the no infectious (PrP^C) and infectious (PrP^Sc) forms are adsorbed by all soils. Only 1% sodium dodecylsulphate (SDS) partially impeded the association of PrP^C, but not that of PrP^Sc with the sandy loam soil. Agents with different interacting properties towards hydrophilic and/or hydrophobic domains failed to extract PrP^Sc from sediments of soil-brain homogenate mixtures. The strong interaction of PrP^Sc with soil favors the accumulation of prions in soils, especially if amended with prion-containing organic fertilizers and/or whenever TSE-affected animal carcasses, placenta, and excreta in general are buried or laid at the soil surface.     

Clinical and pathological features of experimental scrapie in Irish Blackface Mountain sheep

There have been no reports of natural scrapie in Irish Blackface Mountain (BM) sheep which account for approximately 16% of the Irish national sheep flock. The aim of this study was to determine if Irish BM sheep had unusual clinical and/or pathological features of scrapie which would account for failure to diagnose the disease in this breed. BM (n=7), Texel (n=3) and Suffolk sheep (n=1) of scrapie-susceptible PrP genotypes (ARQ/ARQ and VRQ/ARQ) were orally challenged with scrapie-infected brain inoculum. The incubation period, clinical signs, pathology and distribution of disease specific prion protein (PrP(d)) in scrapie-affected BM sheep were similar to scrapie in the Texel and Suffolk sheep. It was concluded that there was no evidence to suggest that scrapie in BM sheep differs clinicopathologically from scrapie in other breeds of sheep.     

表观遗传学的分子机制及其研究进展

表观遗传学(epigenetics)是指不涉及DNA序列改变、可以通过有丝分裂和减数分裂进行遗传的基因表达变化的遗传学分支领域。目前研究主要集中在DNA甲基化、组蛋白密码、染色质重塑和非编码RNA调控等方面。副突变、亲代基因印记、性别相关性基因剂量补偿效应和转基因沉默等都是典型的表观遗传现象。相关研究有利于揭示生物生长发育、多倍体植物基因组进化、杂种优势以及人类疾病等许多生命现象的本质。     

疯牛病发病机理和跨物种传播研究进展

疯牛病（BSE）是严重危害畜牧业和人类健康的一种传染性疾病。该病以侵害中枢神经系统为特征。由于该病的临床症状和组织病理学变化等已为人们所熟知，本文仅就疯牛病病因、发病机理、及跨物种传播作一综述。     

牛海绵状脑病与牛羊源成分的检测

疯牛病不仅给全球畜牧经济造成重大损失,而且还严重危害着人类的健康,而造成疯牛病传播的主要原因则是由于携带有致病因子的牛羊肉骨粉饲料及牛羊制品在各国之间的贸易往来。因而加强对进口饲料产品中牛羊源成分的检测是防止疯牛病流行的重要措施。本文就疯牛病的流行病学、病原学、病理变化与诊断以及牛羊源成分检测方法的研究进展作以简述。     

Increased GH secretion in scrapie,a prion-associated neurodegenerative disease,is not due to suppressed IGF-1 negative feedback

GH secretion is increased in scrapie-diseased sheep. Although the role of the somatotropic axis as a neurotrophic and neuroprotective factor is well documented, no studies have been carried out on the mechanisms and functional significance of somatotropic perturbation in the pathophysiology of prion-associated neurodegenerative disease. The goal of this study was to test the hypothesis that increased GH secretion observed in a natural animal prion disease, scrapie, might reflect a general lack of action of IGF-1 and, more particularly, a suppressed IGF-1 negative feedback. The effect of human recombinant IGF-1 (rhIGF-1) on spontaneous and GHRH-induced secretions was studied in so-called “scrapie-resistant” and “scrapie sensitive” rams in vivo and in vitro on pituitary dissociated cells from both groups. The effect of rhIGF-1 infusion on spontaneous and GHRH-induced GH secretions was evaluated during the preclinical and clinical stages of the disease in vivo. Our results indicated that rhIGF-1 suppressed spontaneous GH secretion but not GHRH-induced secretion in vivo. RhIGF-1 had no effect on spontaneous and GHRH-induced GH secretion from dissociated pituitary cells. Clinical scrapie was associated with a significantly greater rhIGF-1-induced inhibition of GH spontaneous secretion (mean ± S.E.M. inhibition of GH secretion: 31 ± 8% vs. 45 ± 4% in control and scrapie-affected rams, respectively). It can be concluded that the increase in GH secretion in scrapie-affected animals does not reflect a global lack of action of IGF-1. Further investigations are required to determine if other IGF-1 effects and more particularly neuroprotective mechanisms are altered in prion-associated neurodegenerative diseases.