小鼠生后不同发育阶段附睾组织中Crb3的表达

应用HE染色法和免疫荧光组织化学技术分别对小鼠生后不同发育阶段附睾组织结构及Crb3在不同发育时期附睾组织中的定位表达进行了研究。结果显示,附睾在4周龄时上皮为2层～3层的假复层纤毛柱状细胞,顶端纤毛细长;6周龄上皮细胞层数减少至1层～2层;8周龄附睾管上皮厚度增至最大,上皮细胞呈单层高柱状;12周龄后附睾管上皮开始变薄,呈单层柱状。免疫荧光染色结果显示,Crb3蛋白主要分布在附睾上皮柱状细胞的胞膜,在精子尾部有微弱的表达,这提示了Crb3不仅与附睾上皮细胞的极性建立和维持有关,而且对精子活力和血-附睾屏障的形成也可能具有潜在的影响。     

瘦素及其长型瘦素受体在雌性小鼠体内的表达

采用免疫组织化学SP染色法对长型瘦素受体在雌性小鼠生殖周期不同阶段的下丘脑、卵巢、子宫中细胞定位和分布进行了研究,并用ELISA方法对小鼠血清中的瘦素浓度进行了检测。结果显示,下丘脑神经元胞质中有棕褐色阳性颗粒,且阳性细胞数量随妊娠日龄增加逐渐增加,妊娠期与间情期阳性细胞数差异显著(P0.05);在卵巢中,卵母细胞胞质中有长型瘦素受体表达,随卵泡的发育,卵泡细胞中免疫反应阳性细胞数量逐渐增加;在胚泡附植期,瘦素受体在子宫腺和子宫内膜上皮细胞中大量表达。妊娠期血清瘦素浓度均高于间情期,且瘦素浓度从间情期到妊娠4日龄,呈上升趋势,妊娠5日龄稍下降,后随妊娠日龄逐渐增加。结果表明,瘦素及其长型受体能够促进卵泡发育,有利于小鼠胚泡的附植。     

幼鼠与成鼠子宫和输卵管组织形态学比较

本研究选取4周龄和8周龄小鼠,解剖获得子宫和输卵管制作石蜡切片,采用苏木精—伊红(Hematoxylin-Eosin staining,HE)染色法染色.结果显示成鼠子宫内膜褶皱多,内膜变薄,子宫腺增多;同时成鼠纵壁较幼鼠发达,有明显分支,分泌细胞和纤毛细胞数量多,排列紧密.本研究对进一步探索雌性动物生殖器官的生理功能...     

Nupr1 mRNA在小鼠早期妊娠子宫中的表达

试验旨在研究核蛋白1(nuclear protein 1,Nupr1)mRNA在小鼠早期妊娠子宫中的表达,探讨Nupr1与小鼠胚胎着床的相关性。通过建立小鼠早期妊娠模型、假孕妊娠模型、延迟着床及激活模型、人工蜕膜化模型和激素处理模型,采用原位杂交的方法检测Nupr1mRNA在小鼠各种模型子宫组织中的定位表达情况,并应用实时荧光定量PCR法检测早期妊娠模型和假孕妊娠模型中Nupr1mRNA的相对表达量。结果显示,Nupr1mRNA在小鼠早期妊娠第1~4天子宫的腔上皮和腺上皮表达,第5~8天表达于蜕膜区域;假孕妊娠第1~5天,Nupr1mRNA主要表达于小鼠子宫腔上皮和腺上皮;延迟着床模型中信号表达于在小鼠子宫的腔上皮和腺上皮,与正常妊娠第4天结果相似;延迟激活模型中信号表达于蜕膜区,与早期妊娠第5天表达结果相似;人工蜕膜化模型中信号表达于蜕膜区,而蜕膜对照组中信号表达于腔上皮和腺上皮;17β-雌二醇(oestrogen,E2)处理组信号表达于腔上皮和腺上皮,信号增强,孕酮(progesterone,P4)和E2共同处理表达无明显变化;实时荧光定量PCR结果显示,正常妊娠第2天Nupr1mRNA相对表达量较高,假孕妊娠第2天Nupr1mRNA相对表达量也较高。本研究结果表明,Nupr1mRNA在小鼠子宫中的表达与小鼠早期妊娠过程相关,Nupr1mRNA在腔上皮和腺上皮的表达可能受激素调节,在子宫基质中的表达与蜕膜化及活化胚泡相关。     

犬正常发情期与患子宫蓄脓的子宫和卵巢组织结构及乳铁蛋白表达的对比

本研究旨在探明犬子宫和卵巢在正常发情期与患子宫蓄脓时，其组织结构及乳铁蛋白（lactoferrin，LF）表达的变化特征。应用Masson’s、VVG、PAS组织化学染色方法观察乏情期、发情期、患子宫蓄脓时犬子宫及卵巢的组织结构特点，用免疫组织化学SP法观察LF的分布特征。结果显示，正常发情期犬：乏情期、发情期子宫内膜肌层厚度比分别为0.762 0、0.924 3；乏情期子宫固有层中胶原纤维含量大于发情期，发情期卵巢中胶原纤维含量大于乏情期；子宫血管层及卵巢血管内弹性膜清晰完整；子宫腺在乏情期时浅层管腔小，深层较大、腺管上皮为单层柱状上皮、上皮细胞及管腔内PAS阳性反应较强，发情期子宫腺管腔变大、腺管上皮为单层立方上皮、上皮细胞及管腔内有PAS阳性反应；子宫黏膜上皮在乏情期和发情期均为单层柱状上皮，但胞核位置不同，乏情期胞核位于中央，发情期胞核位于顶部。患子宫蓄脓犬：子宫内膜肌层厚度比为1.615 0；子宫固有层和卵巢中胶原纤维含量少于正常发情期；子宫腺管腔大，且形状不规则，管腔内有炎性细胞浸润，腺管上皮为单层立方上皮，有淋巴细胞位于基膜，上皮细胞及管腔内PAS阳性反应较弱；子宫血管层及卵巢血管内弹性膜较正常发情期变薄，且有断裂现象；子宫黏膜上皮为单层柱状上皮，胞核位于基底。LF在乏情期子宫腺上皮和卵巢中的表达水平高于发情期，而在子宫黏膜上皮中发情期的表达水平高于乏情期。患子宫蓄脓时犬子宫和卵巢中LF的表达水平均较低。综上表明，犬正常发情期与患子宫蓄脓时的子宫和卵巢组织结构特点显著不同，LF的表达水平也存在差异。     

瘦素对绵羊子宫内膜上皮细胞增殖、迁移和凋亡的影响

试验旨在明确瘦素在妊娠和非妊娠绵羊子宫内膜组织中的定位特征,探究其对绵羊子宫内膜上皮细胞中胚胎附植的影响。使用免疫组化法检测瘦素及其受体在妊娠和未妊娠绵羊的子宫内膜上皮组织的表达及定位;利用组织块分离培养原代绵羊子宫内膜上皮细胞,CCK8法确定瘦素最适浓度,RT-qPCR和Western blot检测瘦素对绵羊子宫内膜上皮细胞凋亡的作用,细胞划痕试验检测瘦素对绵羊子宫内膜上皮细胞迁移的作用,RT-qPCR检测瘦素对胚胎附植相关因子血管内皮生长因子（VEGF）、白血病抑制因子（LIF）、白细胞介素-6(IL-6)、子宫内膜附植位点黏蛋白（Muc1）、基质金属蛋白9(MMP9)、白细胞介素-1β(IL-1β)、骨桥蛋白（OPN）、泛素样修饰因子（ISG15）等基因mRNA的表达情况。结果显示,瘦素蛋白主要表达在未妊娠绵羊子宫内膜的基质和上皮组织中,主要在妊娠绵羊子宫内膜上皮组织中表达,瘦素受体表达无明显变化;组织块法可成功获得具有典型上皮样细胞特征并表达角蛋白CK18的原代绵羊子宫内膜上皮细胞。250 mg/L瘦素促进绵羊子宫内膜上皮细胞增殖和迁移,并显著上调Bcl-2,下调BAX、Cas...     

八点黑獭兔不同发育时期子宫组织结构变化的研究

观察母兔妊娠20、26、29天和15、30、60、90日龄八点黑獭兔的子宫组织,以探讨八点黑獭兔子宫的形态学发育特点。结果表明：獭兔的子宫随着年龄的增长．其内膜的粘膜上皮由假复层柱状纤毛上皮逐渐分化成紧密规则的单层柱状上皮;固有层在15～30日龄之间出现．内含大量间质细胞;妊娠期固有层发生水肿和充血;子宫肌层在幼龄时分层不明显,各层肌纤维互相交织,随平滑肌细胞分裂增生,3月龄时肌层出现明显的内环、外纵两层,中间形成一层环状排列的血管：獭兔出生后其子宫内膜腺由子宫内膜上皮细胞内陷形成,随后腺上皮细胞经有丝分裂增生,15日龄幼兔已形成单管子宫腺。该研究结果可为兔的解剖学、生殖生理学增添新的内容,也为进一步研究兔生殖系统疾病的治疗与预防提供参考资料。     

BDNF和TRKB蛋白在妊娠期山羊子宫和胎盘的表达

为探讨脑源性神经营养因子(BDNF)及其受体酪氨酸激酶受体B(TRKB)在妊娠期山羊子宫和胎盘中的表达情况,本实验分别于妊娠15、25、65 d采集子宫和胎盘样品,免疫组织化学方法分析BDNF和TRKB蛋白的分布情况。结果表明:妊娠15 d时,BDNF和TRKB蛋白在子宫内膜上皮和腺上皮细胞中强表达,BDNF在肌层弱表达,而TRKB强表达;妊娠25 d和65 d时,2种蛋白在子宫内膜上皮和腺上皮细胞中维持强表达,同时在胎儿绒毛膜滋养层细胞中表达强烈;与妊娠15 d相比,妊娠25 d和65 d时2种蛋白均在浅层子宫腺强烈表达,深层子宫腺弱表达。综上可知,随着妊娠的进行,BDNF和TRKB蛋白的表达情况也发生变化,提示BDNF对山羊妊娠活动有一定的调节作用,与妊娠期山羊子宫和胎盘的组织结构变化相关联,可能通过与其受体TRKB的特异性结合来实现其功能。     

大肠杆菌对奶牛子宫内膜组织合成分泌IL-1β、IL-6和PGE_2的影响

基于奶牛子宫内膜组织体外培养方法,分别采用浓度为1×10~5,1×10~6 CFU/mL的致病性大肠杆菌处理体外培养的子宫内膜组织12,24 h后用HE染色方法评价组织损伤情况,ELISA法检测组织培养液上清中IL-1β,IL-6和PGE_2的分泌变化,RT-qPCR法检测IL-1β和IL-6基因表达变化。结果显示:与对照组相比,大肠杆菌分别处理12,24 h后奶牛子宫内膜组织中子宫内膜上皮细胞和腺上皮细胞脱落、坏死、崩解,并且显著提高奶牛子宫内膜组织IL-1β,IL-6和PGE_2的分泌及IL-1β和IL-6的mRNA的表达量。结果表明:成功建立了体外大肠杆菌型奶牛子宫内膜炎模型,且IL-1β,IL-6和PGE_2在大肠杆菌引起的奶牛子宫内膜组织炎症反应中发挥着重要的调控作用。     

“花山麻鸡”CPT1A基因时空表达规律研究

试验旨在阐明"花山麻鸡"CPT1A基因的组织表达和时序表达规律,以胚胎期到上市日龄的"花山麻鸡"为试验材料,在不同发育时间点采集胸肌、腿肌、心脏、肝脏组织样品,利用实时荧光定量方法分析CPT1A基因在不同组织不同发育阶段表达规律。在相同发育阶段不同组织差异表达结果显示:在胚胎期(9、12、16日胚龄)和上市日龄(9周龄),CPT1A基因在肝脏的表达量显著高于在心脏、胸肌和腿肌的表达量,在出生后1日龄,CPT1A在心脏表达量最高,1周龄时,CPTA在肝脏的表达量最低;5周龄时,CPT1A在腿肌的表达量最高;7周龄时,CPT1A在腿肌和肝脏的表达量显著高于胸肌和心脏。在同一组织不同发育阶段差异表达结果显示,CPT1A在胸肌、腿肌、肝脏和心脏不同发育阶段表达规律与表达量是不同的。研究表明CPT1A基因在胸肌、腿肌、肝脏和心脏相同发育阶段表达量不同,CPT1A基因在胸肌、腿肌、肝脏和心脏织不同发育阶段表达规律与表达量不同。     

Expression of interleukin-18 receptor mRNA in the mouse endometrium

Interleukin-18 (IL-18) is a proinflammatory cytokine involved in chronic inflammation, autoimmune diseases, and a variety of cancers, and is expressed in mouse uteri. Our previous study suggested that IL-18 acts as a paracrine factor, regulating endometrial function. To elucidate the physiological roles of IL-18 in the mouse endometrium, the expression of the IL-18 receptor (IL-18R) alpha subunit was analyzed. IL-18Ralpha mRNA was expressed in several mouse organs in addition to the endometrium. In situ hybridization analysis using a biotin-labeled mouse IL-18Ralpha riboprobe demonstrated that IL-18Ralpha mRNA expression was detected in glandular epithelial cells, stromal cells around uterine glands, and myometrial cells in the mouse uterus, suggesting that these cells are targets for IL-18. The uterine IL-18Ralpha mRNA expression level changed with the estrous cycle. The uterine IL-18Ralpha mRNA levels of estrous mice were higher than those of diestrous mice. In addition, the IL-18Ralpha mRNA levels in uteri at 3 and 14 days after ovariectomy were higher than those at diestrus and decreased following treatment with estradiol-17beta or progesterone. These findings suggest that IL-18Ralpha gene expression is regulated by estrogen and progesterone and that the uterine IL-18 system is involved in the regulation of uterine functions in a paracrine manner.     

Steroid receptor expression and HER-2/neu (c-erbB-2) oncoprotein in the uterus of cats with cystic endometrial hyperplasia-pyometra complex

The presence of oestrogen-alpha receptor (ER), progesterone receptor (PR), and HER-2/neu (c-erbB-2) oncoprotein in the uterine walls of 10 healthy cats and 20 subjects with cystic endometrial hyperplasia-pyometra (CEH-P) were evaluated. Lesions were graded according to the severity of cystic dilation, hyperplasia and inflammation, and were classified as normal, mild uterine hyperplasia and severe uterine hyperplasia. The ER, PR and c-erbB-2 expression in the endometrium, glandular epithelium, stromal fibroblasts and myometrial smooth muscle cells was quantified by immunohistochemistry. The ER, PR and c-erbB-2 staining patterns differed between normal uteri and uteri with CEH-P. The ER expression was tended to be higher in the endometrial surface and glandular epithelium in the severe hyperplasia group (P > 0.05) and significantly lower in the mild hyperplasia cases compared with normal endometrium (P < 0.05), whereas the PR expression in both severe and mild hyperplasia cases tended to be higher in stromal cells and glandular epithelium than those in the normal uteri. C-erbB-2 immunoreactivity was observed only in the endometrial surface and glandular epithelium of the uterine wall and immunostaining was found to be highest in cases with severe hyperplasia. As a conclusion, we suggest that c-erbB-2 oncoprotein may play a role in the pathogenesis of the CEH together with the ER and PR in cats, and that ER does not have a role in the mechanism of pyometra, whereas PR plays a role in the pathogenesis of both CEH and pyometra.     

Normal Postpartum Involution of the Uterus in the Dog

Ninety-eight reproductive tracts from dogs at different postpartum time periods were used to investigate stages of normal involution. Seventy-eight reproductive tracts were obtained from the field, and 20 obtained surgically for gross and microscopic examination. Plasma progesterone was measured in 22 dogs at various times postpartum.

The uterine horns during the first week postpartum were dilated and edematous. The placental sites were 1.5-3 cm in width, rough, granular and covered with mucus and a few blood clots. By the fourth week the placental sites were thick, grayish-tan and nodular with a few blood clots within nodules. The uterine horns during the seventh week were greatly contracted and the placental sites were narrow and light in color. A few nodules were still present on the surface. By the ninth week the uterine horns were uniform in shape and contracted with a narrow lumen. The placental sites appeared as a narrow brown band.

Histologically the placental sites during the first week postpartum were covered by an eosinophilic staining necrotic mass and a few intact epithelial cells scattered on the surface as an interrupted single layer. Under the necrotic mass, large eosinophilic staining cells in moderate number were scattered throughout the lamina propria of the placental site. These cells were considered to be decidual cells. By the fourth week the placental sites were covered by a large lobulated mass of collagen fibers. The uterine glands were greatly dilated and degenerate, and mononuclear cell infiltration in the lamina propria was pronounced. By the seventh week, large masses of collagen fibers were detached from the surface, and endometrial glands were normal in size and shape. By the ninth week surface sloughing was completed. However, regeneration and replacement of the endometrial lining from the mouth of the uterine glands continued until the end of the twelfth week when the involution process was completed.

The progesterone levels were very low for eight weeks postpartum.

     

Structure and Steroidogenesis of the Placenta in the Antarctic Minke Whale (Balaenoptera bonaerensis)

There are few reports describing the structure and function of the whale placenta with the advance of pregnancy. In this study, therefore, the placenta and nonpregnant uterus of the Antarctic minke whale were observed morphologically and immunohistochemically. Placentas and nonpregnant uteri were collected from the 15th, 16th and 18th Japanese Whale Research Programme with Special Permit in the Antarctic (JARPA) and 1st JARPA II organized by the Institute of Cetacean Research in Tokyo, Japan. In the macro- and microscopic observations, the placenta of the Antarctic minke whale was a diffuse and epitheliochorial placenta. The chorion was interdigitated to the endometrium by primary, secondary and tertiary villi, which contained no specialized trophoblast cells such as binucleate cells, and the interdigitation became complicated with the progress of gestation. Furthermore, fetal and maternal blood vessels indented deeply into the trophoblast cells and endometrial epithelium respectively with fetal growth. The minke whale placenta showed a fold-like shape as opposed to a finger-like shape. In both nonpregnant and pregnant uteri, many uterine glands were distributed. The uterine glands in the superficial layer of the pregnant endometrium had a wide lumen and large epithelial cells as compared with those in the deep layer. On the other hand, in the nonpregnant endometrium, the uterine glands had a narrower lumen and smaller epithelial cells than in the pregnant endometrium. In immunohistochemical detection, immunoreactivity for P450scc was detected in most trophoblast cells, but not in nonpregnant uteri, suggesting that trophoblast epithelial cells synthesized and secreted the sex steroid hormones and/or their precursors to maintain the pregnancy in the Antarctic minke whale.     

Mineralised deposits in the uterine glands of mares with chronic endometrial degeneration

Chronic degenerative disease of the mare's endometrium is characterised by changes in the uterine glands, including cystic dilation, hyperplasia and periglandular fibrosis. Endometrial biopsies were taken from 23 mares with different grades of endometrial degeneration. Solid structures were identified within the lumina of the uterine glands and shown to be calcified by histochemical staining. Most of them were not homogenous but composed of a mixture of mineral and organic substances. Further examinations of these mineralised structures by immunohistochemical methods revealed the presence of the non-collagenous matrix proteins osteopontin, osteonectin and bone sialoprotein, which are known to be involved in calcification processes such as urolithiosis. Osteopontin and bone sialoprotein were identified within the calculi, frequently arranged in concentric layers. Osteonectin was the only matrix protein that was also present in the glandular epithelium. Osteocalcin was not found in either the calculi or the glandular epithelial cells.     

Ovine uterine morphogenesis: effects of age and progestin administration and withdrawal on neonatal endometrial development and DNA synthesis

To determine effects of age and administration and withdrawal of a synthetic progestin (P) on endometrial development and DNA synthesis, ewe lambs were ovariectomized on d 0 (birth) and assigned to one of four groups (n = three/group) that provided (by means of hemihysterectomy) the following uterine tissue types: 1) d 0 control, 2) d 13 control, 3) d 26 control, 4) d 13 after 13 d exposure to P (13P) and 5) d 26 after P exposure from d 0 to 13 (26P). Uterine tissues were processed for histology or explanted with [methyl-3 H] thymidine for autoradiography. Labeling index (LI) was determined for stroma and epithelium in caruncular and intercaruncular endometrial areas and for lumenal and glandular epithelium in uteri with glands. Endometrial glands were absent on d 0, evident at d 13 and well developed by d 26. Day 0 LI was greater (P less than .05) in caruncular than in intercaruncular areas, and greater in stromal than in epithelial tissues. Relationships were reversed in d 13 endometrium (day X endometrial area, P less than .07). Caruncular stromal LI was greater on d 0 than later (P less than .02), whereas intercaruncular epithelial LI was greater after d 0 (P less than .05), but decreased from d 13 to 26 (P less than .05). Glandular epithelial LI was higher on d 13 than on d 26 (P less than .03). Administration of P inhibited endometrial gland development and suppressed d 13P intercaruncular LI (P less than .05). Withdrawal of P was followed by endometrial gland development and increased (P less than .01) intercaruncular epithelial LI in d 26P uteri. Ovary-independent initiation of endometrial gland development involves age- and region-specific alterations in DNA synthesis and could involve negative control.