Expression and localization of adiponectin and its receptors in ovarian follicles during different stages of development and the modulatory effect of adiponectin on steroid production in water buffalo

Adiponectin is an adipocyte‐derived hormone regulating energy metabolism, insulin sensitivity and recently found to regulate reproduction. The current study was carried out to investigate gene and protein expression, immunolocalization of adiponectin and its receptors AdipoR1 and AdipoR2 in ovarian follicles of different developmental stages in water buffalo (Bubalus bubalis) and to investigate the effect of adiponectin on steroid production in cultured bubaline granulosa cells. qPCR, western blotting and immunohistochemistry were applied to demonstrate mRNA expression, protein expression and immunolocalization, respectively. The results indicate that adiponectin, AdipoR1 and AdipoR2 were present in granulosa cells (GC) and theca interna (TI) of ovarian follicles and the expression of adiponectin, AdipoR1, AdipoR2 in GC and AdipoR1 and AdipoR2 in TI increased with increase in follicle size (p < .05). Expression of adiponectin was high in small and medium size follicles in TI. The adiponectin and its receptors were immunolocalized in the cytoplasm of GC and TI cells. Further, in the in‐vitro study, GCs were cultured and treated with recombinant adiponectin each at 0, 1 and 10 µg/ml alone or with follicle stimulating hormone (FSH) at 30 ng/ml) or Insulin‐like growth factor I (IGF‐I) at 10 ng/ml for 48 hr after obtaining 75%–80%s confluency. Adiponectin at 10 µg/ml increased IGF‐I‐induced estradiol (E₂) and progesterone (P₄) secretion and FSH‐induced E₂ secretion from GC and also increased the abundance of factors involved in E₂ and P₄ production (cytochrome P45019A1 [CYP19A1] and 3‐beta‐hydroxysteroid dehydrogenase [3β‐HSD]). In conclusion, this study provides novel evidence for the presence of adiponectin and its receptors in ovarian follicles and modulatory role of adiponectin on steroid production in buffalo.     

小鼠卵巢组织定量PCR分析中内参基因的筛选

旨在选择小鼠卵巢组织中合适的内参基因,为研究卵巢发育过程中基因表达提供可靠数据。以不同年龄(0日龄、3周龄、5周龄、8周龄)小鼠卵巢组织为试验材料,Trizol法提取样本总RNA,并合成cDNA,根据已报道的小鼠(Mus musculus)各组织中相对稳定表达的8个常见内参基因(Gapdh、β-actin、β-tubulin、18S rRNA、16S rRNA、H2afz、Ubc、Rpl13a)的GenBank登录序列设计引物,采用实时荧光定量PCR(qRT-PCR)方法构建卵巢cDNA等梯度(1∶10)稀释后的标准曲线;利用geNorm分析候选内参基因的表达稳定性(M);NormFinder筛选表达最稳定的内参基因;BestKeeper计算qRT-PCR结果的标准差(SD)和变异系数(CV),从而预测候选内参基因的稳定性。结果表明,8个内参基因的引物特异性较强,具有良好的线性关系;候选内参基因的表达稳定度排序:Gapdh=β-actin>18S rRNA>Ubc>16S rRNA> H2afz>Rpl13a>β-tubulin;其中Gapdh表达稳定性最好,且标准差及方差系数最小(SD:0.32,CV:1.95),H2afz标准差及方差系数最大(SD>1.0,CV:5.76)。综上表明,本研究成功获得出生后小鼠卵巢发育过程中稳定表达的内参基因(Gapdh和β-actin),可作为其基因表达研究中的最佳候选内参。     

CYSTIC ENDOMETRIAL HYPERPLASIA AND BACTERIAL ENDOMETRITIS ASSOCIATED WITH AN INTRAUTERINE FOREIGN BODY IN A GUINEA PIG WITH OVARIAN CYSTIC DISEASE

A 3-year-old, 920 g intact female guinea pig presented with a 4-month history of nonpruritic hair loss on the ventral abdomen. The physical examination revealed flank and ventral abdominal alopecia, mucoid vulvar discharge, and abdominal distension. Bilateral rounded masses just caudal to the kidneys and structures consistent with enlarged uterine horns were detected on abdominal palpation. Abdominal ultrasound revealed bilateral ovarian cysts, thickened uterine horns, and multiple circular hypoechoic and anechoic structures in the uterine wall. The patient underwent ovariohysterectomy. Gross examination of the uterus revealed a piece of hay in the left uterine horn. A final diagnosis was hormonally active ovarian follicular cysts, cystic endometrial hyperplasia, and purulent bacterial endometritis caused by Escherichia coli, Fusobacterium nucleatum, and Arthrobacter spp. Cystic endometrial hyperplasia is infrequently reported in guinea pigs, and this report describes an associated bacterial endometritis and uterine foreign body with concurrent ovarian cysts.     

香猪卵巢StAR和CYP11A1基因的差异表达研究

为了揭示香猪繁殖调控的分子机制,本试验结合生物信息学分析方法对StAR和CYP11A1基因在香猪卵巢组织中的差异表达进行分析。运用实时荧光定量PCR方法检测高、低产群体6月龄香猪卵巢组织中StAR和CYP11A1基因的表达量变化;从香猪基因组中克隆两个基因的启动子区域,分析两个基因表达差异的分子机制。高通量转录组测序和实时荧光定量PCR检测结果均显示,高产香猪两个基因的表达量较高,启动子区序列长分别为1 144与970 bp。StAR基因启动子区结合转录因子ER、COUP、Sp1等34种72个,转录起始位点上游68~698 bp存在8个变异位点,但不影响转录因子的结合;与CYP11A1基因启动子区相结合的转录因子为Sp1、PU.1、C/EBPdel等30种75个,转录起始位点上游12~593 bp存在5个变异位点,其中69 bp处检测到C-T变异,高产香猪的T等位基因检出频率为17%,低产香猪为33%;对预测结果整合分析,T等位基因丢失了转录因子Sp1的结合位点。高、低产香猪之间StAR和CYP11A1基因的表达存在差异,CYP11A1基因启动子区69 bp处的变异可能是基因表达量差异的原因之一。     

乏情和发情初产母猪卵巢microRNA差异表达分析

为了探索卵巢microRNA （miRNA）在初产母猪生殖调控中的作用,本研究利用Illumina高通量测序技术检测了乏情和发情初产母猪卵巢miRNA的表达谱,并对表达量较高且差异显著的miRNA进行生物信息学分析。结果显示,本研究构建的两个small RNA文库共鉴定出503个miRNAs,其中已知的303个,新预测的200个。在已知的miRNA中,ssc-miR-10b在两个文库中的表达量最高,其次为ssc-miR-143-3p和ssc-miR-26a;在新预测的miRNA中,chr13_2637_mature在两个文库中的表达量最高,其次为chr8_9994_mature。与发情母猪相比,共有145个miRNAs发生显著变化（read counts>10,∣log₂（fold-change）∣>1）,上调114个,下调31个。在进一步筛选的31个表达量较高且差异显著的miRNAs（read counts>1 000,∣log₂（fold-change）∣>1）中,新预测的chr13_2585_mature上调倍数最高,且只在乏情母猪卵巢中表达。31个miRNAs共预测到7 388个靶基因,KEGG信号通路分析显示,有2 788个靶基因注释到了297个KEGG通路,前20个最富集的通路部分与生殖过程或生殖活动调控相关,表明这31个miRNAs参与了初产母猪的生殖调控。本研究结果丰富了猪miRNA数据资源,为进一步深入研究初产母猪的繁殖性能提供了理论依据。     

ULTRASOUND AS AN AID FOR DIAGNOSIS OF OVARIAN ABSCESSES IN TWO MARES

This report describes two mares presented for evaluation of anorexia, fever of unknown origin, and weight loss. Clinical examination, laboratory findings, and transrectal ultrasonographic images suggested ovarian abscessation. One mare was successfully treated medically. Because of financial considerations, the second mare was euthanatized and a postmortem examination was performed. At necropsy, there was an enlarged right ovary with an adhesion to the large colon. Microscopic findings were characteristic of an ovarian abscess.     

中华绒螯蟹成熟卵巢的脂类及脂肪酸组成

中华绒螯蟹成熟卵巢中的脂类以２种形态存在,即脂肪滴（Ｌ）和卵黄体（Ｙ）。脂肪滴为中性脂,卵黄体主要是磷脂。成熟卵巢的总脂含量约占卵巢湿重的１７．７８％,且中性脂（ＮＬ）大于磷脂（ＰＬ）。成熟卵巢的ＮＬ依次主要由甘油三酯（ＴＧ）、胆固醇（ＳＬ）和胆固醇脂（Ｅｓ）组成;ＰＬ主要由卵磷脂（ＰＣ）和脑磷脂（ＰＥ）组成,且ＰＣ含量高于ＰＥ。中华绒螯蟹成熟卵巢的ＮＬ和ＰＬ脂肪酸组成明显不同,ＰＬ短链的饱和与不饱和酸（＜１８个碳）含量显著低于ＮＬ（ｐ＜０．０１）,长链的多不饱和酸（＞１８个碳）含量显著高于ＮＬ（ｐ＜０．０１）,而一烯酸含量在这２种脂类中基本上无大变化。ＰＬ和ＮＬ的主要脂肪酸组成皆为：Ｃ１６∶０、Ｃ１６∶１、Ｃ１８∶１、Ｃ１８∶２、Ｃ２０∶４、Ｃ２０∶５和Ｃ２２∶６。     

雌性牦牛主要生殖器官中DBI的表达与定位分析

为加深对DBI基因功能的探究,揭示其在牦牛生殖生理过程中的作用.本试验采集健康母牦牛(3～6岁)繁殖周期不同阶段(卵泡期、黄体期和妊娠期)的卵巢、输卵管和子宫组织,共分为9组,每组设置3个生物学重复.采用实时荧光定量PCR (qRT-PCR)和蛋白免疫印迹技术(Western blotting,WB)检测牦牛DBI在繁...     

Characteristics and Functional Properties of Ovary from Squid Loligo Formosana

The characteristics and functional properties of the ovary from Loligo formosana were studied. Moisture (72.07 ± 0.24%) was dominant, followed by protein (18.64 ± 0.51%) and carbohydrate (7.44 ± 0.2%). Ash (1.39 ± 0.03%) and lipids (0.46 ± 0.5%) were found as the minor constituents. Albumin (79.02 ± 0.79%) was the major protein of the squid ovary, followed by glutelin-1 (8.31 ± 0.62%) and globulin (6.68 ± 0.08%). Nevertheless, prolamin and glutelin-2 constituted approximately 1% of the total proteins. Based on the electrophoretic studies, albumin had the largest band intensity. The squid ovary was rich in non-essential amino acids (52.26%) and high in hydrophobic amino acids (48.03%). It was also rich in polyunsaturated fatty acid (PUFA, 43.76 ± 0.84%), followed by saturated fatty acid (SFA, 39.36 ± 0.12%) and monounsaturated fatty acid (MUFA, 12.94 ± 0.55%). Ovary lipids had a high amount of docosahexaenoic acid (C22:6) (28.59%). At pH 3, the squid ovary powder (SOP) had the maximum solubility (96.39%), whereas the lowest solubility (38.33%) was observed at pH 9. The foaming capacity and stability of SOP were increased with increasing concentration up to 8% (p < 0.05). The globulin fraction showed the higher foaming capacity, as compared to albumin and glutelin-1 fractions. The squid ovary had good nutritional value and possessed excellent foaming properties. Therefore, the squid ovary could serve as a novel food additive or ingredient.     

Effects of biofloc technology on reproduction and ovarian recrudescence in Nile tilapia

The objectives of this study were to evaluate the reproductive cycle, morphological changes of ovary and mobilization of energy reserves in Nile tilapia reared with biofloc technology (BFT). In general, the growth and reproductive performance were highly similar between BFT and Control system (clear water). Difference between the systems was found in the hepatosomatic index (using mixed‐effects models), which suggested that BFT can alter the energy mobilization in the post‐spawning period. The absolute and relative fecundity, fertilization rate, number of larvae produced per female, gonadosomatic index, proportion of oogenesis cells, number of post‐ovulatory and atretic follicles were similar between the two systems. We also did not detect a reduction in the reproductive cycle length in Nile tilapia reared in BFT. Because there was no evidence of the negative effects of BFT on Nile tilapia reproduction, we concluded that BFT might be used for breeder stocking of this species.