卤虫(Artemia salina)孵化腺细胞分化时相的免疫细胞化学研究(简报)

动物孵化酶(hatching enzyme,HE)是早期胚胎在特定发育阶段由孵化腺细胞产生和分泌的,在动物早期胚胎孵化中具有关键性作用。孵化腺细胞(hatching gland cell,HGC)一般为单细胞腺体,是从胚胎发育到特定阶段(孵化前)出现、至胚胎孵出后的特定时期消失的一时性细胞(transient type ofcells)。完全分化的HGC内充满了低电子密度的酶原颗粒(孵化酶原颗粒),在鱼胚中的分布因物种而异。在大多数鱼中,HGC分布在胚体的外表面和/或卵黄囊中,一般为外胚层来源。如在虹蹲鱼HGC分布在胚体的前表面、卵黄囊、咽部、鳃的内表面及外表面,属于外胚层来源。而日本鳉鱼HGC     

中华大蟾蜍胚胎孵化酶的分离和提纯

研究证明,海胆、鱼、两栖类胚胎孵化主要是由孵化腺细胞分泌的孵化酶(hatching enzyme, HE)作用的结果。研究HE的意义在于:1.研究孵化腺细胞的分化和溶解卵膜的机理。2.搞清HE的性质将有助于提高水生动物的孵化率。我们在研究中华大蟾蜍孵化腺细胞的形成、分化及结构的基础上进一步对孵化酶进行了分离和提纯。     

动物胚胎的孵化

动物的受精卵外都包有卵膜,因此,胚胎发育都是在卵膜内进行的。当胚胎发育到一定的阶段就要脱离其外的卵膜,这过程我们称谓孵化(hatching)。这种情况不仅遍及于卵生动物,并且也发生于胎生动物,其中包括哺乳动物。因此,孵化是所有动物门(单细胞动物除外)一个共同的现象。孵化大体上可分为两类:一类是机械的孵化,即卵膜的破裂由于卵膜胚胎或胚外压力的增加,或者由于胚胎的啄穿或咀嚼;另一类是酶的孵化,即卵膜的破裂是由于胚胎分泌一种酶,先使卵膜软化,加上胚胎肌肉收缩或纤毛运动而使胚胎从卵膜内孵出。前者如爬行类、鸟类和哺乳类;后者如海胆、鱼和两     

河川沙塘鳢孵化腺的发生及孵化酶的分泌

利用光学显微镜和扫描电镜观察了河川沙塘鳢Odontobutis potamophila(Gnther)孵化腺的发生及孵化酶的分泌过程。河川沙塘鳢的孵化腺为单层细胞腺体,发生于外胚层。孵化腺(Hatching gland,HG)最早发生自眼晶体形成期的胚胎,初发生时孵化腺细胞(Hatching gland cells,HGCs)分布于头部腹面及其与卵黄囊连接处。随着胚胎的发育,HG仍以单层细胞分布于胚体和卵黄囊的外表面,HGCs数量急剧增多,细胞体积增大,分布范围更加广泛。至眼黑色素出现期的胚胎,HGCs的数量达到大约900-1200个,广泛分布于胚胎头部两侧、头部腹面及其与卵黄囊连接处、卵黄囊的前腹面。HGCs大多呈椭圆形,短径为5-8μm,长径为7-12μm,在HE染色中呈粉红色。至孵化前期时,孵化酶颗粒自HGCs顶部的开口分泌出来,分泌颗粒呈圆球形,直径为0.5-1.0μm,有的以单体的形式存在,有的粘结成团。孵化酶进入卵周液,对卵膜内层进行消化和降解,使胚胎破膜而出。孵化后2d,HGCs便从表皮中消失。     

鲤胚胎孵化腺细胞

鲤胚胎孵化腺为单细胞腺体,发生于外胚层,可特异地被PAS染色。最早可在眼色素期检验出孵化腺细胞(Hatching gland cell,HGC)它们主要分布在头部腹面及头部与卵黄囊连接处。开始,HGC位于表皮细胞下面,随发育迁移到胚胎表面。根据扫描和透射电镜观察,在分泌孵化酶的前后,HGC区表面细胞呈鸡冠花状和疣状两种突起。前者系HGC处于分泌孵化酶期间;后者系HGC业已完成分泌作用,由于相邻的表皮细胞活动而形成的。HGC内富有粗面内质网、线粒体、核糖体和高尔基体,并由后者合成酶原颗粒。HGC在完成分泌作用后,仍留在表皮中,以后逐渐退化,但在孵化后30h仍可见残留的HGC。     

亲本繁殖和胚胎孵化温度对斑马鱼孵化表现的影响

亲本效应(parental effect)指亲代的表现型及其所经历的环境因素而非基因型对子代表现型差异和适应性的影响,对子代适合度的维持与提高具有重要生态学意义。为探究亲本效应对鱼类胚胎发育可塑性的影响,本研究选取卵生鱼类斑马鱼(Danio rerio)为实验对象,采用2×2双因素设计,测定了不同亲本繁殖温度(22、28℃)和不同胚胎孵化温度(22、28℃)及其交互作用对斑马鱼孵化表现(孵化率、胚胎死亡率、初孵仔鱼畸形率、孵化历时、初始孵化时间、结束孵化时间)的影响。结果表明：亲本繁殖和胚胎孵化温度交互作用对斑马鱼胚胎死亡率、初始孵化时间、结束孵化时间、孵化历时等均有显著影响(P<0.05),亲本繁殖与胚胎孵化温度相一致时胚胎的死亡率更低、孵化时间更短;斑马鱼孵化表现受亲本效应(代际发育可塑性)和子代发育环境(代内发育可塑性)的双重影响,亲本效应对早期生活史阶段鱼类表型特征的塑造有重要作用。     

单独隔离孵化对镇海林蛙蝌蚪生长发育、社交行为和运动表现的影响

两栖类胚胎呈卵群一起孵化在反捕食、维持胚胎间的温度平衡和促进胚胎的同步发育等方面具有重要作用。在实验条件下探究孵化条件(单独隔离孵化和卵群一起孵化)对镇海林蛙(Rana zhenhaiensis)蝌蚪的生长发育、社交行为及其运动表现能力的影响。结果表明,孵出2天后,单独隔离孵化组蝌蚪的体长和发育历期均显著大于卵群一起孵化组。以体长为协变量的协方差分析表明,特定体长的单独隔离孵化组蝌蚪的体宽和体重均显著大于卵群一起孵化组的个体。在蝌蚪的活动行为中,单独隔离孵化组蝌蚪在水体上层和中层出现的频次显著小于卵群一起孵化组,而在水体底层的出现频次显著大于卵群一起孵化组。单独隔离孵化组蝌蚪个体的分布面积率、最近邻个体的距离、个体间的距离均显著大于卵群一起孵化组,但个体间的接触频次显著小于卵群一起孵化组。在运动表现方面,单独隔离孵化组胚胎的摆尾搏动频次显著小于卵群一起孵化组。单独隔离孵化组蝌蚪的运动时长和运动频次均显著小于卵群一起孵化组。本研究结果将为无尾两栖类幼期的生长发育规律提供参考数据。     

温度对中华鳖卵孵化和胚胎发育的影响

在25℃,27．5℃,30℃,32．5℃和35℃条件下,分别研究了中华鳖（Trionyx sinensis)卵的孵化和胚胎的发育。卵黄和卵壳的湿重分别占卵重的85．10％和14．90％,含水量分别为76．63％和21．01％。中华鳖胚胎重随孵化时间呈逻辑斯谛曲线增长,孵化温度越高,胚胎发育速度r越大。25℃－30℃的胚胎发育仅表现为‘S’型生长曲线的前,中段,在孵化后期发育速度未受到明显的抑制,32．5℃和35℃温度条件下,后期发育速度受到了较大的影响。孵出稚鳖湿重和干重均与初始孵重存在极显著的经性相关关系。孵化温度对孵出稚鳖湿重没有显著影响,但对孵出稚鳖干重的影响极显著。随孵化温度的升高,孵出稚鳖干重吾‘U’形变化,在适中温度下,胚胎的生长效率最低。卵黄干重随孵化时间呈指数递减,温度越高,孵黄的利用速度越快。在孵化过程中,卵重,卵干重和卵内水总量都随孵化时间的延长,孵化温度的上升而略有下降。孵化期随温度升高呈双曲线缩短,其相关关系可用方程I＝30．6＋109／（T－22．5）来表示。中华鳖卵的最适孵化温度在32．5℃左右,胚胎孵化后期比早期对温度的需求可能要低一些。     

盐度对条石鲷胚胎发育的影响

为了确定条石鲷胚胎孵化的适宜盐度,在试验水温为23.0℃～25.2℃条件下,设置了8个盐度组进行条石鲷胚胎孵化试验,研究了盐度对条石鲷胚胎发育的影响。结果表明:低盐度和高盐度对胚胎有持续性伤害,均可造成胚胎和卵黄球在卵裂期收缩而死亡,一部分胚胎在原肠期之后收缩死亡;低盐度可造成初孵仔鱼畸形率增加,主要引起仔鱼脊柱"L"形和"C"形弯曲,而高盐度可导致胚体胚孔关闭以后尾部的畸形发育;通过分析各盐度组的孵化率、仔鱼畸形率和盐度之间的关系,得出健康仔鱼比例(PHL)和盐度(S)的关系式为PHL=-0.0018S2-0.1135S-0.8853(R2=0.948),以PHL70%为适宜孵化盐度范围,以PHL90%为最适孵化盐度范围,由此确定在23.0℃～25.2℃条件下,条石鲷的适宜孵化盐度范围为21～42(PHL70%),最适为30～33(PHL90%)。     

对ERP非专业成员有用的遗传术语词汇表（6）

嵌合体 由两种或多种不同表型组织构成的个体。可通过普通杂交或混合不同类型的早期胚胎细胞获得。 条件转基因技术 将基因修饰限定于特定的组织或细胞,或在胚胎、动物发育的不同时间点,或使其被特定的信号（如某一化学物质的注射）激活。 基因构建（或转基因） 一段包含目的DNA序列的人工获得的DNA片段。     

The presence of guanosine 5'-diphospho-5'-guanosine and guanosine 5'-triphospho-5'-adenosine in brine shrimp embryos

Acid-soluble extracts of dormant embryos of the brine shrimp, Artemia salina, contain small amounts of two previously undescribed dinucleotides which we have identified to be guanosine 5'-diphospho-5'-guanosine and guanosine 5'-triphospho-5'-adenosine. These compounds each comprise about 0.03% of the dry weight of the encysted embryos and are related chemically to guanosine 5'-triphospho-5'-guanosine and guanosine 5'-tetraphospho-5'-guanosine which have been shown previously to be major constituents of the nucleotide pool of Artemia cysts. These new dinucleotides were purified from perchloric acid extracts of dormant cysts by ion exchange column chromatography and identified by means of chemical, spectrophotometric, and enzymatic analyses compared to commercially available compounds. The possible role of these new compounds in nucleotide and nucleic acid metabolism in Artemia embryos is discussed.     

mRNA methylation and protein synthesis in extracts from embryos of brine shrimp, Artemia salina.

Cell-free protein-synthesizing extracts prepared from the brine shrimp, Artemia salina, translate methylated mRNAs. Reovirus unmethylated mRNA is inactive as a template when methylation is prevented by the inhibitor, S-adenosylhomocysteine. A salina mRNAs from both undeveloped and developed embryos contain 5'-terminal 7-methylguanosine in an inverted 5'-5' linkage through three phosphate groups to the rest of the polynucleotide chain. Removal of the 7-methylguanosine by beta elimination converts the mRNA from an active form to one inactive in protein synthesis in extracts of A. salina or wheat germ. Extracts of undeveloped and developed embryos methylate reovirus unmethylated mRNA at the 5' ends to form 5'-terminal structures of the type, m7G(5')ppp(5')G and m7G(5')ppp(5')Gm.     

The sequence of the 5S ribosomal RNA of the crustacean Artemia salina

The primary structure of the 5 S rRNA isolated from the cryptobiotic cysts of the brine shrimp Artemia salina is pACCAACGGCCAUACCACGUUGAAAGUACCCAGUCUCGUCAGAUCCUGGAAGUCACACAACGUCGGGCCCGGUCAGUACUUGGAUGGGUGACCGCCUGGGAACACCGGGUGCUGUUGGCAU _OH.     

Keeping Small Mammals in Schools A Note on Maintenance Costs

The brine shrimp, Artemia salina, is a small marine invertebrate. Although not generally used in schools, it has many advantages as an animal for experimental investigation. It is widely available, cheap, and easily cultured. A large number of experiments can be devised to investigate such factors as the importance of salinity on egg hatching success rale, and the effects of temperature and density-dependent competition for food on growth rates. Observations can also be made on development and reproductive behaviour. This paper includes a general account of the biology of Artemia, instructions on how the organism may be cultured, the results of some experiments on the ecology and population biology of the species, and some suggestions for further studies.     

Effects of toxic Alexandrium species on the survival and feeding rates of brine shrimp, Artemia salina

Zooplankton is an important link between phytoplankton and higher consumers in the marine food chain. To investigate the harmful effects of the toxic dinoflagellate Alexandrium species on zooplankton, 4 strains of Alexandrium spp., isolated from the Chinese coast, were used to test the species' effects on the survival and feeding rates of the brine shrimp, Artemia salina. The experiment was designed to assess the response of A. salina in each stage of its life cycle: nauplii, metanauplii, and adult. Each experiment was conducted in a 500 ml treatment that was added. The toxic treatments consisted of single strains of A. minutum, A. catanella, and A. tamarense (Nanhai and Donghai strain), while non-toxic species (dinoflagellate Prorocentrum donghaiense and diatom Chaetoceros minutissimus) were used as control treatments. An additional phytoplankton treatment consisted of, a mixture of A. tamarense (Nanhai strain) and P. donghaiense. Alexandrium spp. species were found to have lethal effects on the brine shrimp at a density of 2000 cells/ml. All the brine shrimps died within 24-168 hours of inoculation with the 4 treatments each containing single toxic Alexandrium species. During the feeding experiment, toxic Alexandrium spp. caused a reduction in the feeding rates in all the three stages of the life cycle of A. salina, whereas this response was not obvious in the treatment involving the nontoxic species P. donghaiense. The body surface of the brine shrimp that were fed on Alexandrium species was consistently covered by a sticky floc. Mortality of A. salina was observed to increase with the occurrence of the floc. The toxicity of the paralytic shellfish poisons (PSP) produced by the Alexandrium species was not significantly correlated with the survival or the feeding rate of the brine shrimp. When A. tamarense was mixed with P. donghaiense, the lethal effect of A. tamarense decreased, as shown by an increase in the survival and the feeding rates of the brine shrimp. A. salina metanauplii were found at the life stage most sensitive to the toxic algae and hunger. In summary, toxic Alexandrium spp. were found to have lethal effects on A. salina and to restrain the brine shrimp's feeding rate. Nontoxic Prorocentrum mitigated the toxicity of Alexandrium to a certain extent. The results also imply that the sticky material on the surface of the body of the brine shrimp may have been an important lethal factor rather than the PSP toxins.     

Effects of toxic Alexandrium species on the survival and feeding rates of brine shrimp, Artemia salina

Zooplankton is an important link between phytoplankton and higher consumers in the marine food chain. To investigate the harmful effects of the toxic dinoflagellate Alexandrium species on zooplankton, 4 strains of Alexandrium spp., isolated from the Chinese coast, were used to test the species' effects on the survival and feeding rates of the brine shrimp, Artemia salina. The experiment was designed to assess the response of A. salina in each stage of its life cycle: nauplii, metanauplii, and adult. Each experiment was conducted in a 500 ml treatment that was added. The toxic treatments consisted of single strains of A. minutum, A. catanella, and A. tamarense (Nanhai and Donghai strain), while non-toxic species (dinoflagellate Prorocentrum donghaiense and diatom Chaetoceros minutissimus) were used as control treatments. An additional phytoplankton treatment consisted of, a mixture of A. tamarense (Nanhai strain) and P. donghaiense. Alexandrium spp. species were found to have lethal effects on the brine shrimp at a density of 2000 cells/ml. All the brine shrimps died within 24-168 hours of inoculation with the 4 treatments each containing single toxic Alexandrium species. During the feeding experiment, toxic Alexandrium spp. caused a reduction in the feeding rates in all the three stages of the life cycle of A. salina, whereas this response was not obvious in the treatment involving the nontoxic species P. donghaiense. The body surface of the brine shrimp that were fed on Alexandrium species was consistently covered by a sticky floc. Mortality of A. salina was observed to increase with the occurrence of the floc. The toxicity of the paralytic shellfish poisons (PSP) produced by the Alexandrium species was not significantly correlated with the survival or the feeding rate of the brine shrimp. When A. tamarense was mixed with P. donghaiense, the lethal effect of A. tamarense decreased, as shown by an increase in the survival and the feeding rates of the brine shrimp. A. salina metanauplii were found at the life stage most sensitive to the toxic algae and hunger. In summary, toxic Alexandrium spp. were found to have lethal effects on A. salina and to restrain the brine shrimp's feeding rate. Nontoxic Prorocentrum mitigated the toxicity of Alexandrium to a certain extent. The results also imply that the sticky material on the surface of the body of the brine shrimp may have been an important lethal factor rather than the PSP toxins.     

Structure of extracellular hemoglobin from the brine shrimp Artemia salina.

1. Hemoglobin from the brine shrimp Artemia salina, purified by ultracentrifugation and preparative gel electrophoresis in non-denaturing medium, gave in sodium dodecyl sulfate-polyacrylamide gel electrophoresis a single band corresponding to a polypeptide chain with Mr 150,000. 2. Crosslinking by glutardialdehyde resulted in the appearance of a band corresponding to a molecular mass twice that of a polypeptide chain. 3. Limited trypsinolysis gave eight proteolytic bands corresponding to submultiples 8/9-1/9 of a polypeptide chain. 4. We conclude that a molecule of Artemia hemoglobin is composed of two single polypeptide chain subunits and that each subunit consists of nine structural units roughly equal in size.