山羊子宫内肽能神经分布及妊娠时的变化

采用免疫组化方法，研究了山羊子宫内含P物质（SP）和含血管活性肠肽（VIP）神经的分布。结果，妊娠及未妊娠山羊子宫颈内有粗细不等的神经束行经于外膜和肌层中，神经分支形成丛状分布于血管壁，子宫颈部未见SP神经元胞体及VIP神经元胞体；未妊娠山羊子宫角内SP神经和VIP神经均呈丛状围绕血管并分布于血管壁；妊娠中期的孕角和非孕角均有胎盘形成，除胎盘内无神经分布外，SP神经及VIP神经同样分支形成丛状分布于血管壁。结果提示，山羊子宫内SP神经和VIP神经主要支配子宫内血管，妊娠时除胎盘内无神经分布外，SP神经及VIP神经的分布无明显变化。     

雌性空怀双峰驼生殖道的形态结构

应用组织学方法观察了雌性空怀双峰驼生殖道的形态结构。结果显示，双峰驼生殖道的基本结构与其他哺乳动物相似，但微细结构有差异。双峰驼输卵管粘膜皱襞极其发达，分支多而呈复杂的网状迷路。皱襞基部的迷路酷似固有膜而存在腺体，迷路网格内常见细胞团块。虽然双峰驼怀孕时胎儿位于左侧子宫角，但左、右子宫角以及子宫体的组织结构基本相同。子宫内膜无肉阜，上皮下陷于固有膜内，形成大量长而弯曲的单管状腺。子宫颈固有膜浅层分布有许多小腺体，深层分布有成群的较大腺体。这些腺体为分支管状腺，腺上皮PAS强阳性。阴道粘膜上皮为复层上皮。从输卵管到阴道，粘膜上皮主要为单层柱状上皮，由纤毛细胞和分泌细胞组成，局部可见假复层柱状纤毛上皮。纤毛细胞由前向后逐渐减少，但在子宫颈仍可见到。粘膜上皮和腺上皮内夹有许多淋巴细胞或中性粒细胞，后段局部甚至见到这些免疫细胞浸润于上皮细胞间。固有膜内分布有大量淋巴细胞、中性粒细胞、肥大细胞、浆细胞和巨噬细胞，有时出现淋巴滤泡。     

棕果蝠雌性生殖器官的大体及组织结构观察

本试验对存在于广东云浮市境内的棕果蝠雌性生殖器官进行了大体及组织学观察。棕果蝠雌性生殖系统由卵巢、输卵管、子宫、阴道、阴道前庭和阴门组成。卵巢一对,位于两侧肾的后方,包于输卵管系膜内,肉眼不易区分。输卵管很短,分为漏斗部、壶腹部和峡部,黏膜上皮为单层柱状上皮,黏膜形成纵向黏膜褶,并具有宽阔的输卵管腺。子宫为双子宫类型,根据子宫内膜上皮类型,每侧子宫分为子宫角和子宫颈,子宫角内膜具有高柱状上皮,并形成类似于输卵管腺的子宫腺。子宫角较粗,外径在1 000μm以上,比输卵管外径大1倍。子宫颈内膜上皮为单层高柱状分泌型细胞。子宫颈内膜形成发达的纵行黏膜皱褶,但不形成宫颈腺或腺样隐窝;两个子宫颈后半部共同形成一个圆锥状子宫颈阴道部,相当发达,顶端向后,其外表面也具有一层高柱状分泌型上皮细胞。两个子宫颈管未能抵达子宫颈后端,而是分别开口于子宫颈阴道部中间段(前后向上)的腹侧面上,分别开口于阴道。阴道穹窿长而深,其最前部形成一对盲管,分别位于左右两宫颈管的外侧。阴道长而宽阔。其前端形成阴道穹窿。阴道黏膜为厚而角化的复层扁平上皮。阴道后腹侧有雌性尿道的开口。阴门呈椭圆孔状,未形成左右两个阴唇。     

淋巴细胞和T淋巴细胞亚型在母山羊生殖道内区域分布：发情周期不同阶段的影响

母畜的生殖道是粘膜系统的一部分，它可保护动物免受病原体的侵害。作者分析了母山羊生殖道内总淋巴细胞、血液细胞(分泌抗体的B细胞)和T淋巴细胞亚型存在和分布的状况。从常规组织学方面估计了发情周期对子宫颈、子宫角内淋巴细胞和血液细胞浓度的影响。借助显微共焦和免疫过氧化酶技术的免疫细胞化学方法研究了淋巴细胞亚型。结果显示，山羊的生殖系统富含淋巴细胞。这些细胞与子宫内膜的上皮细胞混合分布在整个基质上。并在基质上观察到了淋巴细胞总体。随着山羊繁殖阶段的不同而变化的是淋巴细胞数目而不是血细胞数目。激素环境的影响对于子宫颈和子宫角是有差异的。免疫细胞化学试验证实了在子宫颈和子宫角的上皮层和基质上发生CD4 和CD8 抗体免疫反应细胞的存在。这些细胞的数目在子宫颈更多一些，并且发现它们向子宫内膜腺体的鲁米诺上皮细胞渗透。试验结果显示，分布在子宫颈和子宫角的淋巴细胞数量是不同的，并且受繁殖周期不同阶段的影响。总之，CD4 和CD8 T淋巴细胞亚型在山羊的子宫颈和子宫角内膜上都能够被发现。     

马冈鹅肠胚后发育的组织学观察

将72只5~110d马冈鹅(各半)按不同日龄随机分为9组,先后取十二指肠、空肠、回肠和直肠,应用常规组织学技术和Tiger细胞图像分析系统进行观测。结果表明:各部肠管管壁分粘膜、肌层和浆膜三层,无粘膜下层;绒毛既有分支,也有融合现象;肠腺主要是单管状腺,但也有分支管状腺;粘膜上皮中柱状细胞从前向后逐渐减少,而杯状细胞则相应逐渐增多;肠腺中的柱状细胞则由前向后逐渐增多,杯状细胞相应减少;肌层分内纵、中环、外纵三层,内、外两肌层厚度近似,中环肌最厚,内有明显的结缔组织间隔带分布,小肠段呈有规律的"竹节"状;粘膜皱襞不发达,少数皱襞由粘膜、内纵和中环肌层共同构成;肠壁内淋巴组织丰富,在肌层之间及肌层内均可见淋巴小结,肌层内也有神经丛和神经纤维束的断面;绒毛高度发育的绝对生长高峰期在26d以前,但绒毛直径的发育无明显的阶段性。     

发情周期中山羊子宫和输卵管内肥大细胞及组织胺含量的变化

应用改良甲苯胺蓝染色法和透射电镜技术观察了山羊发情周期4个阶段肥大细胞（MC）在子宫和输卵管内的分布、形态及数量变化规律,用荧光测定改良法检测了子宫和输卵管组织中组织胺（HA）含量。结果显示：MC呈圆形、椭圆形或梭形,在子宫内主要分布在肌层,子宫外膜中零星散在,内膜中散在分布于固有层的子宫腺和血管周围,子宫阜和内膜上皮中未发现。输卵管内MC在黏膜皱襞、肌层和外膜中均有分布。MC在发情后期有明显的脱颗粒现象。发情周期中子宫及输卵管MC数量变化为：MC（发情后期）〉MC（发情前期）〉MC（间情期）〉MC（发情期）,子宫肌层MC数量各组间差异极显著（P〈0．01）,子宫固有层和输卵管内无显著变化。子宫角内HA含量在发情后期与其它时期有显著变化（P〈0．05）,子宫体、子宫颈和输卵管的变化不明显（P〉0．05）。     

新生羔羊瘤胃PGP9.5、一氧化氮合成酶和乙酰胆碱酯酶阳性神经元定位分析

应用连续组织切片,采用免疫荧光染色的方法,使用3种抗体:蛋白基因产物9.5(PGP9.5)、一氧化氮合成酶(NOS)和乙酰胆碱酯酶(AChE),分别对新生羔羊瘤胃内的所有神经元、氮能和胆碱能神经元进行定位分析。结果:PGP9.5、NOS和AChE阳性反应广泛分布于神经元胞体和神经纤维中。阳性神经元胞体在黏膜下未发现,但成群出现在肌间神经节内。阳性神经元胞体数目PGP9.5最多,其次是胆碱能,氮能最少。阳性神经纤维成簇分布在神经节内和节间形成神经束,并且延伸到环纵肌内。阳性神经纤维密度PGP9.5最多,其次是氮能,胆碱能最少。另外,一些阳性神经纤维经常围绕在血管周围,甚至延伸入瘤胃上皮。PGP9.5、NOS和AChE神经元和纤维在新生羔羊瘤胃内有广泛的分布,该结果提示其分布模式可能与功能的适应直接相关。     

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。     

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。     

母鸡输卵管子宫部副交感节后神经元支配的逆行追踪研究

将CB－HRP溶液注入鸡输卵管子宫部浆膜下，取泄殖腔神经节和双侧阻部内神经以及肠神经直肠段，制成50μm厚度的边疆冰冻切片，TMB反应，明视野观察。结果发现：鸡输卵管子宫部受泄殖腔神经节和双侧阻部内神经节的副交感节后神经元支配；双侧阴部内神经节的标记细胞无显著差异；说明尽管母鸡输卵管子宫部是单侧发育器官，但其受双侧阴部内神经的融交感节后神经元的支配。鸡输卵管子宫部还受肠神经的节后神经元的支配。另外本实验对汇殖腔神经节、阴部内神经及肠神经直肠段进行了大体解剖观察。     

Crb1在小鼠生后不同发育阶段子宫组织中的表达

应用HE染色方法和免疫荧光组织化学技术对小鼠生后不同发育阶段子宫组织结构的发育以及极性调控蛋白Crb1的定位表达进行了研究,结果显示,随个体发育,子宫管腔及子宫腺腔不断扩大,子宫内膜上皮持续增厚、固有层内子宫腺逐渐发达;4周龄时子宫腔面出现纤毛,随发育进程越来越发达;在各发育阶段的小鼠子宫组织中均有Crb1的表达,随个体发育表达呈现先增强后减弱的趋势,在8周龄时达到最强;Crb1主要定位于子宫内膜上皮细胞和子宫腺上皮细胞的胞膜和胞质,提示Crb1可能与小鼠子宫内膜上皮细胞和腺上皮细胞的极性建立和维持有关。     

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.     

Matrix Metalloproteinase (MMP)-2 and MMP-9 Activity in the Canine Uterus Before and During Placentation

The aim of the present study was to demonstrate the presence and localization of MMP-2 and -9 by means of RT-PCR and immunohistochemistry (IHC) within the canine uterus from the pre-implantation stage until mid-gestation and to determine MMP-2 and -9 activities by means of zymography. For this purpose, samples of the uterus and salpinx from bitches were obtained after ovariohysterectomy. Pre-implantation stages (5-12 days after mating, n = 11) were determined by verifying embryos after flushing the uterus. Further groups were determined as implantation (15-19 days after mating, n = 9), post-implantation (20-30 days after mating, n = 9) and placental stages (30-45 days after mating, n = 3). A non-pregnant group (17-30 days after mating, n = 4) served as control. MMP-2 and -9 positive cells were detected in all specimens from pregnant and nonpregnant bitches, however, with different distributions. MMP-2 was present in endothelium and smooth muscles of blood vessels and the myometrium of pregnant and nonpregnant bitches, additionally in the surface epithelium of the oviduct. The latter also stained positive for MMP-9. During placentation, MMP-2 was detected mainly in fetal blood vessels and trophoblastic cells. Higher MMP-2 activity was observed in the endometrium and myometrium of all pregnant groups compared with the nonpregnant group (p < 0.05). The pregnant groups did not differ significantly from each other (p > 0.05). MMP-9 was present in blood vessels, smooth muscle cells and epithelia, such as maternal surface epithelial cells, uterine crypts and glands. During placentation, the deep uterine glands and the epithelium of the glandular chambers were immunoreactive to MMP-9. Highest MMP-9 activities were reached in the endometrium of the pre-implantation group (23.2% of total MMP-9) and placental parts (33.3%).     

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.     

Distribution and Heterogeneity of Mast Cells in Female Reproductive Tract and Ovary on Different Days of the Oestrus Cycle in Angora Goats

The physiological distribution of mast cells (MCs) in the reproductive tract and ovary of 12 Angora goats was determined using light microscopic histochemical techniques. Uterus (corpus uteri and cornu uteri), uterine cervix, uterine tubes (isthmus and ampulla) and ovary samples were obtained by laparatomy from groups of animals during metoestrus, dioestrus and proestrus (days 5, 10 and 16 of the oestrous cycle). Tissues were fixed in Mota's fixative (basic lead acetate) for 48 h and embedded in paraffin. Six-micrometre-thick sections were stained with toluidine blue in 1% aqueous solution at pH 1.0 for 5 min and alcian blue-Safranin at pH 1.0 for 30 min. MCs were generally associated with blood vessels in all reproductive organs. In the uterus, they were concentrated mainly in the close of the uterine gland and deep stroma in the endometrium. Higher MC numbers were observed by toluidine blue staining in the uterus, uterine cervix and uterine tubes on days 10 (corpus uterine: 4.7 ± 3.8 and cornu uterine: 4.9 ± 3.5) and 16 (corpus uterine: 5.9 ± 4.5 and cornu uterine: 5.4 ± 2.4) of the oestrous cycle compared with day 5 (p < 0.05). Mast cells were not observed in the follicles, the corpus luteum and the underside of the surface epithelium of the ovarian cortex, but were observed in the interstitial cortical stroma and the ovarian medulla. In the ovary, MC numbers were significantly higher on day 16 of the oestrous cycle (cortex: 3.4 ± 2.4 and medulla: 5.7 ± 4.5, p < 0.05). Safranin-positive connective tissue MCs were not observed in the uterine tube on any occasion. These results indicate oestrous cycle-related changes in the number and location of MCs in goat reproductive organs.     

Expression of Inhibin/Activin Subunits in the Equine Uteri during the Early Pregnancy

The establishment of equine pregnancy is a unique and long process during which a series of physical and possibly biochemical interactions are required between the conceptus and uterus. In this study, we investigated the expression pattern of inhibin/activin subunits in the uterus during early pregnancy. The uteri from four adult mares on cyclic day 13 or pregnancy day 25 were obtained. Immunohistochemical experiments suggested that inhibin/activin subunits were immunolocalized in the luminal and glandular epithelium on pregnancy day 25. In addition, the inhibin α and inhibin/activin β_B subunits were not detected, and inhibin/activin β_A subunit was detected, in the luminal and glandular epithelium on cyclic day 13. Real‐time polymerase chain reaction and Western blotting results for the inhibin/activin subunits suggested a significant increase in the expression of inhibin/activin subunit β_B and a significant decrease in the expression of inhibin/activin subunit β_A on pregnancy day 25 compared with those on cyclic day 13. Enzyme‐linked immunosorbent assays suggested a significant decrease in the concentration of activin A in endometrium extracts from cyclic day 13 to pregnancy day 25. These results suggest that inhibins or activins synthesized in the uterus, as endocrine factors and necessary nutriments, have different expression patterns and may play different, important roles during early embryonic development of the equine.     

FZD5在山羊妊娠早期的表达及激素调控

实验旨在研究雌性山羊早期妊娠和发情周期中 Frizzled-5(FZD5)蛋白在子宫中的表达,以及类固醇激素对 FZD5 的表达调控。选取发情周期、早期妊娠及雌激素(50 μg/mL)与孕酮(50 ng/mL)处理山羊的子宫组织,采用免疫组织化学染色、荧光定量 PCR和 Western blot检测 FZD5在山羊子宫中的表达规律。结果表明:FZD5 蛋白在山羊妊娠早期的子宫腔上皮和腺上皮中表达;FZD5 mRNA 和蛋白在胚胎与子宫的黏附前(D6)和黏附中(D16)表达较高,在黏附后(D19)和胎盘形成早期(D25)呈下降趋势;孕酮处理导致FZD5表达降低,表明山羊子宫中孕酮下调 FZD5的表达。研究提示FZD5可能在山羊胚胎着床过程中发挥作用。     

Different patterns of vasoactive intestinal polypeptide (VIP)-immunoreactive and acetylcholinesterase (AChE)-positive innervation in the internal carotid artery and cerebral arterial tree of the quail

The innervation pattern of vasoactive intestinal polypeptide-immunoreactive (VIP-IR) nerves in the quail internal carotid artery (ICA) and cerebral arterial tree was investigated and compared with that of acetylcholinesterase-positive (AChE-P) nerves. The supply of VIP-IR nerves to the two arterial systems was distinctly richer than that of AChE-P nerves. It was focused mainly on the walls from the distal ICA to the caudal half of the anterior ramus (AR) through the cerebral carotid artery (CCA). Indeed, double staining clearly showed that numerous VIP+/AChE-axons were distributed over these arterial regions where VIP+/AChE+ or AChE+/VIP- axons were sporadic or often lacking. The finding that nerve bundles accompanying the ICA within the carotid canal contained abundant VIP+/AChE- nerve cells suggests that cerebrovascular VIP-IR nerves in the quail have their major source at these neurons and enter the cranial cavity through the CCA. Another significant finding was that a small number of nerve cells, which were mostly stained for AChE alone and occasionally for VIP alone or both, occurred in the major arteries located more rostral than the middle AR. Thus, the quail cerebral arterial tree, at least the rostral segment of the anterior circulation, is multiply innervated by these three distinct categories of the extracranial and intracranial VIP-IR and AChE-P neurons.