Juvenile hormone III produced in male accessory glands of the longhorned beetle,Apriona germari,is transferred to female ovaries during copulation

We report on juvenile hormone (JH) biosynthesis in vitro by male accessory glands (MAGs) in the longhorned beetle, Aprionona germari, accompanied by the transfer of JH from males to females during copulation. JH was extracted from the MAGs and separated by reversed‐phase high‐performance liquid chromatography. JH III was identified as the major JH by gas chromatography–mass spectrometry. A radiochemical assay and a non‐radioactive method were used to measure the in vitro rate of JH biosynthesis by the MAGs. After 4 h of incubation with ³H‐methionine in the medium, the radioactivity in the MAGs substantially increased. In a separate assay, incubation of the MAGs with non‐radioactive methionine for 4 h resulted in a 39% increase in JH III. Seven‐day‐old males were injected with medium 199 containing ³H–methionine and 24 h later they were mated with virgin females. Hemolymph and the MAGs were collected from the mated males and hemolymph, ovaries and eggs were collected from the mated females for assaying radioactive JH. The radioactivity incorporated into JH in the MAGs was transferred to the females during copulation and later transferred into their eggs. Assayed 1 h after copulation, JH III level in the MAGs decreased 42% and the content of JH III in the male hemolymph did not change, whereas the content of JH III in the female hemolymph and ovaries both increased. © 2010 Wiley Periodicals, Inc.     

Growth of the male accessory gland in adult locusts: Roles of juvenile hormone,JH esterase,and JH binding proteins

The participation of juvenile hormone (JH) in the regulation of growth and protein synthesis in the accessory reproductive gland of male Locusta migratoria has been investigated. After elimination of endogenous JH with ethoxyprecocene, the accessory gland failed to grow, but growth was restored by a single application of the JH analog, pyriproxyfen. Pyriproxyfen appeared to stimulate total protein synthesis by 3 h, with a significant effect by 12 h, in contrast to 24 h observed in fat body. The dose curve for stimulation of protein synthesis 12 h after applying pyriproxyfen gave an ED₅₀ of 0.1 μg; the dose curve for gland growth at 72 h was biphasic, with steps at about 0.01 μg and 10 μg, suggesting two phases in JH action. SDS-PAGE analysis showed several components that were stimulated by pyriproxyfen, the effect being strongest in an 11 kDa band. A 5 kDa component was enhanced in the soluble and reduced in the particulate fraction after precocene treatment. The accessory gland contained JH esterase activity at levels about 100 times those in fat body or hemolymph, and was higher in precocene treated locusts. Binding activity for [³H]10R -JH III was high in cytosolic and nuclear fractions, and was identified immunologically as due to the previously described hemolymph JH binding protein. The results indicate that the mode of action of JH in the accessory gland may differ from that in the fat body. The presence of intracellular JH binding protein suggests a direct action of JH within the gland, that may be modulated by JH esterase. © 1995 Wiley-Liss, Inc.     

Protein production in components of the accessory gland complex of male Melanoplus sanguinipes (Insecta: Orthoptera)

Protein production during sexual maturation or after allatectomy (followed by juvenile hormone replacement therapy) has been examined in the long hyaline glands, short hyaline glands, white glands, and seminal vesicles, which make up the accessory gland complex of male Melanoplus sanguinipes. During maturation, the amount of protein in the long hyaline glands increases about 14-fold, and in each of the other components between 5- and 6-fold. Most protein accumulates between days 3 and 5, and this is reflected in high levels of incorporation of radiolabelled leucine in this period.

The components show differential sensitivity to the effects of allatectomy. After this operation, the protein content of, and incorporation of radiolabel into, the long hyaline glands remain near the day 0 level. In the white glands and short hyaline glands, allatectomy also has a marked, though less severe, effect on protein synthesis and accumulation. The seminal vesicles are least affected by allatectomy and continue to accumulate protein (though more slowly) to about 60% of the normal level by day 10. Juvenile hormone compounds applied topically to allatectomized insects on day 2 restored the ability of the gland components to accumulate proteins, though to differing degrees. JHI is the most effective compound, stimulating synthesis and accumulation of protein to near normal levels by day 10, whereas application of JHIII or Stauffer's synthetic JH led to only partial restoration of protein synthesis in the glands.     

棕尾别麻蝇雄性附腺分泌物的生理功能

棕尾别麻蝇Boettcherisca peregrina Robinean-Desvoidy雄性附腺的分泌物中包含许多生物活性分子,将成熟的雄性附腺分泌物注射到未交配的雌虫体内,发现其能影响雌虫生殖活性的很多方面:如降低雌虫的再次交配率;增加卵巢内的含卵量;缩短交配后雌虫的寿命。另外,还证明附腺分泌物中存在有可抑制革兰氏阳性菌的物质的存在。     

蜱雄性附腺分泌物及其功能的研究进展

交配是蜱类繁殖过程的关键环节,诱发雌蜱发生一系列的生理变化,并最终产卵。蜱雄性附腺分泌物在交配过程中发挥着重要作用,具有保护、活化精子,促进受精、卵巢发育和卵黄发生的功能,并对雌蜱的生殖生理行为等产生影响,如诱导雌蜱快速吸血和加速产卵。本文在简要分析蜱雄性附腺结构和分泌物生化特性基础上,系统阐述了蜱雄性附腺分泌物中各种功能因子的研究现状,着重论述其在精子获能、诱导雌蜱吸血、促进雌蜱卵巢发育和卵黄发生等方面的进展,并对未来研究提出了展望,以期为此领域的研究拓展思路。     

Juvenile hormone stimulation of sperm activator production in male monarch butterflies

A sperm activating substance is demonstrable in the male Monarch butterfly ejaculatory duct, but only during reproductively active stages. Experiments using neck-ligatures and allatectomies, with or without injections of juvenile hormone isomers, show that the post-eclosion production of sperm activator is stimulated by juvenile hormone.     

Nuclear binding sites for juvenile hormone III in the male accessory reproductive glands of Melanoplus sanguinipes

Summary

We have identified a potential nuclear juvenile hormone (JH) receptor in the long hyaline tubules (LHT), part of the male accessory reproductive gland (MARG) of M. sanguinipes. The MARG was incubated in vitro with [³H]JH III, and the distribution of the [³H]JH III among the cellular fractions of the LHT was determined. Some 37±4% of the radioactivity was associated with the crude nuclear pellet, while the cytosolic, microsomal and mitochondrial fractions contained 30±3%, 23±2% and 10±1%, respectively. The bound JH III was measured in nuclear extracts of LHT from males up to 15 days post-eclosion. These results revealed that JH binding increased in an age-dependent manner up to day 7, then levelled off to day 12, to increase again on day 14. The nuclear-binding component in the LHT had a very strong affinity for JH III, with a K_D value of 0.8 nM. Our observations are considered in relation to the potential site and mode of action of JH.     

The role of the male accessory gland fluid in stimulating vitellogenesis in Aedes taeniorhynchus

Injection of 20-hydroxyecdysone (20-OH-ecdysone) at high concentrations (5.0 μg) into intact or decapitated female Aedes taeniorhynchus induced vitellogenin synthesis, whereas low concentrations (5.0 ng) were ineffective. Injections of male accessory gland fluid (MAGF), however, at a concentration that was equivalent to 0.25 of the content of a pair of accessory glands, into intact or decapitated A. taeniorhynchus induced viteliogenin synthesis only in intact females. Ovariectomized mosquitoes did not synthesize vitellogenin after MAGF injection or blood feeding. Females that were first injected with MAGF and decapitated 12 h later synthesized viteliogenin at a rate that was 80% of intact controls. Egg development neurosecretory hormone (EDNH) activity in the heads of ovariectomized or intact females injected with MAGF was 9.0 pmol/min/head and 2.5 pmol/min/head, respectively, indicating that MAGF does not stimulate the corpus cardiacum (CC) to release EDNH. Incubation of MAGF and EDNH with fat bodies failed to induce vitellogenin synthesis. These results indicate that in A. taeniorhynchus the MAGF induces the ovary to release corpus cardiacum stimulating factor, which then signals CC to release stored EDNH.     

烟夜蛾雄蛾性附腺因子对雌蛾性信 息素合成的抑制作用

烟夜蛾Helicoverpa assulta处女蛾在交配后1 h,其性信息素滴度即显著降低,72 h内未见恢复。生测结果表明,烟夜蛾性信息素合成抑制因子主要来源于雄蛾性附腺。不同日龄雄蛾性附腺提取物的抑制活性无显著差异。光暗期对其活性具显著影响,暗期中雄蛾的性附腺物质对雌蛾性信息素合成具有较强抑制作用,而光期中雄蛾的性附腺物质不具抑制活性。在暗期的不同时间处理,对处女蛾性信息素合成的抑制作用无显著差异。雄蛾性附腺提取物对雌蛾性信息素合成的抑制作用与注射剂量有明显的相关性,0.2 ME（雄蛾当量）是产生显著抑制作用的最小剂量。对交配雌蛾注射性信息素生物合成激活神经肽（PBAN）提取物后,其性信息素合成又可恢复,这说明雌蛾交配后,性信息素滴度降低的原因是由于缺少了PBAN的调控。     

Ultrastructure and nature of secretory proteins in the male accessory gland of Drosophila funebris

The ultrastructural differentiation of the secretory cells and the nature of secretory proteins in the male accessory gland of Drosophila funebris have been studied by electron-microscopic and immunological methods. (1) In the pupae at $\text{1}\text{1}\text{2}$ days before eclosion, secretory products can be detected in the lumen, even though most glandular cells are at the initial phase of differentiation. At the time of eclosion both main and secondary cells are fully differentiated, but the whole set of five immunologically active proteins are detectable only on the second to third day of adult life. (2) The secondary cells contain giant protein granules, the so-called filamentous bodies, which become partially fused and the filaments assume a twisted form. Randomly dispersed filaments and closely packed filament bundles are also visible in the gland lumen. Antigenic labelling of ultrathin sections and immunoreplica electrophoresis yielded no evidence for the microtubular nature of these filaments. The secretion stored in the lumen contains in addition a large quantity of flocculent proteins which have their origin in the main cells. (3) During the period of high secretory activity in the 7-day-old male flies no vacuolization and disintegration of either the main or secondary cells have been observed. We conclude that both types of cells have the merocrine secretory mechanism. (4) Ultrastructural alterations in the glandular cells confirmed our previous observation that copulation stimulates RNA and protein synthesis.     

Male accessory gland substances and the control of sexual receptivity in female Culex tarsalis

ABSTRACT. Homogenates of accessory glands from male Culex tarsalis Coquillet (Diptera; Culicidae) were fractionated using gel filtration column chromatography. A component of low molecular weight capable of suppressing female sexual receptivity was obtained and partially characterized. Proteins present in homogenates of isolated accessory glands formed aggregates with other proteins in extracts of whole mosquitoes. Consequently, previous reports of the molecular weight of biologically active components in homogenates from accessory glands from other mosquito species may be inaccurate.
When female Cx. tarsalis were mated with ³H-leucine labelled males. radioactivity could be found throughout the body of the female 24 h after mating. Autoradiography showed that the radioactivity was incorporated into the cells of various organs, and was not specifically localized in a particular target tissue. Approximately 30% of the total radioactivity transferred to the female was found in the head and thoracic regions, and proteins from the head had a specific activity twice that of the thoracic proteins. Bioassay of isolated body parts from mated and virgin females showed that injections of head tissues from mated females into virgin females suppressed sexual receptivity. These results suggest that the head is involved in the physiological mechanisms resulting in refractory mating behaviour.     

Juvenile hormone regulation of male accessory gland activity in the red flour beetle, Tribolium castaneum

Male accessory gland proteins (Acps) act as key modulators of reproductive success in insects by influencing the female reproductive physiology and behavior. We used custom microarrays and identified 112 genes that were highly expressed in male accessory glands (MAG) in the red flour beetle, Tribolium castaneum. Out of these 112 identified genes, 59 of them contained sequences coding for signal peptide and cleavage site and the remaining 53 contained transmembrane domains. The expression of 14 of these genes in the MAG but not in other tissues of male or female was confirmed by quantitative real-time PCR. In virgin males, juvenile hormone (JH) levels increased from second day post adult emergence (PAE), remained high on third day PAE and declined on fourth day PAE. The ecdysteroid titers were high soon after adult emergence but declined to minimal levels from 1 to 5 days PAE. Feeding of juvenile hormone analog, hydroprene, but not the ecdysteroid analog, RH-2485, showed an increase in size of MAGs, as well as an increase in total RNA and protein content of MAG. Hydroprene treatment also increased the expression of Acp genes in the MAG. RNAi-mediated knock-down in the expression of JHAMT gene decreased the size of MAGs and expression of Acps. JH deficiency influenced male reproductive fitness as evidenced by a less vigor in mating behavior, poor sperm transfer, low egg and the progeny production by females mated with the JH deficient males. These data suggest a critical role for JH in the regulation of male reproduction especially through MAG secretions.     

Age-specific energetics of reproduction in Nucula turgida (Leckenby and Marshall) a bivalve with lecithotrophic larval development

Summary

Nucula turgida is a protobranch bivalve of the shallow sublittoral which produces, for a bivalve, unusually large eggs with a high lipid content to sustain the lecithotrophic development of the larvae. Males and females have similar reproductive investments, and the most noticeable feature of their output is the proportional increase in reproductive output as the animals age. Reproduction, as a proportion of the assimilation (Rep/C), was around 0.17 at population level but ranged from 0.055 in the 3 + year class to 0.54 in the 7 + year class. Similarly, C(=1 ? [(A ? G/R]) where A is assimilation, G gonad production and R is respiration, which appeared to be almost perfectly balanced at population level (C= ?0.05) in fact varied from very conservative (C= ?1.63) at 3 + to very “reckless” at 7+ (C=0.57). Using age-specific data, more accurate estimates can be made of future recruitment, and by back-calculation the survival rate from spawning to appearance in the benthos can be calculated.     

The accessory gland and metathoracic scent gland function in Oncopeltus fasciatus

The paper provides further clues as to the physiological function and biological significance of the ‘accessory gland’ in the metathoracic scent apparatus of the lygaeid Oncopeltus fasciatus. From various lines of evidence (gas chromatographic, cytochemical) it is concluded as probable that the accessory gland secretes small quantities of a mucopolysaccharide secretory product together with water. The difficulty of reconciling these data with other data indicating that the accessory gland is involved in the biosynthesis of the scent aldehydes is discussed. It is suggested that the water secreted into the median scent reservoir by the accessory gland provides O. fasciatus with a means of volumetric compensation for a diminished output of scent repellent.     

Molecular evolution and population genetics of duplicated accessory gland protein genes in Drosophila

To investigate the potential importance of gene duplication in D. melanogaster accessory gland protein (Acp) gene evolution we carried out a computational analysis comparing annotated D. melanogaster Acp genes to the entire D. melanogaster genome. We found that two known Acp genes are actually members of small multigene families. Polymorphism and divergence data from these duplicated genes suggest that in at least four cases, protein divergence between D. melanogaster and D. simulans is a result of directional selection. One putative Acp revealed by our computational analysis shows evidence of a recent selective sweep in a non-African population (but not in an African population). These data support the idea that selection on reproduction-related genes may drive divergence of populations within species, and strengthen the conclusion that Acps may often be under directional selection in Drosophila.     

Juvenile hormone-binding protein and juvenile hormone esterase activity in hemolymph from last-instar nymphs of Leucophaea maderae

The concentration of the juvenile hormone-binding protein (JHB) in hemolymph was determined throughout the last nymphal instar. It was found to be 3.9 μM at the molt to the instar, rising to 13 μM by mid-instar, and dropping to 6.7μM the day before emergence. Endocrine control of its production during the last nymphal instar could not be established. The apparent juvenile hormone esterase (JHF) activity was low at the molt to the last instar, but rose about fivefold by mid-instar, and then modestly declined. On the day of emergence, JHF activity rose to the highest level observed. A four- to fivefold increase in absolute JHF activity was determined during the first half of the last nymphal instar. This increase is not regulated by JH. Removal of the JHB from hemolymph samples by precipitation with a polyclonal specific antibody increased the JHF activity up to 1,000-fold. Thus, changes in the concentrations of JHB can affect the apparent activity of JHE, which is unrelated to the production or degradation of the JHF.     

优雅蝈螽雄性附腺的超微结构及其腺管提取物对精子束的作用

为阐明优雅蝈螽Gampsocleis gratiosa Brunner von Wattenwyl雄性附腺的结构与功能的关系, 本文利用透射电镜(transmission electron microscope, TEM)技术研究了优雅蝈螽雄性附腺的超微结构, 利用微分干涉相差显微镜(differential interference contrast microscope, DIC)技术并结合雄性附腺匀浆提取物与精子束在体外的短暂培养, 研究了优雅蝈螽雄性附腺对精子束的作用。结果表明： 优雅蝈螽雄性附腺3类腺管组织结构相似, 腺管管壁为单层上皮细胞, 缺少内表皮, 说明其来源于中胚层。上皮细胞富含粗面内质网、 线粒体、 高尔基体, 具有分泌细胞的特点。腺管管腔中分泌物有4种形态, 即电子透明的物质、 电子致密的颗粒物质、 细纤维状物质以及绒球状物质。上皮细胞的分泌方式主要有2种, 即顶质分泌和局部分泌。乳白短腺管的匀浆提取物参与了帽状精子束解聚的过程, 乳白长腺管和透明腺管的匀浆提取物有维持精子束活性的作用。本研究结果为进一步阐明螽斯雄性附腺的生理功能奠定了基础。     

Ecdysterone antagonism,mimicry, and synergism of juvenile hormone action on the Monarch butterfly reproductive tract

Previous research on Monarch butterflies has shown that juvenile hormone (JH) stimulates the development of the ovary and certain reproductive glands of both sexes. Ecdysterone injections into intact Monarchs demonstrate that low doses of this hormone inhibit ovarian development, and higher doses stimulate the male and female reproductive glands. In addition, experiments using neckligatured adults show that ecdysterone stimulates the reproductive glands of both sexes, in the apparent absence of JH, with the most pronounced effect being observed on the female colleterial gland. Other studies with neck-ligatured animals demonstrate that ecdysterone also synergizes with JH on the female gland and all three male glands. The feasibility of using Monarch reproductive glands for studies on the mode of action and interaction of JH and ecdysterone, and the possibility of a rôle of ecdysterone in the normal regulation of Monarch oögenesis, are discussed.