Comparative expression profiling of E. coli and S. aureus inoculated primary mammary gland cells sampled from cows with different genetic predispositions for somatic cell score.
Comparative expression profiling of E. coli and S. aureus inoculated primary mammary gland cells sampled from cows with different genetic predispositions for somatic cell score.
复制标题
DOI：
10.1186/1297-9686-43-24
复制
发表时间：
2011-06-24
期刊：
影响因子：
--
通讯作者：
Schwerin M
中科院分区：
文献类型：
--
作者：
Brand B;Hartmann A;Repsilber D;Griesbeck-Zilch B;Wellnitz O;Kühn C;Ponsuksili S;Meyer HH;Schwerin M
近十年来，在牛中发现了许多影响乳腺炎发病率的数量性状位点（QTL）和乳腺炎相关性状（如体细胞评分（SCS））。然而，对影响乳腺炎易感性的QTL的分子结构以及引起乳腺炎易感性的潜在生理机制和基因知之甚少。本研究通过全基因组表达分析，分析了牛牛autosome 18 （BTA18）上SCS特异性QTL影响乳腺炎易感性的分子机制。因此，对乳腺上皮细胞影响感染过程的遗传决定机制进行了初步研究。采用标记辅助选择策略，结合BTA18端粒区已确认的SCS QTL的QTL和分子标记信息，从乳腺炎高易感性和低易感性奶牛的乳腺薄壁组织中选取原代牛乳腺上皮细胞（pbMEC）。细胞培养后分别接种热灭活乳腺炎病原菌大肠杆菌和金黄色葡萄球菌。在1、6和24 h后，收集细胞并使用微阵列表达芯片技术分析mRNA表达谱的差异，这些差异归因于遗传易感性、接种和细胞培养。共表达谱的比较分析清楚地表明，遗传有利QTL等位基因“Q”的低易感动物的pbMEC在病原体攻击后的反应比遗传不利QTL等位基因“Q”的高易感动物的pbMEC更快、更强。此外，研究结果强调了RELB是一个功能和位置的候选基因，而相关的非规范Nf-kappaB信号传导是受QTL影响的功能机制。然而，在两组中，接种导致与独创性途径“树突状细胞成熟”和“急性期反应信号传导”相关的基因上调，而细胞培养影响“细胞发育”的生物过程。结果表明，从具有可选QTL等位基因的奶牛中提取的病原体挑战pbMEC的复杂表达谱适合于体外研究乳腺上皮细胞中乳腺炎易感性的遗传决定分子机制，并揭示QTL效应最可能的功能途径和候选基因。
During the past ten years many quantitative trait loci (QTL) affecting mastitis incidence and mastitis related traits like somatic cell score (SCS) were identified in cattle. However, little is known about the molecular architecture of QTL affecting mastitis susceptibility and the underlying physiological mechanisms and genes causing mastitis susceptibility. Here, a genome-wide expression analysis was conducted to analyze molecular mechanisms of mastitis susceptibility that are affected by a specific QTL for SCS on Bos taurus autosome 18 (BTA18). Thereby, some first insights were sought into the genetically determined mechanisms of mammary gland epithelial cells influencing the course of infection. Primary bovine mammary gland epithelial cells (pbMEC) were sampled from the udder parenchyma of cows selected for high and low mastitis susceptibility by applying a marker-assisted selection strategy considering QTL and molecular marker information of a confirmed QTL for SCS in the telomeric region of BTA18. The cells were cultured and subsequently inoculated with heat-inactivated mastitis pathogens Escherichia coli and Staphylococcus aureus, respectively. After 1, 6 and 24 h, the cells were harvested and analyzed using the microarray expression chip technology to identify differences in mRNA expression profiles attributed to genetic predisposition, inoculation and cell culture. Comparative analysis of co-expression profiles clearly showed a faster and stronger response after pathogen challenge in pbMEC from less susceptible animals that inherited the favorable QTL allele 'Q' than in pbMEC from more susceptible animals that inherited the unfavorable QTL allele 'q'. Furthermore, the results highlighted RELB as a functional and positional candidate gene and related non-canonical Nf-kappaB signaling as a functional mechanism affected by the QTL. However, in both groups, inoculation resulted in up-regulation of genes associated with the Ingenuity pathways 'dendritic cell maturation' and 'acute phase response signaling', whereas cell culture affected biological processes involved in 'cellular development'. The results indicate that the complex expression profiling of pathogen challenged pbMEC sampled from cows inheriting alternative QTL alleles is suitable to study genetically determined molecular mechanisms of mastitis susceptibility in mammary epithelial cells in vitro and to highlight the most likely functional pathways and candidate genes underlying the QTL effect.
登录
查看更多内容
影响因子：
2.9
作者：
Brand B;Baes C;Mayer M;Reinsch N;Kühn C
通讯作者：
Kühn C
影响因子：
12.3
作者：
Gentleman RC;Carey VJ;Bates DM;Bolstad B;Dettling M;Dudoit S;Ellis B;Gautier L;Ge Y;Gentry J;Hornik K;Hothorn T;Huber W;Iacus S;Irizarry R;Leisch F;Li C;Maechler M;Rossini AJ;Sawitzki G;Smith C;Smyth G;Tierney L;Yang JY;Zhang J
通讯作者：
Zhang J
影响因子：
3.5
作者：
Brand, B.;Baes, C.;Kuehn, Ch.
通讯作者：
Kuehn, Ch.
影响因子：
3.5
作者：
Griesbeck-Zilch, B.;Meyer, H. H. D.;Wellnitz, O.
通讯作者：
Wellnitz, O.
影响因子：
3.5
作者：
Hinrichs, D;Stamer, E;Kalm, E
通讯作者：
Kalm, E