Transcriptome‑wide mapping of milk somatic cells upon subclinical mastitis infection in dairy cattle

Background Subclinical intramammary infection(IMI)represents a significant problem in maintaining dairy cows’health.Disease severity and extent depend on the interaction between the causative agent,environment,and host.To investigate the molecular mechanisms behind the host immune response,we used RNA-Seq for the milk somatic cells(SC)transcriptome profiling in healthy cows(n=9),and cows naturally affected by subclinical IMI from Proto-theca spp.(n=11)and Streptococcus agalactiae(S.agalactiae;n=11).Data Integration Analysis for Biomarker discov-ery using Latent Components(DIABLO)was used to integrate transcriptomic data and host phenotypic traits related to milk composition,SC composition,and udder health to identify hub variables for subclinical IMI detection.Results A total of 1,682 and 2,427 differentially expressed genes(DEGs)were identified when comparing Prototheca spp.and S.agalactiae to healthy animals,respectively.Pathogen-specific pathway analyses evidenced that Proto-theca’s infection upregulated antigen processing and lymphocyte proliferation pathways while S.agalactiae induced a reduction of energy-related pathways like the tricarboxylic acid cycle,and carbohydrate and lipid metabolism.The integrative analysis of commonly shared DEGs between the two pathogens(n=681)referred to the core-mastitis response genes,and phenotypic data evidenced a strong covariation between those genes and the flow cytometry immune cells(r2=0.72),followed by the udder health(r2=0.64)and milk quality parameters(r2=0.64).Variables with r≥0.90 were used to build a network in which the top 20 hub variables were identified with the Cytoscape cyto-hubba plug-in.The genes in common between DIABLO and cytohubba(n=10)were submitted to a ROC analysis which showed they had excellent predictive performances in terms of discriminating healthy and mastitis-affected animals(sensitivity>0.89,specificity>0.81,accuracy>0.87,and precision>0.69).Among these genes,CIITA could play a key role in regulating the animals’response to subclinical IMI.Conclusions Despite some differences in the enriched pathways,the two mastitis-causing pathogens seemed to induce a shared host immune-transcriptomic response.The hub variables identified with the integrative approach might be included in screening and diagnostic tools for subclinical IMI detection.

刺激下丘脑对猫小脑皮层神经元电活动的影响

在戊巴比妥钠麻醉的猫上观察了电刺激下丘脑外侧区（ＬＨＡ）、背侧区（ＤＨＡ）、后侧区（ＰＨＡ）和腹内侧核（ＶＭＮ）对小脑皮层第Ⅵ和Ⅶ小叶两类不同神经元（浦肯野细胞和非浦肯野细胞）电活动的影响．结果如下：（１）刺激ＬＨＡ、ＤＨＡ、ＰＨＡ和ＶＭＮ可引起小脑皮层神经元三种不同类型的反应，但以抑制反应为主（８０．８％，８４／１０４）；反应的潜伏期为１０—９０ｍｓ，并且半数细胞（５１％）对刺激呈现＜２０ｍｓ的短潜伏期反应．（２）刺激ＬＨＡ、ＤＨＡ、ＰＨＡ和ＶＭＮ均可影响小脑皮层神经元的放电活动，但刺激ＬＨＡ、ＤＨＡ和ＰＨＡ引起细胞反应的比率均明显高于ＶＭＮ，有统计学差异．（３）刺激ＬＨＡ、ＤＨＡ、ＰＨＡ和ＶＭＮ主要影响小脑皮层中非浦肯野细胞的活动和浦肯野细胞的复杂锋电位活动．（４）静脉注射组胺Ｈ１受体阻断剂扑尔敏可以阻断刺激下丘脑对小脑皮层神经元放电活动的抑制作用．这些结果提示：下丘脑－小脑投射纤维的激活可以影响到小脑皮层神经元网络中不同神经元，特别是中间神经元的活动．推测下丘脑－小脑投射纤维可能在机体的躯体－内脏功能整合中发挥重要作用．

刺激猫下丘脑LHA和VMN对小脑皮层神经元电活动的影响

在戊巴比妥钠麻醉的猫,电刺激下丘脑外侧区(LHA)和腹内侧核(VMN)可引起小脑皮层第Ⅵ和Ⅶ小叶浦肯野细胞和非浦肯野细胞抑制性、兴奋性和抑制-兴奋性的电活动,但以抑制性活动为主;这些反应的潜伏期多数为10-20ms,但也可长达90ms;一般刺激LHA比刺激VMN更有效。刺激下丘脑所激发的小脑神经元抑制性电活动可为静脉注射组胺H_1受体阻断剂扑尔敏所阻断。本文对上述观察的可能作用作了讨论。