鳗鲡肝脏,脾脏显微与超微结构

经光镜和电镜观察发现：鳗鲡肝脏的肝小叶不规则。肝细胞胞质内翕含多种细胞器及包含和,胆小管由２－４个肝细胞围成,相邻肝细胞间有连接复合体封闭胆小管。血窦为有孔型,孔处无隔膜,内有巨噬细胞。窦周隙明显,未见贮脂细胞。肝猾胆小管腔与窦周隙面伸出许多指状微绒毛。脾脏内白髓中淋巴细胞聚集成群,无未见明显脾小结,淋巴鞘,红髓由脾索与脾窦组成,动脉分支末端（壁厚的毛细血管）可开放于红髓,无明显巨噬细胞中心,脾窦     

欧洲鳗鲡外周血细胞的显微和超微结构

利用光镜和透射电镜技术研究了欧洲鳗鲡（Anguilla anguilla）外周血细胞的显微和亚显微结构。结果表明：在外周血细胞中可以区分出红细胞、单核细胞、大淋巴细胞、小淋巴细胞、嗜中性粒细胞和血栓细胞;嗜中性粒细胞内的特殊颗粒包括A、B、C三型;嗜中性粒细胞分为Ⅰ型粒细胞（内含A、B、C三种特殊颗粒）、Ⅱ型粒细胞（内含A型和C型两种特殊颗粒）和Ⅲ型粒细胞（内含C型特殊颗粒）。还见到幼稚的和正在分裂的红细胞和单核细胞,提示示幼稚的红细胞能直接进入外周血流中,嗜酸性粒细胞和嗜碱性粒细胞无论在血涂片上或超薄切片上均未见到。描述了上述各种血细胞在光镜和电镜观察下的形态和细微结构。血栓细胞体积最小,嗜中性粒细胞体积最大;淋巴细胞数目最多,嗜中性粒细胞数目最少。     

唐鱼肝脏显微和超微结构观察

应用光镜和透射电镜对繁殖期间唐鱼Tanichthys albonubes肝脏组织的显微和超微结构进行了观察。结果显示,唐鱼肝细胞具单核,中央核仁显著;细胞质内分布着粗面内质网、线粒体、糖原颗粒和脂滴等细胞器和内含物。胆小管由2～3个相邻肝细胞质膜凹陷围成,而肝血窦则由内皮细胞的胞质、成纤维细胞等参与构成。肝细胞与周边细胞通过3种不同方式进行联系:肝细胞之间的紧密连接;与血窦的间接连接;与胆小管的邻接。这些联系方式显示了肝脏具有内分泌腺和外分泌腺功能的特点。研究还发现雌性唐鱼肝脏具有"暗"细胞和"淡"细胞两种类型。本文还讨论了唐鱼肝脏与其他硬骨鱼类肝脏一般组织结构和超微结构的异同点。     

铜锈环棱螺肝脏的显微与超微结构观察

应用组织切片、光学显微镜及透射电子显微镜技术,对铜锈环棱螺肝脏进行观察研究。结果表明,铜锈环棱螺肝脏为复管泡状腺,由许多肝小叶组成,每个肝小叶管壁由单层上皮细胞构成,分3种细胞：消化细胞、排泄细胞、钙细胞。光镜显示,肝小叶多球形或近球形,横切面管腔呈星射状裂隙,纵切面可见上皮细胞呈柱状,核靠近基膜,上皮细胞极性明显。消化细胞构成腺上皮的主体,染色较淡。排泄细胞和钙细胞染色较深。肝腺管之间有不规则圆形的网状细胞,结缔组织填充在肝小叶之间及小叶与导管之间。电镜显示,消化细胞和排泄细胞顶端都具微绒毛,消化细胞主要特征为具有溶酶体囊泡系统,参与细胞内消化过程。排泄细胞中粗面内质网、线粒体等细胞器含量丰富,细胞内还散布着大量脂滴。钙细胞锥体形,主要特征为细胞内具钙粒子,也含有大量细胞器。     

鳗鲡繁殖生物学研究——Ⅲ．鳗鲡性腺发育组织学和细胞学研究

鳗鲡精巢发育可划分为6个时期,即精原细胞前增殖期,精原细胞后增殖期,精母细胞生长、成熟期,精子开始出现期,精子完全成熟期和精子退化吸收期。卵细胞的发育可划分为6个时相,即卵原细胞时相,卵母细胞单层滤泡时相,卵母细胞出现脂肪泡时相,卵母细胞卵黄充满时相,卵母细胞核极化时相和卵母细胞退化时相。以卵细胞发育6个时相在卵巢中组成的差异,也可把卵巢划分为相应的6个时期。对鳗鲡性腺发育的分期,卵黄积累方式,产卵类型等问题进行了讨论。     

植物血凝素对兴国红鲤头肾和脾脏的比较组织学研究

PHA注射前后兴国红鲤头肾和脾脏结构基本相同。红鲤头肾有被膜,为淋巴样组织,由许多血管,血窦和淋巴索组成,脾脏是实质性器官,淋巴细胞聚集成团,有弥散的胰腺组织渗入,注射PHA后头肾和脾脏内的大淋巴细胞,小淋巴细胞,巨噬细胞以及原始型细胞显著增加,而粒细胞数量变化不明显。     

饲料中添加胆汁酸对欧洲鳗鲡幼鱼肝脏脂肪代谢的影响

文章旨在研究饲料中添加胆汁酸对欧洲鳗鲡(Anguilla anguilla)幼鱼肝脏脂肪代谢的影响。将养殖欧洲鳗鲡幼鱼的9口水泥池[初始规格(141.5±1.9) g/尾; 初始鱼重(682±23) kg/池]随机分为3个处理组, 每组3口池, 分别投喂基础饲料(对照组)、基础饲料添加500 mg/kg胆汁酸(BA1组)、基础饲料添加1000 mg/kg 胆汁酸(BA2组)的饲料。实验期为15周。实验结果表明, BA组比对照组欧洲鳗鲡幼鱼肝脏组织中脂肪空泡数量明显减少, 肝脏粗脂肪水平显著降低(P<0.05)。BA组欧洲鳗鲡幼鱼肝脏脂肪酸合成酶水平显著降低(P<0.05), 仅BA1组乙酰辅酶A羧化酶水平显著降低(P<0.05), BA组肝脂酶、脂蛋白酯酶和总脂酶活性均显著升高(P<0.05); BA组间上述酶活性或水平均无显著差异(P>0.05)。BA1组欧洲鳗鲡幼鱼肝脏磷脂酰胆碱和溶血磷脂酰胆碱水平上调, 主要是甘油磷脂代谢和甘油酯代谢增强。由此可见, 欧洲鳗鲡幼鱼饲料中添加胆汁酸可通过降低脂肪合成有关酶水平及增加脂肪分解有关酶活性, 主要增强甘油磷脂代谢和甘油酯代谢, 从而可减少肝脏脂肪蓄积。     

产后鳗鲡生殖生理学的研究

产卵后的鳗鲡（Ａｎｇｕｉｌｌａ ｊａｐｏｎｉｃａ）可分为顺产鳗鲡（Ｓｍｏｏｔｈｌｙ ｏｖｕｌａｔｅｄ ｅｅｌｓ,ＳＯＥ）和部分产鳗鲡（Ｐａｒｔｌｙ ｏｖｕｌａｔｅｄ ｅｅｌｓ,ＰＯＥ）两种类型,经组织学检查发现,ＳＯＥ性腺中第Ⅳ＾＋＋＋和第Ⅴ时相的卵细胞较多,而ＰＯＥ性腺中第Ⅳ＾＋＋和第Ⅵ时相的卵细胞较多。ＳＯＥ脑部的ｍＧｎＲＨ、血清ＧｔＨ和雌二醇（Ｅ２）的含量均显著高于ＰＯＥ,但ＳＯＥ脑部ｃＧｎＲＨ－Ⅱ及血清睾酮（Ｔ）含量与ＰＯＥ无显著差异。     

次声暴露对大鼠脾、肝脏某些酶活性的影响

目的:观察8 Hz,130 dB次声暴露不同时间对大鼠脾、肝脏某些酶活性的影响.方法:35只SD大鼠随机分为5组,即对照组,1周,2周,3周,4周组.每天次声暴露1次,每次2 h.实验后,观察大鼠脾、肝脏组织中MAO,GSH-px,SOD活性和MDA含量的变化.结果:大鼠脾脏MAO活性1周,2周时显著增高(P＜0.01),3周下降,4周时又显著增加(P＜0.05).肝脏组织MAO活性变化不明显(P＞0.05).脾脏组织中GSH-px活性在4周时明显增高(P＜0.05),肝脏组织中GSH-px活性在1周时就有显著性增高(P＜0.05).脾脏SOD活性在1周至4周均有显著性增高(P＜0.05).肝脏组织在实验期变化不明显(P＞0.05).脾脏组织中MDA含量在3周至4周时有显著性增高(P＜0.05).肝脏组织在1至2周时有非常显著的增高(P＜0.01),在3周时下降,到4周时又显著高于对照组(P＜0.05).结论:8Hz,130 dB次声暴露,大鼠脾、肝脏组织活性氧自由基、脂质过氧化物增高,抗氧化能力降低,造成对组织的损伤.     

日本鳗鲡肝脏表达抗菌肽2基因的克隆与表达

为探讨鱼类抗菌肽基因的生物学功能,研究应用RACE方法克隆获得了日本鳗鲡 (Anguilla japonica) 肝脏表达抗菌肽2基因 (Liver-Expressed Antimicrobial Peptide 2,LEAP-2),即AJLEAP-2的cDNA序列,全长为450 bp,开放阅读框编码89个氨基酸。其成熟肽含有LEAP-2保守基序C-X5-C-X4-C-X4-C。AJLEAP-2基因组结构与其他脊椎动物LEAP-2相同,都包含有三个外显子。利用荧光定量PCR检测了AJLEAP-2在日本鳗鲡不同组织/器官中的表达,发现其转录子在肝脏中表达量最高,是内参基因 (-actin) 的6倍; 其次是肠道,但其表达量仅为肝脏的1/130。此外,还检测了AJLEAP-2在日本鳗鲡玻璃鳗(Glass eel)阶段的转录表达水平,结果显示,玻璃鳗中AJLEAP-2的转录表达量仅低于黑仔期的肝脏,为黑仔鳗肠道表达量的2倍。LPS和迟缓爱德华菌 (Edwardsiella tarda) 刺激能显著上调鳗鲡血液中AJLEAP-2的转录表达,刺激16h后上调倍数最高,分别为对照组的86倍和12倍。此外,LPS刺激72h和E. tarda 刺激8h后,肠道中AJLEAP-2显著上调表达(P0.05),为对照组的8倍。Poly I:C刺激24h后,血液中AJLEAP-2转录表达显著下调。结果表明,AJLEAP-2在日本鳗鲡抗细菌感染过程中起重要的作用。     

Inhibitory effect of cholesterol on the uptake of liposomes by liver and spleen

The effect of cholesterol content of small unilamellar (SUV) and reverse phase (REV) liposomes on blood clearance and tissue distribution has been studied. [¹⁴C]Inulin has been used as an aqueous marker of liposomes to represent the uptake of intact liposomes in tissues. The blood clearance of the intravenously-injected SUV and REV liposomes depends on the cholesterol content of liposomes. The cholesterol-free (0 mol%) liposomes are cleared more readily from the circulation than the cholesterol-poor liposomes (20 mol%) and the cholesterol-poor are cleared more rapidly than the cholesterol-rich (46.6 mol%) liposomes. This clearance pattern of liposomes from the circulation is not attributed to the change of size of liposomes due to the increase in cholesterol content of liposomes. However, poor stability of cholesterol-free or cholesterol-poor liposomes in the circulation is partly responsible, but the predominant factor responsible for the observed blood clearance pattern is the inhibitory effect of cholesterol on the uptake of liposomes by reticuloendothelial-rich tissues liver and spleen. Uptake of liposomes by these organs is decreased with increasing cholesterol content of vesicles. It is suggested that to produce liposome preparations with a long circulating half life in vivo it is necessary to inhibit their uptake by liver and spleen.     

A WXW Motif Is Required for the Anticancer Activity of the TAT-RasGAP317–326 Peptide

TAT-RasGAP_317–326, a cell-permeable 10-amino acid-long peptide derived from the N2 fragment of p120 Ras GTPase-activating protein (RasGAP), sensitizes tumor cells to apoptosis induced by various anticancer therapies. This RasGAP-derived peptide, by targeting the deleted in liver cancer-1 (DLC1) tumor suppressor, also hampers cell migration and invasion by promoting cell adherence and by inhibiting cell movement. Here, we systematically investigated the role of each amino acid within the RasGAP_317–326 sequence for the anticancer activities of TAT-RasGAP_317–326. We report here that the first three amino acids of this sequence, tryptophan, methionine, and tryptophan (WMW), are necessary and sufficient to sensitize cancer cells to cisplatin-induced apoptosis and to reduce cell migration. The WMW motif was found to be critical for the binding of fragment N2 to DLC1. These results define the interaction mode between the active anticancer sequence of RasGAP and DLC1. This knowledge will facilitate the design of small molecules bearing the tumor-sensitizing and antimetastatic activities of TAT-RasGAP_317–326.     

鳗鲡寄生伪指环虫(单殖吸虫)的危害性、生物学和治疗试验

本文研究了人工放养池内引起鳗鲡鳃病的寄生虫是秉伪指环虫和鳗鲡伪指环虫。影响秉伪指环虫纤毛幼虫孵化的因素,与温度和寄主粘液密切相关,而与光照可能无关。在实验室里,分别用药物处理带有病原体的鳗苗,表明:用0.3—0.5ppm90％,晶体敌百虫,可达全部杀灭病原体的良好效果,1％氯化钠长期药浴也有显著效果。     

Cholinoceptor-mediated control of catecholamine release from chromaffin cells in the American eel,Anguilla rostrata

The cholinergic agonist-induced secretion of catecholamines from chromaffin cells in the American eel, Anguilla rostrata, was assessed using a salineperfused posterior cardinal vein preparation. Direct membrane depolarization with 60 mmol·l^-1 K⁺ caused a significant release of catecholamines (adrenaline + noradrenaline) into the perfusate which was unaffected by pre-treatment with the ganglion blocker, hexamethonium (final concentration = 10^-3 mol · l^-1). The nicotinic receptor agonist, 1,1-dimethyl-4-phenylpiperazinium iodide, evoked catecholamine release in response to several doses exceeding 10^-7 mol; at 10^-5 mol the response was abolished by pre-treatment with the ganglion blocker, hexamethonium (final concentration = 10^-3 mol · l^-1). The muscarinic receptor agonist, pilocarpine, did not elicit catecholamine release in response to any of the doses administered (10^-8–10^-4 mol). A single injection of the mixed nicotinic/muscarinic cholinoceptor agonist, carbachol (10^-5 mol), caused the release of catecholamines which was abolished by pre-treatment with hexamethonium but which was unaffected by pre-treatment with the muscarinic receptor antagonist atropine (final concentration = 10^-5 mol · l^-1). The results of this study indicate that the process of cholinergic agonist-induced catecholamine secretion from the chromaffin cells in the American eel is mediated exclusively by activation of nicotinic receptors with no involvement of the muscarinic receptor.Abbreviations DMPP 1,1-dimethyl-4-phenylpiperazinium iodide - MS222 ethylaminobenzoate - HPLC high-performance liquid chromatography - PCV posterior cardinal vein - SEM standard error of the mean     

Fine structure of the lateral-line organ of the common eel,Anguilla japonica

Summary The sensory epithelium of the lateral line organ of the common eel consists of two types of cells, (sensory and supporting). The sensory cell bears a kinocilium together with about 40 to 60 stereocilia on its surface. The kinocilium is situated either at rostral or at caudal margin of this cilial group. Such polarity of the cilial group of one cell is inverse to that of an adjacent cell.Two types of crystal-like inclusions exist in the sensory cells, consisting of granules 100 Å in diameter. Granules in one type are arranged regularly whereas those in the other rather irregularly.Two types of nerve endings exist at the base of sensory cells: one is predominant in number and contains few vesicles, accompanied by a dense spherical body surrounded by small vesicles in the sensory cell and the other is rare in number and contains many vesicles, accompanied by a small flat sac just beneath the plasma membrane of the sensory cell.The supporting cells contain numerous mitochondria, a well developed Golgi apparatus and rough-surfaced endoplasmic reticulum, and surround a sensory cell completely. Physiologic significance of some of these components is discussed.     

Spectrum of variations in dog-1/FANCJ and mdf-1/MAD1 defective Caenorhabditis elegans strains after long-term propagation

Background

Whole and partial chromosome losses or gains and structural chromosome changes are hallmarks of human tumors. Guanine-rich DNA, which has a potential to form a G-quadruplex (G₄) structure, is particularly vulnerable to changes. In Caenorhabditis elegans, faithful transmission of G-rich DNA is ensured by the DOG-1/FANCJ deadbox helicase.

Results

To identify a spectrum of mutations, after long-term propagation, we combined whole genome sequencing (WGS) and oligonucleotide array Comparative Genomic Hybridization (oaCGH) analysis of a C. elegans strain that was propagated, in the absence of DOG-1 and MDF-1/MAD1, for a total of 470 generations, with samples taken for long term storage (by freezing) in generations 170 and 270. We compared the genomes of F₁₇₀ and F₄₇₀ strains and identified 94 substitutions, 17 InDels, 3 duplications, and 139 deletions larger than 20 bp. These homozygous variants were predicted to impact 101 protein-coding genes. Phenotypic analysis of this strain revealed remarkable fitness recovery indicating that mutations, which have accumulated in the strain, are not only tolerated but also cooperate to achieve long-term population survival in the absence of DOG-1 and MDF-1. Furthermore, deletions larger than 20 bp were the only variants that frequently occurred in G-rich DNA. We showed that 126 of the possible 954 predicted monoG/C tracts, larger than 14 bp, were deleted in unc-46 mdf-1 such-4; dog-1 F₄₇₀ (JNC170).

Conclusions

Here, we identified variants that accumulated in C. elegans’ genome after long-term propagation in the absence of DOG-1 and MDF-1. We showed that DNA sequences, with G₄-forming potential, are vulnerable to deletion-formation in this genetic background.

Electronic supplementary material

The online version of this article (doi:10.1186/s12864-015-1402-y) contains supplementary material, which is available to authorized users.