牦牛皱胃组织结构及黏膜免疫相关细胞研究

为揭示牦牛皱胃组织结构和黏膜免疫相关细胞的分布与数量变化的规律,采用组织化学法、图像分析法及透射电镜技术,对牦牛皱胃组织结构及上皮内淋巴细胞、浆细胞和肥大细胞变化进行了研究.结果表明:牦牛皱胃胃壁由黏膜层、黏膜下层、肌层和浆膜构成.皱胃幽门腺区胃小凹深度最深、腺体最长、肌层最厚;3个腺区肌层厚度、腺体长度之间差异极显著(P＜0.01);幽门腺区与胃底腺区、贲门腺区之间胃小凹差异极显著(P＜0.01),胃底腺区与贲门腺区之间差异不显著(P＞0.05).牦牛皱胃黏膜上皮内淋巴细胞数量和浆细胞数量,3个腺区之间差异不显著(P＞0.05);肥大细胞数量以胃底腺区最多,幽门腺区最少,两者之间差异极显著(P＜0.01),贲门腺区与胃底腺区和幽门腺区之间差异不显著(P＞0.05).皱胃各腺区固有层中均有大量的弥散淋巴细胞和孤立淋巴小结.电镜观察表明,胃小凹柱状上皮细胞排列紧密.幽门腺区固有层中有大量的黏液细胞,黏液细胞呈高柱状或锥体状,核位于基底部,在细胞顶端常聚集有较多的电子密度较高的颗粒.胃底腺区和贲门腺区有大量的壁细胞和主细胞.牦牛皱胃的组织结构和其他反刍动物基本相似,但各层有其明显特点.牦牛皱胃各腺区固有层中均有大量弥散淋巴细胞和孤立淋巴小结,使牦牛皱胃具有比其他反刍动物更强的黏膜免疫功能.     

在继代培养中贡蕉胚性悬浮细胞的分化能力和染色体数目的变化

将贡蕉[Musa acuminata cv.Mas(AA)]胚性悬浮细胞通过不同时间的培养后,对其体胚发生能力和染色体数目进行了分析。结果表明,随着培养时间的延长,贡蕉胚性悬浮细胞的体胚发生能力下降,继代培养1.5a的悬浮细胞体胚发生能力为1.76×104个/mLPCV(packed cell volume,细胞密实体积)胚性悬浮细胞,继代培养3a后下降到0.85×104个/mL PCV胚性悬浮细胞。整个胚性细胞悬浮系为混倍体,细胞的染色体数目变化从3个到70个不等,既有含染色体数目为整倍体的细胞,也含有大量染色体数目为非整倍体的细胞;继代培养1.5a时,含正常二倍体染色体数目的细胞比例为15.8%,继代培养3.0a时下降到9.7%。     

哺乳动物生殖细胞冷冻保存对其DNA甲基化影响的研究进展

哺乳动物生殖细胞和胚胎冷冻保存在辅助生殖技术中得到了广泛的应用,同时在保存物种的多样性方面发挥着巨大的作用。但是,冷冻引起的DNA甲基化对其印记基因缺陷性疾病的发生和后代生理功能的差异会有很大的影响。作者就冷冻保存技术对配子（精子和卵子）和胚胎的DNA甲基化的影响以及冷冻引起的DNA甲基化对后代的影响进行了综述,以期对配子的冷冻保存和辅助生殖技术的发展提供参考。     

Time after seawater transfer influences immune cell abundance and responses to SAV3 infection in Atlantic salmon

Pancreas disease (PD) caused by salmonid alphavirus (SAV) severely affects salmonid aquaculture during the seawater phase. To characterize immune cells in target tissues for SAV infection, heart, pancreas and pyloric caeca were analysed from two groups of fish adapted to seawater for 2 and 9 weeks. The sections were scored for the relative abundance of cells expressing MHC class II, IgM, CD3, CD8 or neutrophil/granulocyte markers using immuno‐histochemical techniques. In general, necrosis of tissue was more severe in fish infected at 2 weeks post‐seawater transfer (wpt) compared with those infected at 9 wpt. At 9 wpt, there were higher numbers of MHC II⁺ cells in heart, pancreas and pyloric caeca, IgM⁺ cells in heart and pancreas, and CD3⁺ cells in pancreas compared to those infected at 2 wpt. The majority of the immune cells infiltrating PD‐affected tissues were MHC II⁺ and CD3⁺ cells suggesting that antigen‐presenting cells and T lymphocytes are the main types of immune cells responding to SAV infection. All the investigated cell types were also observed in pyloric caeca of infected fish, suggesting that this tissue may play a role in the immune response to SAV.     

富铁酵母对家兔红细胞免疫功能的影响

为探讨富铁酵母对家兔红细胞免疫功能的影响,将32只家免随机分成富铁酵母高、低剂量组,FeSO4组及空白对照组,分别灌服富铁酵母(Fe元素8 mg/kg、4 mg/kg)、FeSO4(Fe元素4 mg/kg)及空白生理盐水;连续5 d后,检测红细胞C3g受体花环率和复合物(IC)花环率.结果:富铁酵母较FeSO4及空白组能显著提高红细胞C3h受体花环和红细胞复合物(IC)花环的百分率,但高、低剂量间无显著性差异.结论:富铁酵母能显著提高红细胞免疫功能.     

Gonadal morphogenesis and sex differentiation in cultured chub mackerel,Scomber japonicus

The timing of primordial germ‐cell (PGC) migration with regard to the gonadal anlagen, gonad formation and sex differentiation was examined histologically in the chub mackerel (Scomber japonicus) at 5–190 days post hatching (dph). At 5 dph, PGCs appeared on the peritoneal epithelium surface or in the mesentery, on the dorsal side of the abdominal cavity. By 10 dph, stromal cells around the PGCs proliferated. The gonadal primordium was formed by 15 dph. The gonadosomatic index was 0.01% at 30 dph and increased thereafter (0.32% in females and 0.04% in males at 160 dph). Ovarian differentiation occurred at 30–40 dph, indicated by ovarian cavity formation (elongation and fusion of the upper and lower ovarian edges). Meiosis was subsequently initiated. A few meiotic oocytes surrounded the cavity at 50 dph; most were in the perinucleolus stage at 60 dph and attained a diameter of 60–70 μm at 190 dph. Testicular differentiation occurred at 30 dph, indicated by the formation of the sperm duct primordium. Spermatogonia gradually proliferated, developing into spermatocytes at the chromatin–nucleolus stage (after 90 dph) and subsequently into spermatids and spermatozoa (160 dph). These data could aid the development of seeding and cell‐engineering technologies for scombrid fish.     

低温驯化下斑马鱼LINE1的表达检测

长散布核元件-1(Long spread nuclear element-1,LINE1)是跳跃基因。前期比较基因组研究发现,南极鱼经历漫长的低温适应进化后,与南极圈外的同亚目鱼类相比较,在基因水平上LINE1的扩增效率高达8~300倍,但LINE1的扩增与鱼类抵御寒冷之间的关系尚未明了。本实验对斑马鱼(Danio rerio)胚胎成纤维细胞ZF4进行了不同时间梯度的低温处理(18℃、5 d和18℃、30 d),同时对斑马鱼成鱼也进行了不同时间的低温处理(10℃,3 h、6 h、1 d、3 d、5 d)。采用RT-qPCR检测了LINE1的mRNA水平,并克隆了斑马鱼LINE1基因启动子区,利用Luciferase双荧光报告系统,在ZF4细胞中验证LINE15’UTR在低温压力下的生物活性。结果显示,短时间低温处理下,ZF4细胞中LINE1 mRNA水平有所降低,而在长时间低温处理中,LINE1的mRNA水平显著升高。在成鱼中,短时间低温处理下,LINE1 mRNA水平降低;长期低温处理下,LINE1 mRNA水平显著升高。在ZF4细胞中发现,LINE15’UTR具有生物活性。在低温处理(18℃,3 d)下,报告基因信号减弱,间接表明LINE1启动子活性减弱。研究结果表明,低温压力会影响LINE1在鱼类中的表达。本研究为进一步探究LINE1在鱼类适应低温环境中的作用机制奠定了基础。     

Characterization and expression of SLO1 potassium channels during spermatogenesis in Eriocheir sinensis

SLO1 potassium channels are pivotal to many aspects of spermatogenic cell. Experiments were conducted to assess physiology and function of SLO1 potassium channels in different developmental stages of spermatogenic cell in Eriocheir sinensis. First, the expression of SLO1 protein was examined via Western blot, RT‐PCR, immunohistochemistry and immunofluorescence assays. The results showed that the expression of the SLO1 protein was not uniform in the spermatogenic cells of E. sinensis. Second, whole‐cell patch clamp technique was used to record the potassium current of spermatogenic cells and to analyse the electrophysiological characteristics of the potassium channels with the aid of inhibitors. It is proved that the potassium current in E. sinensis germ cells is associated with intracellular Ca²⁺, and the calcium‐activated potassium channel mediated by SLO1 protein is mainly large‐conductance Ca²⁺‐activated K⁺ channels (BK_Ca). Based on these researches, the cDNA of SLO1 from testis was cloned and sequenced. The SLO1 protein contained domains bound to calcium ions, and the spatial structure formed by its tetramers constituted potassium channels. Phylogenetic analysis revealed that SLO1 was much closer to Scylla paramamosain than other examined species. Finally, iberiotoxin (IbTX) and CdCl₂ were used to inhibit the acrosome reaction (AR) induced by A23187 and to explore the role of SLO1 potassium channels in the AR of E. sinensis. The experimental results showed that SLO1 potassium channels were expressed in E. sinensis spermatogenic cells and played an important role in the AR of crab sperm (SP).     

胚胎干细胞的研究进展及应用前景

干细胞(Stem Cell)是一类具有分化潜能和自我复制的早期未分化细胞。胚胎干细胞(Embryonic stem cells,ES细胞)是一种早期胚胎内细胞团(inner cell mass,ICM)或原始生殖细胞(primordial germ cell,PGC)经体外分化抑制培养,分离和克隆得到的具有发育全能性的高度未分化细胞。本文综述了胚胎干细胞的形态学、生长特性、免疫学鉴定方法,ES细胞抑制分化和诱导分化的机理以及饲养层、血清和细胞因子等影响胚胎干细胞分离克隆的因素,并进一步阐述了胚胎干细胞的应用前景以及存在的问题。