Induced expression of c-fos in the diencephalon and pituitary gland of goats following transportation

To identify regions of the caprine diencephalone and pituitary gland related to transportation stress, the expression of c-fos protein was examined immunohistochemically as an indicator of neural activation. Ten castrated Shiba goats (Capra hircus), five transported and five controls, were used. Transported goats were trucked for 1 h and killed by transcardiac perfusion 1 h after the end of transportation. Control goats were housed in single pens killed in the same manner and at the same time as the transported goats. The diencephalon and the pituitary gland were removed after perfusion and used for immunostaining. Plasma cortisol concentrations during and after transportation also were investigated. During transportation, plasma cortisol concentrations increased (P < 0.05) compared with those in the controls. In the diencephalon, c-fos immunoreactive cells were detected in the subcallosa, the lateral septal area, the bed nucleus of stria terminalis (BNST), the preoptic hypothalamic area (POA), the suprachiasmatic nucleus (SCN), the supraoptic nucleus, the paraventricular hypothalamic nucleus parvocellular (PVNp), the paraventricular hypothalamic nucleus magnocellular (PVNm), the arcuate nucleus (ARC), the paraventricular thalamic nucleus, and the stria medullaris in both control and transported goats. The numbers of c-fos immunoreactive cells were increased (P < 0.05) by transportation in the PVNm, the PVNp, the BNST, the POA, the ARC, and the SCN (P < 0.10). In the anterior pituitary gland, the number of c-fos immunoreactive cells in transported goats was 4 to 30 times as much as in control goats; however, there were no differences in the intermediate and posterior lobes between control and transported goats. This study has identified regions in the caprine diencephalon and pituitary gland that show transport-induced increases in c-fos immunoreactive cells. In conclusion, the PVNm, the PVNp, the BNST, the POA, the SCN in the diencephalons, and the anterior lobe of pituitary gland may be involved in the stress responses of goats to transportation.     

纳洛酮对大鼠大脑皮质c-fos mRNA表达的影响

为研究纳洛酮对小型猪特异性麻醉剂(XFM)麻醉下大鼠大脑皮质c-fos mRNA表达的影响,探讨纳洛酮催醒XFM麻醉大鼠与大脑皮质c-fos基因的关系.将78只SD纯种大鼠随机分为空白对照组、XFM对照组、XFM+纳洛酮组、XFM+生理盐水组,采用实对定量PCR技术检测大脑皮质内c-fos mRNA表达量.结果表明,腹腔注射XFM后引起大鼠大脑皮质c-fos mRNA高效表达,各时期c-fos mRNA表达与空白对照组比较差异极显著(P＜0.01);纳洛酮注射后引起XFM麻醉大鼠大脑皮质c-fos mRNA表达量减少,与XFM对照组比较差异显著(P＜0.01或P＜0.05).结果提示,大脑皮质c-fos基因参与了纳洛酮颉颃XFM的麻醉作用,纳洛酮抑制XFM诱导大鼠大脑皮质c-fos基因表达,可能是纳洛酮催醒XFM麻醉大鼠的重要机理之一.     

隆朋对替来他明/唑拉西泮诱导的大鼠不同脑区c-fos基因表达的影响

分离麻醉药替来他明和苯二氮卓类安定药唑拉西泮按照1:1的质量比组成的Zoletil(中文商品名为舒泰),目前被广泛应用于兽医领域.隆朋是α2肾上腺能受体激动剂,具有一定的镇痛、镇静和肌松作用,并且与舒泰有明显的协同作用[1].原癌基因c-fos参与细胞内的信息传递,被称为神经元的第三信使,它参与重要脑功能的信号转导和调控过程,是一种脑功能活动形态学定位标记物[2].当机体受到某种刺激时,神经系统内与此机能有关的神经元均处于活动状态,而激发相应神经元内的c-fos基因表达,位于细胞浆内的c-fos mRNA很快进入细胞核转录形成Fos蛋白[3].本试验的目的是通过研究隆朋对替来他明／唑拉西泮诱导大鼠不同脑区Fos蛋白表达的影响,探讨隆朋对替来他明／唑拉西泮的作用机理.     

阿替美唑对舒泰/噻啦嗪诱导大鼠丘脑和大脑皮质c-fos基因表达的影响

24只SD大鼠被随机分成对照组、麻醉组和催醒组,对照组注射二甲基亚砜,麻醉组注射舒泰(13.81mg/kg)和噻啦嗪(5.21mg/kg),催醒组在给予麻醉剂10min后注射阿替美唑(0.522mg/kg),试验组大鼠分别于给药1h(即麻醉期)即刻,断头处死,冰面上分离丘脑和大脑皮质,利用免疫印记技术测定Fos蛋白表达量。结果显示,舒泰联合噻啦嗪注射后引起麻醉期大鼠丘脑和大脑皮质脑区Fos蛋白表达量显著高于对照组(P<0.01),阿替美唑注射后显著地抑制了舒泰联合噻啦嗪诱导大鼠丘脑和大脑皮质脑区Fos蛋白表达(P<0.01)。结果表明,阿替美唑可抑制舒泰联合噻啦嗪麻醉引起大鼠中枢脑区Fos蛋白的表达,对神经损伤起到一定的保护作用。     

旋毛虫Serpin基因在不同发育时期的mRNA表达水平变化及虫体表达特性

为研究旋毛虫丝氨酸蛋白酶抑制剂(Serpin),基因在不同发育时期的mRNA表达水平变化,本研究应用实时荧光定量PCR技术对不同发育时期的虫体的Serpin在mRNA表达水平进行检测。同时利用鼠抗重组蛋白血清对Serpin在旋毛虫成虫、新生幼虫、成囊前期幼虫及肌幼虫中进行定位,以鉴定该Serpin基因的表达特性。研究结果显示,在成虫时期,3 d时表达量明显高于6 h和5 d的表达量(P﹤0.05);成囊前期表达量很低,其中18 d时表达量最低(P﹤0.05);肌幼虫时期在26 d、38 d和48 d均有大量表达(P﹤0.05)。免疫荧光染色结果表明,Ts Serpin蛋白在5 d成虫期的体表表达;在新生幼虫和14 d成囊前期幼虫的体内体表均有表达;38 d肌幼虫时期明显可见表皮及体内均有表达。本实验为探究旋毛虫在宿主体内免疫逃避机制奠定基础。     

缺氧与肉鸡心肌细胞c-fos和c-myc基因表达的关系

利用细胞培养、原位杂交、免疫组化、图像分析等技术研究缺氧与肉鸡心肌细胞癌基因c-fos和c-myc表达的关系,进一步阐明缺氧与以右心肥大、衰竭为特征的肉鸡腹水综合征的关系。结果显示,缺氧显著引起心肌细胞内癌基因c-fos和c-mycmRNA的表达(c-fos mRNA:常氧149.6&#177;22.4;缺氧205.4&#177;57.6,P&lt;0.01;c-myc mRNA:常氧155.2&#177;25.7;缺氧197.4&#177;26.2,P&lt;0.01)。缺氧显著引起心肌细胞内癌基因c-fos和c-myc蛋白的表达(c-fos蛋白:常氧159.1&#177;37.1,缺氧240.3&#177;49.6;c-myc蛋白:常氧149.6&#177;24.9,缺氧236.3&#177;55.4;P&lt;0.01)。结果表明缺氧显著诱发肉鸡心肌细胞内癌基因c-fos、c-myc的转录和表达,是启动心肌细胞肥大的重要原因。因此本研究进一步阐明缺氧是以右心肥大衰竭为特征的肉鸡腹水综合征的主要诱因。     

草地早熟禾NAC基因鉴定及非生物胁迫下表达模式分析

草地早熟禾(Poa pratensis)是世界上广泛应用的草坪草种,其抗逆基因的挖掘对抗逆新品种的选育具有重要意义.NAC基因家族是植物特有的转录因子之一,在植物逆境响应中起到重要作用.以二穗短柄草(Brach ypodium distach yon)的NAC蛋白序列为请求序列,从草地早熟禾转录组数据库序列比对分析得到15个草地早熟禾PpNACs基因的cDNA全长序列,具有完整的开放阅读框.运用生物信息学方法预测早熟禾NAC家族成员的理化性质和保守结构域,发现15个PpNACs转录因子都是酸性蛋白,除PpNAC16外都属于不稳定蛋白,疏水性蛋白,具有相似的结构,通过聚类分析可将15个PpNACs成员分为3类.荧光定量PCR结果显示,在重金属胁迫下显著上调表达的基因有Pp-NAC10、PpNAC18、PpNAC19和PpNAC24,在盐胁迫过程下显著上调表达的基因有PpNAC10和PpNAC24,在干旱胁迫下显著上调表达的基因有PpNAC1、PpNAC6、PpNAC10和PpNAC13,在低温胁迫下显著上调表达的基因有PpNAC1、PpNAC10和PpNAC18,在高温胁迫下显著上调表达的基因有PpNAC20.其中,PpNAC10在重金属、盐、干旱和低温胁迫下均被显著诱导上调表达,Pp-NAC18在重金属、干旱和低温胁迫下均被显著诱导上调表达,PpNAC10和PpNAC18可作为草地早熟禾逆境胁迫响应的候选基因.     

Sympathetic activation of hepatic and splenic IL-1beta mRNA expression during oscillation stress in the rat

Interleukin (IL)-1beta mRNA expression in the liver and spleen was examined after subjection to oscillation stress in the rat. Thirty-minute subjection to oscillation stress increased IL-1beta mRNA expression in the both organs. Prior treatment of rats with gadolinium chloride, which eliminates macrophages, prevented the stress-induced IL-1beta expression. Either adrenalectomy or treatment of guanethidine, a blocker of norepinephrine release in the sympathetic nerve endings, partially attenuated the stress-induced response, but the combined treatment completely blocked it. Injection of beta-adrenergic antagonist (propranolol) also suppressed the stress-induced response. These results suggest that oscillation stress induces IL-1beta mRNA expression in the liver and spleen, probably in Kupffer cells and splenic macrophages, and that stress-induced IL-1beta expression is elicited by catecholamines released from sympathetic nerve terminals and the adrenal gland.     

Functional MRI activity in the thalamus and occipital cortex of anesthetized dogs induced by monocular and binocular stimulation.

The neuroanatomy of the mammalian visual system has received considerable attention through electrophysiological study of cats and non-human primates, and through neuroimaging of humans. Canine neuroanatomy, however, has received much less attention, limiting our understanding of canine vision and visual pathways. As an early step in applying blood oxygenation level dependant (BOLD) functional magnetic resonance imaging (fMRI) for veterinary use, we compared visual activity in the thalamus and occipital cortex of anesthetized dogs presented with binocular and monocular visual stimuli. Activity in the left and right thalamus and occipital cortex during monocular stimulation was also compared. Six beagles were presented with a vertical grating visual stimulus and scanned at 4 Tesla. Each dog was scanned twice under each of 3 anesthetic protocols (isoflurane, propofol, and fentanyl/midazolam). We found: 1) significant BOLD activation in the lateral geniculate nucleus (LGN) of the thalamus and the occipital cortex; 2) a significantly larger area of activation in the LGN during monocular stimulation than during binocular stimulation; and 3) that activity in the hemisphere contralateral to the stimulus was not significantly greater than that ipsilateral to it.     

猪SOCS1基因的克隆、生物信息分析及热应激条件下的表达

本试验克隆了土杂猪SOCS1基因cDNA序列,发现SOCS1基因cDNA编码220个氨基酸的前体蛋白。该蛋白相对分子质量24 370,等电点(PI)10.98,与GenBank中登录的普通猪、人、牛、羊和小鼠SOCS1序列同源性分别为99.9%、97.1%、96.1%、94.3%和93.7%。具有1个中央SH2域(Scr-homology 2)、1个长度可变的N端结构域和1个位于C端包含40个氨基酸残基的保守序列(SOCS盒)。荧光定量PCR结果显示,脾脏、小肠和胸腺中SOCS1mRNA的表达在热应激条件下显著上调(P≤0.05),且呈时间依赖性。但在肠系膜淋巴结中,热应激第3、6、9天显著下调(P≤0.05)。结果表明,SOCS1表达量在热应激条件下的土杂猪发生明显改变,并呈组织和时间依赖性。     

荣昌猪和长荣猪肌肉生长抑制素基因表达规律的研究

为了研究荣昌猪和长荣猪胴体瘦肉率和骨骼肌肌肉生长抑制素(MSTN)基因表达量的变化规律,试验采用2×2因子试验设计,荣昌猪和长荣猪(两种基因型猪)分别饲喂按中国瘦肉型猪饲养标准和荣昌猪饲养标准配制的日粮,共4个处理,每个处理6个重复,在20 kg、35 kg、50 kg、80 kg和110 kg时每个重复屠宰1头。结果表明:荣昌猪和长荣猪的瘦肉率随体重的增加呈降低的趋势,背最长肌中MSTN基因的表达量呈上升的趋势;在相同体重条件下,荣昌猪的胴体瘦肉率显著(P<0.05)或极显著(P<0.01)地低于长荣猪,其背最长肌中MSTN基因表达均明显高于长荣猪,而日粮营养水平及品种×营养水平交互作用对猪胴体品质和MSTN基因的表达量没有明显影响(P>0.05)。     

The expression pattern of TIR8 is conserved among vertebrates

IL-1R/TLRs play an important role in the inflammatory responses. Their signaling pathway is tightly regulated to keep inflammation under control. The regulation involves both extracellular and intracellular mechanisms. TIR8, also known as SIGIRR (Single Immunoglobulin IL-1R-Related molecule), is an orphan receptor belonging to the IL-1R/TLRs super-family. Recent studies suggest its role in modulating the inflammatory response in the gastrointestinal tract in mice, acting as an IL-1 receptor family antagonist. We identified the sequence and analyzed the expression of TIR8 in a wide panel of organs and tissues in different animals. There was high homology of the cDNA sequence among human, mouse and the domestic species (74–85%). The pattern of expression of this receptor was similar in all the species examined (high levels in kidney and gastrointestinal tract) and similar to the human. These results demonstrate that TIR8 is conserved, in evolutionary terms, both with regard to sequence and pattern of expression, a finding consistent with a key function of this molecule in modulating inflammation in the gastrointestinal tract.     

The expression pattern,polymorphisms and association analyses of the porcine NREP gene

NREP (neuronal regeneration related protein homolog) plays a role in the transformation of neural, muscle, and fibroblast cells and in smooth muscle myogenesis. The NREP gene was selected for detailed study as an expressional and functional candidate gene on the basis of data from the expression microarray, which detected the differences in gene expression between Czech Large White pigs and wild boars in the longissimus lumborum et thoracis and biceps femoris muscle tissues. Quantitative real-time PCR results confirmed that porcine NREP was expressed in both skeletal muscles and significantly overexpressed in Czech Large White pigs compared with wild boars (14.5- and 11.6-fold; p < .05). We identified 9 polymorphic sites in the genomic DNA of NREP. Six of these polymorphisms were in complete linkage disequilibrium, and therefore, only 4 loci were informative. The associations of the HF571253:g.103G>A, HF571253:g.134G>A, HF571253:g.179T>C and HF571253:g.402_409delT polymorphisms with backfat thickness, lean meat content and average daily gain were assessed in Czech Large White pigs. The GG genotypes HF571253:g.103G>A and HF571253:g.134G>A, the TT genotypes HF571253:g.179T>C and 67 HF571253:g.402_409delT genotypes had favourable effects on the studied traits. Our results indicate the possibility of utilizing the variability of the NREP gene in marker-assisted selection in order to improve meat production in pigs.     

Ganglioglioma in the thalamus of a puppy

A solitary brain mass of a 4-month-old miniature dachshund showing seizure-like neurological signs was examined histopathologically. At necropsy a white tumor mass, replacing the thalamus, approximately 1.5 cm in diameter, was found. There was cystic space filled with yellowish pale fluid in the central area of the tumor mass. Histopathological examination revealed that the mass consisted of irregularly arranged well-differentiated neuronal and glial cells, and multifocal mineral deposits. The neuronal cells had a large clear nucleus and various amount of Nissl substances in the cytoplasm. Some neural cells were bi-nucleated. Neither mitotic figures nor proliferating cell nuclear antigen (PCNA)-positive nuclei was found in the neuronal cells. Immunostaining for glial fibrillary acidic protein (GFAP) revealed diffuse proliferation of GFAP-positive glial cells and their processes, while these glial cells did not show apparent cellular atypism, mitotic activity, or PCNA-immunoreactivity. Accordingly, the present tumor was diagnosed as ganglioglioma, and hamartomatous histogenesis might be possible.     

猪各组织器官内肌生成抑制素基因mRNA表达谱的研究

用Trizol试剂法提取军牧一号猪的骨骼肌、心肌、肝、肾、肺、脾、胰、脂肪的总RNA做RT-PCR和嵌套PCR,结果在军牧一号猪骨骼肌、心肌和脂肪组织内检测到了肌生成抑制素mRNA的表达,而在其他组织中未检测到.     

Effect of simulated transport stress on the rat small intestine: A morphological and gene expression study

The present study investigated the effects of simulated transport stress on morphology and gene expression in the small intestine of laboratory rats. Sprague Dawley rats were subjected to 35 °C and 0.1×g on a constant temperature shaker for physiological, biochemical, morphological and microarray analysis before and after treatment. The treatment induced obvious stress responses with significant decreases in body weight (P < 0.01), increases in rectal temperature, serum corticosterone (CORT), serum glucose (GLU), creatine kinase (CK) and lactate dehydrogenase (LDH) levels (P < 0.01), as well as expression of Hsp27/70/90 mRNA (P < 0.05; P < 0.01). The rat jejunum was severely damaged and apoptotic after mimicking transport stress, which may mainly be related to cell death, oxidation reduction and hormone imbalance determined by microarray analysis. The bioinformatics analysis from the present study would provide insight into the potential mechanisms underlying transport stress-induced injury in the rat small intestine.     

Dynamics of mast cells in lymph node following antigenic stimulation

Dynamics of mast cells in rat cervical lymph nodes were examined using conventional histological techniques after injection of Salmonella paratyphi B-H antigen. There was no significant change in the number of mast cells at sixth hour and on the first day of stimulation compared with the controls. The number of mast cells was increased in all lymph node compartments on the second day of stimulation, which continued in the following 3 days. On the eighth day of stimulation, although the mast cell number decreased in the subcapsular area, it was still high in the paracortical area and medullary sinuses of the lymph nodes. On the second day of stimulation, the mast cell number was apparently increased in the subcapsular area than those of the other compartments. In the following days of stimulation, the highest number of mast cells was seen in the medullary sinuses. The highest paracortical mast cell number was determined on the third day of stimulation and some mast cells were observed near the high endothelial venules (HEVs). The changes of mast cell number among the lymph node compartments after antigenic stimulation support the hypothesis that the migration of mast cells occurred. This migration pattern indicates that mast cells enter the lymph node via afferent lymphatics and migrate to the lymph node compartments following antigenic stimulation.