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1.
Summary

Concurrent morphological, anatomical and physiological changes took place during the first reproductive cycle in the Australian red-claw crayfish Cherax quadricarinatus, which prepared the female for spawning and holding of the newly deposited eggs. The endopod became longer and wider than the exopod and developed a mixture of plumose and long thin simple (ovigerous) setae. Small oocytes (0.24±0.05 mm) were present in the immature ovary. The growing ovary contained two distinct oocyte populations: one consisted of small (0.55±0.07 mm), barely growing oocytes, while the other consisted of large oocytes, which increased in size continuously (0.73 to 2.55 mm) until egg laying took place. A gradual change in the relative abundance of ovarian polypeptides occurred until the late vitellogenic stage (large oocytes < 1.8 mm). Three predominant female-specific, SDS-PAGE separated, polypeptides were observed (103, 78 and 73 kDa) that may represent vitellin subunits. The most abundant carotenoid in the ovary was astaxanthin, while β-carotene was present at a lower concentration. The strong correlation between the increasing diameter of the oocyte and the concentration of astaxanthin in the ovary and in the hemolymph suggested an association of astaxanthin with vitellin and vitellogenin.  相似文献   

2.
Estrogen levels in the gonads of marine bivalves, the Pacific oyster Crassostrea gigas and scallop Patinopecten yessoensis were determined by high performance liquid chromatography (HPLC) using an electrochemical detector. Estrone (E1), estradiol-17β (E2), and a small amount of estriol (E3) were identified in the ovary, while only E2 was found in the testis. The level of E2 in the ovary was consistently higher than E1 and it increased with sexual maturation. These results indicate that E2 may play a role in the reproductive events of the oyster and scallop. In vitro experiments demonstrated the presence of 17β-hydroxysteroid dehydrogenase (17β-HSD) in the ovaries of both bivalves. The activity of 17β-HSD in the ovary was lower in the postspawning stage than in the early differentiating stage. The evidence for the presence of aromatase activity in the scallop ovary was obtained by 3H-water assay. The immunoreactivity against 3β-hydroxysteroid dehydrogenase (3β-HSD), P450 aromatase and E2 was detected in the cells along the outside of germinal acini of the scallop ovary. It is concluded that estrogens can be synthesized in the gonad, that their levels vary with the reproductive cycle, and that they have a role in the development of gametes.  相似文献   

3.
SYNOPSIS. This paper describes factors which regulate the corporaallata (CA) during the reproductive cycle in the viviparouscockroach Diploptera punctata. Experiments, designed to alterthe rate of juvenile hormone (JH) synthesis by the CA, are performedon whole animals and their effects are demonstrated by an invitro radiochemical assay for JH synthesis by the CA. The CAis both inhibited and stimulated. Axonal pathways from the brainto the CA are required for inhibition of the CA in virgin femalesbut not in young pregnant females. Humorally transmitted inhibitionof the CA is caused in vivo by JH (in high titers) and ecdysteroidhormone. JH in low titers also stimulates the CA. Since thesehormones do not affect the CA in vitro we suggest that theyexert their effect via the central nervous system. The ovary,a possible source of ecdysteroids in the hemolymph, is necessaryboth for maintenance of JH synthesis and for inhibition in therate of JH synthesis at the end of egg growth.  相似文献   

4.
龟纹瓢虫卵黄蛋白的分子特性及发生动态   总被引:1,自引:1,他引:0  
李恺  张天澍  张丽莉  王斌  王群 《昆虫学报》2007,50(10):975-980
研究了龟纹瓢虫Propylea japonica (Thunberg) 卵黄蛋白的基本特性以及卵黄发生过程中卵黄蛋白的动态变化。PAGE和SDS-PAGE实验表明,龟纹瓢虫卵黄蛋白分子量为294.81±40.70 kD,并由分子量分别为144.68±0.03 kD和51.23±0.27 kD的两种亚基组成。对卵黄蛋白的氨基酸组成和含量分析发现,其必需氨基酸总量占57.48%,略高于非必需氨基酸,其中谷氨酸(Glu)含量最高,为15.26%;色氨酸(Trp)和蛋氨酸(Met)含量较低,分别为0.50%和0.11%。采用间接竞争ELISA法,系统测定了龟纹瓢虫成虫期脂肪体、血淋巴和卵巢中卵黄蛋白的动态变化,结果表明:脂肪体是卵黄原蛋白合成的场所,卵黄原蛋白的合成始于羽化后第2天;脂肪体、血淋巴和卵巢中卵黄原蛋白的滴度在羽化后第4天开始迅速上升,至成虫期的第8天左右达到高峰期。  相似文献   

5.
Summary

The parthenogenetic ovaries of the black bean aphid, Aphis fabae, contain developing embryos. When reared at 15°C in long days (LD 16:8) oocyte development begins within the ovaries of the largest embryos of a fourth instar mother 24–48 hr after her ecdysis from the third instar. Starvation, decapitation and precocene III treatment inhibit embryonic oocyte development; juvenile hormone treatment reverses this inhibition. A method for the in vitro culture of embryos is described and under these conditions juvenile hormone again stimulates oogenesis. Embryogénie growth in vivo, as measured by the increase in length of the oldest daughter embryos, is also stimulated by juvenile hormone treatment. The results are discussed in relation to other roles proposed for juvenile hormone in aphid development.  相似文献   

6.
Ion-exchange chromatography of crude ovarian extracts of the primitive insect Thermobia domestica allowed the separation, in native conditions, of major and minor vitellins of molecular weights of 300,000 and 430,000, respectively. Their polypeptide subunits were determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunotransfer using an antiserum prepared against major vitellin. This protein was resolved into large (Mr 166,000–212,000) and small (around Mr 50,000) polypeptides. Minor vitellin, on the other hand, exclusively contained small polypeptides that are immunologically different from those of the major vitellin. Vitellogenin polypeptides from the hemolymph of mature females exhibited electrophoretic mobilities and immunological properties similar to vitellin polypeptides. Pulse-chase experiments showed that the female fat body synthesizes radioactive and immunoprecipitable proteins, whose polypeptide pattern is close to that of the major vitellogenin. However, part of the primary vitellogenic polypeptides, at Mr 210,000 and 212,000, is rapidly processed to Mr 176,000 and 182,000 subunits. These two polypeptides, as well as the precursors, enter into the composition of the major hemolymph vitellogenin. Finally, processing of the still uncleaved 210,000–212,000 polypeptides takes place in the ovary, which performs the same step of vitellogenin maturation as the fat body.  相似文献   

7.
Abstract. The concentrations of fat body and haemolymph vitellogenin and ovarian vitellin during the first gonadotropic cycle of the cockroach Blattella germanica (L.) (Dictyoptera, Blattellidae) have been studied. For these purposes, a polyclonal antibody against B. germanica vitellogenin and vitellin has been obtained, and an ELISA to quantify these proteins has been developed. Ovarian vitellin levels follow a pattern which parallels those of basal oocyte growth and Juvenile Hormone production by the corpora allata. This suggests that Juvenile Hormone regulates vitellogenin uptake into oocytes. Fat body and haemolymph vitellogenin levels give cyclic and parallel patterns. However, the cycle of Juvenile Hormone appears delayed with respect to that of vitellogenin. We suggest that the production of Juvenile Hormone, although cyclic in profile, does not modulate alone the cycle of vitellogenin. At least a supplementary mechanism, apparently independent of Juvenile Hormone, may be involved in the decline of vitellogenin production at the end of the vitellogenic cycle.  相似文献   

8.
Gonadal development and plasma levels of sex steroids were investigated in female Arctic charr at 3-week intervals over a 12-month period. Circulating levels of oestradiol-17β (E2), testosterone (T) and 17,20β-dihydroxy-4-pregnen-3-one (17,20β-P) were measured by radioimmunoassay, and gonadal status assessed through histological examination and measurement of gonadosomatic index (GSI) and frequency distribution of oocyte size-classes. Gonadal recrudescence during March-July was characterized by modest but insignificant increases in plasma levels of E2 (2–4 ng ml?1) and T (2–5 ng ml?1) and recruitment of oocytes into yolk accumulation. Only a small and insignificant rise in GSI and no apparent increase in oocyte diameter occurred during this period, indicating that the rate of yolk formation and oocyte growth was low. Following transformation from stage V (peripheral yolk granule stage) to stage VI (yolk granule migration stage) in late July, the vitellogenic oocytes entered a phase of rapid growth which resulted in a marked rise in GSI until ovulation commenced in late September. Gonadal growth during this period was accompanied by increases in plasma levels of E2 and T which peaked at 11 ± 1 (mid-August) and 71 ± 5ng ml?1 (late September), respectively. The levels of both steroids dropped rapidly during final maturation and ovulation, followed by a surge in plasma levels of 17,20β-P which peaked at an average of 74 ± 17 ng ml?1 in early October. All three steroids returned to basal levels within a month after ovulation, and all steroids, except E2, remained low until March of the following year. A slight increase in E2 detected in February and March during the second season may have been associated with recruitment into vitellogenesis of a new generation of oocytes. It is suggested that the abrupt increase in vitellogenesis in late July may reflect a condition-dependent decision to proceed with maturation, once the energy reserves have been repleted during spring-early summer.  相似文献   

9.
Summary

Reproductive effort in terms of fecundity and energy allocation was studied in the iteroparous and long lived polychaete Marphysa sanguinea. Both measures show great variability. Fecundity varied from 8500 to 24300 oocytes; no linear relationship was found between oocyte number and jaw length whereas a direct relationship was established between oocyte number and wet body weight. The energy content of germinal and somatic tissues was determined by differential scanning calorimeter (DSC). The reproductive effort for a single reproductive event was calculated according to the formula: RE = EG/(EG + ES) where EGis the total energy of the germinal tissues and ES is the total energy of the somatic tissues. The lack of correlation between reproductive effort and size index strongly suggests that reproductive allocation does not increase with age. The reproductive effort ranged from 0.04 to 0.19 with a mean value of 0.120.  相似文献   

10.
Methyl (2E,6E)-10,11-epithio-3,7,11-trimethyl-2,6-dodecadienoate (the thiirane analog of JH III), 6,7-epithiogeranyl 4-methylphenyl ether and 6,7-epithiogeranyl 3,4-methylenedioxyphenyl ether were synthesized. An infrared absorption band at ~1090 cm?1 was attributable to the thiirane group. The biological activity of these three sulfur-containing JH mimics was tested on Culex pipiens, Aedes aegypti and Spodoptera litura to reveal weak or no JH-like activity.  相似文献   

11.
12.
Mouse oocyte development in vitro with various culture systems   总被引:7,自引:0,他引:7  
These experiments were designed to determine whether or not hormones are required for the growth of mouse oocytes and to assess the possible role of companion granulosa cells in oocyte growth. To approach these problems, four systems for the culture of oocytes, either alone or in association with granulosa cells, were utilized: (1) isolated oocyte culture, (2) isolated oocyte-ovarian cell coculture, (3) isolated follicle culture, and (4) ovarian organ culture. Oocytes from 8-day-old B6D2F1 mice failed to grow in isolated oocyte culture. Addition of follicle-stimulating hormone (FSH), 17β-estradiol (E2), or serum to the medium failed to prevent oocyte degeneration or to promote oocyte growth. On the other hand, oocytes in isolated follicle culture or in organ culture grew significantly in defined medium. The results showed that oocytes grown in isolated follicle culture under defined conditions and in the absence of gonadotropins resemble oocytes grown in vivo in terms of their ultrastructural characteristics, with the exception of enlarged mitochondria. In addition, these oocytes were shown to exhibit some normal functional characteristics in terms of their increased levels of CO2 evolution from exogenous pyruvate, and the ability of the fully grown oocytes to initiate meiotic maturation when freed from granulosa cells. It was concluded that gonadotropins are not necessary for oocyte growth and that gonadotropins are not required to potentiate the spontaneous meiotic maturation of oocytes which occurs after their isolation from granulosa cells. The results indicated that association of granulosa cells and oocytes was necessary for oocyte growth. However, isolated oocytes in coculture with ovarian cells failed to grow. Addition of FSH or E2 to the cocultures failed to promote oocyte growth or delay oocyte degeneration. It was concluded that, under the culture conditions used, granulosa cells must be in contact with the oocyte, perhaps by means of specialized cell junctions, for oocyte growth to occur.  相似文献   

13.
A polyclonal antibody specific to an egg protein of Suminoe oyster Crassostrea ariakensis was previously developed in our laboratory to assess the reproductive life cycle of the oyster. The present study was undertaken to investigate vitellin of C. ariakensis (CAVt). Vitellin is an essential component of egg proteins in marine invertebrates as it provides energy and nutrients to the embryo and larvae. CAVt was purified from eggs of the oyster using ammonium sulfate precipitation followed by affinity chromatography with Concanavalin A-agarose. Native polyacrylamide gel electrophoresis (PAGE) and sodium dodecyl sulfate PAGE showed that CAVt is a high molecular weight [532 kiloDaltons (kDa)] protein, with multiple subunits. Similar to other vitellin proteins, it is a phospholipoglycoprotein composed of phospholipids (12.06%), carbohydrates (mannose, 10.08% or glucose, 9.84%), and alkali-labile phosphates (4.16%). Affinity chromatography, enzyme-linked immunosorbent aasay (ELISA) and western blot analysis revealed that CAVt is only present in the ovary, and two subunits of CAVt (72 and 35 kDa) are believed to be incorporated from the hemolymph into the oocyte. The antibody specific to CAVt (anti-CAVt), raised in rabbit, strongly cross reacted with the egg proteins of oyster species and scallops, suggesting that the antigenic epitopes are highly conserved among species. Our results suggest that the anti-CAVt antibody can be used to develop a tool similar to ELISA or western blotting for investigation of the effect of microorganisms on reproduction as well as the effect of chemicals on the endocrine system in C. ariakensis.  相似文献   

14.
Vitellin, the major egg yolk protein, and vitellogenin, the hemolymph precursor of egg yolk protein, have been purified to apparent homogeneity from the mosquito Aedes aegypti. The purification procedure included chromatography on ion exchange, hydrophobic, and gel filtration columns. Vitellin and vitellogenin have a similar molecular weight (Mr 300,000) on gel filtration columns. However, the molecular weights of vitellin and vitellogenin, as determined from SDS electrophoresis, were 393,000 and 337,000, respectively. Vitellin in sodium dodecyl sulfate released six subunits of molecular weight 116,000, 83,000, 75,000, 54,000, 36,000, and 29,000, whereas vitellogenin released only three subunits (155,000, 120,000, and 62,000). The average molecular weights of vitellin and vitellogenin after gel filtration and SDS electrophoresis were 346,000 and 318,000, respectively. Vitellin has a high content of aspartic acid and glutamic acid, and a low content of histidine, methionine, cysteine, and tryptophan. Vitellin also contains 0.9% mol of glucosamine and no galactosamine. The isoelectric points of vitellin and vitellogenin are at pH 6.4 and 6.3, respectively. Aedes aegypti fat bodies incubated for short intervals in tissue culture medium in the presence of [3H]valine showed incorporation by radio-immunoprecipitation and SDS electrophoresis into three primary vitellogenin polypeptides of molecular weights (± SEM) 156,000 ± 4,000, 114,000 ± 5,000, and 62,000 ± 400 inside the fat body and 162,000 ± 3,000, 118,200 ± 2,000, and 63,000 ± 300 in the medium. These results suggest that the molecular weight of vitellogenin synthesized inside the fat body (Mr 332,000) remains unchanged when secreted into the hemolymph (Mr 343,000). The three vitellogenin subunits are processed by the ovary into six subunits which are then deposited in the yolk granules as vitellin.  相似文献   

15.
Vitellin from the cabbage butterfly Pieris rapae L. was purified and characterized by electrophoresis. Vitellin from P. rapae is a phosphorylated glycolipoprotein of 380,000 ± 10,000 molecular weight as determined by nondenaturing polyacrylamide gel electrophoresis. Two subunits with an Mr of 150,000 and 40,000 were obtained from vitellin. The native molecule is thought to be a tetramer composed of two molecules of each of these subunits. The isoelectric point, as determined by isoelectric focusing on polyacrylamide gels, is 6.10. Vitellin and vitellogenin were indistinguishable by immunological methods such as double diffusion and tandem-crossed immunoelectrophoresis. Vitellogenin from the hemolymph and vitellin from the ovary were quantified by rocket immunoelectrophoresis. Vitellogenin and vitellin were first detected in 6-day-old pupae, and their levels increased continuously during ovarian development. Vitellogenin synthesis by the fat body in 4-day-old female pupae could be induced by juvenile hormone I.  相似文献   

16.
Sex steroid hormones are important for reproduction in all vertebrates, but few studies examine inter-individual, temporal, and population-level variations, as well as environmental influences on circulating steroid levels within the same species. In this study we analyzed plasma 11-ketotoestosterone (11-KT) and 17β-estradiol (E2) levels in the oyster toadfish to test for 1) individual and temporal variations by serially sampling the same individuals during the reproductive and post-reproductive period, 2) variations in steroid levels among toadfish obtained from different sources or maintained under different holding conditions, and 3) correlations with environmental parameters. Results from serial sampling showed marked inter-individual variations in male 11-KT levels in two separate groups of toadfish, but no temporal differences from June to September. Females also showed inter-individual variations in E2 concentrations, but most had elevated levels late in the reproductive season coincident with oocyte growth prior to winter quiescence. E2 concentration, but not 11-KT, was positively correlated with water temperature, and negatively correlated with daylength and lunar phase. Maricultured toadfish held under constant conditions had elevated levels of E2 and 11-KT that should be considered when using these fish for experimentation. This study provides important comparative information on the relationship between individual variations in steroid levels, and how they relate to physiological and environmental correlates in a model marine teleost.  相似文献   

17.
The molecular properties and roles of luteinizing hormone (Lh) and its receptor (Lhcgrbb) have not been studied for the medaka (Oryzias latipes), which is an excellent animal model for ovulation studies. Here, we characterized the medaka Lh/Lhcgrbb system, with attention to its involvement in the ovulatory process of this teleost fish. In the medaka ovary, follicle-stimulating hormone receptor mRNA was expressed in small and medium-sized follicles, while lhcgrbb mRNA was expressed in the follicle layers of all growing follicles. Experiments using HEK 293T cells expressing medaka Lhcgrbb in vitro revealed that gonadotropin from pregnant mare’s serum and medaka recombinant Lh (rLh) bound to the fish Lhcgrbb. The fish gonadotropin subunits Gtha, Fshb, and Lhb were essentially expressed at fairly constant levels in the pituitary of the fish during a 24-h spawning cycle. Using medaka rLh, we developed a follicle culture system that allowed us to follow the whole process of oocyte maturation and ovulation in vitro. This follicle culture method enabled us to determine that the Lh surge for the preovulatory follicle occurred in vivo between 19 and 15 h before ovulation. The present study also showed that oocyte maturation and ovulation were delayed several hours in vitro compared with in vivo. Treatment of large follicles with medaka rLh in vitro significantly increased the expression of Mmp15, which was previously demonstrated to be crucial for ovulation in the fish. These findings demonstrate that Lh/Lhcgrbb is critically involved in the induction of oocyte maturation and ovulation.  相似文献   

18.
U. Kutschera  P. Schopfer 《Planta》1986,167(4):527-535
Plastic and elastic in-vitro extensibilities (E pland E el ) of cell walls from growing maize (Zea mays L.) coleoptile segments were measured by stretching frozen-thawed tissue, pre-extended to its in-vivo length, at constant force (creep test) in a custom-buildt extensiometer, equipped with a linear-displacement transducer. The indole-3-acetic acid (IAA)-induced change of E pl (E pl ) is strictly correlated with the growth rate for a period of 3–4 h. Subsequently, E plremains constant while the growth rate is slowing down. Since this discrepancy can be accounted for by a growth-dependent reduction of osmotic pressure, it is concluded that E plrepresents quantitatively the relative increase of in-vivo extensibility (cell wall loosening) involved in IAA-mediated cell growth over a much longer time. On the other side it is argued that the growth rate may not be strictly correlated with wall extensibility during long-term growth. Abscisic acid (ABA) inhibits segment growth induced by auxin, fusicoccin, or exogenous acid, and this effect can be quantitatively attributed to an ABA-mediated reduction of cell wall extensibility as determined by the E plmeasurement. Both, IAA and ABA have no effect on total protein synthesis, RNA synthesis, and amount of osmotic solutes. Fusicoccin-induced proton excretion is only slightly inhibited by ABA. In contrast to ABA, growth inhibition by cycloheximide (CHI) is always much larger than the concomitant reduction of E pl , indicating that a further growth parameter is also involved in the inhibition of cell growth by CHI. E el is not affected by either IAA, ABA, or CHI. It is concluded that E pl as determined by the applied method, represents a relative measure of the actual in-vivo extensibility of the growing cell wall at the very moment when the tissue is killed, rather than an average extensibility accumulated over some immediate-past period of time as suggested by Cleland (1984, Planta 160, 514–520). Hence, we further draw the conclusion that IAA and ABA control of cell growth can entirely be attributed to a modulation of cell wall extensibility by these hormones in maize coleoptiles.Abbreviations ABA ±abscisic acid - CHI cycloheximide - E el , Epl elastic and plastic in vitro extensibilities, respectively (E el+Epl=Etot>) - FC fusicoccin - IAA indole-3-acetic acid  相似文献   

19.
To understand the changes of serum levels of sex steroids in the wild Japanese eel Anguilla japonica during silvering process, eels collected from the Kaoping River of Taiwan from August 2000 through June 2001 were examined. The maturational stages of female eels before and during silvering were divided into four stages: juvenile, sub-adult, pre-silver and silver stages based on skin coloration and oocyte diameter. Male eels were investigated only in the silver stage. Radioimmunoassays were employed to measure serum levels of estradiol-17β (E2) and testosterone (T). The mean liver mass of the female eels increased significantly during silvering, but the mean hepatosomatic index remained constant. In contrast, mean ovarian mass and gonadosomatic index increased significantly during silvering. Serum concentrations of E2 in females increased significantly during silvering (P<0.05), while E2 was undetectable in silver males. The mean serum T concentrations increased significantly in females (P<0.05) during silvering, with lowest mean values in the juvenile stage and highest mean value in the silver stage. The mean serum T level in the silver males was significantly lower than in silver females (P<0.05). In conclusion, both serum E2 and T concentrations increased with ovarian development of wild Japanese eels during silvering, while serum E2 was undetectable in the silver male eels. The findings support the idea that androgen, but not estrogen, plays a major role in silvering process of the eels in both sexes.  相似文献   

20.
Elucidation of a profile of scallop vitellin formation associated with oogenesis and its endocrine control, and identification of a vitellogenin synthesizing site were immunologically undertaken by using anti-scallop Vn serum. Vn content increased during ovarian growth and accounted for more than 80% of the water soluble protein of the ovary at the mature stage. In vivo injection of estradiol-17 beta (E(2)) resulted in an increase in Vn content in the ovary. In vitro accumulation of Vn in the ovarian tissue was promoted with E2 and a vitellogenesis promoting factor (VPF) from cerebral plus pedal ganglion which was heat stable, less than MW 10,000 and trypsin/chymotrypsin resistant. Estrogen receptor (ER)-like immunoreactivity was found in the growing oocyte and the auxiliary cell in close contact with growing oocytes, in which Vn immunoreactivity was also found. It is suggested that the vitellogenin synthesis occurred inside the ovary, especially in the auxiliary cell, and is controlled by E2 and VPF via ER.  相似文献   

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