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1.
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。  相似文献   

2.
为了研究鸡脑神经干细胞(neural stem cells,NSCs)体外的分离、培养及诱导分化等生物学特性,试验采用机械吹打法分离鸡脑神经干细胞,通过免疫荧光对体外培养鸡脑神经干细胞进行表面标记检测,并在适宜的诱导条件下向神经元细胞、星形胶质细胞和少突胶质细胞三个方向进行诱导,检测其多向分化潜能。结果表明:体外分离培养的鸡NSCs为目的细胞,并可向神经元细胞、星形胶质细胞和少突胶质细胞诱导分化。说明鸡脑NSCs具有良好的体外增殖能力和多向分化潜能。  相似文献   

3.
乌鸡小脑的解剖学及神经肽Y神经元分布的研究   总被引:3,自引:0,他引:3  
为了探明乌鸡小脑的形态结构及其细胞构筑和神经肽Y免疫反应神经元在小脑内的分布情况 ,采用石蜡切片HE染色和免疫组化SABC染色技术 ,对 1 0只泰和乌鸡的小脑进行了研究 ,并与北京鸭、肉鸽、本地鸡及非洲鸵鸟等禽类的相关结构进行了比较。结果表明 :①泰和乌鸡小脑皮质分 3层 ,由外至内依次为分子层、蒲肯野氏细胞层和颗粒层 ;②小脑分 1 0叶 ,Ⅰ、Ⅱ和Ⅹ为单叶 ,其余都有2个~ 3个小叶 ;③小脑中央核的神经元数量稀少 ;④神经肽Y免疫阳性神经元主要存在小脑皮质的蒲肯野氏细胞层 ,分子层、颗粒层 ,在小脑中央核未见阳性反应细胞 ;⑤泰和乌鸡小脑的形态结构及其细胞构筑与其它家禽相似 ;⑥小脑内NPY阳性神经元在蒲肯野氏细胞层的分布与鸡、肉鸽、大鼠的大体相似 ,在小脑白质内与肉鸽的有一定差异  相似文献   

4.
类器官被定义为三维(three-dimensional, 3D)多细胞体外自组装组织结构,在细胞类型、结构和功能方面能模拟还原相应的体内器官,具有明显的组织特异性,可作为新型体外模型。随着技术的发展,肠类器官已被应用于器官发育、器官功能、器官修复与移植、疾病诊断和药物筛选等多个研究领域。目前除了小鼠肠类器官外,关于其他种属动物肠类器官培养的研究较少。但在临床中,其他动物,特别是家畜动物仍存在多种传染性或非传染性的肠道疾病,亟待新型的体外模型用于揭示致病机理或开发新的治疗方案。为此,本文对近十年动物肠类器官培养的研究进展进行综述,总结了动物肠类器官培养的优、缺点并对动物肠类器官培养进行了小结与展望,以期为动物肠类器官的培养提供参考。  相似文献   

5.
支持细胞对维持精子形成过程中的微环境起决定作用,它可以通过分泌功能、细胞间连接形成的血睾屏障功能以及吞噬功能等来促进精子的形成过程,其发育异常会导致不同程度的雄性生殖缺陷。基于支持细胞在雄性动物生殖过程中的作用,体外培养高纯度支持细胞可成为研究睾丸两大核心功能-精子发生和性激素分泌功能相关调节机制重要的细胞模型。此外,体外培养睾丸支持细胞也可作为生殖毒理学等新兴热点领域的细胞模型,为评估和研究环境因素对雄性生殖的影响提供便利。因此,作者系统地归纳、总结了目前关于动物支持细胞生物功能的研究及常用的体外分离纯化、培养及鉴定方法,以期为利用动物支持细胞开展雄性生殖领域的研究提供参考。  相似文献   

6.
奶牛子宫内膜细胞体外培养及应用研究概况   总被引:1,自引:0,他引:1  
子宫内膜细胞体外培养技术在研究动物子宫内膜生理功能、繁殖障碍疾病机制及治疗药物筛选等过程中具有重要的作用和意义。论文就国内外有关奶牛子宫内膜细胞体外培养技术中常用的分离、纯化和鉴定等方法以及该细胞体外模型相关应用进行综述,为探索简便、高效的奶牛子宫内膜细胞体外培养方法及国内有关奶牛子宫内膜生理病理的分子细胞学研究提供参考。  相似文献   

7.
疯牛病 1 概念:“疯牛病”有多种名称,目前世界上广泛采用的病名是“传染性海绵状脑病”(TranSmissible spongifolm encephaiopathies TSE),简称“牛海绵状脑病”(BSE)。由于它是由“朊病毒”(又称“朊粒”)引起,所以,有的又叫“朊病毒病”或“朊粒病”(Prion diseases)。除此之外还有称为“传染性变性脑病”(TDE)、“传染性脑淀粉样变性”(TCA)、“亚急性海绵状脑病”(SSE)、“亚急性海绵状病毒性脑病”(SSVE)等的。严格意义上的“TSEs”应译为“可传递性海绵状脑病”。该病是人和动物共患、呈亚急性、渐进…  相似文献   

8.
血脑屏障是隔离外周体循环和中枢神经系统的屏障结构.它可以选择性允许血液中的一些物质通过脑微血管内皮细胞进入大脑中.除了脑微血管内皮细胞之外,血脑屏障的组成主要还有星型胶质细胞、周细胞、神经元和细胞外基质.本试验通过原代培养猪脑微血管内皮细胞和星型胶质细胞,以Transwell细胞板为载体构建猪体外血脑屏障的共培养模型.经过4h渗漏试验和TEER电阻的测定试验显示所构建的猪体外血脑屏障的模型具备了血脑屏障基本的生物学特性,可以用于猪血脑屏障相关疾病尤其是人兽共患性疾病致病机制的研究.  相似文献   

9.
通过建立输卵管上皮细胞的体外培养和传代的方法,进行形态学观察及冻存。取雌性绵羊输卵管上皮细胞,用胶原酶消化和不锈钢网过筛法分离输卵管内膜基质细胞和上皮细胞,进行单细胞原代及传代培养。在输卵管上皮细胞的培养可持续培养4-6周,传3-4代。原代培养细胞7d长成单层,传代细胞4d长成单层。该研究旨在建立一套输卵管上皮细胞体外培养和传代的方法,为进一步研究β-防御素在雌性生殖道的作用方面提供很好的研究模型。  相似文献   

10.
大鼠脑皮质微血管内皮细胞的分离培养及鉴定   总被引:1,自引:0,他引:1  
本研究目的在于建立一种简易可行、获得细胞纯度较高的体外分离、培养大鼠脑皮质微血管内皮细胞的方法,为进一步研究其功能打好基础。新生SD大鼠,分离大脑皮质,经匀浆、两步过滤法获得大鼠脑微血管段,用胶原酶消化后,在37℃、5%CO2培养箱中进行原代细胞培养。通过差速消化和差速贴壁方法并结合倒置显微镜下形态学的观察进行传代和纯化,利用第Ⅷ因子相关抗原免疫荧光检测法对培养的细胞进行鉴定。结果显示:成功分离培养了大鼠脑微血管内皮细胞,倒置显微镜下单个细胞呈多角形,互相融合后呈"铺路石"样,单层贴壁生长;免疫荧光检测第Ⅷ因子相关抗原呈阳性。说明本研究建立的大鼠脑微血管内皮细胞的体外培养方法,程序简单,获得的细胞纯度较高。  相似文献   

11.
Feline spongiform encephalopathy (FSE) is a transmissible spongiform encephalopathy associated with the consumption of feedstuffs contaminated with tissue from bovine spongiform encephalopathy-affected cattle and characterized by the accumulation in the central nervous system of an abnormal isoform of the prion protein (PrP(sc)). Clinically, it presents as a progressive fatal neurologic syndrome that is not easily distinguished from other feline neurologic conditions. Most cases of FSE have been reported in England, where it was first detected in 1990, but a few cases have been reported from other European countries. To identify possible cases of FSE in Italy, the Italian Ministry of Health funded a 2-year surveillance project during which the brains from 110 domestic cats with neurologic signs were evaluated histologically for spongiform encephalopathy and immunohistochemically to detect PrP(sc). Although no cases of FSE were found, the study proved useful in monitoring the Italian cat population for other neurologic diseases: neoplasia (21.8%), toxic-metabolic encephalopathy (18.2%), granulomatous encephalitis (15.5%), suppurative encephalitis (4.6%), trauma (3.6%), circulatory disorders (3.6%), degeneration (2.7%), nonsuppurative encephalitis (2.7%), and neuromuscular diseases (1.8%). No histologic lesions were found in 20% of the brains, and samples from 5.5% of the cats were rejected as unsuitable.  相似文献   

12.
Monitoring of transmissible spongiform encephalopathy (TSE) in Swiss sheep and goats is based on the examination of animals from different sources. In this study, frequencies and proportions of the different diagnoses were compared between routinely submitted sheep and goats, notified scrapie suspects as well as fallen stock. Meningitis/ encephalitis cases were significantly more frequent (OR = 2.2) in the scrapie suspect group when compared to the routine submissions. Metabolic-toxic encephalopathy was seen more frequently within the fallen stock. Rare neurological diagnoses were more frequent among scrapie suspects and routine submissions when compared to fallen stock. Listeriosis was diagnosed equally frequent among the scrapie suspects and routine submissions but less frequent in fallen stock. Scrapie prevalence among the fallen stock and the routine submissions was 0 (zero), with 95% certainty that prevalence is < 1%. The examined animals are representative for most of the Swiss regions with considerable sheep and goat production. Continuation of the detailed neuropathological examination of small ruminants from these three groups, substituted by actively testing a sufficiently large sample of fallen stock and possibly also healthy-slaughtered adult sheep and goats for transmissible spongiform encephalopathies would ensure a good surveillance within the small ruminant population.  相似文献   

13.
Many studies have been undertaken in rodents to study the pathogenesis of transmissible spongiform encephalopathies (TSE). Only a few studies have focused on the pathogenesis of bovine spongiform encephalopathy (BSE) and scrapie in their natural hosts. In this review, we summarize the most recent insights into the pathogenesis of BSE and scrapie starting from the initial uptake of TSE agents and crossing of the gut epithelium. Following replication in the gut-associated lymphoid tissues (GALT), TSE agents spread to the enteric nervous system (ENS) of the gut. Infection is then carried through the efferent fibers of the post-ganglionic neurons of the parasympathetic and sympathetic nervous system to the pre-ganglionic neurons in the medulla oblongata of the brain and the thoracic segments of the spinal cord. The differences between the pathogenesis of BSE in cattle and scrapie in sheep are discussed as well as the possible existence of additional pathogenetic routes.  相似文献   

14.
Tropical Animal Health and Production - Bovine spongiform encephalopathy (BSE) of the cattle is the outstanding disease among other transmissible spongiform encephalopathy (TSEs). It can be...  相似文献   

15.
A juvenile raccoon was euthanized because of severe neurologic signs. At postmortem examination, no significant gross lesions were present. Histologic evaluation demonstrated nonsuppurative encephalitis in thalamus, brainstem, and hippocampus, cerebellar Purkinje cell loss, as well as poliomyelitis and demyelination of the spinal cord. Parvovirus antigen–specific immunohistochemistry revealed immunopositive neurons in the brainstem, cerebral cortex, and hippocampus. A few Purkinje cells were also immunopositive. DNA extracted from formalin-fixed, paraffin-embedded brain tissue (thalamus, hippocampus, cerebral cortex) yielded a positive signal using PCR targeting both feline and canine parvovirus. Sequencing analyses from a fragment of the NS1 gene and a portion of the VP2 gene confirmed the presence of DNA of a recent canine parvovirus variant (CPV-2a–like virus) in the cerebellum. Our case provides evidence that a recent canine parvovirus (CPV) strain (Carnivore protoparvovirus 1) can infect cerebral and diencephalic neurons and cause encephalitis in an otherwise healthy raccoon. Parvovirus-induced encephalitis is a differential diagnosis of rabies and canine distemper in raccoons with neurologic signs.  相似文献   

16.
Scrapie is the longest known and most widely spread of the transmissible spongiform encephalopathies and remains the model for much of the research regarding these diseases. Because scrapie is a reportable disease and the subject of an active eradication program in the United States, veterinary practitioners should have a basic understanding of the disease.  相似文献   

17.
Samples of brain and lymphoid tissues from 1107 meat and bone meal-fed, culled adult pigs from 24 Irish farms were examined for evidence of transmissible spongiform encephalopathy (TSE) by histopathological, immunohistochemical and Western blotting techniques. No evidence of deposits of abnormal prion protein suggesting the presence of TSE was found. Neuropil vacuolation was apparent in the rostral colliculus in 64 per cent of the brains examined and neuronal vacuolation was present in the dorsal vagal nucleus in 15.4 per cent of the brains. However, similar lesions have been described in pigs used as controls in a bovine spongiform encephalopathy challenge experiment. Age-related changes were also observed, including spheroids in the funicular nucleus of 24.5 per cent of the pigs, deposits of lipofuscin in the trigeminal neurons of 13.75 per cent, and mineral deposits in the walls of vessels in the dorsal vagal nucleus of 0.6 per cent. Low-grade non-suppurative inflammatory changes of uncertain origin were observed in 4 per cent of the animals.  相似文献   

18.
Scrapie and bovine spongiform encephalopathy are fatal neurodegenerative diseases caused by the accumulation of a misfolded protein (PrPres), the pathological form of the cellular prion protein (PrPC). For the last decades, prion research has greatly progressed, but many questions need to be solved about prion replication mechanisms, cell toxicity, differences in genetic susceptibility, species barrier or the nature of prion strains. These studies can be developed in murine models of transmissible spongiform encephalopathies, although development of cell models for prion replication and sample titration could reduce economic and timing costs and also serve for basic research and treatment testing. Some murine cell lines can replicate scrapie strains previously adapted in mice and very few show the toxic effects of prion accumulation. Brain cell primary cultures can be more accurate models but are difficult to develop in naturally susceptible species like humans or domestic ruminants. Stem cells can be differentiated into neuron‐like cells and be infected by prions. However, the use of embryo stem cells causes ethical problems in humans. Mesenchymal stem cells (MSCs) can be isolated from many adult tissues, including bone marrow, adipose tissue or even peripheral blood. These cells differentiate into neuronal cells, express PrPC and can be infected by prions in vitro. In addition, in the last years, these cells are being used to develop therapies for many diseases, including neurodegenerative diseases. We review here the use of cell models in prion research with a special interest in the potential use of MSCs.  相似文献   

19.
The transmissible spongiform encephalopathies of domesticated animals, scrapie in-sheep and bovine spongiform encephalopathy (BSE), and transmissible mink encephalopathy are more than a scientific curiosity; under certain circumstances their impact on commercial activities can be calamitous. Knowledge of their causation and pathogenesis is still rudimentary, but many consider than an unconventional agent, the prion (a brain protein, PrP), that is not associated with nucleic acid is involved in both. Others believe that conventional viruses, which replicate by virtue of their nucleic acid-defined genes, are involved in the causation and progression of the encephalopathies but that technical problems have prevented their identification. Others postulate even more exotic causative agents. While this paper will particularly address the possibility of a viral aetiology for these diseases, it is also emphasized that our knowledge of the state of the immune system in animals with encephalopathy needs broadening. There are remarkable gaps in our knowledge of the histopathology of these diseases, particularly the nature of the characteristic vacuoles. Much further work is needed on the biochemical changes in the brain and the serum, particularly of the latter as it could lead to an additional means of recognizing clinical cases without waiting for the animal to die with subsequent examination of the brain for characteristic lesions and the presence of protease-K-resistant PrP.Abbreviations AI artificial insemination - BSE bovine spongiform encephalopathy - CJD Creutzfeldt-Jakob disease - ET embryo transfer - GSSD Gerstmann-Sträussler-Scheinker disease - HDV hepatitis delta virus - MCF mink cell focus - PK proteinase K - PrP prion protein - PrPSc scrapie prion protein - PrP-C the proteinase-K sensitive homologue in normal brain - SAF scrapie-associated fibrils - TME transmissible mink encephalopathy  相似文献   

20.
During active surveillance for transmissible spongiform encephalopathies (TSEs) in sheep, an initial reactor was detected using a rapid test on a brain sample. Immunohistochemistry confirmed an atypical TSE presentation that closely resembled the previously described Nor98 cases. Sequencing of the prnp gene confirmed the ARQ/AHQ genotype with the L141F mutation at codon 141 associated with this phenotype. The head, including the brain and cranial lymphoid tissues, was sampled and examined thoroughly. Non-purulent encephalitis, with ectopic lymphoid follicle formation within the brain, was diagnosed concomitant to the TSE. When scrapie-associated prion protein (PrP(sc)) deposition was studied by immunohistochemistry there was a noticeable lack of lymphotropism. The distribution of PrP(sc) in the brain differed considerably from that of classical scrapie cases. Astrogliosis and microgliosis were demonstrated by histochemical procedures.  相似文献   

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