Origin of the Regeneration Blastema in Polychaete Annelids

Since the early part of this century, controversy has surroundedthe origin of the cells ot the regeneration blastema in annelids.Cells of the epidermis and reserve mesodermal cells have eachbeen thought to form the entire blastema. With increasing frequency,it is being suggested that in numerous polychaetes each layerof tissue in the regenerate arises from the corresponding oldgerm layer. Evidence presented here from experiments utilizingtechniques of radioautography, histochemistry, X-irradiation,and electron microscopy in addition to histological observationlargely supports the last hypothesis. In the light of recentexperiments, it appears that the new epidermis and gut arisefrom the corresponding old tissue. The formation of the mesodermcannot yet be generalized, but reports indicate that it mayarise from dedifferentiated muscle cells, cells of the peritonealepithelium, and free coelomocytes.     

蜱类卵黄发生研究进展

蜱是一类特殊的节肢动物,有关其生殖生理的研究远远落后于昆虫。卵黄发生直接影响蜱类的繁殖力,一直是蜱类生理学一个十分活跃的研究领域。目前,对蜱类卵黄发生的研究主要包括卵黄蛋白的纯化与鉴定,卵黄原蛋白的合成与调控,卵母细胞对卵黄原蛋白的摄取以及转变为卵黄蛋白的分子过程。蜱类卵黄发生受激素调控,激素的作用与作用激素的种类在硬蜱和软蜱中表现出较大差别。对蜱类卵黄发生了解得尚少,在许多方面需深入研究。     

Vitellogenin concentrations in the haemolymph and ovaries of Ixodes scapularis ticks during vitellogenesis

The vitellogenin and vitellin concentrations in the haemolymph and ovaries of Ixodes scapularis females were determined using a double sandwich enzyme-linked immunosorbent assay. The level of vitellogenin in the haemolymph began to increase just prior to tick detachment from the host and continued to increase until 2 days after detachment. There was a slight decrease in the vitellogenin level 4 days after detachment, but a second peak was observed approximately 5 days after oviposition. Subsequent to oviposition, the vitellogenin levels in the haemolymph quickly decreased. The concentration of vitellogenin in the haemolymph ranged from 1.55 to 11.48 g l^-1 during the period after dropping from the host through oviposition. The concentration of vitellin in the ovaries began to increase as the female began rapid engorgement (0.03 mg per female) and declined after oviposition (0.1 mg per female).     

Development of monoclonal antibodies for monitoring Aedes atropalpus vitellogenesis

Monoclonal antibodies to a mixture of Aedes atropalpus and A. aegypti soluble yolk proteins were produced by hybridomas between the fusion of P3X63.653 myeloma cells and splenocytes of immunized BALB/c mice. Ascites fluid collected from mice innoculated with cloned hybridoma cells contained high specificity and affinity to the soluble yolk proteins of both Aedes species. Seven different hybridoma lines produced antibodies with specificity to both A. atropalpus and A. aegypti and one cell line produced antibodies monospecific to A. aegypti soluble yolk proteins. Monoclonal antibodies specific to A. atropalpus vitellin and vitellogenin were characterized by a combination of gel electrophoresis, western blotting and immunohistochemical staining. An indirect double antibody sandwich enzyme-linked immunosorbent assay was developed using a mixture of the seven hybridoma antibodies to A. atropalpus vitellin for monitoring vitellogenin levels in individual mosquito haemolymph samples. With this procedure, the peak period of vitellogenin synthesis in A. atropalpus was found to be 18 to 30 h after adult eclosion.     

福建漳江口红树林和盐沼湿地的多毛类动物群落

为了比较漳江口4种植物生境之间多毛类动物群落的差异性,2010年对漳江口潮间带秋茄、桐花树、白骨壤和互花米草4种植物生境的多毛类动物进行4个季度的定量取样.共获得15种多毛类动物,4个季度在4种植物生境中均出现三角洲双须虫、溪沙蚕、拟突齿沙蚕、凿贝才女虫、小头虫和加州中蚓虫.多毛类动物栖息密度、生物量、丰度指数、均匀度指数和多样性指数的季节变化不明显;但4种植物生境之间多毛类动物栖息密度、生物量、丰度指数、均匀度指数和多样性指数有显著差异,且互花米草生境与3种红树林生境之间多毛类动物优势种不同.Pearson相关分析表明,漳江口红树林和盐沼湿地除了多毛类物种数与泥温显著相关外,多毛类动物栖息密度、生物量、丰度指数、均匀度指数和多样性指数均与泥温、盐度、总有机碳、总氮无显著相关关系,其原因是漳江口4种植物生境多毛类动物常见种小头虫、加州中蚓虫和溪沙蚕均是广温、广盐及耐高有机质含量的种类.     

Effects of Copper on the Burrowing Behavior of Estuarine and Coastal Invertebrates,the Polychaete Nereis diversicolor and the Bivalve Scrobicularia plana

Alterations of the burrowing behavior of two benthic invertebrates living in intertidal mudflats, the polychaete Nereis diversicolor and the bivalve Scrobicularia plana, were studied in individuals exposed to soluble copper. The design of the contamination procedure took into account the results assessing the influence of some natural factors potentially able to influence burrowing (artificial vs. natural sediment, lighting, size of the shell for bivalves). Animals were exposed for 4 d to concentrations ranging from 25 to 150 μ g Cu l^{? 1}. At the end of exposure, the burrowing kinetics in clean sediment were determined after 1 and 2 d, then the animals were frozen until acetylcholinesterase (AChE) activity determination. Even at the lowest tested concentrations, copper caused hypoactivity in organisms belonging to both species studied. Behavioral impairments were not related to AChE activity inhibition. Metabolical or physiological disturbances could be the cause of these impairments. Concentrations affecting burrowing behavior were below those responsible for lethality in these species.     

成虫饲料中糖与奶粉不同配比对家蝇繁殖力及卵黄蛋白发生的影响

本文研究了成蝇饲料中糖与奶粉不同配比对家蝇繁殖力及卵黄蛋白发生的影响,旨在为家蝇Musca domestica大规模高效养殖提供技术支持。结果表明,随着成蝇饲料中奶粉比例提高,单雌产卵量逐渐提高,且差异显著,但当饲料中奶粉比例达到和超过80%以后,单雌产卵量又显著减少,其中以60%奶粉+40%白糖饲养雌蝇产卵量最高(1935.83粒/雌),用100%白糖饲养产卵量最低(328.17粒/雌)。当成蝇饲料中白糖的添加比例为20%~60%时,雌虫寿命和产卵期差异均不显著,但用100%奶粉或80%以上的白糖饲养,雌虫寿命和产卵期显著缩短;雄虫寿命则随饲料中白糖比例的增加而延长;奶粉与白糖配比的变化对雌蝇产卵前期无显著影响。卵孵化率随饲料中白糖比例的增加而下降,尤其当饲料中白糖比例超过60%以后,卵孵化率下降更为明显。进一步测定卵黄蛋白的结果表明,雌蝇取食含糖量越高的饲料,其每日合成的卵黄蛋白及羽化后15 d内合成的总卵黄蛋白也越少,尤其是雌蝇完全摄取白糖,其每日合成的卵黄蛋白均维持在一个较低的水平,而蛋白质补充过多(如奶粉在饲料中的比例超过60%),雌蝇每日合成的卵黄蛋白及羽化后15 d内合成的总卵黄蛋白,基本不随饲料中奶粉的增加而增加。综合以上结果,作者认为用60%奶粉+40%白糖作为家蝇成虫的营养补充料,其不仅能保持雌蝇良好的繁殖力,同时也能有效降低成蝇养殖成本。     

Polychaete zonation and its relation to algal distribution down a vertical cliff in the western Mediterranean (Italy): a structural analysis

The zonation of polychaetes down a vertical cliff (0–5 m in depth) at Cape Romito, Leghorn, Italy, was investigated. The structural organization of the community changed with depth. A community typical of photophilic environments was found in the superficial zone (0–2 m), while in the deeper zone (3–5 m), a more diversified community characterized by species typical of sciaphilic environments was present. At the surface (0 m) the community was very poor in species and individuals and thus probably corresponds to the midlittoral fringe. In fact, some species typical of the midlittoral zone were found. This distribution pattern was constant for samples taken in February and August 1985. The distribution of polychaetes was closely related to that found for macroalgae. It is hypothesized that algae condition the substratum and that this is the main factor responsible for the zonation of polychaetes.     

Temporal events of gypsy moth vitellogenesis and ovarian development

Abstract The vitellogenic period of gypsy moth ovarian development starts on day 3 of the pupal stage and continues through adulthood. During this period, rapid increases occur in follicle size, protein content, and wet weight of the ovary. Patency is observed on day 3 of the pupal stage.
Pre-vitellogenic follicles are formed in the last larval stadium. Newly formed follicles detach from the germarium on day 4, and increase rapidly to 140 per ovariole at the end of the last larval stadium. The pre-vitellogenic follicles are uniformly around 50 um in diameter. No vitellogenin is incorporated into the oocytes until the pupal stage.
Polyacrylamide gel electrophosesis (PAGE) in the presence of sodium dodecylsulphate (SDS) analysis of male and female haemolymph samples and vitellogenic ovaries demonstrates the presence of two female-specific subunits of vitellogenin of 180 kD and 160 kD. These proteins are detected only in haemolymph and ovarian extracts of vitellogenic females. The molecular weight of the native protein determined by size exclusion chromatography is approximately 400–420 kD.
A highly sensitive double antibody sandwich enzyme-linked immunosorbent assay (ELISA) was developed to monitor the temporal changes in vitellogenin titre in haemolymph. Vitellogenin production starts on day 2 of the last larval stadium, reaching a maximum level by day 6 of the last larval stadium, and decreasing in the late pupal stage as vitellogenin was internalized into the oocytes. This is the first report of vitellogenin production occurring in the larval stage of a holometabolous insect. The fact that vitellogenin production and uptake occur during different stages of development in the gypsy moth, opens up some interesting questions concerning the underlying regulatory mechanisms controlling each process.     

Ovarian development and vitellogenesis in the sawfly,Athalia rosae ruficornis Jakovlev (Hymenoptera,Tenthredinidae)

Summary

Ovarian development in Athalia rosae ruficornis Jakovlev (Hymenoptera: Tenthredinidae) is described. Number of nurse cells per egg chamber is most often around 60 (close to 63 according to the 2ⁿ–1 rule), but in many cases it deviates from this number significantly. Two major yolk proteins [vitellins: large (apparent molecular weight 160–170 kD) and small (48–50 kD] were identified by SDS-PAGE. Western blotting and immunochemical detection using polyclonal antibodies prepared against each of the vitellins revealed that adult female but not male (both haploid and diploid) hemolymph contains vitellogenins corresponding to these vitellins. Vitellogenins become detectable in the hemolymph of late pupae, and vitellins one day later in the oocytes of adults. Transplantation of immature ovaries into the adult male abdomen caused not only significant accumulation of vitellins in the oocyte but also appearance of small amounts of hemolymph vitellogenins in host males. Injection of homogenate of immature ovaries also caused appearance of small amounts of hemolymph vitellogenins in host males.     

Effects of starvation on the vitellogenesis,ovarian development and fecundity in the ectoparasitoid,Nasonia vitripennis （Hymenoptera： Pteromalidae）

A female-specific protein, vitellogenin （Vg）, and its corresponding egg vitellin （Vt） are identified in the ectoparasitic wasp Nasonia vitripennis. Both native Vt and Vg have a molecular mass of about 350 kDa, which is composed of two subunits of approximately 190 kDa and 165 kDa under reducing and denaturing conditions （sodium dodecyl sulfate-polyacrylamide gel electrophoresis）. An indirect sandwich enzyme-linked immunosorbent assay developed with both monoclonal and polyclonal antibodies against N. vitripennis Vt. Vg was first detected in the hemolymph on the 10th day after parasitism, and was first observed in oocytes on the 12th day. In adults deprived of food, the highest hemolymph Vg level occurred at the time of adult eclosion and the highest level of Vt in ovaries was found at 30 h after eclosion. In contrast, feeding adults with 20% sucrose resulted in the reduction of Vt uptake by ovaries and the extension of life span, but had little effect on Vg production. Deprived of hosts, starvation of female wasps had no significant effects on ovariole growth and oocyte maturation before the wasps died. However, starvation of female wasps supplied with hosts accelerated the wasps laying progeny into hosts, but resulted in a decrease of total progeny production by comparison with wasps fed with 20% sucrose.     

Effects of starvation on the vitellogenesis,ovarian development and fecundity in the ectoparasitoid,Nasonia vitripennis (Hymenoptera: Pteromalidae)

A female‐specific protein, vitellogenin (Vg), and its corresponding egg vitellin (Vt) are identified in the ectoparasitic wasp Nasonia vitripennis. Both native Vt and Vg have a molecular mass of about 350 kDa, which is composed of two subunits of approximately 190 kDa and 165 kDa under reducing and denaturing conditions (sodium dodecyl sulfate—polyacrylamide gel electrophoresis). An indirect sandwich enzyme‐linked immunosorbent assay developed with both monoclonal and polyclonal antibodies against N. vitripennis Vt. Vg was first detected in the hemolymph on the 10th day after parasitism, and was first observed in oocytes on the 12th day. In adults deprived of food, the highest hemolymph Vg level occurred at the time of adult eclosion and the highest level of Vt in ovaries was found at 30 h after eclosion. In contrast, feeding adults with 20% sucrose resulted in the reduction of Vt uptake by ovaries and the extension of life span, but had little effect on Vg production. Deprived of hosts, starvation of female wasps had no significant effects on ovariole growth and oocyte maturation before the wasps died. However, starvation of female wasps supplied with hosts accelerated the wasps laying progeny into hosts, but resulted in a decrease of total progeny production by comparison with wasps fed with 20% sucrose.     

Phase polymorphism in Schistocerca gregaria: Assessment of juvenile hormone synthesis in relation to vitellogenesis

Juvenile hormone (JH) synthesis by the corpora allata of gregarious and solitarious phase females of Schistocerca gregaria was determined in vitro during the penultimate and last stadia as well as during the first gonotrophic period of adults. Generally, the corpora allata of solitarious females showed higher rates of JH synthetic activity. In addition, in adult females there was a temporal difference between the corpora allata activities of gregarious and solitarious locusts, the latter exhibiting relatively higher rates of JH synthesis early in the first gonotrophic period. The corpus allatum volumes of solitarious females were also generally larger than those of their gregarious counterparts; there was no synchrony between fluctuations in JH synthetic activity and changes in corpus allatum volume in either phase.The early onset of relatively high JH synthetic rates in solitarious females was correlated with the early detection, by rocket immunoelectrophoresis, of vitellogenin in the haemolymph and vitellin in the oöcytes. Vitellogenin appeared in the haemolymph on day 4 in solitarious females and on day 6 in gregarious females and vitellin appeared in the oöcytes on days 6 and 8 respectively. Oöcyte length at which vitellogenesis was first detected was 1.8 mm for gregarious and 1.3 mm for solitarious females. However, despite the accelerated onset of both vitellogenin synthesis and uptake, oöcyte maturation time of solitarious females was longer. In both gregarious and solitarious females, vitellogenin titres increased until oöcytes reached a length of about 4 mm and declined thereafter. Vitellin content of ovaries increased proportionately to oöcyte growth until they attained a length of 5.0 mm. The subsequent increase in length of oöcytes to maturity is attributed to postvitellogenic growth, possibly by hydration.     

Evolutionarily conserved aspects of animal nutrient uptake and transport in sea anemone vitellogenesis

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温度对家蝇广州种群成虫繁殖力及卵黄蛋白发生的影响

研究了不同温度对家蝇(Musca domestica L.)广州种群繁殖力及卵黄蛋白发生的影响,旨在为利用家蝇广州种群大规模高效养殖提供技术支持。选同一天刚羽化的健康雌、雄蝇,分别置于20、24、28、32和36℃环境下单对饲养,重复6次。每天测定成虫日产卵量、卵孵化率及死亡数等,统计家蝇单雌产卵量、卵孵化率、成虫寿命、产卵前期和产卵期;同时自雌蝇羽化第一日起,连续15 d,每天每个温度处理下取雌蝇3头,采用间接酶联免疫(indirect enzyme-linked immunosorbentassay,ID-ELISA)法测定5种温度下雌蝇每日卵黄蛋白合成量及羽化后前15 d卵黄蛋白合成总量。结果表明,温度对家蝇成虫繁殖力影响显著。产卵量以24℃环境下最高(1 921.17粒/雌),24～28℃时产卵量显著下降。28℃时卵孵化率最高(93.48%),高或低于此温度,卵孵化率显著下降,当温度达36℃时,孵化率仅为60.78%。雌蝇在20～28℃范围内产卵期差异不显著,32℃及以上温度下,雌蝇产卵期显著缩短。产卵前期以20℃最长(9.83 d),28℃最短(3.00 d),高于28℃产卵前期又显著延长。雌、雄蝇寿命均随着温度升高而显著缩短。温度对广州种群雌蝇羽化后前15 d的卵黄蛋白合成影响显著,24℃下前15 d合成的总卵黄蛋白最高(1 683.15μg/雌),其次为28℃(1 591.95μg/雌),20℃最低(232.81μg/雌)。家蝇广州种群在24～28℃繁殖效率最高。     

Native vitellins are modified during ovarian development in the stick insect Carausius morosus (Br.)

Vitellins from ovarian follicles and newly laid eggs of the stick insect Carausius morosus were examined by ion exchange chromatography on a HPLC Mono Q column. Under these conditions, vitellins from newly laid eggs resolved as two distinct peaks, referred to as VtA and VtB, that eluted at 8.5 and 12.0 min, respectively. On native gels, both VtA and VtB separated into two different variant forms (VtA′ and VtA′, VtB′ and VtB′). By two-dimensional gel electrophoresis, VtA′ and VtA′ were shown to contain polypeptides A₁, A₂ and A₃. On the other hand, VtB′ and VtB′ appeared to comprise polypeptides B₁ and B₂ and B₁, A₁, A₂, B₂ and A₃*, respectively. A similar Vt polypeptide composition was also observed by size-exclusion chromatography of vitellins from newly laid eggs. Vitellins from early vitellogenic ovarian follicles resolved into a single chromatographic peak at 7.5 min that coeluted with a major peak from the hemolymph of egg-laying females. Ovarian follicles progressively more advanced in development exhibited a more complex chromatographic profile, consisting of three separate peaks. By two-dimensional gel immunoelectrophoresis, vitellins from ovarian follicles appeared to consist of two closely related, immunologically cross-reacting antigens that gradually shifted apart as ovarian development proceeded to completion. By size-exclusion chromatography, each Vt from ovarian follicles was shown to consist of a unique set of polypeptides different from those listed above. Single ovarian follicles were fractionated into yolk granules and yolk fluid ooplasm and tested by immunoblotting against Mab 12. Under these conditions, VtA variant forms in yolk granules and yolk fluid ooplasm reacted differently. Sections from ovarian follicles in different developmental stages were exposed to Mab 12 and stained with a peroxidase-conjugated, goat anti-mouse antibody. Regardless of the developmental stage attained, staining for peroxidase was restricted to free yolk granules, suggesting that native vitellins in stick insects are structurally modified upon fusion into the yolk fluid ooplasm. Arch. Insect Biochem. Physiol. 36:335–348, 1997. © 1997 Wiley-Liss, Inc.     

Carcinogenesis: Evolution of concepts

Cancer is considered as an unintended consequence of internal imperfection of multicellular organisms: Darwinian evolution “does not foresee the future and does not plan for it”, it is forced to handle only anything that it has at a given moment “at hand”, which makes inevitable compromises and restrictions. In this case, there are a number of founding dogmas including mutagenesis as the main driving force of carcinogenesis; the environment as the main source of mutagenic effects; tumor monoclonality; cancer cell multistage transformation as Darwinian process of successive mutation—selection cycles. Recent discoveries complicate, supplement, and sometimes transform into an opposite fixed concepts. As a result, a new “image” of carcinogenesis is formed as a biological phenomenon whose conservation is indicative of its evolutionary utility.     

Cypermethrin induction of vitellogenesis and ovarian development in unfed adult female Ornithodoros moubata (Acari: Argasidae)

Summary

Vitellogenesis in ticks is known to be induced by engorgement and mating. In this paper, the synthetic pyrethroid cypermethrin (CyM) is shown to induce production of yolk protein precursor, vitellogenin (Vg), and ovarian development in unengorged mated adult female Ornithodoros moubata. The levels of Vg found in the hemolymph and ovarian development induced by CyM were dose-dependent. i.e., CyM doses of more than 0.2 and 1.0 μg/tick were needed for significant increase of Vg titer in the hemolymph and yolk deposition in oocytes, respectively. Immunological and electrophoretical analyses of Vg and Vitellin (Vn) induced by CyM were identical with those induced by engorgement. Vg titer induced by CyM in unengorged females followed approximately the same time course as that in the normal engorged females. However, Vg titer induced by CyM continued to increase after day 8 and reached a maximum (95 μg/μ1) on day 10 after treatment, while Vg titer induced by engorgement decreased again after reaching a maximum (60 μg/μ1) on day 6, correlated with yolk Vn deposition in oocytes. Ovarian development induced by even high doses (10 or 20 μg/tick) of CyM was slow compared to normal development stimulated by engorgement. Oviposition was not observed in females treated with CyM.