镜湖不同湖区沉积物中轮虫休眠卵萌发的比较研究

实验室内对镜湖沉积物中的轮虫休眠卵进行了萌发,共孵出轮虫47种,隶属于10科19属;其中大湖区沉积物中萌发出的轮虫40种,小湖区41种,两湖区共同种类34种.每毫升大湖区沉积物中休眠卵萌发出的轮虫平均数量为(1.1±0.1)个,显著高于小湖区的(0.4±0.0)个.在两湖区沉积物中,以3种孵化方式所萌发出的平均密度超过4.0ind./200mL的轮虫种类总计有6种,它们是多须伪前翼轮虫(Proalides tentaculates)、裂痕龟纹轮虫(Anuraeopsis fissa)、螺形龟甲轮虫(Keratella cochlearis)、裂足臂尾轮虫(Brachionus diversicornis)、臂三肢轮虫(Filinia brachiata)和萼花臂尾轮虫(Brachionus calyciflorus);其中前三种轮虫为两湖区所共有.共有的3种轮虫的孵出情况在两湖区沉积物间也存在着差异.在小湖区沉积物中,3种孵化方式下孵出的平均密度大于0.05ind./200mL的轮虫属是臂尾轮属、伪前翼轮属、龟甲轮属和龟纹轮属,而大湖区沉积物中孵出的平均密度较高的轮虫属还包括三肢轮属、巨头轮属(Cephalodella)和异尾轮属(Trichocerca);水体营养程度较高的大湖区沉积物中孵出的臂尾轮属、伪前翼轮属和龟甲轮属轮虫的密度均高于营养程度较低的小湖区.     

不同培养条件下萼花臂尾轮虫休眠卵的萌发

为确定萼花臂尾轮虫休眠卵的最适形成条件 ,或为提高特定培养条件下形成的休眠卵的萌发率 ,对不同温度、食物、光照和 pH条件下萼花臂尾轮虫形成的休眠卵进行了萌发研究。发现不同培养条件下形成的休眠卵在不同萌发条件下的萌发历时均为实验开始后的第 1～ 7天 ,萌发率高峰均出现在第 1～ 3天。在萌发温度为15℃～ 30℃范围内 ,2 0℃下形成的休眠卵在 2 0℃的萌发温度下累积萌发率最高 ,为 5 2 5 0± 6 89% ;母体以小球藻为食物时 ,其所产休眠卵的平均累积萌发率为 2 7 92 % ,显著大于以斜生栅藻或蛋白核小球藻和斜生栅藻所组成的混合藻为食物时 ;休眠卵形成和萌发时的光照条件对其累积萌发率无显著的影响 ;pH6 5和 7 5条件下形成的休眠卵的累积萌发率较高 ,分别为 32 5 0± 7 0 7%和 38 75± 15 5 3%。本研究和已报道的研究结果表明 :不同培养条件下形成的休眠卵在不同条件下萌发时均呈同步萌发型式 ;2 0℃和pH7 5分别是武汉东湖萼花臂尾轮虫休眠卵形成的最适温度和最适 pH条件。     

Asian medaka fishes offer new models for studying mechanisms of seawater adaptation

Japanese medaka (Oryzias latipes) is a freshwater (FW) teleost that is popular throughout the world for laboratory use. In this paper, we discuss the utility of Japanese medaka and related species for studying mechanisms of seawater (SW) adaptation. In addition to general advantages as an experimental animal such as their daily spawning activity, transparency of embryos, short generation time and established transgenic techniques, Japanese medaka have some adaptability to SW unlike the strictly stenohaline zebrafish (Danio rerio). Since other species in the genus Oryzias exhibit different degrees of adaptability to SW, comparative studies between Japanese medaka, where molecular-biological and genetic information is abundant, and other Oryzias species are expected to present varying approaches to solving the problems of SW adaptation. We introduce some examples of interspecies comparison for SW adaptabilities both in adult fish and in embryos. Oryzias species are good models for evolutionary, ecological and zoogeographical studies and a relationship between SW adaptability and geographic distribution has been suggested. Medaka fishes may thus deliver new insights into our understanding of how fish have expanded their distribution to a wide variety of osmotic environments.     

Consequences of sex-specific growth on sibling competition in black-headed gulls: a sexually-size dimorphic species with scramble competition

Biased mortality of the larger sex during the early developmental period has been reported for a number of size-dimorphic bird species. This can partly be explained by the fact that growing to larger size renders the larger sex more vulnerable to food shortage. However, since sibling rivalry is often size-dependent, chicks of the larger sex should have a competitive advantage. This raises the question as to why the larger sex does not always benefit from its size in sibling competition. We studied sibling competition in the black-headed gull (Larus ridibundus), a sexually-size dimorphic species with male-biased mortality. We manipulated the natural brood sex ratio and placed one male chick in direct competition with one female chick while concurrently controlling for differences in age, size and laying order. Male chicks outgrew their female siblings by 15% in asymptotic body mass and did not suffer from enhanced mortality. Female chicks tended to be more alert when the parents returned to the nest and were more persistent in gull-typical begging displays. Females were more likely to get the first food item, but they did not get more food, possibly due to a size-mediated dominance over the non-monopolizable regurgitated food. Thus, it is unlikely that sex differences in competitiveness significantly contribute to male-biased mortality in black-headed gulls. The previously reported male-biased mortality is more likely due to a disadvantage of a higher food demand and a higher sensitivity towards low egg quality, as has been shown in previous studies.     

Cloning of a quail homologue of hatching enzyme: its conserved function and additional function in egg envelope digestion

The aim of the present study was to reveal molecular entities participating in the digestion of the egg envelope in the Japanese quail, Coturnix japonica. We isolated a 1,510-bp cDNA from extraembryonic tissues of developing embryos and designated it quail hatching enzyme (QHE) cDNA. The QHE cDNA was found to code a protein molecule comprising an astacin protease domain in the N-terminal half and a complement subcomponents C1r/C1s, Uegf, Bmp1 (CUB) domain in the C-terminal half. A phylogenetic analysis showed that QHE belonged to the hatching enzyme group and was distinct from other proteases in the astacin family. Northern blotting and in situ hybridization demonstrated that expression of the QHE mRNA occurred twice during the development: first in ectodermal cells of the yolk sac on days 0–5, then in those of the albumen sac on days 8–13. Zymography revealed that proteolytic activity in extracts of days 3–4 and 9–12 embryos appeared at the position of 40 kDa. Immunoblotting tests showed that anti-QHE antiserum stained a 40-kDa molecule in extracts of day 3 area vitellina. Anti-QHE antibody stained the ectodermal cells of the area opaca on days 0–1, those of the area vitellina of the yolk sac on days 2–5, and those of the albumen sac on days 9–12. The temporal and spatial expression pattern of QHE mRNA was closely associated with digestion of the vitelline membrane occurring on days 1–4, and with that of the egg white on days 9–12.     

Exogenous administration of octanoic acid accelerates octanoylated ghrelin production in the proventriculus of neonatal chicks

Ghrelin is modified by fatty acid at the third serine residue. In this study, derivation of fatty acid for acylation of ghrelin was investigated using a hatchling chicken model. We first studied ghrelin gene expression and production in the neonatal chick proventriculus and then investigated the effect of exogenous octanoic acid (OA) administration on acylated ghrelin production. In a free-feeding condition on day 2.5 after hatching, the density of ghrelin mRNA-expressing (ghrelin-ex) cells was greater than that of ghrelin-immunopositive (ghrelin-ip) cells, but no difference was found between those densities in adult chickens. Intraperitoneal or oral administration of OA for a few days significantly increased the density of ghrelin-ip cells without any changes in ghrelin-ex cells and elevated only octanoylated ghrelin levels in the proventriculus. The results indicate that fatty acid absorbed from food is directly utilized in acylated ghrelin production in the chicken.     

非洲爪蟾孵化酶分子对卵黄膜作用方式的研究

非洲爪蟾的孵化液对卵黄膜和二甲基酷蛋白具有降妥活性。用非洲爪蟾孵化酶的特异性抗ＧＳＴ－ＵＶ．２抗体进行Ｗｅｓｔｅｒｎ杂交的结果表明,孵化液中出现一种分子量为６０ｋＤ的大组分,有时也会出现一种分子量为４０ｋＤ的小组分。     

Successful sperm cryopreservation of the brown-marbled grouper,Epinephelus fuscoguttatus using propylene glycol as cryoprotectant

This study developed the cryopreservation of brown-marbled grouper spermatozoa for practical application. We examined 32 cryodiluents, developed from four types of cryoprotectants [propylene glycol (PG), dimethyl-sulphoxide (Me₂SO), dimethyl-acetamide (DMA) and ethylene glycol (EG)] at four concentrations of 5, 10, 15 and 20% in combination with two extenders [Fetal bovine serum (FBS) and artificial seminal plasma (ASP). Cooling rates were examined by adjusting the height of straws (2.5–12.5 cm) from the liquid nitrogen (LN) vapor and cooled for 5 min before immersion into LN. DNA laddering was used to detect DNA damage in cryopreserved sperm. In fertilization trials, 0.5 g of eggs was mixed with cryopreserved sperm stored for 30 days in LN. The best motility of post-thaw sperm was achieved using 15% PG + 85% FBS (76.7 ± 8.8%); 10% PG + 90% FBS was also effective as cryodiluent. Generally, FBS gave better post-thaw motility compared to ASP. The optimum cooling rate was at 17.6 °C min⁻¹ obtained by freezing at the height of 7.5 cm surface of LN. The results obtained showed that cryopreserved sperm of brown-marbled grouper suffered slight DNA fragmentation, which resulted in significantly lower motility. However, the fertilization (90.9 ± 0.5%), hatching (64.5 ± 4.1%) and deformity rates (3.8 ± 0.2%) obtained from cryopreserved sperm showed no significant difference with fresh sperm. These findings show that the developed protocol for cryopreservation of brown-marbled grouper sperm was viable and will be useful for successful breeding and seed production of brown-marbled grouper.     

Elongated microvilli support the sea urchin embryo concentrically within the perivitelline space until hatching

Summary The early sea urchin embryo is supported in a concentric position within the perivitelline space by elongated microvilli which are attached to the fertilization envelope by extracellular matrix fibers. This “attachment complex,” of microvillus tip: extracellular matrix fibers: fertilization envelope, was revealed by two methods: the use of pronase or calcium-free sea water to dissolve the extracellular matrix fibers, thus causing the eggs to lose their concentric location, and the visualization of the “attachment complex” using video-enhanced differential interference contrast microscopy and transmission electron microscope images. The presence of the “attachment complex” helps in understanding two types of early developmental events: (1) the apparently continual change in microvillus length during cleavage stages which retains the embryos in their concentric position and (2) the hatching process.     

歪水蚤两种生理型卵的季节变化及其温度效应

实验室观测表明,3种歪水蚤的卵皆覆以半透明外鞘,仅特氏歪水蚤生产一定数量的不孵的无鞘卵。特氏歪水蚤夏卵与滞育卵的外鞘形态不同,前者较窄(≤15μm)且折皱均匀,后者多宽而折皱不均匀,随其外鞘加宽,活力明显提高。在钳形歪水蚤,两种卵的形态无明显差异;而右突歪水蚤尚未发现能够生产滞育卵。歪水蚤卵型的复杂多变,可能与其能够生产两种不同生理型卵的遗传特性正相适应。在短至12小时和6天的驯养期内,右突歪水蚤所产卵的卵径和鞘宽均分别与温度呈显著负相关及正相关,并随季节而异;这进一步表明动物对本海区温度季节性变化的适应。