Molecular evolution of the androgenic hormone in terrestrial isopods

In crustaceans, the androgenic gland (AG), thanks to the synthesis of the androgenic gland hormone (AGH), controls the differentiation of the primary and secondary male sexual characters. In this study, we amplified 12 new AGH cDNAs in species belonging to five different families of the infra-order Ligiamorpha of terrestrial isopods. Putative essential amino acids for the production of a functional AGH protein exhibit signatures of negative selection and are strictly conserved including typical proteolytic cleavage motifs, a putative N-linked glycosylation motif on the A chains and the eight Cys positions. An insulin-like growth factor motif was also identified in Armadillidium AGH sequences. The phylogenetic relationships of AGH sequences allowed one to distinguish two main clades, corresponding to members of the Armadillidiidae and the Porcellionidae families which are congruent with the narrow specificity of AG heterospecific grafting. An in-depth understanding of the regulation of AGH expression would help deciphering the interaction between Wolbachia, widespread feminizing endosymbiotic bacteria in isopods, and the sex differentiation of their hosts.     

Endocrine and genetic control of sex differentiation in the Malacostracan Crustacea

Summary

Sex differentiation in Malacostraca is controlled by hormone secreted from the androgenic glands. Experimentally induced sex inversions in isopods and amphipods proved that the genetic female and male possess primordia of the androgenic glands, gonads, and gonoducts, along with sexual characteristics of both sexes. During the sensitive period, the presence or absence of androgenic gland hormone (AGH) affects the differentiation of these primordia.

Genetic control of the development of androgenic gland primordium seems to be brought about assuming of the following: 1. Both genetic female and male possess gene(s) (AGH-G) responsible for the AGH-synthesis situated on the homologous loci of the sex chromosomes and/or on the autosomes. 2. The gene(s) are activated spontaneously with the lack of inhibition of the major sex factor carried by the W or X chromosome. The W and X factors are hypostatic to major sex factor carried by the Y chromosome. The Z factor does not seem to influence sex differentiation. Sufficient allochthonous AGH seems to render the W and X factors ineffective.     

The effect of partial brood loss on male desertion in a cichlid fish: an experimental test

There is little experimental evidence testing whether currentbrood size and past brood mortality influence mate desertion.In the cichlid Aequidens coeruleopunctatus both parents initiallydefend offspring. In a field study, all experimental broods,irrespective of initial brood size (222.9 ± 60.4, mean± SD), were manipulated to a size of 100 fry. Neitherthe duration nor investment of females in parental care differed between control and brood reduced pairs, even though care seemedcostly. On average, females lost 5.1 ± 4.8% of initialweight while guarding a brood until independence. In contrast,males with experimentally reduced broods guarded fry for significantlyfewer days before deserting their mate than did males fromcontrol pairs with natural-sized broods (20.5 ± 7.5 vs. 14.2 ± 6.2 days). In at least 20% of cases (n = 9/45),the deserting male immediately mated with another female. Maleswith experimentally reduced broods also spent less time guardingfry before deserting and attacked fewer brood predators thandid males with control broods. For broods manipulated to have100 fry, there was a significant negative relationship betweenthe days until male desertion and the proportion of the initialbrood removed. This indicates that male assessment of the futuresuccess of the current brood (hence its reproductive value)is based on past mortality and/or that there is variation amongmales in the expected size of future broods. Both current broodsize and brood size relative to initial brood size are thereforepredictors of male, but not female, parental behavior and matedesertion. Female care may be unaffected by brood reductiondue to limited breeding opportunities and partial compensationfor reduced male care.     

Characterization and cDNA cloning of androgenic gland hormone of the terrestrial isopod Armadillidium vulgare.

The sex differentiation in crustaceans is known to be controlled by a peptide hormone called androgenic gland hormone (AGH). AGH was extracted and purified from the androgenic glands (AGs) of the male isopod Armadillidium vulgare by high-performance liquid chromatography. AGH consisted of two peptide chains and their N-terminal amino acid sequences were determined. A cDNA encoding AGH was cloned by PCR and sequenced. The cDNA had an open reading frame of 432 bp, which encoded a preproAGH consisting of a signal peptide (21 residues), B chain (44 residues), C peptide (46 residues), and A chain (29 residues). Through processing, the A and B chains might form a heterodimer interlinked by disulfide bonds. The A chain possessed a putative N-linked glycosylation site. A Northern blot analysis using the cDNA as a probe detected a hybridization signal with 0.8 kb in the RNA preparation only from the AGs.     

几种甲壳动物促雄性腺素结构预测分析

利用生物信息学方法对目前已知的3种甲壳动物促雄性腺素前体(AGH precursor)和5种类胰岛素促雄性腺因子(insulin-like AG factor)进行分析,探讨了促雄性腺素前体的氨基酸理化特性、信号肽、跨膜结构域、二级结构、motif等,并利用Phyre软件对其三级进行同源性收索。结果显示:促雄性腺素前体包含信号肽,存在跨膜结构域,并和信号肽同位。PDB库中没有找到匹配的motif。3种促雄性腺素前体的二级结构有比较高的相似性,比如都包含两个中心螺旋区。Phyre搜索显示,与8种蛋白的三级结构匹配的均为胰岛素家族的蛋白,这也进一步证实了促雄性腺素前体和胰岛素原的相似性。     

Why do sperm-depleted parasitoid males continue to mate?

Insect males of several parasitoid species have limited amountof sperm at emergence and experience sperm depletion when mateacquisition rate is high, suggesting that sperm production andstorage could be a limiting factor for male's fitness. Sperm-depletedmales of the parasitoid wasp Trichogramma evanescens continueto mate, and the impact of this behavior on sperm storage byfemales has been studied. Virgin females T. evanescens stored50.6 ± 13.1 (Mean ± SD) sperm when mating firstwith a virgin male. However, when these females mated firstwith a sperm-depleted male and then with a virgin male, theystored only 18.9 ± 7.8 sperm, indicating that matingwith a sperm-depleted male has a cost and limits sperm acquisitionfrom fertile males. Following a mating with a sperm-depletedmale, females had to mate with three virgin males to restoretheir sperm supply confirming that additional mating enabledthe female to store a limited number of sperm. According toour results, by continuing to mate, sperm-depleted males donot prevent mates from copulating again but they decrease theirability to store other male's sperm. Continuing to mate couldbe a strategy to increase the relative fitness of sperm-depletedmales.     

Reduced urea flux across the blood-testis barrier and early maturation in the male reproductive system in UT-B-null mice

A urea-selective urine-concentrating defect was found in transgenic mice deficient in urea transporter (UT)-B. To determine the role of facilitated urea transport in extrarenal organs expressing UT-B, we studied the kinetics of [¹⁴C]urea distribution in UT-B-null mice versus wild-type mice. After renal blood flow was disrupted, [¹⁴C]urea distribution was selectively reduced in testis in UT-B-null mice. Under basal conditions, total testis urea content was 335.4 ± 43.8 µg in UT-B-null mice versus 196.3 ± 18.2 µg in wild-type mice (P < 0.01). Testis weight in UT-B-null mice (6.6 ± 0.8 mg/g body wt) was significantly greater than in wild-type mice (4.2 ± 0.8 mg/g body wt). Elongated spermatids were observed earlier in UT-B-null mice compared with wild type mice on day 24 versus day 32, respectively. First breeding ages in UT-B knockout males (48 ± 3 days) were also significantly earlier than that in wild-type males (56 ± 2 days). In competing mating tests with wild-type males and UT-B-null males, all pups carried UT-B-targeted genes, which indicates that all pups were produced from breeding of UT-B-null males. Experiments of the expression of follicle-stimulating hormone receptor (FSHR) and androgen binding protein (ABP) indicated that the development of Sertoli cells was also earlier in UT-B-null mice than that in wild-type mice. These results suggest that UT-B plays an important role in eliminating urea produced by Sertoli cells and that UT-B deletion causes both urea accumulation in the testis and early maturation of the male reproductive system. The UT-B knockout mouse may be a useful experimental model to define the molecular mechanisms of early puberty. urea transporter; Sertoli cell; testis; male sexual function; spermatogenesis     

Female preference in complex acoustical environments in the midwife toads Alytes obstetricans and Alytes cisternasii

Gravid females of Alytes obstricans and Alytes cisternasii weretested with synthetic calls in seven-speaker playback tests.A first, "mean-centered" test presented calls with frequenciesrepresenting an array of different calls spanning over the rangeof the population (±2.25 SD). In this test, females ofboth species approached a synthetic call that was lower thanthe average call frequency of the male population although thedifference was significant only for A. obstetricans. The regressionbetween female weight and size and preferred frequency was notsignificant in either species. These results confirm the reportedtrends of females preferring lower frequency calls (correspondingto larger males) based on two-speaker playback tests for A.obstetricans. For A. cisternasii, the lack of significance ofthe seven-speaker test suggests that the preference trend previouslyfound in two-speaker tests may be obscured in more complex acousticalenvironments. A second "supernormal stimulus" test presentedfemales with calls ranging from the lowest frequency valuesof the male population (–2.25 SD) and lower, up to –6.75SD beyond the range. In both species females preferentiallyapproached calls higher than the mean frequency of the stimuluspresented. This result suggests that in both cases selectionfor low frequencies is not open ended, and that the preferredfrequency is within the range of the male population.     

Do male golden egg bugs carry eggs they have fertilized? A microsatellite analysis

In the golden egg bug, Phyllomorpha laciniata (Heteroptera,Coreidae), both males and females carry eggs on their back.Although females cannot carry their own eggs, males may carryeggs that they have fertilized. If males carry eggs they havefertilized, their behavior may be interpreted as paternal care.In this article, we provide genetic data for paternity assignmentof eggs carried by 40 males collected from the field. The malesand the eggs were typed by using four highly polymorphic microsatelliteDNA markers. Out of the 247 eggs typed, 87% were excluded fromthe father-offspring relationship based on single-locus (leastconservative exclusion) mismatches. Under the more conservative(exclusion by at least two single locus mismatches) method,78% of the eggs were nonpaternal. Relatedness estimates furthersupported our paternity analyses. The average relatedness ofthe eggs to the carrying males was low (b_em = -0.052 ±0.024 SE). Within the population, males were unrelated to eachother (b_mm = -0.004 ± 0.0002 SE), as were the eggs carriedby individual males (b_eggs = -0.004 ± 0.0001 SE). Thisfirst genetic study on the breeding system of the golden eggbugs did not find any support for the claim that egg carryingfunctioned as paternal care, nor did it support kin selectionas explanation for conspecific egg carrying.     

Mechanisms of parasite-induced sex reversal in Gammarus duebeni

The amphipod Gammarus duebeni is host to the feminising microsporidian parasite Nosema granulosis that converts males into functional females. To test the hypothesis that the parasite acts through endocrine disruption we compared the morphology of the gonad and activity of the androgenic gland, which coordinates male sexual differentiation, in infected and uninfected animals. Male gonad consisted of testis, seminal vesicle and vas deferens that was anchored to the genital papilla on segment 7. The androgenic gland was associated with the distal end of the vas deferens. In female and intersex animals the bi-lobed ovary opened into the oviduct at segment 5, vestigial vas deferens and vestigial androgenic gland were retained. The majority of parasitised individuals (38/39) were either phenotypic females or intersexes with fully developed ovaries and an undifferentiated androgenic gland. Our data suggest that the parasite prevents differentiation of the androgenic gland. In further support of this hypothesis, mass spectrometry of a single androgenic gland from males revealed a dominant molecular ion with a mass/charge ratio of 4818.4+H, corresponding to a peptide of androgenic gland hormone from Armadillidium vulgare. In contrast the vestigial androgenic gland from parasitised and unparasitised females showed only low intensity peaks. Our observations demonstrate that the parasite manipulates host sex by preventing androgenic gland differentiation, androgenic gland hormone production and consequently male differentiation. This is in agreement with observations of A. vulgare with inherited Wolbachia infection, suggesting that phylogenetically distant feminisers manipulate hosts through a common mechanism. The high frequency of infection in intersexes (89.3%) suggests that this phenotype results from incomplete feminisation by the parasite.     

Direct evidence for the function of crustacean insulin-like androgenic gland factor (IAG): Total chemical synthesis of IAG

Insulin-like androgenic gland factor (IAG) is presumed to be a sex differentiation factor so-called androgenic gland hormone (AGH) in decapod crustacean, although the function of IAG peptide has not yet been reported. In this study, we synthesized IAG from the prawn, Marsupenaeus japonicus, and its function was assessed by an in vitro bioassay. As a result, IAG with the insulin-type disulfide bond arrangement showed biological activity, whereas its disulfide isomer did not. These results strongly suggest that the native IAG peptide has an insulin-type disulfide, and it is the decapod AGH.     

Male polyterritoriality and imperfect female choice in the pied flycatcher, Ficedula hypoleuca

Polyterritoriality, the occupation of two spatially separateterritories by male birds, has been suggested to enable malesto conceal their pairing status. The idea that males deceivefemales has recently been questioned with arguments that femaleswould easily be able to detect polyterritorial males and thatthey mate in the second territories because better breedingoptions are not available close by. We arranged for pied flycatcher(ficedula hypoleuca) females a choice between neighboring unmatedand already-mated males. Females were unable to discriminateagainst already-mated males, even though their breeding successthereby would have been increased. These results allow rejectionof any polygyny models that assume females use male pairingstatus as a criterion for mate choice. Females would reducethe probability of mating polygynously by choosing dull, yearlingmales with high song rate, even though in most cases it is difficultto differentiate already-mated and unmated males. In fact, femalesdid not prefer colorful males, opposing a general pattern foundin other bird species. However, females did not specificallyprefer dull males nor did they respond to slight differencesin song rates. The seemingly nonadaptive, imperfect female behaviorsuggests that the evolution of female selectivity is constrained,allowing deception by males to be successful in the evolutionary‘arms race’ between the sexes. [Behav Ecol 1990,-1:171–177]     

The Crustacean Androgen: A Hormone in an Isopod and Androgenic Activity in Decapods

The androgenic gland has been described in a variety of crustaceanspecies—isopods, amphipods and decapods. It has been shownto play a role in the regulation of male differentiation andin the inhibition of female differentiation. Upon its applicationfor endocrine manipulation, it inhibits female characteristics.Recently, the androgenic hormone from the isopod Armadillidiumvulgare was purified and characterized on the basis of a morphologicalbioassay. The hormone is a glycosylated protein composed oftwo peptide chains connected each to the other by two disulfidebridges. The pro-hormone consists of the same two chains connectedby a third peptide in a complex that resembles the insulin superfamily hormones. The study of the androgenic gland in decapodslags behind that in the isopods, and a decapod androgenic hormonehas yet to be identified. In this review, five decapod speciesare described as models, in which the androgenic gland exertsmorphological, anatomical, physiological and behavioral effects.These models could serve as the basis of possible bioassaysfor the study of the structure and mode of action of the androgenichormone in decapod crustaceans.     

Male attractiveness covaries with fighting ability but not with prior fight outcome in house crickets

In many species males that tend to win fights against othermales are more attractive to females. There are three ways inwhich male fighting ability and attractiveness may be associated:(1) attractiveness and fighting ability are influenced by thesame underlying traits (e.g., body size), (2) females prefermales that have directly observed winning fights, or (3) winningprevious fights indirectly improves a male's chance of beingpreferred by females. The last possibility may arise as a consequenceof the "loser effect"; in many species when a male loses a fighthis probability of losing subsequent fights increases. Thereare, however, no studies testing whether such a "loser effect"also influences male attractiveness. Here we show that maleattractiveness and fighting ability are positively correlatedin the house cricket, Acheta domesticus. Our experiment wasdesigned so that females could not directly observe the outcomeof fights, thus eliminating possibility (2) above. We then testedbetween possibilities (1) and (3) by making use of the factthat in some cricket species the "loser effect" can be eliminatedexperimentally by ‘shaking’ a male and stimulatingthe motor program for flying. We showed that in A. domesticus‘shaking’ does affect the outcome of subsequentfights. Males that had won two previous fights were less likelyto win a fight after being ‘shaken’ than when subjectto a control treatment. In contrast, males that had lost twoprevious fights were more likely to win a fight after being‘shaken’ than when they were not shaken. There was,however, no effect of ‘shaking’ on male attractiveness.We conclude that the "loser effect" does not alter the tendencyfor large, dominant males to be attractive to females. Instead,it appears that there are traits correlated with both fightingability and attractiveness. One such trait is body size. Fightwinners were significantly larger than losers and attractivenesswas positively correlated with male body size.     

Heterogeneity of Odorant-binding Proteins in the Antennae of Bombyx mori

Different odorant-binding proteins (OBPs) were isolated fromtotal antennal homogenates of male and female Bombyx mori. Proteinswere separated according to their isoelectric point by usingpreparative fast-flow isoelectrofocusing. Odorant-binding proteinswere identified in immunoblots by antisera raised against thepheromone-binding protein (anti-PBP) and the general odorant-bindingprotein (anti-GOBP2) of Antheraea polyphemus. Four proteinscross-reacting with anti-PBP were detected in males and twoin females, while three proteins cross-reacting with anti-GOBP2were found in males and five in females. Both anti-PBP and anti-GOBP2cross-reacting proteins had an apparent molecular weight of15–16 kDa. In parallel, the same two antisera were usedin immunocytochemical studies in order to determine the distributionof these proteins within the various subtypes of olfactory sensilla.The presence of multiple odorant-binding proteins within onemoth species as well as their complex distribution pattern supportthe suggestion that soluble OBPs might have a function in odorantdiscrimination. Chem. Senses 22: 503–515, 1997.     

Male and female cooperatively breeding fish provide support for the "Challenge Hypothesis"

The idea that territorial aggression is regulated by androgensand that aggression itself can modulate androgen levels is wellestablished in males. In many species, females also displayaggressive behavior, yet little work has been conducted on theeffects of female aggression on hormone levels. In this study,we compared the effects of a simulated territory intrusion (amethod for testing the Challenge Hypothesis) on males and femalesof the fish, Neolamprologus pulcher. This cichlid fish fromLake Tanganyika is a particularly useful species to examinesex differences in the behavioral mediation of hormones as breedingpairs remain in a territory year round and both sexes defendthis territory against conspecific and heterospecific intruders.In our study, both sexes indeed aggressively defended theirterritory against a simulated territory intruder. In responseto intruders, both males and females displayed elevated levelsof circulating 11-ketotestosterone, but only females exhibitedincreases in testosterone. Neither aggressing male nor femalefish showed changes in estradiol levels compared to control(nonaggressing) fish. Residents were more aggressive than theintruders and won most of the interactions. However, residents(or winners) did not show higher hormone levels than intruders(or losers). We suggest that aggression commonly modulates androgenlevels in both male and female teleost fish.     

Sexual conflict over breeding substrate causes female expulsion and offspring loss in a cichlid fish

Females of the Lake Tanganyika cichlid Lamprologus callipterusexclusively breed in empty snail shells that males collect intheir territories. Male–male competition for shells issevere, leading to frequent shell stealing and territory takeover.As a consequence, males have breeding females in their shellsthat spawned with competitors. In this field study, we investigatedboth naturally occurring and experimentally induced encountersof territorial males with females that had spawned with othermales. We found that the breeding success of females that weretaken over by a different male was significantly reduced. Behavioralobservations after experimental shell relocation further showedthat males recognized females that they had not spawned with:males directed more exploration and manipulation behavior towardsuch shells compared with controls. Reoccupation rate of emptiedexperimental shells was significantly higher than that of unmanipulatedempty shells. This indicates that shell stealing and nest takeover,followed by female expulsion, contribute to the reproductivesuccess of L. callipterus males. We also found that female matechoice reduces expulsion risk: females preferred to mate withlarge males, and male size correlated with dominance. We concludethat the limited availability of breeding substrate is a keydeterminant of both intrasexual competition and intersexualconflict in this species.