Histomorphological and immunohistochemical study of angiogenesis and angiogenic factors in the ovary of the mare

Cyclical ovaries of 18 mares were examined histologically and immunohistochemically for vascular endothelial growth factor A and B (VEGF A; VEGF B), angiopoietin1 and 2 (Ang1; Ang2), vascular endothelial growth factor receptor 1 and 2 (VEGF-R1; VEGF-R2), angiopoietin receptor (Tie2) and von Willebrand factor. The most intensive coexpression of the examined factors and receptors was detected in the periovulatory period, when a distinctive ovarian angiogenesis takes place, being essential for tertiary follicle maturation and for the endocrine function of the Corpus luteum. Based on the immunohistochemical results, VEGF A, Ang2, VEGF-R2 and Tie2 in particular seem to play a significant role on angiogenesis during follicular and luteal development in the mare, while Ang1 supports vessel stabilisation. The findings of luteal regression and follicular atresia showed that, in the absence of VEGF A, Ang2 and its receptor Tie2 contribute substantially to vessel regression and therefore to luteolysis and follicular atresia.     

长期碳酸盐碱度胁迫对脊尾白虾生长及卵巢发育的影响

为探讨碳酸盐碱度对脊尾白虾（Exopalaemon carinicauda）生长及卵巢发育的影响。本研究设置3 mmol/L（对照组）、5 mmol/L和8 mmol/L 3种碳酸盐碱度梯度养殖脊尾白虾60天,比较不同碳酸盐碱度胁迫对脊尾白虾生长性能、组织结构、酶活性以及卵巢发育的影响,结果显示：5 mmol/L组与对照组体长增长率、体重增长率以及成活率差异不显著（P>0.05）,8 mmol/L组体长增长率、体重增长率以及成活率显著低于对照组（P<0.05）。肝胰腺组织切片显示,5 mmol/L组中部分B细胞体积增大,部分转运泡内出现颗粒物质,8 mmol/L组管腔的形态结构严重变形。鳃组织切片显示,5 mmol/L组出现轻微鳃丝肿大现象,角质层略有变形,且角质层下间隙扩张变长,上皮细胞与支柱细胞排列紊乱;8 mmol/L组鳃丝肿大,排列不规则,出现血细胞肿胀现象,鳃丝上皮细胞严重破坏,毛细血管网结构形态改变,角质层下间隙变形。肌肉组织酶活性测定结果显示,5mmol/L组的碳酸酐酶活性显著高于对照组（P<0.05）,Na+/K+-ATP酶活性在肌肉中差异不显著（P>0.05）;肝胰腺组织酶活性测定结果显示,碳酸酐酶在肝胰腺组织中差异不显著（P>0.05）,8 mmol/L组Na+/K+-ATP酶活性显著低于其他组（P<0.05）,鳃组织中,5 mmol/L和8 mmol/L组鳃组织中和Na+/K+-ATP酶活性均显著高于对照组（P<0.05）,且两组间差异显著。卵巢发育统计结果显示,三组脊尾白虾卵巢均能发育,其发育至Ⅱ期的百分比分别为20.51%、10.52%和6.25%,其中5 mmol/L组与对照组间无显著差异（P>0.05）,8 mmol/L组显著低于对照组（P<0.05）;3 mmol/L组和5 mmol/L组均有卵巢发育至Ⅲ期的脊尾白虾,但两组差异不显著（P>0.05）,8 mmol/L组未见有卵巢发育至Ⅲ期的脊尾白虾。卵巢组织切片显示,8 mmol/L组卵原细胞细胞排列疏松,周围滤泡细胞排列疏松且数量较少。综上所述,脊尾白虾可以在低于8 mmol/L的碳酸盐碱度中生长发育,但高碳酸盐碱度可能会造成脊尾白虾鳃和肝胰腺组织损伤,导致其生长受到影响,同时猜测高碳酸盐碱度可能也会影响卵巢的发育速度。     

牛精子不同获能方法对体外受精效果的影响

为了提高牛体外受精率,探讨牛冷冻精液不同获能方法对体外受精效果的影响,对牛冷冻解冻精液进行了两次离心获能法与一次离心加上游法获能对体外受精效果影响试验.结果表明,两次离心组受精率卵裂率(75.6-±4.5)％与一次离心加上游法组受精卵裂率(76.4±1.9)％无显著差异(P＞0.05);二次离心组囊胚率(35.7±4.1)％与一次离心加上游法组(36.3±2.7)％差异不显著(P＞0.05).而两次离心组却可以大大节省了体外获能时间.     

Multiple molecular forms of inhibin in buffalo (Bubalus bubalis) ovarian follicular fluid

Inhibin is a heterodimeric glycoprotein hormone involved in the regulation of FSH release from the anterior pituitary gland and it has been characterized from various animals. Although, multiple molecular forms of inhibin have been reported from different species, however, the molecular nature of inhibin has not been studied in buffaloes. In the present study, attempts were made to identify inhibin in buffalo ovarian follicular fluid. Buffalo ovaries were obtained from the local abattoir and follicular fluid was aspirated from surface follicle (with diameter ?5 mm). A combination of techniques (viz., gel filtration, SDS-PAGE, Western blot etc.) was employed for identification and isolation of inhibin(s). Inhibin bands were detected at 129 and 63 kDa by Western blot analysis in non-reducing conditions. In reduced SDS-PAGE, 63 kDa fraction produced a single band while 129 kDa fraction resolved into four components of 63, 43, 29 and 20 kDa. Out of them only 29, 63 and the native 129 kDa fractions produced bands on Western blot analysis. In total five fractions (63, 54, 39, 29, 25 kDa) were obtained by trypsin digestion of 129 kDa form. However, only 63 and 29 kDa fractions showed immunoreactivity. In this study, for the first time, we have identified two major forms of inhibin (129 and 63 kDa) with little proteolytic cleavage/processing of the large precursor in the buffalo follicular fluid.     

玉米未授粉子房离体培养及植株再生

探讨了不同材料、不同培养基对玉米未授粉子房的影响。确定了未授粉子房最佳培养时期,筛选出一批用于玉米未授粉子房诱导频率高的培养材料。     

芥菜型油菜和黑芥与诸葛菜属间杂种的获得及其特性

属间远缘杂交是创造新种质、解决育种材料短缺的一种有效方法。为了使诸葛菜的优良基因转入芥菜型油菜和黑芥中, 采用子房培养技术, 进行芥菜型油菜和黑芥与诸葛菜的正反属间杂交。在6种不同的培养基中培养子房。结果表明, 不同的杂交组合、培养基和母本的属间杂交可交配性不同。通过子房培养, 获得了10株属间杂种, 其中9株在形态上近似母本, 或介于父母本之间, 尤嫩斯与诸葛菜杂交组合的1株杂种表现出完全的雄性不育, 并在分枝类型和叶型方面表现出明显的变异; 形态学和SSR分子生物学鉴定表明10株杂种都是真正的属间杂种。     

山羊转t-PA突变体基因的体细胞核移植研究

比较了用转t-PA突变体基因的胎儿成纤维细胞和卵丘细胞作供体时核移胚的发育情况。结果表明：卵丘细胞作供体时,核移胚的卵裂率（82.9%）和桑葚胚率（55.9%）均高于胎儿成纤维细胞卵裂率（75.7%）和桑葚胚率（48.7%）,但差异不显著（P＞0.05）;卵丘细胞作供体时的囊胚率（19.7%）则高于胎儿成纤维细胞（11.0%）,差异显著。(P<0.05)。比较了饥饿处理与否对胎儿成纤维细胞作供体时转基因核移胚的发育情况。结果表明：供体细胞饥饿后,其核移胚的卵裂率（75.6%）与供体不饥饿的卵裂率（75.8%）差异不显著;供体细胞饥饿后,其核移胚的桑葚胚率（46.5%）和囊胚率（11.0%）高于供体不饥饿的桑葚胚率（39.3%）和囊胚率（9.0%）,但差异不显著。比较了SOFaa和CR1aa分别添加10%FBS和bFF后,转基因核移胚的发育情况。结果表明：用CR1aa液培养时,添加10%bFF,其囊胚率（9.1%）高于添加10%FBS（4.2%）（P＞0.05）;用SOF液培养时,添加10%bFF,其囊胚率（11.2%）高于添加10%FBS（5.5%）（P＞0.05）。说明用SOF液优于CR1aa液。     

Dysgenesis atypica ovarü bei einer XY-Kalbin. Histologische, biochemische, genetische und Rezeptor-Befunde

Inhalt: Die Phänomene der gonadalen Dysgenese weisen eine erhebliche Bandbreite au Die hier vorgestellte XY-Kalbin hatte ein geringgradig hypoplastisches, weibliches Genitale, in den dysgenetischen Gonaden lagen 2 Primordialrestfollikel vor. Ihre inter-stitiellen Zellen waren hyperplastisch, doch gab es keine Anzeichen für eine tumoröse Entartung, in Art eines Gonadenblastoms oder Dysgerminoms. Das Tier hatte zur Behandlung der bestehenden primären Amenorrhoe keine Hormonbehandlung erfahren. Die Konzentrationen uon Sexualsteroiden (Testosteron, Epitestosteron, Dehydroepian-drosteron) glichen solchen, wie sie im Zyklus des Rindes auftreten, ein Konzentrations-anstieg nach Pregnylapplikation war nichtzu erzielen. Nucleare Rezeptoren für Dihydro-testosteron waren nicht nachzuweisen. Die Persistenz von einzelnen Follikeln und von Keimzellen in den postpartalen resp. postpubertären Gonaden wurde in den letzten Jahren beim Menschen gelegentlich beschrieben, ist jedoch fur die reine gonadale Dysgenese nicht charakteristisch. Contents: Dysgenesis atypica ovarii in a XY-heifer The phenomena of gonadal dysgenesis show a wide range of variations. The XY-heifer described in this article showed low grade of hypoplasia of the inner female genital organes. Two primordial residual follicles were detected in serial sections of the dysgenetic gonads. The interstitial cells were hyperplastic, but there was no indication of tumorous degeneration, such as gonadoblastoma or dysgerminoma. No hormones had been given to treat primary amenorrhoe. Levels of sexual steroids (testosterone, epitestosterone, dehydroepiandrosterone were similar to those occurring in normal bovine cycle; Pregnyl failed to produce any rise in the concentration of sexual steroids. No evidence was found of nuclear receptors for dihydrotestosterone. Persistence of singular follicles and germinal cells in the postpartal and postpubertary gonads resp. of XY females has been sometimes observed (Cussen & Mac Mahon, 1979; Russel et al., 1980; Warner et al., 1985), but this is — by no means —characteristic for pure gonadal dysgenesis.     

芸薹属种间F1与羽衣甘蓝回交亲和性及子房培养研究

以大白菜与羽衣甘蓝CMS种间杂种F1为母本,以羽衣甘蓝自交系为父本进行回交,并对杂交子房进行了离体培养.结果表明:上述亲本间存在较强的生殖隔离,花期杂交不结实,蕾期重复授粉提高了亲和力.授粉后9d的杂交子房进行挽救培养效果最佳,离体条件下结籽率10%.其次为授粉后12d及6d的子房,结籽率分别为5%和4.4%.外源激素及其配比对子房离体结实具有极显著影响,以BA2 NAA0.1最佳,结籽率为9%;其次为BA2 NAA0.2,结籽率为3%;BA2 NAA0.3及无激素MS培养基上均未获得饱满种子.     

西伯利亚百合子房的组织培养及离体快繁研究

以子房为外植体,对西伯利亚百合离体快繁进行研究.结果表明:试验浓度范围内适合不定芽诱导的培养基以MS+6-BA 1.0 mg/L+NAA 1.5 mg/L为佳,芽增殖最佳培养基为MS+6-BA 1.0 mg/L+NAA 0.2 mg/L,适合生根的培养基为MS0,生根率达100%,且根系发达,长势粗壮,练苗3～5 d后移栽,长势良好,移栽成活率达90%以上.