Multiple skin tumours in a Doberman Pinscher with colour dilution alopecia

Abstract An 11-year-old Doberman Pinscher with clinical and histological skin features consistent with colour dilution alopecia had a long history of skin disease. The animal had hundreds of papules affecting the ‘blue’ haired áreas of the hair coat. The predominant skin lésions included keratinizing infundibular cysts, keratin horns, perifollicular dermatitis, and sebaceous hyperplasia; these lésions were often accompanied by fürunculosis and suppurative panniculitis. Over a 9-month period, 32 lésions were examined histologically, which included two lipomas, one infiltrative lipoma, one squamous papilloma, two mast cell tumours, four cavernous haemangiomas, one haemangiosarcoma, and three melanomas. It is unknown at present whether the colour dilution phenotype confers increased risk for tumours of the skin and or subcutaneous tissues. Résumé— Un doberman de 11 ans avec des manifestations cutanées cliniques et histologiques compatibles avec le díagnostic d'alopécie des couleurs diluées avait une longue histoire de problèmes cutanés. L'animal présentait des centaines de papules affectant les zones de couleur bleue. Les lésions cutanées prédominantes consistaient en kystes infundibulaires kératinisants, cornes cutanées, dermatite périfolliculaire et hyperplasie sébacée; ces lésions s'accompagnaient souvent de füronculose et de panniculite suppurative. Sur une période de 9 mois, 32 lésions ont fait l'objet d'un examen histologique, dont deux lipomes, un lipome infiltrant, un papillome, deux mastocytomes, quatre hémangiomes caverneux, un hémangiosarcome, et trois mélanomes. On ne sait actuellement pas si le phénotype couleur diluée constitue un facteur de risque accru pour les tumeurs de la peau et des tissus sous cutanés. [Madewell, B. R., Ihrke, P. J., Griffey, S. M. Multiple skin tumours in a Doberman Pinscher with colour dilution alopecia. (Tumeurs cutanées multiples chez un doberman présentant une alopécie des couleurs diluées.) Veterinary Dermatology 1997; 8 : 59–62.] Resumen Un doberman pinscher de 11 años con caracteristicas clinicas e histológicas compatibles con alopecia de color diluido tenia una larga historia de enfermedad cutanea. El animal presentaba centenares de pápulas afectando las áreas de pelo “azul”. Las lesiones prédominantes incluian quistes infundibulares queratinizados, cuernos de queratina, dermatitis perifolicular e hiperplasia sebácea; estas lesiones se acompañaban de fürunculosis y paniculitis supurativa. Durante un periodo de 9 meses, 32 lesiones fueron examinadas histológicamente, incluyendo 2 lipomas, un lipoma infiltrativo, un papiloma escamoso, dos mastocitomas, cuatro hemangiomas cavernosos, un hemangiosarcoma y tres melanomas. Se desconoce aún si el fenotipo de color diluido aumenta el riesgo para la aparición de tumores en tejidos cutáneos y subcutáneos. [Madewell, B. R., Ihrke, P. J., Griffey, S. M. Multiple skin tumours in a Doberman Pinscher with colour dilution alopecia. (Multiples tumores cutáneos en un Doberman Pinscher con alopecia de color diluido.) Veterinary Dermatology 1997; 8 : 59–62.] Zusammenfassung— Ein elfjähriger Dobermann mit klinischen und histologischen Befunden, die bei Farbmutantenalopezie auftreten, wies eine lange Vorgeschichte mit Hautkrankheiten auf. Das Tier zeigte Hunderte von Papeln, die die “blau” behaarten Bezirke des Felles betrafen. Die vorherrschenden Hautveränderungen bestanden in keratinisierenden infundibulären Zysten, Keratinhörnern, perifollikulärer Dermatitis und Talgdrüsenhyperplasie; diese Veränderungen wurden oft von Furunkulose und eitriger Pannikulitis begleitet. Über eine Zeitspanne von 9 Monaten wurden 32 Veränderungen histologisch untersucht, wovon zwei in Lipomen bestanden, eine in einem infiltrativen Lipom, eine in einem squamösen Papillom, zwei in Mastzelltumoren, vier in kavernösen Hämangiomen, eine in einem Hämangiosarkom und drei in Melanomen. Es ist zur Zeit nicht bekannt, ob der Phänotyp mit der Farbverdünnung mit einem erhöhten Risiko für Tumoren der Haut und/oder des subkutanen Gewebes einhergeht. [Madewell, B. R., Ihrke, P. J., Griffey, S. M. Multiple skin tumours in a Doberman Pinscher with colour dilution alopecia. (Multiple Hauttumoren bei einem Dobermann mit Farbmutantenalopezie.) Veterinary Dermatology 1997; 8 : 59–62.]     

Development of a method for simultaneously genotyping multiple horse coat colour loci and genetic investigation of basic colour variation in Thoroughbred and Misaki horses in Japan

In order to develop a genotyping method that can be used in the registration procedure for Thoroughbreds, we developed a method for simultaneously genotyping multiple coat colour genes on the basis of single nucleotide polymorphism typing by using the SNaPshot^TM technique. This method enabled precise and reasonable detection of causal mutations; it was effective for genotyping of MC1R, ASIP, and SLC45A2 at the Extension (E), Agouti (A), Cream dilution (C) loci, and the possibility of identification of rare variants of MC1R, EDNRB and KIT at the E, Overo (O) and Sabino 1 (SB1) loci, respectively, was also indicated. It was considered that this genotyping method would provide information not only for the registration of Thoroughbreds but also for the preservation of phenotypic characters, such as coat colour, of endangered Misaki native horses in Japan. Therefore, genetic variations at the five coat colour loci were investigated in 1111 Thoroughbred and 99 Misaki native horses. Allele frequencies at the polymorphic E and A loci were estimated, and the proportions of basic coat colours that could be expected in the Thoroughbred population were bay, 0.662; black, 0.070; chestnut, 0.268. In the Misaki population, they were bay, 0.792; black, 0.129; chestnut, 0.080. The data presented were the first of its kind on genetic coat colour variation, and will be important with regard to the registration of Thoroughbreds and the management of Misaki horses.     

小鼠骨骼肌Desmin启动子的克隆及表达活性分析

为了研究在体细胞中小鼠Desmin基因启动子是否具有表达活性,试验根据NCBI中小鼠Desmin启动子的序列,利用生物软件Primer Premier 6.0设计序列上、下游引物,克隆小鼠基因组中Desmin启动子片段,将克隆到的Desmin启动子片段与pAcGFP1-N1载体中的CMV启动子进行置换以构建真核表达载体pAcGFP1-N1-Des,然后将pAcGFP1-N1-Des依次转染CHO-K1、COS-7、293GP和NIH-3T3等细胞后,观察绿色荧光蛋白表达活性。结果表明:克隆得到的Desmin基因上游启动子(841 bp)与NCBI序列(NT_039173.7)比对同源性为99.76%;克隆得到的启动子中含有CAAT-box、GC-box核心调控区以及MyoD等多种转录因子结合位点;所构建的pAcGFP1-N1-Des经转染COS-7、CHO-K1和Hela细胞后可观察到呈现表达活性的绿色荧光。说明克隆获得了在COS-7、CHO-K1以及Hela细胞中的具有表达活性的小鼠Desmin基因启动子。     

Genetic parameters of the thick-to-thin albumen ratio and egg compositional traits in layer-type chickens

ABSTRACT

1. Generating a robust egg albumen is one of the major factors contributing to interior egg quality owing to its nutritive value and superior appearance. However, the genetic factors regulating the proportion of thick albumen are poorly understood.

2. In this study, 1330 eggs were collected from 450 Rhode Island White layers, aged 40 weeks, to measure egg compositional traits for three successive days. The restricted maximum likelihood method was applied to estimate genetic parameters for the thick-to-thin albumen ratio and other egg compositional traits. A univariate animal model was fitted to calculate heritability for each trait.

3. The heritabilities of egg weight, yolk weight, albumen height, Haugh units, percentages of yolk, thick albumen, thin albumen and the thick-to-thin albumen ratio were 0.32, 0.34, 0.28, 0.47, 0.61, 0.39, 0.31, and 0.45, respectively. The percentage of thick albumen was negatively correlated genetically with all traits, and percentage of thin albumen was negatively correlated genetically with all traits except for Haugh units. The thick-to-thin albumen ratio was positively correlated genetically with egg weight, albumen height and Haugh units, with correlations ranging from 0.21 to 0.54.

4. The results indicated that the percentage of thick albumen and the thick-to-thin albumen ratio were found to be moderately to highly heritable, and selection for the thick-to-thin albumen ratio could be conducive to the improvement of egg albumen quality.     

鸡Wnt3a基因组织表达差异与SNPs对鸡毛囊密度性状的遗传效应

Wnt家族成员3a （Wnt family member 3a,Wnt3a）基因是Wnt信号通路的重要成员,Wnt信号通路在动物皮肤毛囊的发生发育中具有重要作用。试验以Wnt3a为候选基因,旨在探讨鸡Wnt3a基因的组织表达差异与单核苷酸多态性（single nucleotide polymorphisms,SNPs）位点对鸡皮肤毛囊密度性状的遗传效应。用实时荧光定量PCR检测Wnt3a基因在鸡不同组织中的表达差异;用PCR扩增直接测序法对Wnt3a基因的SNPs位点进行筛查,验证和分析不同SNPs位点基因型背部毛囊密度的差异;用实时荧光定量PCR技术检测不同品种鸡皮肤组织中Wnt3a基因mRNA表达差异。结果表明,Wnt3a mRNA在皮肤组织中表达量最高,显著高于其他组织（P<0.05）,其次是肝脏、睾丸、卵巢和下丘脑,心脏、胸肌、垂体和脾脏等组织表达量较低。在Wnt3a基因第2和3外显子区各筛选到1个SNP位点：g.2587569 G>A和g.2555812 T>C;在第3内含子区筛选到1个SNP位点：g.2555377 T>C。卡方检验结果显示,3个位点均极显著偏离Hardy-Weinberg平衡。群体遗传参数分析发现,g.2555812 T>C位点为中度多态,有效等位基因数为1.7,遗传变异程度较g.2587569 G>A和g.2555377 T>C位点高。SNP位点分型与毛囊密度的关联分析结果表明,g.2555377 T>C位点CC基因型个体的毛囊密度显著高于TT和CT基因型个体（P<0.05）。在毛囊密度具有显著差异的不同品种鸡的背部皮肤组织中,Wnt3a mRNA表达差异显著（P<0.05）。该研究结果为进一步深入解析Wnt3a在鸡皮肤毛囊生长发育中的作用、筛选与毛囊密度性状相关的辅助育种标记提供了基础资料。     

家蚕胚胎各发育时期酚氧化酶原基因的转录活性分析

酚氧化酶在昆虫体液免疫反应中起着非常重要的作用。利用GenBank上登录的家蚕酚氧化酶原基因的cDNA序列设计特异引物．通过半定量RT—PCR技术检测了家蚕胚胎各发育时期（从未受精卵到产卵后第9d）酚氧化酶原基因的转录活性,发现该基因的转录表达具有时空特异性。我们的试验结果为进一步研究家蚕酚氧化酶原基因的功能提供了新的线索。     

Eye-drop DNA can induce IgA in the tears and bile of chickens

DNA vaccines protect chickens against lethal virus infections but whether they induce local antibody which is associated with preventing viral entry, is unknown. We were able to show how avian DNA vaccines can induce local IgA. 65 μg plasmid DNA encoding the reporter protein beta-galactosidase induced antigen-specific IgA in the tears of 6/10 birds, IgA in the bile of 4/10 birds and IgG in the serum of 2/10 birds. Giving the DNA by the intramuscular route, as is more usual, induced lacrimal IgA in 2/8 birds, biliary IgA in no birds and serum IgG in 4/8 birds. Eye-drop DNA therefore favoured local IgA whereas intramuscular DNA favoured serum IgG. Further to this preliminary work eye-drop DNA should be improved by adjuvants and cytokines as a way of inducing protective IgA at the mucosal surfaces of the alimentary and respiratory tracts.     

Association of SNPs in exon 2 of the MHC B-F gene with immune traits in two distinct chicken populations: Chinese Beijing-You and White Leghorn

Abstract

Antibody titers raised for vaccinations against avian influenza (AI) and Newcastle disease (ND) were higher in Chinese Beijing-You (BJY) than in White Leghorn (WL) (P<0.001), but there was no breed difference in titers for sheep red blood cells (SRBC). Genotyping by PCR-SSCP identified seven haplotypes in WL and 17 in BJY. After sequencing PCR products (35 and 85, respectively), 43 (WL) and 47 (BJY) single nucleotide polymorphisms (SNPs) were found in the 264 bp of exon 2. In WL chickens, significant associations were found with antibody responses to AI (two SNPs), ND (six SNPs), and SRBC (one SNP), while in BJY there was association with responses to ND (two SNPs) and SRBC (two SNPs), but none with AI. These results indicate that the genomic region bearing exon 2 of the major histocompatibility complex B-F gene has significant effects on antibody responses to SRBC and vaccination against AI and ND. Different SNPs affected antibody titers for each of the antigens and they differed between these very distinct breeds.     

鸡传染性贫血免疫对肝微粒体抗氧化酶活性的影响

目的是观察鸡传染性贫血多次免疫对肝微粒体中抗氧化酶活性的影响.对20只SPF鸡随机分为2组,每组10只,免疫组鸡用鸡传染性贫血弱毒苗免疫4次,每次间隔2周,对照组鸡注射同剂量的生理盐水.最后一次免疫后10d取肝脏制备微粒体,利用测试盒测定肝微粒体中的GSH-Px活性、SOD活性、CAT活性和MDA含量.结果与对照组相比,免疫组肝微粒体中GSH Px活性、SOD活性和CAT活性都显著提高(P＜0.05),MDA含量显著减低(P＜0.05).结论为鸡传染性贫血多次免疫可提高鸡体抗氧化能力.     

ADSL基因启动子区遗传变异与鸡肉冻藏新鲜度的相关性分析

试验旨在探究鸡腺苷琥珀酸裂解酶（adenylosuccinate lyase,ADSL）基因启动子区多态性及其与鸡肉冻藏新鲜度的相关性。以161只青脚麻鸡为研究对象,通过DNA混池测序法检测ADSL基因启动子区遗传变异。采用等位基因特异性PCR完成个体基因型分析,并分析了各基因型与鸡胸肌肉色、体重及胸肌冷冻60 d后新鲜度K值和pH的相关性。结果显示,在ADSL基因启动子区ATG上游1 670 bp处发现1个SNP位点（c.-1670 C>A）,其AA基因型个体肌肉冷冻60 d后新鲜度K值（23.61%）极显著低于CA（33.00%）和CC（33.56%）基因型（P<0.01）,即冻藏60 d后AA基因型个体肌肉较CA和CC基因型新鲜,CA和CC基因型间新鲜度K值差异不显著（P>0.05）;各基因型间鸡体重、胸肌肉色及胸肌冷冻60 d后pH均无显著差异（P>0.05）。生物信息学预测发现,c.-1670 C>A导致ADSL基因启动子区转录因子结合位点的显著改变,该突变位点可能造成ADSL基因的表达差异。综上,ADSL基因启动子区c.-1670 C>A与青脚麻鸡胸肌冷冻60 d后新鲜度K值具有显著相关性,该位点可作为青脚麻鸡肉品新鲜度的候选分子遗传标记。     

鹅MyoG基因启动子活性及转录调控元件分析

旨在分析鹅MyoG基因启动子活性区域和转录因子,探究该基因的转录调控机制.本研究首先通过PCR扩增泰州鹅MyoG基因5'侧翼区序列1 245 bp并对其进行测序和生物信息学分析,其次,构建4个不同缺失片段的双荧光素酶报告载体,转染C2C12细胞系.进一步利用在线软件预测核心启动子区关键转录因子,对转录因子结合位点HNF...     

Polymorphisms in the ovoinhibitor gene (OIH) and their association with egg quality of Xinhua E-strain chickens

1. Chicken ovoinhibitor has been reported to prevent contamination and affect storage for eggs. The objective of the present study was to conduct an association analysis between ovoinhibitor gene (OIH) polymorphisms and egg quality traits in the population of Xinhua E-strain chickens and explore its expression characteristics in different tissues.

2. Three single nucleotide polymorphisms (SNPs) were identified and genotyped, in which one synonymous mutation was located in exon16 (G9810A), one in intron7 (A4363G) and one in intron14 (C8937G). The A4363G and C8937G polymorphisms signi?cantly in?uenced Haugh unit (HU) and albumen height (AH), while HU and AH in AG and CT heterozygous birds were signi?cantly higher than in birds with homozygous genotypes. Diplotype association analysis showed individuals with an H1H4 diplotype had the highest AH and HU values in the investigated population.

3. RT-qPCR results showed that the expression level of the OIH gene was higher in the oviduct and liver than other tissues. During the development of liver at different phases, a drastic decrease occurred during the period of first ovulation event, which suggests the regulation of some unknown factors effected it.

4. Our results indicated that OIH plays a vital role in egg quality. The combinations of A4363G and C8937G might be potential advantageous molecular markers for improving HU of chicken eggs.     

摩拉水牛和尼里-拉菲水牛PRKAA2基因SNPs检测及遗传多样性分析

为研究水牛蛋白激酶AMP活化的催化亚基α2（protein kinase AMP-activated catalytic subunit alpha 2,PRKAA2）基因多态性,本试验以摩拉水牛和尼里-拉菲水牛基因组DNA为模板,扩增PRKAA2基因外显子4及内含子3部分序列,通过常规测序法检测其SNP并进行遗传多样性分析。结果发现,PRKAA2基因外显子4内存在1个SNP位点（c.462 G>A）,PRKAA2基因内含子3部分序列存在3个SNPs位点（IVS3.557 T>C、IVS3.560 C>T和IVS3.565 G>A）。经遗传多样性分析表明,在c.462 G>A位点的野生纯合型和杂合型比突变纯合型更有优势,IVS3.557 T>C和IVS3.560 C>T位点的突变纯合型为非优势基因型,IVS3.565 G>A位点杂合型为优势基因型。IVS3.565 G>A位点在摩拉水牛群体中处于Hardy-Weinberg非平衡状态;c.462 G>A位点在尼里-拉菲水牛群体中处于Hardy-Weinberg非平衡状态。4个SNPs位点在摩拉水牛群体中均为中度多态;c.462 G>A、IVS3.557 T>C位点在尼里-拉菲水牛群体中为低度多态,IVS3.560 C>T、IVS3.565 G>A位点为中度多态。IVS3.557 T>C位点在两个水牛群体中杂合度较低。说明摩拉水牛IVS3.565 G>A位点和尼里-拉菲水牛c.462 G>A位点的基因型频率和基因频率遗传状态不平衡,尼里-拉菲水牛群体中IVS3.557 T>C位点遗传变异小,选择潜力不高。4个多态位点可以构建5种单倍型,其中T-C-G-G是摩拉水牛群体和尼里-拉菲水牛群体的优势单倍型。综上,本研究检测的摩拉水牛和尼里-拉菲水牛PRKAA2基因上4个SNPs位点可为水牛标记辅助选择育种提供参考。     

牛SCD1基因C878T位点遗传多态性与中国西门塔尔牛部分脂肪相关性状的效应分析

为探索8个中国牛群SCD1基因多态性与屠宰和肉质性状的相关性,选取中国西门塔尔牛、雷琼牛、云南高峰牛、BMY牛、闽南黄牛、鲁西黄牛、渤海黑牛和中国南方荷斯坦牛等8个群体共682头个体为研究对象,采用PCR-SSCP法分析SCD1基因遗传多态性。结果表明,在878bp处发现1个碱基C→T的突变(C878T),导致蛋白质肽链中丙氨酸(alanine)突变为缬氨酸(valine)。C878T位点在所研究群体中表现为CC、CT和TT3种基因型,其中,中国西门塔尔牛中TT基因型频率较高(0.114),鲁西黄牛和渤海黑牛中较低(0.050/0.063),4个热带群体中未发现TT基因型。采用GLM对SCD1基因C878T位点与132头中国西门塔尔屠宰牛的部分脂肪相关性状进行关联分析。结果,CC基因型个体肌间脂肪含量和肠系膜油质量显著高于TT型个体(P<0.05),背膘厚极显著低于TT型个体(P<0.01),其他性状间差异不显著(P>0.05)。结果表明,SCD1基因C878T位点突变对中国西门塔尔牛脂肪相关性状有较大的遗传效应,可用于其部分屠宰与肉质性状的分子标记辅助选择。     

利用鳕鱼皮制备饲用胶原肽添加剂及其抗氧化研究

以鳕鱼皮为原料,采用双酶复合法提取胶原多肽作为饲用添加剂。以水解度为指标,通过正交试验确定酶解的最适条件。结果表明:碱性蛋白酶和木瓜蛋白酶混合水解,加酶量分别为4%和4.5%,温度55℃p、H值8,时间11 h,蛋白质水解度可达到25.61%。分离得到的多肽组分,Mr<3 500 Da的占82%以上,抗氧化试验结果表明,水解产物有较好的抗氧化特性,有望作为高档水产饲料添加剂使用。     

鹅p21基因克隆、启动子分析及转录调控研究

【目的】 分析鹅p21基因的结构和启动子活性,探讨p21基因的转录调控机制。【方法】 以泰州鹅为试验对象,通过同源克隆、RACE和生物信息学分析等方法获得鹅p21基因全长序列和5′-侧翼区序列特征;构建6个不同缺失片段的启动子区双荧光素酶报告载体并分析其荧光素酶活性,进而确定p21基因核心启动子区;对核心启动子区转录因子结合位点生肌决定因子(MyoD)(+25~+36 bp)进行定点突变,并构建突变报告基因载体,在C2C12细胞系内初步鉴定鹅p21基因核心转录调控因子。【结果】 鹅p21基因cDNA全长1 943 bp,CDS区大小为453 bp,编码151个氨基酸,蛋白序列包含高度保守的CDI家族结合位点。系统进化树分析表明,鹅p21基因与鸭亲缘关系最近,与鸡和火鸡有较强的进化关系。鹅p21基因5′-侧翼区包含启动子元件,—35~+37 bp是核心启动子区,发挥正向调控作用,结合定点突变技术初步鉴定MyoD是鹅p21基因核心转录调控元件。【结论】 本研究获得了鹅p21基因完整的cDNA序列和启动子区域,MyoD是p21基因核心转录调控因子,为探究p21基因在鹅胚胎期肌肉发育过程中的调控机制提供理论依据。