Humoral autoimmune response to rat male accessory glands. Comparative specificity studies of auto and heteroantibodies

A comparison of specific antibodies induced by a saline extract of chemically modified rat male accessory glands (MRAG) in isologous (male and female rats) and heterologous animals (rabbits and a goat) was made. This study was focused on the specificity of the antibodies confronted with saline extract of rat male accessory glands (RAG), their Sephadex G-100 fractions and autoantigen fragments. Specificity studies with Sephadex G-100 fractions of RAG showed that whereas in male rats only the antibodies reactive with Fraction 1 containing autoantigens were detected, one additional population of antibody with different specificity was revealed in female sera. In contrast, antibodies against several other macromolecules were present in heterologous sera. When analyzed for the serum specificity against three enzymatic fragments of autoantigen, the main specificities of male and female sera were different.     

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。     

桑天牛雄性附腺内容物组分分析

测定桑天牛Apriona germaric(Hope)雄性附腺内容物中各组分含量及变化情况。结果表明,内容物中可溶性蛋白占总腺管鲜重的8.39%;总糖占总鲜重的4.21%;海藻糖和游离氨基酸分别占总鲜重的0.50%、0.25%。随虫龄的增大,内容物中各组分含量不断降低。交配过程中,雄性附腺内容物部分转移到雌虫,交配后附腺内容物中总糖、海藻糖、游离氨基酸等组分含量均降低,蛋白质组分含量先升高再降低,48h即恢复可到交配前水平。大、小附腺内各组分含量变化有差异。     

BMP-regulated exosomes from Drosophila male reproductive glands reprogram female behavior

Male reproductive glands secrete signals into seminal fluid to facilitate reproductive success. In Drosophila melanogaster, these signals are generated by a variety of seminal peptides, many produced by the accessory glands (AGs). One epithelial cell type in the adult male AGs, the secondary cell (SC), grows selectively in response to bone morphogenetic protein (BMP) signaling. This signaling is involved in blocking the rapid remating of mated females, which contributes to the reproductive advantage of the first male to mate. In this paper, we show that SCs secrete exosomes, membrane-bound vesicles generated inside late endosomal multivesicular bodies (MVBs). After mating, exosomes fuse with sperm (as also seen in vitro for human prostate-derived exosomes and sperm) and interact with female reproductive tract epithelia. Exosome release was required to inhibit female remating behavior, suggesting that exosomes are downstream effectors of BMP signaling. Indeed, when BMP signaling was reduced in SCs, vesicles were still formed in MVBs but not secreted as exosomes. These results demonstrate a new function for the MVB–exosome pathway in the reproductive tract that appears to be conserved across evolution.     

Correlated evolution of female neoteny and flightlessness with male spermatophore production in fireflies (Coleoptera: Lampyridae)

The beetle family Lampyridae (fireflies) encompasses ～100 genera worldwide with considerable diversity in life histories and signaling modes. Some lampyrid males use reproductive accessory glands to produce spermatophores, which have been shown to increase female lifetime fecundity. Sexual dimorphism in the form of neotenic and flightless females is also common in this family. A major goal of this study was to test a hypothesized link between female flight ability and male spermatophore production. We examined macroevolutionary patterns to test for correlated evolution among different levels of female neoteny (and associated loss of flight ability), male accessory gland number (and associated spermatophore production), and sexual signaling mode. Trait reconstruction on a molecular phylogeny indicated that flying females and spermatophores were ancestral traits and that female neoteny increased monotonically and led to flightlessness within multiple lineages. In addition, male spermatophore production was lost multiple times. Our evolutionary trait analysis revealed significant correlations between increased female neoteny and male accessory gland number, as well as between flightlessness and spermatophore loss. In addition, female flightlessness was positively correlated with the use of glows as female sexual signal. Transition probability analysis supported an evolutionary sequence of female flightlessness evolving first, followed by loss of male spermatophores. These results contribute to understanding how spermatophores have evolved and how this important class of seminal nuptial gifts is linked to other traits, providing new insights into sexual selection and life-history evolution.     

Kallikrein localization in rodent salivary glands and kidney with the immunoglobulin-enzyme bridge technique.

Kallikrein has been localized in rodent kidney and salivary glands by means of an immunoglobulin-enzyme bridge technique. In sections of kidney, anti-kallikrein antibodies bound to the apical region of certain distal tubule segments in the cortex, to reabsorption droplets of proximal convoluted tubules, and to certain duct segments in the papilla. In salivary glands of both male and female rats and mice, and apical rim of most striated duct cells of submandibular, parotid and sublingual glands and granular tubules of submandibular glands exhibited immunoreactivity. Granular intercalated duct cells in female submandibular glands also displayed immunostaining for kallikrein. Phenylephrine administration resulted in loss of immunoreactive granules from the granular convoluted tubule cells of male mouse submandibular gland. This response was paralleled by a biochemically demonstrable decrease in kallikrein-like tosylarginine methyl ester (TAME) esterase activity.     

Effects of olive, corn, sesame or peanut oil on the body weights and reproductive organ weights of immature male and female rats.

Olive, corn, sesame or peanut oil which have been used as vehicles in the immature rat uterotrophic assay or Hershberger assay, for detection of endocrine disrupting effects of environmental chemicals, was administered to ten immature female rats by subcutaneous injection from postnatal day (PND) 21 for 3 or 7 days, and each oil was also administered to ten male rats from PND 21 for 7 and 10 days. The body weights, and the weights of sex and sex accessory organs in female and male rats were measured. There were no significant differences in body weights of female rats between each oil group and the control group, while the body weight of male rats in the group given peanut or olive oil was significantly increased from 8 or 9 days after administration. There were no changes in the sex and sex accessory organ weights of female or male rats related to the endocrine disrupters. The results of the body weights and organ weights demonstrate that each oil is a suitable vehicle for the immature rat uterotrophic assay. However, each oil is suggested to be unsuitable for the Hershberger assay, because the analysis of changes of sex accessory organ weights in this assay might be confused by the increased body weights.     

Impaired 3H-testosterone uptake and metabolism in the accessory sex glands of diabetic castrated male rats.

The uptake and retention of 1,2-3H-testosterone in accessory sex glands, muscle and liver of streptozotocin diabetic castrated male rats, insulin-treated diabetic castrated rats and non-diabetic castrated control rats were studied at various time intervals after an intravenous injection. Diabetes reduced the retention of 3H-testosterone in the prostate, the preputial gland and the epididymis. Exogenous insulin slightly increased the retention of 3H-testosterone in these tissues of diabetic rats. No significant differences in the radioactivity in the rectus abdominis muscle, the coagulating glands and the seminal vesicles were found between the various experimental groups. Ventral prostate homogenates obtained from diabetic and control rats were incubated with 3H-testosterone in vitro. The steroids were extracted and thin-layer chromatographs were scanned for radioactivity. In prostatic homogenates taken from diabetic rats, testosterone transformation to dihydrotestosterone was reduced. The results indicate that the impaired function and androgen retention of the accessory sex glands of diabetic male rats is at least partly due to the reduced formation of dihydrotestosterone from testosterone.     

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.     

Sexual behavior in developing male rats

During normal development, the onset of reproductive behavior in male rats was not preceded by any change in plasma testosterone (T) levels. Implantation of Silastic capsules containing T in 14-day-old male rats advanced the onset of all parameters of sexual behavior by 20 days. Implantation of Silastic capsules containing estradiol in 14-day-old male rats stimulated precocious mounting and intromitting, but not ejaculation. Implantation of dihydrotestosterone-filled Silastic capsules in 14-day-old male rats completely inhibited the development of sexual behavior. All hormones suppressed plasma LH levels. These findings in immature male rats are similar to previous findings in adult males. Immature male rats were behaviorally less responsive to T than adult males, and it was suggested that, during development, male rats become progressively more sensitive to the behavior-stimulating effects of circulating T. No effects of copulatory experience on plasma concentration of T or on the weights of testes, penes, or accessory sexual glands were detected.     

Spleen cells derived from male non-obese diabetic mice are capable of suppressing the autoantigen-specific production of interferon-gamma of female cells in vitro

Non-obese diabetic (NOD) mice spontaneously develop type 1 diabetes with a strong female predilection. Using co-culture systems with both female and male spleen cells, we found that spleen cells derived from male NOD mice suppress autoantigen (glutamic acid decarboxylase) induced production of interferon-gamma of female cells in vitro. In addition, this suppression appeared to be mediated by a soluble factor(s) produced by male cells in response to the same antigen. Our experimental systems might be useful for further understanding sex differences in autoimmunity as well as in the basic immune response.     

Transfer and fate of male ejaculate in female Queensland fruit flies

Abstract Insect seminal fluid commonly comprises a complex cocktail of proteins and other biochemical components that migrate away from the female reproductive tract to sites elsewhere in the female body and elicit changes in female reproductive behaviour. The transfer of male seminal fluid molecules to reproductive and somatic tissues of the female Queensland fruit fly (‘Q‐fly’) Bactrocera tryoni is examined and some putative target sites identified. Male Q‐flies are fed a diet containing radiolabelled (³⁵S) amino acids, which are incorporated into male accessory gland products. Radioactivity diminishes within the accessory glands and increases in all assessed parts of the female body during copulation, indicating the transfer of these products into the female soma via the reproductive tract. There are significant changes in the absolute and proportional radioactivity profiles among female tissues over the next 22 h, with substantial reductions in the thorax and increases in the head. This is consistent with accumulation of behaviour‐modifying male products at binding sites in the female head. Parallels can be drawn between the data in the present study and seminal fluid distribution profiles and receptor binding documented in other insects.     

A male accessory gland peptide that regulates reproductive behavior of female D. melanogaster

The adult male accessory glands of D. melanogaster synthesize and secrete a peptide that represses female sexual receptivity and stimulates oviposition. Normally, this peptide is transferred to females during copulation; however, the peptide shows the same biological activity after purification and subsequent injection into the abdominal cavity of female virgins. Amino acid sequencing of the purified peptide and oligonucleotide-directed cDNA cloning established that the peptide consists of 36 amino acids. It appears to be synthesized as a precursor with a hydrophobic signal sequence of 19 residues at its N-terminal end. The precursor peptide is encoded by a short mRNA that accumulates exclusively in the male accessory gland. The gene has been localized by in situ hybridization to polytene chromosomes at 70A.     

Potentiation of autoimmune response in rats infected with Toxoplasma gondii. Effect of the infection route

The aim of this report is to describe the influence of the way of infection with 3 X 10(3) trophozoites of Toxoplasma gondii on the auto- and heteroimmune responses of rats immunized with chemically modified rat male accessory glands (MRAG) and human serum albumin (HSA) 6 and 36 days after infection. The delayed hypersensitivity (DTH) response to MRAG was potentiated in rats infected in thymus proximity (p less than 0.02) when compared with rats only immunized. The parasites introduced intraperitoneally did not modify the cellular autoimmune response. The titers of hemagglutinating antibodies to MRAG were increased in animals infected by both ways (p less than 0.05). The response to HSA did not show significant difference between the infected and uninfected rats. Thymic involution and proliferation of lymphocytes and plasma cells were observed at 6 and 52 days post-infection only in rats infected in thymus proximity (p less than 0.05). A correlation between the potentiation of cellular autoimmune response and the injection of parasites in thymus proximity was observed.     

Phenotype characteristics of transgenic male mice expressing human aromatase under ubiquitin C promoter

To study the significance of the increased ratio of the estrogen/androgen concentration for the male reproductive functions, we have generated transgenic mice expressing human P450 aromatase under a promoter providing ubiquitous and permanent transgene expression (AROM+ mice). AROM+ male mice are characterized by elevated serum estradiol and prolactin (Prl) concentrations, combined with markedly reduced testosterone levels. The mice are present with a multitude of structural and functional alterations in the reproductive organs such as cryptorchidism, Leydig cell hyperplasia, disrupted spermatogenesis and infertility. Furthermore, the mice develop infravesical obstruction associated with the rhabdosphincter atrophy and rudimentary accessory sex glands. Interestingly, the mammary gland in AROM+ males undergo a ductal and alveolar development morphologically resembling terminally differentiated female mammary glands, and express several signaling proteins typical for female mammary glands. Some of the abnormalities seen in AROM+ mice are similar to those described in both mice and humans exposed to diethylstilbestrol (DES) in utero. The importance of the AROM+ model may lie in its predictability, i.e. the model suggests which abnormalities of the human reproductive functions may be associated with the increased ratio of estrogen/androgen concentrations in early life and at adult age as well.     

Interrelationship between adrenal cortex and genital apparatus of the male rat]

Interactions between gonadal hormones and adrenocortical function have been studied in the male rat. After injection (im) of testosterone, the accessory reproductive organs are more stimulated in the castrated rats than in the castrated adrenalectomized rats. Gonadectomy increases lipids, cholesterol and corticosterone in adrenal glands and plasma corticosterone levels. Testosterone reverses these effects.     

On the functional significance of juvenile hormone in the accessory sex glands of male Heliothis virescens

The storage of large quantities of juvenile hormone (JH) in male abdomens is a phenomenon known from some species of moths. Juvenile hormone, stored in male accessory sex glands (ASG), may be transferred to the female during copulation, but the physiological significance of the JH transfer remains unclear. Here, using the moth Heliothis virescens as a model, we show that JH transferred from male to the promiscuous female promotes JH synthesis and egg development in the female. We propose that this explains the functional significance of JH transfer in species that exhibit last male sperm precedence, and that this hormone acts as a bioactive substance which the first male to mate uses for co-opting and regulating the female's gonadotropic mechanisms, thereby ensuring that despite last male sperm precedence he will sire a significant number of viable offspring.     

日本弓背蚁雄蚁生殖系统结构及其化学组成研究

【目的】雄蚁是蚂蚁巢群的重要成员,主要参与种群的繁殖。本文对日本弓背蚁Camponotus japonicus雄蚁的生殖系统结构及其化学组成进行了研究,拟为进一步开展雄蚁的生殖行为研究提供基础信息。【方法】利用光学和扫描电镜观察了雄蚁的生殖系统结构,并利用GC-MS方法分析了其化学组成。【结果】(1)雄蚁生殖系统包括1对附腺、1对输精管和射精管3部分,睾丸退化消失;附腺最为发达,肾形,乳白色;输精管简单细长,与附腺基部1/3处相连。扫描电镜下发现附腺表面平坦,包裹有发达的网状肌肉组织。(2)利用GC-MS方法检测到13种化学物质,主要以醛类、烃类和酸类为主,分别占29.31%、21.87%和20.56%,还有一些醇类(12.71%)和酯类(15.05%);含量最高的3种成分分别为十八烯醛(15.86%)、11-十六烯醛(13.45%)和9,12,15-十八碳三烯酸-2,3-二羟基丙酯(11.78%)。【结论】雄蚁生殖系统主要由附腺、输精管和射精管三部分组成,其中附腺最为发达,输精管细长,睾丸退化消失;生殖系统中含有多种化学成分,但其功能还有待于进一步的研究。     

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.     

Female accessory reproductive gland activity in the yellow dung fly Scathophaga stercoraria (L.)

The role of the female accessory reproductive glands has been investigated in relatively few insects. Gland secretion has a number of potential functions, including lubrication during copula, involvement in fertilization and protection of eggs. Female yellow dung flies (Scathophaga stercoraria) have large paired accessory glands whose function(s) prior to this study were unknown. Our study indicated glands were involved in copulation and egg laying. The volume of secretion remaining in glands was negatively associated with copulation duration, and this effect was most pronounced in non-ovipositing females. Gland volume and secretion volume remaining in the glands were significantly smaller in females which were allowed to oviposit. In addition, there was a significant interaction between male size, female size and whether or not females were allowed to oviposit which affected the volume of the secretion remaining in the glands, with changes in secretion volume being greatest when males were large. Sperm were found in the accessory glands of some females and this was apparently not related to age, mating history of either sex, to female nutrition or male size. Our results indicate that either large males stimulate greater secretory responses from females or that females alter their responses based on male size.