PDGF及其受体mRNA在鹅不同等级前卵泡中的表达差异

为探讨血小板衍生生长因子(Platelet-derived growth factor,PDGF)及其受体(PDGFR)mRNA在鹅等级前卵泡中的表达差异及生物学意义。选取健康的产蛋前期和产蛋期籽鹅各3只为试验材料,利用实时荧光定量PCR方法检测PDGF和PDGFR mRNA在产蛋前期和产蛋期鹅等级前卵泡(分为SYF、LWF、MWF、SWF、PE 5个等级)中的表达差异。结果表明:产蛋期PDGF及其受体mRNA在5个等级前卵泡中的表达量均高于产蛋前期(P0.05),说明PDGF及其受体mRNA对卵泡发育有促进作用;产蛋前期和产蛋期PDGF及其受体mRNA表达量均在初级卵泡(PE)中最高,说明初级卵泡是主要表达部位,其中颗粒细胞形态由扁平变成立方形后开始大量增殖,通过促进颗粒细胞的增殖进而促进卵泡发育;在初级卵泡向小白卵泡(SWF)发育过程中,PDGF及其受体mRNA表达量有明显下降(P0.05),推测这可能与PDGF抑制类固醇的合成进而抑制卵泡发育有关,使优势卵泡更有序的进行筛选。     

低温对四川华鳊卵巢发育的影响及调控

四川华鳊（Sinibrama taeniatus）是长江上游特有的小型经济鱼类,在野外其一年可以繁殖两次,但在26℃条件下进行人工养殖时其繁殖周期仅为14天,其中低温是限制其在野外繁殖的重要因素。为探究低温对四川华鳊卵巢发育的影响及调控方式,本研究对其产卵后雌鱼进行11℃的低温处理（LT组）,并与26℃条件下（OT组）卵巢发育状况进行对比。通过形态学和组织学观察发现LT组卵巢发育被抑制,发生退化,卵母细胞明显发育不良,出现典型闭锁卵泡的特征。对雌鱼血清中部分生殖激素及卵巢中相应受体基因的表达水平进行测定,发现低温使FSH和DHP水平显著下降,并严重抑制激素受体基因fshr,lhcgr,pgr,esr1,ar的表达。通过转录组测序和分析发现,在低温处理过程中类固醇激素合成,细胞生长增殖以及凋亡和自噬相关通路被显著富集,与低温的影响显著相关。低温下调了类固醇激素合成和卵母细胞发育相关基因StAR,cyp17a1,hsd3b,cyp19a2,hsd17b1,vtg1,zp4,mmp9,mmp15以及细胞生长增殖和抗凋亡相关基因ccni,cdk16,igf1r,egfr,nobox,bcl2的表达,上调了促凋亡基因bax,tnf,tp53的表达。上述结果表明,低温导致卵泡闭锁的原因一方面可能是低温抑制生殖激素的合成和受体基因的表达,使卵巢发育和卵母细胞生长增殖受限;另一方面,可能是低温促使卵泡发生自噬凋亡,最终导致卵泡发生退化闭锁。     

Preovulatory serum estradiol concentration is positively associated with oocyte ATP and follicular fluid metabolite abundance in lactating beef cattle

Cattle induced to ovulate a small, physiologically immature preovulatory follicle had reduced oocyte developmental competence that resulted in decreased embryo cleavage and day 7 embryo quality compared with animals induced to ovulate a more advanced follicle. RNA-sequencing was performed on oocytes and their corresponding cumulus cells approximately 23 h after gonadotropin-releasing hormone (GnRH) administration to induce the preovulatory gonadotropin surge suggested reduced capacity for glucose metabolism and oxidative phosphorylation in the cumulus cells and oocytes from follicles ≤11.7 mm, respectively. We hypothesized that induced ovulation of a small, physiologically immature preovulatory follicle results in a suboptimal follicular microenvironment and reduced oocyte metabolic capacity. We performed a study with the objective to determine the impact of preovulatory follicle diameter and serum estradiol concentration at GnRH administration on oocyte metabolic competence and follicular fluid metabolome profiles. We synchronized the development of a preovulatory follicle and collected the follicle contents via transvaginal aspiration approximately 19 h after GnRH administration in lactating beef cows (n = 319). We determined ATP levels and mitochondrial DNA (mtDNA) copy number in 110 oocytes and performed ultra-high-performance liquid chromatography–high resolution mass spectrometry metabolomic studies on 45 follicular fluid samples. Intraoocyte ATP and the amount of ATP produced per mtDNA copy number were associated with serum estradiol concentration at GnRH and time from GnRH administration to follicle aspiration (P < 0.05). mtDNA copy number was not related to follicle diameter at GnRH, serum estradiol concentration at GnRH, or any potential covariates (P > 0.10). We detected 90 metabolites in the aspirated follicular fluid. We identified 22 metabolites associated with serum estradiol concentration at GnRH and 63 metabolites associated with follicular fluid progesterone concentration at the time of follicle aspiration (FDR < 0.10). Pathway enrichment analysis of significant metabolites suggested altered proteinogenesis, citric acid cycle, and pyrimidine metabolism in follicles of reduced estrogenic capacity pre-gonadotropin surge or reduced progesterone production by the time of follicle aspiration.     

发酵饲料对蛋鸡生产性能、蛋品质、输卵管壁厚及卵泡分级的影响

试验旨在探究饲粮添加发酵饲料对蛋鸡生产性能、蛋品质、输卵管壁厚及卵泡分级的影响.取鸡舍环境条件趋于一致的580日龄海兰褐蛋鸡960只,随机分为4组,每组8个重复,每个重复30只鸡.各组饲粮分别添加0(A组)、2.0％(B组)、2.5％(C组)、3.0％(D组)发酵饲料.预试期1w,正式试验期6w.结果 显示,与对照组相...     

MIF在不同直径猪卵泡的表达变化规律

为分析巨噬细胞迁移抑制因子(Macrophage migration inhibitory factor,MIF)在不同直径健康猪卵泡上的表达变化规律,将收集到的90个健康母猪卵巢,通过ELISA检测不同直径卵泡液中MIF质量浓度,通过实时荧光定量PCR和Western blot检测颗粒细胞中MIF基因mRNA和蛋白的表达水平,采用免疫荧光检测MIF在颗粒细胞中的分布情况。结果显示:1)中等直径卵泡(3～4mm)液中MIF的质量浓度(54.17±7.86ng/mL)显著高于大卵泡(6mm,32.29±1.47ng/mL)和小卵泡(1～2mm,38.89±0.98ng/mL)(P0.05);2)中等直径卵泡颗粒细胞中MIF基因mRNA和蛋白质表达量也显著高于大卵泡和小卵泡的表达量(P0.05);3)MIF也在不同直径卵泡颗粒细胞中的细胞核和细胞质表达,但多集中于细胞质中。综上,随着猪卵泡发育过程,MIF在中等直径卵泡中表达量最高。本研究为母猪卵泡发育和排卵提供新的线索。     

应用磁珠免疫亲和层析法从猪卵泡液中提纯抑制素

采集猪卵泡液 ,用 70 %饱和度硫酸铵溶液沉淀后 ,分别过SephadexG 2 0 0柱和SephadexG 10 0柱 ,最后用磁珠免疫亲和层析法进一步提纯 ,使抑制素纯度从 12 4 μg·mg-1升至 39 4 μg·mg-1。PAGE电泳分析证明 ,提纯物只出现一条相对分子质量为 32 0 0 0的带 ,SDS PAGE电泳分析发现有两条相对分子质量分别为 14 0 0 0～ 15 0 0 0和 170 0 0～ 180 0 0的带 ,即β、α亚基。     

Effect of aglepristone (RU534) administration during follicular phase on progesterone,estradiol-17β and LH serum concentrations in bitches

Aglepristone was administered in bitches during the follicular phase to evaluate its effects on progesterone, estradiol-17β and LH serum concentrations. Ten German Shepherds were divided into two groups (treated n = 5; control n = 5). Treated bitches received 10 mg/kg BW of aglepristone subcutaneously during the early follicular phase, 24 hr after and then 7 days later. The control group was injected, at the same time periods, with saline solution (0.3 ml/kg BW). For the steroid evaluations, blood was collected daily from the onset of proestrus until the first day of cytological dioestrus. For LH base-line serum determination, blood was also collected every 20 min for 2 hr at the onset of proestrus. For LH surge identification, blood was collected daily (every 6 hr) starting from the day of the first administration of aglepristone or saline solution until the first day of dioestrus. All animals ovulated but the treated group presented longer ovulation-dioestrus intervals than the control group (5.2 ± 2.2 days p < .05). Serum concentrations of the evaluated hormones were similar between experimental animals except for serum LH. Indeed, no LH peaks were detected in the treated group while LH surges were clearly observed in the control group (9 ± 1 days after the beginning of proestrus. In particular, the area under the curve for LH was significantly lower in treated than control animals (12 ± 4 ng/ml x Day; p = .01). In conclusion, administrations of aglepristone during the follicular phase of the bitch does not affect the steroid hormone patterns but does prevent the occurrence of a LH surge. This work raises significant questions and opens perspectives concerning the mechanisms of ovulation in bitches.     

Abscessation following intralesional formalin treatment of congenital parotid salivary duct atresia

A 5-month-old colt with a history of nonpainful left-sided facial swelling present since birth was presented for evaluation and treatment. A diagnosis of congenital parotid salivary duct atresia was made, and surgical correction was attempted but was unsuccessful. Follow-up treatments consisted of repeat intralesional injection of 10% neutral-buffered formalin. The horse developed ventral mandibular abscessation with multiple draining tracts secondary to this treatment; however, no other significant complications occurred and a cosmetic outcome was achieved.     

小鼠卵泡卵母细胞的一步冷冻研究

通过对冷冻液和平衡时间的筛选,研究了裸露小鼠卵泡卵母细胞的一步冷冻。结果表明:当冷冻液含25％的1,2—丙二醇和0.25mol/L的蔗糖,平衡时间为10min时,冷冻效果最为有效,解冻后存活率和形态正常率分别达66.2％和58.1％.解冻后形态正常的卵泡卵母细胞可以发育成熟,成熟率为22.7％。     

表皮生长因子对胎鼠卵巢卵泡体外发育和类固醇激素分泌的影响

表皮生长因子（EGF）是参与卵巢功能调节的一个重要肽类因子，关于EGF对小鼠胚胎期卵巢卵泡发育和类固醇激素发育的 今未见报道。本研究以单个卵巢的放泡数、生长卵泡数和5个最大卵母 平均直径为卵泡发育优劣的衡量指标，利用我们早期研究建立的无血清培养体系研究了EGF对胎鼠卵巢卵泡发育和类固醇激素分泌的作用。结果发现：①EGF处理组胎鼠卵巢总卵泡数、生长卵九和卵巢内5个最大卵母细胞平均直径从培养第7天起即显著高于ITS对照组直至培养结束；②EGF在培养后期显著促进胎鼠卵巢分泌睾酮，但对雌二醇的基础分泌无影响；③EGF处理组胎鼠卵巢在培养过程的后期引起贴壁的卵巢边缘脱离培养板底壁发生卷边现象，丧失卵母细胞皮持－髓质生长模式。结果提示：EGF在胎鼠卵巢原始卵泡生长起始中起重要作用， 同时EGF促进胎鼠卵巢小腔前卵泡／卵母细胞进一步生长、发育和体外存活；EGF促卵泡发作用与雌二醇的作用无关。