家蚕性附腺发育与核酸及利它素蛋白变化的关系

对不同发育时期家蚕Bombyx mori雌性性附腺核酸和蛋白质含量测定的结果表明, 从家蚕化蛹后第6天到成虫羽化当天的性附腺内总蛋白质含量不断增加,至羽化当天为最高,达860±70 μg/对。不同时期内能引诱野蚕黑卵蜂识别寄主的利它素在总蛋白中所占的比例差异明显,从化蛹后第6天的10%增加到羽化后的58%。性附腺的总RNA含量从化蛹第6天到成虫羽化前1天几乎是直线增加,但羽化后下降很快。不同时期分泌部中总RNA含量变化与贮存部明显不同。含量最高时,分泌部RNA可占整个性附腺RNA的90%以上,而贮存部总RNA含量则远少于分泌部,羽化当天约为分泌部的十分之一。分泌部总RNA中的18S亚基含量远高于28S亚基,明显不同于贮存部的。从分泌部总RNA中分离的mRNA存在明显的条带分布,这预示着高丰度的mRNA的存在与总蛋白中高含量利它素的存在有着特殊对应关系。     

Isolation,partial characterization,and localization of the A and B proteins from the tubular accessory gland of male Tenebrio molitor

In its fully differentiated state, the tubular accessory gland of the male mealworm beetle, Tenebrio molitor, synthesizes five groups of proteins (A, B, C, D₁ and D₂) which are easily distinguished from one another on two-dimensional pI-SDS polyacrylamide gels (Black et al., 1982: Devl Biol. 94, 106–115). In the present work, the A and B proteins have been isolated by preparative gel electrophoresis and Amicon ultrafiltration. The isolation procedure provided two fractions of interest: one contained a mixture of A and B proteins (A/B) and the other consisted of only B proteins. The major proteins in the A class have a molecular weight of 17,900 while those of the B class are 19,000 daltons in size.Antibodies have been produced to the A/B mixture and to the B fraction. Ouchterlony immunodiffusion and straight line immunoelectrophoresis show that the A and B proteins share common immunological characteistics. The proteins from the tubular accessory gland were displayed on one dimensional SDS gels and electrophoretically blotted onto nitrocellulose paper. The antibodies to the A/B mixture recognize A and B bands on these gels. In addition, these antibodies show affinity for C proteins and another band of lower molecular weight.Using the anti-A/B with techniques of immunodiffusion, straight line immunoelectrophoresis, and immunoblotting, we have identified the A and B protein in extracts of soluble proteins in the spermatophore. Furthermore, the A/B proteins have been localized by immunohistochemical techniques within the apical portions of the secretory cells of the tubular gland and also in the lumen of the spermatophore.     

Development of the accessory reproductive glands and genital ducts in female Schistocerca gregaria

The accessory reproductive glands of female S. gregaria are tubular extensions of the paired genital ducts, which in the mature female contain large amounts of a proteinaceous secretion used in the formation of the egg pod. In the 4th and 5th-instar female the glands are indistinguishable from the remainder of the mesodermally derived genital ducts. Towards the end of the 5th stadium, however, the accessory gland region only acquires characteristic convolutions which persist throughout the adult stages. At this time the epithelium of the entire ducts becomes reorganized into a unicellular epithelium. Only one cell type occurs throughout the length of the glands, and also in the egg calyces and lateral oviducts. The cells are inactive immediately after final ecdysis and remain in this state until the level of juvenile hormone in the haemolymph rises. The hormone acts directly on the cells triggering a rapid proliferation of organelles associated with protein secretion, and thereby increasing the volume of apical cytoplasm. Microvilli develop at the luminar plasma membrane, while irregular infoldings form at the base of the cells. As the gland matures the major organelle, the rough endoplasmic reticulum, changes from the lamellar to the vesicular form. Secretion is released into the lumen by the ‘microapocrine’ method.     

Development of functionally competent cabbage looper moth sex pheromone glands

Unlike some moths, pheromone production in Trichoplusia ni is not regulated by a pheromone activating neuropeptide. Rather, competency to produce pheromone apparently is linked with changes in the ecdysteroid titer that occur late in metamorphosis. In contrast to adult pheromone glands, glands from pharate adults 2 days before eclosion were non-competent, and (1) had undetectable levels of the pheromone, (Z)-7-dodecenyl acetate, and pheromone-specific intermediates, (2) showed little or no conversion of radiolabeled substrate to product in enzyme assays of fatty acid synthetase, Δ11 desaturase, and acetyltransferase, and (3) failed to incorporate radiolabeled acetate into pheromone in gland culture. Glands 1 day before adult eclosion exhibited low titers of pheromone and the intermediate, (Z)-11-hexadecenoate, and showed low levels of radiolabeled acetate incorporation into pheromone in gland culture. By the time of adult eclosion, the gland was fully competent. Precocious development of pheromone gland competency was induced by removing the head and thorax from pupae 2 days before adult eclosion. This effect appears to result from the reduction of ecdysteroid, since it was blocked by the administration of 20-hydroxyecdysone. This ability to manipulate the development of the pheromone gland was restricted to a critical period, since removal of head and thorax from younger pupae did not induce pheromone gland competency, and administration of 20-hydroxyecdysone to older pupae did not block its onset. In addition to differences in competency, early pharate and adult glands exhibited dissimilarities with respect to (1) the types of proteins synthesized in gland culture, and (2) the types of proteins translated from mRNA in vitro.     

Histology and histochemistry of the accessory reproductive glands in the male hairy-nosed wombat (Lasiorhinus latifrons)

Summary The accessory male reproductive glands of the hairy-nosed wombat, Lasiorhinus latifrons, are a prostate and three pairs of Cowper's glands. Component units of all are branched tubular structures of varying epithelial makeup and secretory content. The prostate has the carrotlike shape and three consecutive regions commonly found in marsupials. The regions differ in their tubular histology and histochemistry: all contain secretory globules in glandular lumina. Cowper's glands A and B are histologically identical except for the absence of interstitial mast cells from gland B: gland C is characterized by narrower tubules and larger epithelial cells. Histochemical tests for protein, carbohydrate and iron indicate that glycogen is a major secretory product of the prostate (largely posterior region), iron is also secreted (mainly posterior region) and a small quantity of acid mucin is produced (mainly central region). Glycogen is a feature also of anterior prostatic glandular epithelium and of the capping cells of the urethral transitional epithelium. Cowper's gland A has considerable protein in its secretion, gland B a neutral glycoprotein and gland C a sialomucin: the latter two also exhibit cytoplasmic glycogen in their secretory cells.     

Trehalase from the bean-shaped accessory glands and the spermatophore of the male mealworm beetle,Tenebrio molitor

Summary InTenebrio molitor, male adults transfer sperm to the female via a spermatophore or sperm sac. The spermatophore is formed from secretions of the bean-shaped accessory glands (BAGs) and the tubular accessory glands (TAGs) of the male beetle. Trehalase is found in the adult BAGs. During the pupal stage, the activity in the BAGs was very low. After adult ecdysis, the total activity increased 100-fold from 0 days to 6 days and reached maximum levels at 9 days. The specific activity increased 20-fold from the time of ecdysis to 6 days thereafter. In the 10 day adult, trehalase levels in testes, seminal vesicles, vas deferens, TAGs, or ejaculatory ducts, were lower by two orders of magnitude than in the BAGs. However, the specific activity in the spermatophore was similar to that in the BAGs. Trehalases in the BAGs and the spermatophores showed very similar properties (soluble, optimum pH of 5.75 andK _m value of 5.4 mM for trehalose). Thus trehalase appears to be secreted from the BAGs and becomes incorporated into the spermatophores.Abbreviations BAG bean-shaped accessory gland - TAG tubular accessory gland     

Ultrastructure of the male reproductive accessory glands in the medfly Ceratitis capitata (Diptera: Tephritidae) and preliminary characterization of their secretions

The morphology and the ultrastructure of the male accessory glands and ejaculatory duct of Ceratitis capitata were investigated. There are two types of glands in the reproductive apparatus. The first is a pair of long, mesoderm-derived tubules with binucleate, microvillate secretory cells, which contain smooth endoplasmic reticulum and, in the sexually mature males, enlarged polymorphic mitochondria. The narrow lumen of the gland is filled with dense or sometimes granulated secretion, containing lipids. The second type consists of short ectoderm-derived glands, finger-like or claviform shaped. Despite the different shape of these glands, after a cycle of maturation, their epithelial cells share a large subcuticular cavity filled with electron-transparent secretion. The ejaculatory duct, lined by cuticle, has epithelial cells with a limited involvement in secretory activity. Electrophoretic analysis of accessory gland secretion reveals different protein profiles for long tubular and short glands with bands of 16 and 10 kDa in both types of glands. We demonstrate that a large amount of accessory gland secretion is depleted from the glands after 30 min of copulation.     

Histology and histochemistry of the accessory reproductive glands in the male hairy-nosed wombat (Lasiorhinus latifrons)

The accessory male reproductive glands of the hairy-nosed wombat, Lasiorhinus latifrons, are a prostate and three pairs of Cowper's glands. Component units of all are branched tubular structures of varying epithelial makeup and secretory content. The prostate has the carrotlike shape and three consecutive regions commonly found in marsupials. The regions differ in their tubular histology and histochemistry: all contain secretory globules in glandular lumina. Cowper's glands A and B are histologically identical except for the absence of interstitial mast cells from gland G: gland C is characterized by narrower tubules and larger epithelial cells. Histochemical tests for protein, carbohydrate and iron indicate that glycogen is a major secretory product of the prostate (largely posterior region), iron is also secreted (mainly posterior region) and a small quantity of acid mucin is produced (mainly central region). Glycogen is a feature also of anterior prostatic glandular epithelium and of the capping cells of the urethral transitional epithelium. Cowper's gland A has considerable protein in its secretion, gland B a neutral glycoprotein and gland C a sialomucin: the latter two also exhibit cytoplasmic glycogen in their secretory cells.     

Morphological changes in the male accessory glands and testes in Vespula vulgaris (Hymenoptera,Vespidae) during sexual maturation

Abstract. The present study documents the pace of accessory gland and testes degeneration in the wasp Vespula vulgaris by means of a histological and metric approach, that has not been carried out for social wasps so far. To a certain extent, comparison is made with the degenerative processes of the mucus glands of the honeybee drone. In V. vulgaris, no generative tissue is left by the end of 9 d of age, and so degeneration is a fast process. The three different parts of the accessory glands (muscle layer, gland epithelium, and lumen) change with respect to age. The secretory cells of the epithelium reach their maximum activity during the first days of adult life, which results in a maximally filled gland lumen by 9 d. We also provide, for the first time, a histological study of testes degeneration for this species. At eclosion, well‐defined cystic structures are still visible, whereas at 9 d, it is no longer possible to distinguish different cystic structures. The diameter of the testes decreases with respect to age.     

Cell cycles in the male accessory glands of mealworm pupae

During the pupal stage of Tenebrio molitor, the accessory reproductive glands of males grow by cell division. Within the secretory epithelium of the bean-shaped accessory glands (BAGs), cell numbers triple. In the tubular accessory glands (TAGs), the increase is 14-fold. There are two mitotic maxima in each gland. The first maximum occurs at 1-2 days while the second is at 4-5 days. The second maximum coincides with the major ecdysteroid peak described by Delbecque et al. [Dev. Biol. 64, 11-30 (1978)]. Nuclei were isolated from TAGs during the pupal mitotic bouts and during mitotic inactivity in the adult. After Feulgen or propidium iodide staining, the DNA content of these nuclear populations was measured by absorption cytophotometry or by fluorescence flow cytometry, respectively. The proportion of cells in each phase of the cycle was calculated using an iterative model. After mitoses have ended in the late pupa, the cells were arrested in G2. [3H]Thymidine was injected into 1- and 4-day pupae to pulse-label cells of the TAGs. After allowing various periods from 4 to 60 hr for cells to progress through G2 to reach mitosis, fractions of labelled mitoses were determined by autoradiography. From the combined cytometric and autoradiographic data, the duration of each phase of the cell cycle was calculated assuming the population was in exponential growth. Cell cycles in 4-day pupal TAGs take 48 hr. G1, S, G2, and, M lasted 13, 14, 17, and 4 hr, respectively.     

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.     

S-Adenosylmethionine:Juvenile hormone acid methyltransferase in male accessory reproductive glands of Hyalophora cecropia (L.)

The accessory reproductive glands of the adult male Hyalophora cecropia contain S-adenosylmethionine:juvenile hormone acid methyltransferase. The enzyme is soluble and can be found in the gland epithelium as well as in the glandular secretion, but not in any other part of the genital tract of the unmated male. The appearance of this enzyme activity in the pharate adult precedes the formation of a measurable pool of its substrate, juvenile hormone acid, and the onset of the juvenile hormone accumulation in the accessory reproductive glands. The accessory reproductive glands of Antherea pernyi and Manduca sexta, species which do not accumulate juvenile hormone, lack methyltransferase activity. It is concluded that the methyltransferase is an essential component of the juvenile hormone accumulation mechanism in H. cecropia.     

Morphology and protein patterns of honey bee drone accessory glands

We used light and transmission electron microscopy to examine the morphology of the accessory glands of immature and mature adult males of Apis mellifera L. We also made an electrophoretic analysis of the protein content of the mature gland. The glands of the immature male actively secrete a mucous substance that can be seen in the lumen of the gland of the mature male. This secretion stains with mercury bromophenol blue and with periodic acid-Schiff reaction, which stain glyconjugates. The protein content was higher in the lumen secretion than in the gland wall extracts. The electrophoresis patterns of the wall extracts were different from those of the secretion found in the gland lumen.     

The transparent accessory reproductive gland secretes a polypeptide into the hemolymph of male Rhodnius prolixus

The transparent accessory reproductive gland of Rhodnius prolixus synthesizes and accumulates a variety of polypeptides. Ouchterlony immunodiffusion demonstrates that the hemolymph contains proteins which react with polyclonal antibodies against extracts of transparent accessory glands. Accessory glands and hemolymph contain a 170 kDa polypeptide with similar mobility on SDS-polyacrylamide gel electrophoresis. This polypeptide reacts with antibodies against extracts of accessory glands. Surgical removal of the accessory glands prevents the appearance of the 170 kDa polypeptide in the hemolymph. In vivo labeling of accessory gland proteins with a mixture of [¹⁴C]amino acids demonstrates that the newly synthesized TARG polypeptide appears in the hemolymph between days 2 and 3 after feeding. It is concluded that a specific polypeptide which is synthesized in the transparent accessory gland is exported to the hemolymph.     

Ultrastructure of male reproductive accessory glands and ejaculatory duct in the Queensland fruit fly,Bactrocera tryoni (Diptera: Tephritidae)

Ultrastructure of male reproductive accessory glands and ejaculatory duct in the Queensland fruit fly (Q-fly), Bactrocera tryoni, were investigated and compared with those of other tephritid flies. Male accessory glands were found to comprise one pair of mesodermic glands and three pairs of ectodermic glands. The mesodermic accessory glands consist of muscle-lined, binucleate epithelial cells, which are highly microvillated and extrude electron-dense secretions by means of macroapocrine transport into a central lumen. The ectodermic accessory glands consist of muscle-lined epithelial cells which have wide subcuticular cavities, lined with microvilli. The electron-transparent secretions from these glands are first extruded into the cavities and then forced out through small pores of the cuticle into the gland lumen. Secretions from the two types of accessory glands then flow into the ejaculatory duct, which is highly muscular, with epithelial cells rich in rough endoplasmic reticulum and lined with a thick, deeply invaginated cuticle. While there are some notable differences, reproductive accessory glands of male Q-flies generally resemble those of the olive fruitfly, Bactrocera oleae, and to a lesser extent the Mediterranean fruit fly, Ceratitis capitata.     

Ultrastructure and cytochemistry of salivary glands of the predator Podisus nigrispinus (Hemiptera: Pentatomidae)

Podisus nigrispinus Dallas (Hemiptera: Pentatomidae) is a zoophytophagous insect with a potential for use as a biological control agent in agriculture because nymphs and adults actively prey on various insects by inserting mouthparts and regurgitating the contents of the salivary glands inside the prey, causing rapid paralysis and death. However, the substances found in saliva of P. nigrispinus that causes the death of the prey are unknown. As a first step to identify the component of the saliva of P. nigrispinus, this study evaluated the ultrastructure and cytochemistry of the salivary glands of P. nigrispinus. The salivary system of P. nigrispinus has a pair of principal salivary glands, which are bilobed with a short anterior lobe and a long posterior lobe, and a pair of tubular accessory glands. The principal gland epithelium is composed of a single layer of cells enclosing a large lumen. Epithelial cells of the principal salivary gland vary from cubic to columnar shape, with one or two spherical and well-developed nuclei. Cells of the anterior lobe of the principal salivary gland have an apical surface with narrow, short, and irregular plasma membrane foldings; apical and perinuclear cytoplasm rich in rough endoplasmic reticulum; and mitochondria with tubular cristae. The basal portion of the secretory cells has mitochondria associated with many basal plasma membrane infoldings that are short but form large extracellular canals. Secretory granules with electron-dense core and electron-transparent peripheral are dispersed throughout the cytoplasm. Cells of the posterior lobe of the principal salivary gland are similar to those of the anterior lobe, except for the presence of mitochondria with transverse cristae. The accessory salivary gland cells are columnar with apical microvilli, have well-developed nucleus and cytoplasm rich in rough endoplasmic reticulum, and have secretory granules. Cytochemical tests showed positive reactions for carbohydrate, protein, and acid phosphatase in different regions of the glandular system. The principal salivary glands of P. nigrispinus do not have muscle cells attached to its wall, suggesting that saliva-releasing mechanism may occurs with the participation of some thorax muscles. The cytochemical and ultrastructural features suggest that the principal and accessory salivary glands play a role in protein synthesis of the saliva.     

The architecture of the accessory reproductive glands of the male desert locust. 5: ultrastructure during maturation

The accessory reproductive glands of the male desert locust were studied with the electron microscope from the time of adult emergence until full sexual maturity was attained (10 days). Observations on the changes in the nucleus and the cytoplasm of the glandular epithelium, particularly those organelles involved in the elaboration of cell secretions, were made during the maturation of the accessory glands. Cells of gland 16 (the functional seminal vesicle) exhibit numerous ribosomal aggregates, have a concentration of mitochondria along their luminal surface, and show some secretory activity, even at the time of adult emergence. On the other hand, all other accessory glands show very little development of the secretory apparatus (rough-surfaced endoplasmic reticulum, polyribosomes, Golgi elements, elaborate mitochondria, etc.) until after the fifth adult day. These changes parallel changes in the secretory activity of the adult corpus allatum; and it is suggested that the corpus allatum hormone regulates the maturation of the truly glandular accessory glands of the locust, but not that of the functional vesicle.     

B-mode ultrasonographic examination of the accessory sex glands of boars

Thorough examinations of the reproductive system of boars are generally not performed on normal boars to be used for breeding; only boars with problems undergo a form of a breeding soundness examination. In order for veterinarians to identify pathological conditions, the normal architecture of the accessory sex glands needs to be described. The purpose of this study was to use B-mode ultrasonography to describe the accessory sex glands in the boar and to see if transrectal ultrasonography would be a viable option in which to obtain this data. Initially, cross-sectional saline bath examinations of accessory sex glands were performed on crossbred boar reproductive tracts (n = 4) using B-mode ultrasonography equipped with a 5 MHz dual frequency linear array transducer. In situ examinations were also performed on terminal line crossbred boars (n = 16) ranging in age from 10 to 23 months old using the same ultrasound methodology; four boars were under general anesthesia and the remaining 12 were standing in crates. Eight boars were abstinent for 2 days and the other eight had ejaculates collected 2 h prior to examination. The paired bulbourethral glands are best described as a long oval gland with a uniformly echogenic appearance with a large anechoic space in the center of the gland extending most of its length. The walls of the vesicular glands were found to be thin, with the parenchyma having multiple small echolucent areas that appeared to merge and form a central canal. The prostate gland was best identified as a pecan-sized gland with a uniform echogenic appearance. Visualization of the prostate gland was accomplished with more proficiency using the saline bath ultrasonography as compared to in situ examinations. All of the accessory sex glands could be examined using both methodologies of ultrasonographic examination with a 5 MHz frequency linear array transducer. It was determined that each accessory sex gland could be recognized, and differences between ejaculated and nonejaculated boars could be identified. The results of this study demonstrate that transrectal ultrasonography can be used as a diagnostic aid in assessing the accessory sex glands of boars.     

Ability of male Queensland fruit flies to inhibit receptivity in multiple mates, and the associated recovery of accessory glands

Mating success of male insects is commonly determined by their ability to find and copulate with multiple females, but is also determined by their ability to transfer an effective ejaculate. In order to succeed in these tasks, males must first succeed in replenishing the necessary reproductive reserves between mating opportunities. We here investigate the ability of male Queensland fruit flies ('Q-fly') to recover from their first matings in time to both mate again the following day and to induce sexual inhibition in successive mates. We have previously found that accessory gland fluids (AGFs) transferred in the ejaculate of male Q-flies are directly responsible for induction of sexual inhibition in their mates. We here investigate changes in male accessory gland, testis and ejaculatory apodeme dimensions that are likely to reflect depletion and recovery of contents. We found no differences between virgin and previously mated males in their ability to obtain matings or to induce sexual inhibition in their mates, indicating a full recovery of the necessary reproductive reserves between mating opportunities. Whereas no changes were detected in testis or ejaculatory apodeme size following mating, the recovery of male ability to inhibit female remating was closely reflected in the mesodermal accessory gland dimensions; these accessory glands greatly diminished in size (length and area) immediately after mating, with recovery commencing between 5.5 and 11 h after mating. The accessory glands then expanded to reach their original size in time to mate the following day and induce sexual inhibition in the next mate.