Genome-wide DNA methylation profiles in Tibetan and Yorkshire pigs under highaltitude hypoxia

Background: Tibetan pigs, which inhabit the Tibetan Plateau, exhibit distinct phenotypic and physiological characteristics from those of lowland pigs and have adapted well to the extreme conditions at high altitude.However, the genetic and epigenetic mechanisms of hypoxic adaptation in animals remain unclear.Methods: Whole-genome DNA methylation data were generated for heart tissues of Tibetan pigs grown in the highland(TH, n = 4) and lowland(TL, n = 4), as well as Yorkshire pigs grown in the highland(YH, n = 4) and lowland(YL, n = 4), using methylated DNA immunoprecipitation sequencing.Results: We obtained 480 million reads and detected 280679, 287224, 259066, and 332078 methylation enrichment peaks in TH, YH, TL, and YL, respectively. Pairwise TH vs. YH, TL vs. YL, TH vs. TL, and YH vs. YL comparisons revealed6829, 11997, 2828, and 1286 differentially methylated regions(DMRs), respectively. These DMRs contained 384, 619,192, and 92 differentially methylated genes(DMGs), respectively. DMGs that were enriched in the hypoxia-inducible factor 1 signaling pathway and pathways involved in cancer and hypoxia-related processes were considered to be important candidate genes for high-altitude adaptation in Tibetan pigs.Conclusions: This study elucidates the molecular and epigenetic mechanisms involved in hypoxic adaptation in pigs and may help further understand human hypoxia-related diseases.     

Genome-wide DNA methylation and transcriptome analyses reveal the key gene for wool type variation in sheep

Background Wool fibers are valuable materials for textile industry. Typical wool fibers are divided into medullated and non-medullated types, with the former generated from primary wool follicles and the latter by either primary or secondary wool follicles. The medullated wool is a common wool type in the ancestors of fine wool sheep before breeding. The fine wool sheep have a non-medullated coat. However, the critical period determining the type of wool follicles is the embryonic stage, which l...     

Surgical denervation of porcine ovaries during the middle luteal phase of the oestrous cycle changes their morphology and steroidogenic activity

Changes in both the morphology and the steroidogenic activity of porcine ovaries denervated surgically on day 12 of the oestrous cycle were studied. Neurectomy of the plexus and the superior ovarian nerves caused a dramatic reduction in the number (or even a disappearance) of dopamine-beta-hydroxylase- and/or neuropeptide tyrosine-immunoreactive nerve terminals. On day 20 of the subsequent oestrous cycle, the number of small follicles increased (P < 0.01) and that of large follicles decreased (P < 0.05) in the denervated ovaries, as compared to the controls. Neurectomy led to a decrease in the level of progesterone (P4; P < 0.001) and androstenedione (A4; P < 0.01) in the fluid from small follicles, A4 (P < 0.001) and testosterone (T; P < 0.05) in the fluid from medium-sized follicles, as well as in the content of all these steroids in the fluid from large-sized follicles (P < 0.001 for P4 and P < 0.05 for A4 and T). Denervation also caused a decrease in the content of A4 (P < 0.01) and T (P < 0.001) in the wall of follicles. Neurectomy resulted in a significant increase in the immunoexpression of cholesterol side-chain cleavage cytochrome P450 in the follicles and a decrease of 33-hydroxysteroid dehydrogenase. After denervation, plasma levels of LH, P4, A4, T, oestrone and oestradiol-17beta were lower (P < 0.05-0.001) on the particular days of the study than in the control group. Our data revealed that the denervation of ovaries during the middle luteal phase of the oestrous cycle in gilts caused distinct changes in both the morphology and the steroidogenic activity of the organ, confirming an important role of the peripheral nervous system in the control of the gonad in this species.     

Oxytocin receptors in dioestrous and anoestrous canine uteri

The aim of the study was to localize oxytocin receptors (OTR) and measure mRNA expression of OTR in the canine uterus with and without the influence of progesterone. Uterine samples were taken from nine anoestrous and eight dioestrous bitches during ovariohysterectomy. Histological changes were evaluated in haematoxylin and eosin (HE)‐stained samples. Purified polyclonal antibody for OTR was used in immunohistochemistry to localize receptors in uterine layers. Relative mRNA concentration of OTR was evaluated with real‐time PCR from full‐thickness uterine samples taken from the middle horn and the body. Myometrial smooth muscle cells, endometrial luminal epithelium (LE) and deep and superficial glandular epithelium were positively stained for oxytocin receptors in non‐pregnant animals. No significant difference in staining intensity was detected between uterine middle horn and body. However, the staining intensity of LE was significantly higher in dioestrous than in anoestrous uteri (p < .05). Leucocytes and endothelium of blood vessels were also positively stained for OTR. Real‐time PCR showed no significant differences in OTR mRNA expression between the middle horn and the body of the uterus, or between anoestrous and dioestrous uterus. No correlation was noted between OTR mRNA expression and blood progesterone concentration. In conclusion, despite the apparent inactivity, the uterus of the non‐pregnant bitch expresses OTR. The distribution or relative expression of OTR does not differ between uterine horn and body in dioestrus or anoestrus except in LE. LE may have more oxytocin‐dependent activity during dioestrus than anoestrus.     

山羊关节炎-脑炎核酸疫苗研究进展

山羊关节炎脑炎病毒（Caprine arthritis-encephalitis virus，CAEV）是反转录病毒科慢病毒属的成员之一，与其同属于一科的还有马传染性贫血、人免疫缺陷病、猴免疫缺陷病、牛免疫缺陷病、猫免疫缺陷病以及梅迪-维斯纳病毒，山羊关节炎脑炎病毒与梅迪-维斯纳病毒极为相似，二者在血清学上有多种交叉反应。山羊关节炎脑炎病毒主要侵染宿主的单核／巨噬细胞系的细胞，引起以成年山羊的关节炎以及羔羊的脑炎症状为主的慢性持续性感染。该病潜伏期长，可通过母羊哺乳及呼吸道和消化道广泛传播，又没有有效的药物和疫苗对其进行防制，使CAEV的感染呈世界性分布。     

Shape changes in caprine erythrocytes exposed to chlorpromazine and lysolecithin

Stomatocytic and echinocytic transformations of caprine erythrocytes were studied in vitro using chlorpromazine as a stomatocyyic agent and lysolecithin as an echinocytic agent. Morphologic changes in erythrocytes generally varied with the cell shape and the concentration of the substances used. Discoytic, triangular, and pear-shaped red cells common to normal goats, exhibited classical stomatocytic and echinocytic changes with formation of spherostomatocytes, sphero-echinocytes, and spherocytes. In comparison, fusiform and spindle-shaped red cells found in certain Angora goats seemed less prone to shape changes and required greater concentrations of the inducing agents to effect such changes. Chlorpromazine at higher concentrations also inflicted localized membrane damage in form of tiny pits and lysolecithin likewise induced formation of fragile smooth or beaded filaments.     

Genetic differences in oestrous signs and oestrogen metabolism‐related genes between Chinese Mi and European Landrace‐Large White pigs

Oestrous signs affect timely mating and reproductive efficiency in swine breeding herds. To study the genetic difference of oestrous signs between Chinese and European pigs, 100 Landrace‐Large White (LLW) cross gilts and 50 Chinese Mi gilts were assessed for oestrous signs and the concentrations of serum estradiol‐17β and progesterone were determined. The genotype of 39 single nucleotide polymorphisms (SNPs) in 11 oestrogen metabolism and function‐related genes was determined by Sequenom iPLEX platform. Compared with LLW gilts, Mi gilts had longer time of standing reflex (p < .001), higher scores of vulva reddening (p = .001) and greater serum estradiol‐17β concentration (p < .01). Gilts with greater serum estradiol‐17β concentrations also had greater (p < .05) scores for oestrous signs. Genetic polymorphisms of nine genes in oestrogen metabolism pathways had significant differences (p < .05) between LLW and Mi gilts. There were three and six haploblocks of SNPs in LLW and Mi, respectively. Compared with LLW, the distribution of haplotypes was more centralized in Mi pigs. Genetic polymorphisms of oestrogen metabolism‐related genes have considerable differences between Chinese Mi and European LLW pigs. Because of the important roles of oestrogen during the oestrus, some genes of oestrogen metabolism pathway could be considered as candidate genes for oestrous signs.     

Ultrastructural changes in caprine lungs infected with Mycoplasma capricolum subspecies capripneumonia

Ultrastructural examination of the lungs from goats with natural infections with Mycoplasma capricolum subspecies capripneumonia, the causative agent of contagious caprine pleuropneumonia, revealed extensive hyperplasia of type II pneumocytes. Typically, these type II cells contained abundant numbers of osmiophilic lamellar bodies that had lost most of their characteristic lamellar ultrastructure. These findings were absent in healthy control goats.     

不孕奶牛卵巢异常的B超形态学

应用便携式实时B超诊断仪,采用直肠探测法探查武汉市几个奶牛场的37头长期不发情或屡配不孕奶牛的卵巢形态结构。采集到大量典型的奶牛异常卵巢B超声像图,包括卵巢囊肿、幼稚型卵巢、卵巢静止、卵巢萎缩、卵巢炎等。试验利用超声探查技术观察了不孕奶牛卵巢形态结构的变化特点,为奶牛不孕疾病诊断提供参考。     

Age related changes in the capillary index of the caprine hypophysis cerebri

The capillary index of the hypophysis cerebri of goats, ranging in age from one day to over 6 years of age. Observations in rostral, middle and caudal regions were made and comparisons are made with other species.     

犏牛雄性不育的DNA甲基化

采用Western blot法检测犏牛、牦牛睾丸组织中DNA甲基转移酶3a(Dnmt3a)蛋白的相对表达量,用甲基化定量试剂盒(荧光法,Epigentek公司)检测其全基因组DNA甲基化的水平,分析犏牛与牦牛在这2个表达量上的差异。结果显示:犏牛、牦牛睾丸组织中Dnmt3a蛋白的相对表达量分别为(0.853±0.015)和(0.651±0.016),犏牛的显著高于牦牛(P<0.05);犏牛、牦牛睾丸组织的全基因组DNA甲基化水平分别为(0.950±0.013)%和(0.808±0.016)%,犏牛的显著高于牦牛(P<0.05)。可见,犏牛睾丸组织中Dnmt3a蛋白表达水平与其全基因组DNA甲基化水平均显著高于牦牛,这可能是犏牛雄性不育的DNA甲基化机制之一,也将为犏牛雄性不育的表观遗传学研究奠定基础。     

Dexamethasone-induced changes in sympathetic innervation of porcine ovaries and in their steroidogenic activity

The aim of the present study was to determine changes in the density of sympathetic nerves in porcine ovaries with dexamethasone (DXM)-induced cysts and the alterations in steroidogenic activity and amounts of catecholamines in the affected gonads. Cystic ovaries were supplied by numerous sympathetic nerve fibers. The amount of noradrenaline in the cysts (fluid, wall) was significantly higher than in the large follicles of the control group. After DXM injections, the amounts of noradrenaline and adrenaline significantly increased in the walls of small and medium-sized follicles. In the cysts (fluid, wall) the levels of androgens and estrogens were significantly lower, whereas progesterone was higher in the cystic wall. DXM administration led to a significant increase in the estrone content in the fluid of small follicles. Moreover, a decrease in the amounts of progesterone and androgens was found in the follicular fluid and walls of medium-sized follicles. DXM injections resulted in a significant increase in the immunoexpression of P450(scc) and 3beta-HSD in the cysts, a significant increase of P450(scc) in the follicles, and a decrease of 3beta-HSD and P450(arom). The present study shows that the DXM treatment leads to an increase in the density of intraovarian sympathetic nerves, paralleled by the amount of catecholamines, and that it is capable of changing the steroidogenic activity of porcine ovary bearing cysts. Thus, it appears possible that these events may be, at least partly, involved in the pathogenesis of this disorder.     

狼山鸡下丘脑和卵巢组织全基因组DNA甲基化分析

为了获得狼山鸡性腺轴组织基因组DNA甲基化水平和模式等表观遗传信息,试验采用全基因组重亚硫酸盐测序（whole genome bisulfite sequencing,WGBS）技术检测狼山鸡下丘脑和卵巢组织基因组DNA甲基化状态,分析两组织DNA甲基化水平及特异甲基化模式。结果表明,狼山鸡下丘脑和卵巢基因组整体甲基化水平分别为4.35%和3.48%,差异显著（P<0.05）;下丘脑和卵巢中分别检测到6 150 000和10 320 000个甲基化胞嘧啶（mC）位点,其中mCG类型位点分别占69.99%和87.88%,下丘脑中非mCG位点占比约为卵巢中的2.5倍;与各染色体不同,两组织线粒体基因组中mCHH位点占比最高,其次是mCHG位点;卵巢基因组启动子区DNA甲基化水平极显著低于内含子和外显子区（P<0.01）,极显著高于基因间区（P<0.01）;下丘脑基因组启动子区DNA甲基化水平与内含子和外显子区相比差异不显著（P>0.05）,却显著高于基因间区（P<0.05）;下丘脑基因组各功能元件DNA甲基化水平均显著或极显著高于卵巢基因组（P<0.05;P<0.01）。综上,狼山鸡下丘脑和卵巢组织具有不同的DNA甲基化模式和特征,下丘脑中较高的非mCG位点比例可能在中枢神经系统发育中发挥重要作用,本研究结果为进一步分析鸡卵巢和下丘脑基因组DNA甲基化对其繁殖性能调控机制提供参考依据。     

Repeated pregnant mare serum gonadotropin‐mediated oestrous synchronization alters gene expression in the ovaries and reduces reproductive performance in dairy goats

This study aimed to elucidate the effects of repeated pregnant mare serum gonadotropin (PMSG) treatment for oestrous synchronization (ES) on ovarian gene expression and reproductive parameters in Xinong Saanen dairy goats, the dominant breed of dairy goat in China. The experiment was carried out at the Research Station of Northwest A&F University (NWAFU), China (34°16′N, 108°4′E). Forty‐one does were randomly assigned to groups receiving ES treatments thrice every fortnight (3‐PMSG group; n = 19), or ES treatment only once simultaneously with the third ES treatment in the 3‐PMSG group (1‐PMSG group; n = 22) during middle of the breeding season from late July (14 hr light) until late September (12 hr light). ES treatment was performed via intravaginal insertion of a controlled internal drug release (CIDR) device impregnated with 300 mg progesterone (P4), followed by 300 IU PMSG injections 48 hr before CIDR withdrawal. Oestrus was monitored using vasectomized bucks. Ovaries of three goats in oestrus from both groups were harvested for morphological examination and RNA sequencing (RNA‐Seq). Then, all the oestrous goats in the 1‐PMSG (n = 21) and 3‐PMSG (n = 11) groups were artificially inseminated twice. The 3‐PMSG group showed reduced oestrous rate (57.89%), pregnancy rate (31.58%) and litter size (1.17) compared, respectively, with 95.45%, 68.18% and 1.67 for 1‐PMSG group (p < 0.05). However, no differences were found in the ovarian morphology between the 1‐PMSG and 3‐PMSG groups (p > 0.05). RNA‐Seq revealed 114 differentially expressed genes (DEGs) in the ovaries of the 3‐PMSG group, among which GCG, FSTL3, TET3 and AQP3 were deemed novel and promising candidate genes for regulating fertility. The present study indicates that the three‐time PMSG treatment dysregulated several ovarian genes, thereby reducing reproductive performance.     

Expression levels of mRNA-encoding PDGF receptors in goat ovaries and the influence of PDGF on the in vitro development of caprine pre-antral follicles

The aims of this study were to investigate the expression levels of mRNA for platelet-derived growth factor (PDGF) receptors (PDGFR-α and -β) in caprine follicles at different developmental stages and to evaluate the influence of PDGF on the in vitro development of pre-antral follicles. For this, goat primordial, primary and secondary follicles, as well as small (1-3 mm) and large (3-6 mm) antral follicles, were obtained, and PDGFR-α and -β mRNA levels were quantified by real-time PCR. Furthermore, pre-antral follicles (≥ 200 μm) were isolated from goat ovaries and cultured for 18 days in α- minimum essential medium supplemented with PDGF at 50 or 100 ng/ml, containing or not FSH. Real-time PCR showed highest PDGFR-α mRNA levels in secondary follicles, while PDGFR-β mRNA levels were highest in primary follicles onwards. Both receptors showed higher mRNA levels in granulosa/theca cells from small and large antral follicles than in their corresponding cumulus-oocyte complexes. In culture, the percentage of antrum formation was significantly higher in 100 ng/ml PDGF compared with the same PDGF concentration associated with FSH. After 18 days, PDGF in both concentrations associated with FSH promoted follicular growth significantly higher than the control. Moreover, the addition of FSH to 50 ng/ml PDGF positively influenced the follicular growth when compared with the same PDGF concentration in the absence of FSH. In conclusion, PDGF is important for early goat folliculogenesis, because the presence of PDGFR-α and -β mRNA was detected in all follicular categories, and PDGF associated with FSH stimulated the growth of goat pre-antral follicles isolated and cultured in vitro.