Gene expression alterations in Rocky Mountain elk infected with chronic wasting disease

Chronic wasting disease (CWD) is an invariably fatal neurologic disease that naturally infects mule deer, white tailed deer and elk. The understanding of CWD neurodegeneration at a molecular level is very limited. In this study, microarray analysis was performed to determine changes in the gene expression profiles in six different tissues including brain, midbrain, thalamus, spleen, RPLN and tonsil of CWD-infected elk in comparison to non-infected healthy elk, using 24,000 bovine specific oligo probes. In total, 329 genes were found to be differentially expressed (> 2.0-fold) between CWD negative and positive brain tissues, with 132 genes upregulated and 197 genes downregulated. There were 249 DE genes in the spleen (168 up- and 81 downregulated), 30 DE genes in the retropharyngeal lymph node (RPLN) (18 up- and 12 downregulated), and 55 DE genes in the tonsil (21 up- and 34 downregulated). Using Gene Ontology (GO), the DE genes were assigned to functional groups associated with cellular process, biological regulation, metabolic process, and regulation of biological process. For all brain tissues, the highest ranking networks for DE genes identified by Ingenuity Pathway Analysis (IPA) were associated with neurological disease, cell morphology, cellular assembly and organization. Quantitative real-time PCR (qRT-PCR) validated the expression of DE genes primarily involved in different regulatory pathways, including neuronal signaling and synapse function, calcium signaling, apoptosis and cell death and immune cell trafficking and inflammatory response. This is the first study to evaluate altered gene expression in multiple organs including brain from orally infected elk and the results will improve our understanding of CWD neurodegeneration at the molecular level.     

The ecology of chronic wasting disease in wildlife

Prions are misfolded infectious proteins responsible for a group of fatal neurodegenerative diseases termed transmissible spongiform encephalopathy or prion diseases. Chronic Wasting Disease (CWD) is the prion disease with the highest spillover potential, affecting at least seven Cervidae (deer) species. The zoonotic potential of CWD is inconclusive and cannot be ruled out. A risk of infection for other domestic and wildlife species is also plausible. Here, we review the current status of the knowledge with respect to CWD ecology in wildlife. Our current understanding of the geographic distribution of CWD lacks spatial and temporal detail, does not consider the biogeography of infectious diseases, and is largely biased by sampling based on hunters' cooperation and funding available for each region. Limitations of the methods used for data collection suggest that the extent and prevalence of CWD in wildlife is underestimated. If the zoonotic potential of CWD is confirmed in the short term, as suggested by recent results obtained in experimental animal models, there will be limited accurate epidemiological data to inform public health. Research gaps in CWD prion ecology include the need to identify specific biological characteristics of potential CWD reservoir species that better explain susceptibility to spillover, landscape and climate configurations that are suitable for CWD transmission, and the magnitude of sampling bias in our current understanding of CWD distribution and risk. Addressing these research gaps will help anticipate novel areas and species where CWD spillover is expected, which will inform control strategies. From an ecological perspective, control strategies could include assessing restoration of natural predators of CWD reservoirs, ultrasensitive CWD detection in biotic and abiotic reservoirs, and deer density and landscape modification to reduce CWD spread and prevalence.     

牛痘病毒感染人巨噬细胞基因表达谱的RNA-Seq分析

[目的]利用RNA-Seq,分析人巨噬细胞在牛痘病毒(VACV)感染前后基因表达的变化,探索牛痘病毒与宿主细胞相互作用的机制.[方法]用牛痘病毒感染人巨噬细胞,采用RNA-Seq比较感染组和对照组的差异表达基因,并进行KEGG、GO以及STRING网络分析.[结果]感染组与对照组相比,筛选出显著性差异表达基因4796个...     

Interaction among deer in a chronic wasting disease endemic zone

Although it is known that chronic wasting disease (CWD) can be transmitted by both direct animal-to-animal contact and contact with contaminated environments, the relative role of each mechanism in the spread of CWD in free-ranging populations has yet to be defined. We investigated patterns of interaction between mule deer (Odocoileus hemionus) in order to understand how factors such as season and landscape may influence patterns of disease spread in these populations. Using location data from male and female Global Positioning System (GPS)-collared mule deer in 5 study areas located in and around a CWD-endemic zone in southern Saskatchewan, Canada, we quantified close proximity events, or events involving both spatial and temporal overlap of individuals. We defined close proximity events as occurrences in which 2 deer were located <25 m apart at the same point in time. We looked at seasonal variation in the probability of close proximity events, as well as landscape factors associated with these events when compared to areas of shared space use, or spatial overlap alone. Overall probability of an individual GPS-collared deer being located in close proximity to another GPS-collared deer was 0.092 (n = 107). The early gestation (16 Dec–31 Mar) and late gestation (1 Apr–15 May) seasons had the highest probability of close proximity events occurring, and same-sex pairs were more likely to be found in close proximity than between-sex pairs during all seasons aside from the rut (1 Nov–15 Dec). High probability of close proximity events during the gestation seasons agrees with the tendency of mule deer to aggregate into large groups during late winter and suggests that this may be an important time period for CWD transmission to occur. Close proximity events occurred more in cropland and wetland than expected based on availability, whereas close proximity events occurred less than expected in grassland. The opposite was true for spatial overlap between individuals, which occurred more than expected in areas of low elevation and rugged terrain and in grassland or shrub–wood habitats. These results suggest that cropland may be a higher risk habitat for direct and indirect CWD transmission between individuals and that, although coulees and other areas of rugged topography are less likely to be associated with close proximity events, those areas may be more likely to contain environmental contamination in CWD-affected areas due to common use by multiple deer. © 2011 The Wildlife Society.     

Applicability of current bovine spongiform encephalopathy (BSE) diagnostic procedures for chronic wasting disease (CWD)

Chronic wasting disease (CWD) in cervids is one of the transmissible spongiform encephalopathies ; however, its risk to humans is still obscure. An increase in number of diseased deer in North America has raised concerns regarding the CWD risk to humans. We demonstrated that the con-firmatory procedures and the commercial diagnostic kits for bovine spongiform encephalopathy (BSE) can be adopted for the diagnosis of CWD. No CWD case was confirmed in the surveillance of 558 cervids that were examined between 2003 and 2006 in Japan.     

鸡胚公母性腺差异表达基因的基因芯片杂交筛选

采用Affymetrix公司鸡基因组芯片对9日龄鸡胚公母性腺总RNA进行了芯片杂交, 并对基因表达谱进行了分析。统计结果显示, 9日龄母鸡性腺表达基因数19 368个, 公鸡性腺表达基因数19 493个; 公母性腺绝对差异表达基因,即公鸡性腺表达而母鸡性腺不表达基因145个, 母鸡性腺表达而公鸡性腺不表达基因189个。绝对差异表达基因功能分类结果显示, 参与细胞组成、细胞加工和分子结合基因占多数, 部分基因参与细胞器组成、代谢加工、生物学调控以及催化反应和细胞信号转导等。值得注意的是, 本研究发现了一些已经报道同性别决定和分化有一定关联的基因, 如ASW、CHD1和SOX9等, 同时也发现了一些未知其同性腺分化和发育有关联的基因和编码假想蛋白的表达序列。进一步分析这些基因和表达序列的生物学功能和表达模式, 将对鸟类性别决定和分化机制的了解提供有益参考。     

Prion protein gene sequence and chronic wasting disease susceptibility in white-tailed deer (Odocoileus virginianus)

ABSTRACT

The sequence of the prion protein gene (PRNP) affects susceptibility to spongiform encephalopathies, or prion diseases in many species. In white-tailed deer, both coding and non-coding single nucleotide polymorphisms have been identified in this gene that correlate to chronic wasting disease (CWD) susceptibility. Previous studies examined individual nucleotide or amino acid mutations; here we examine all nucleotide polymorphisms and their combined effects on CWD. A 626 bp region of PRNP was examined from 703 free-ranging white-tailed deer. Deer were sampled between 2002 and 2010 by hunter harvest or government culling in Illinois and Wisconsin. Fourteen variable nucleotide positions were identified (4 new and 10 previously reported). We identified 68 diplotypes comprised of 24 predicted haplotypes, with the most common diplotype occurring in 123 individuals. Diplotypes that were found exclusively among positive or negative animals were rare, each occurring in less than 1% of the deer studied. Only one haplotype (C, odds ratio 0.240) and 2 diplotypes (AC and BC, odds ratios of 0.161 and 0.108 respectively) has significant associations with CWD resistance. Each contains mutations (one synonymous nucleotide 555C/T and one nonsynonymous nucleotide 286G/A) at positions reported to be significantly associated with reduced CWD susceptibility. Results suggest that deer populations with higher frequencies of haplotype C or diplotypes AC and BC might have a reduced risk for CWD infection – while populations with lower frequencies may have higher risk for infection. Understanding the genetic basis of CWD has improved our ability to assess herd susceptibility and direct management efforts within CWD infected areas.     

NGAL基因在永生化食管上皮细胞恶性转化中过表达的研究

为研究NGAL（neutrophil gelatinase-associated lipocalin）基因在永生化食管上皮细胞恶性转化中的表达情况,以永生化食管上皮细胞系SHEE和食管癌细胞系SHEEC互为对照,用cDNA微列阵进行筛选,用RNA印迹和RT-PCR进行鉴定,cDNA克隆测序后与GenBank进行BLAST分析比较.结果表明NGAL基因在SHEEC中出现显著差异过表达,其cDNA序列与小鼠24p3、大鼠NRL（neu-related lipocalin）、人中性粒细胞NGAL和卵巢癌NGAL具有较高的相似性.这提示NGAL基因在永生化食管上皮细胞恶性转化中可能发挥着重要作用,可能是一种新的癌基因或促癌基因.     

Prevalence of chronic wasting disease and bovine tuberculosis in free-ranging deer and elk in South Dakota

Heads of hunter-harvested deer and elk were collected throughout South Dakota (USA) and within established chronic wasting disease (CWD) surveillance areas from 1997-2002 to determine infection with CWD and bovine tuberculosis (TB). We used immunohistochemistry to detect CWD-infected individuals among 1,672 deer and elk sampled via geographically targeted surveillance. A total of 537 elk (Cervus elaphus nelsoni), 813 white-tailed deer (Odocoileus virginianus), and 322 mule deer (O. hemionus) was sampled for CWD. Estimated overall prevalence and associated confidence intervals (95%) in white-tailed deer was 0.001% (0-0.007%). Similarly, estimated overall prevalence in elk and mule deer was 0.0% (0-0.004%) and 0.0% (0-0.011%), respectively. A total of 401 elk, 1,638 white-tailed deer, and 207 mule deer was sampled for TB. Estimated overall prevalence of infection with TB in elk harvested in South Dakota was 0.0% (0-0.009%). Similarly, estimated overall prevalence of TB in white-tailed deer and mule deer harvested throughout South Dakota was 0.0% (0-0.002%) and 0.0% (0-0.018%), respectively.     

Implications of chronic wasting disease,cougar predation,and reduced recruitment for elk management

Emerging diseases and expanding carnivore populations may have profound implications for ungulate harvest management and population regulation. To better understand effects of chronic wasting disease (CWD) and cougar (Puma concolor) predation, we studied mortality and recruitment of elk (Cervus elaphus) at Wind Cave National Park (WICA) during 2005–2009. We marked 202 elk (83 subadult M and 119 subadult and ad F) with Global Positioning System (GPS) collars, observed 28 deaths during 74,220 days of monitoring, and investigated 42 additional deaths of unmarked elk found dead. Survival rates were similar for males and females and averaged 0.863 (SE = 0.025) annually. Leading causes of mortality included hunting (0.065, SE = 0.019), CWD (0.034, SE = 0.012), and cougar predation (0.029, SE = 0.012). Marked elk killed by hunters and cougars typically were in good physical condition and not infected with CWD. Effects of mortality on population growth were exacerbated by low rates of pregnancy (subadults = 9.5%, SE = 6.6%; ad = 76.9%, SE = 4.2%) and perinatal survival (0.49, SE = 0.085 from 1 Feb to 1 Sep). Chronic wasting disease, increased predation, and reduced recruitment reduced the rate of increase for elk at WICA to approximately λ = 1.00 (SE = 0.027) during the past decade. Lower rates of increase are mitigating effects of elk on park vegetation, other wildlife, and neighboring lands and will facilitate population control, but may reduce opportunities for elk hunting outside the park. © 2011 The Wildlife Society     

Chronic wasting disease management in ranched elk using rectal biopsy testing

Chronic wasting disease (CWD) is a transmissible spongiform encephalopathy (TSE) affecting members of the cervid species, and is one of the few TSEs with an expanding geographic range. Diagnostic limitations, efficient transmission, and the movement of infected animals are important contributing factors in the ongoing spread of disease. Managing CWD in affected populations has proven difficult, relying on population reduction in the case of wild deer and elk, or quarantine and depopulation in farmed cervids. In the present study, we evaluated the effectiveness of managing endemic CWD in a closed elk herd using antemortem sampling combined with both conventional and experimental diagnostic testing, and selective, targeted culling of infected animals. We hypothesized that the real-time quaking-induced conversion (RT-QuIC) assay, a developing amplification assay, would offer greater detection capabilities over immunohistochemistry (IHC) in the identification of infected animals using recto-anal mucosa associated lymphoid tissue (RAMALT). We further sought to develop a better understanding of CWD epidemiology in elk with various PRNP alleles, and predicted that CWD prevalence would decrease with targeted culling. We found that RT-QuIC identified significantly more CWD-positive animals than IHC using RAMALT tissues (121 vs. 86, respectively, out of 553 unique animals), and that longstanding disease presence was associated with an increasing frequency of less susceptible PRNP alleles. Prevalence of CWD increased significantly over the first two years of the study, implying that refinements in our management strategy are necessary to reduce the prevalence of CWD in this herd.     

Transmission of prions from mule deer and elk with chronic wasting disease to transgenic mice expressing cervid PrP

We generated mice expressing cervid prion protein to produce a transgenic system simulating chronic wasting disease (CWD) in deer and elk. While normal mice were resistant to CWD, these transgenic mice uniformly developed signs of neurological dysfunction approximately 230 days following intracerebral inoculation with four CWD isolates. Inoculated transgenic mice homozygous for the transgene array developed disease after approximately 160 days. The brains of sick transgenic mice exhibited widespread spongiform degeneration and contained abnormal prion protein and abundant amyloid plaques, many of which were florid plaques. Transmission studies indicated that the same prion strain caused CWD in the analyzed mule deer and elk. These mice provide a new and reliable tool for detecting CWD prions.     

Gene expression profile associated with Asmt knockout-induced depression-like behaviors and exercise effects in mouse hypothalamus

Sleep disorder caused by abnormal circadian rhythm is one of the main symptoms and risk factors of depression. As a known hormone regulating circadian rhythms, melatonin (MT) is also namely N-acetyl-5-methoxytryptamine. N-acetylserotonin methyltransferase (Asmt) is the key rate-limiting enzyme of MT synthesis and has been reportedly associated with depression. Although 50–90% of patients with depression have sleep disorders, there are no effective treatment ways in the clinic. Exercise can regulate circadian rhythm and play an important role in depression treatment. In the present study, we showed that Asmt knockout induced depression-like behaviors, which were ameliorated by swimming exercise. Moreover, swimming exercise increased serum levels of MT and 5-hydroxytryptamine (5-HT) in Asmt knockout mice. In addition, the microarray data identified 10 differentially expressed genes (DEGs) in KO mice compared with WT mice and 29 DEGs in KO mice after swimming exercise. Among the DEGs, the direction and magnitude of change in epidermal growth factor receptor pathway substrate 8-like 1 (Eps8l1) and phospholipase C-β 2 (Plcb2) were confirmed by qRT-PCR partly. Subsequent bioinformatic analysis showed that these DEGs were enriched significantly in the p53 signaling pathway, long-term depression and estrogen signaling pathway. In the protein–protein interaction (PPI) networks, membrane palmitoylated protein 1 (Mpp1) and p53-induced death domain protein 1 (Pidd1) were hub genes to participate in the pathological mechanisms of depression and exercise intervention. These findings may provide new targets for the treatment of depression.     

Species barriers for chronic wasting disease by in vitro conversion of prion protein

Chronic wasting disease (CWD) is a transmissible spongiform encephalopathy that can affect North American cervids (deer, elk, and moose). Using a novel in vitro conversion system based on incubation of prions with normal brain homogenates, we now report that PrP^CWD of elk can readily induce the conversion of normal cervid PrP (PrP^C) molecules to a protease-resistant form, but is less efficient in converting the PrP^C of other species, such as human, bovine, hamster, and mouse. However, when substrate brain homogenates are partially denatured by acidic conditions (pH 3.5), PrP^CWD-induced conversion can be greatly enhanced in all species. Our results demonstrate that PrP^C from cervids (including moose) can be efficiently converted to a protease-resistant form by incubation with elk CWD prions, presumably due to sequence and structural similarities between these species. Moreover, partial denaturation of substrate PrP^C can apparently overcome the structural barriers between more distant species.     

Gene expression profiling of cardiovascular disease models

Recent development of gene expression profiling technologies has enabled the large-scale analysis of gene expression changes during disease progression. Frequently, cardiovascular diseases involve complex interactions of multiple cell types over prolonged periods of time. A better understanding of the pathology of cardiovascular diseases and the potential identification of underlying genetic defects are currently being explored by using profiling methodologies in a number of animal and tissue-culture models.