Seasonal variations in sperm production and sperm quality in male winter flounder, Pleuronectes americanus: the effects of hypophysectomy, pituitary replacement therapy, and GnRH-A treatment

The pattern of sperm release and changes in sperm quality were investigated throughout the normal spawning season of male winter flounder, Pleuronectes americanus (Walbaum), caught in Conception Bay, Newfoundland, Canada in April and November 1994. In addition, the impact of hormonal manipulation on sperm production and sperm quality was examined through gonadotropin-releasing hormone analogue (GnRH-A) treatment, hypophysectomy, and pituitary extract injection – pituitary replacement therapy (PRT). While a lengthy period of spermiation, lasting ∼6 months (December to July), can be detected in some males, the major period for sperm release occurred in May to July in the summer spawning season. The spawning period started with peak levels of plasma testosterone (T) and 11 ketotestosterone (11-KT). By late July sperm production fell rapidly along with a notable deterioration in sperm motility. While GnRH-A treatment advanced the period of sperm release, and the milt became diluted and increased in volume, the treatment did not increase total sperm output over the spawning season. In hypophysectomized male flounder, by contrast, milt was more concentrated and sperm production declined along with reduced plasma androgen levels during the spawning season. Increased plasma T and 11-KT levels in hypophysectomized males followed PRT in December and January. Moreover, PRT appeared to be responsible for advancement of spermiation in a large number of prespawning males, which released an increased amount of milt. Finally, these studies indicated that both sperm motility and egg fertilization rates remained unchanged after these hormonal treatments. Received: 5 September 1997 / Accepted: 6 January 1999     

Plasma sex steroid profiles and the seasonal reproductive cycle in male and female winter flounder,Pleuronectes americanus

The pattern of seasonal gonadal development and variations in plasma sex steroids were investigated in adult male and female winter flounder, Pleuronectes americanus (Walbaum), from Conception Bay Newfoundland beginning August 1987 to December 1988. The winter flounder reproductive cycle can be divided into five consecutive phases of relative reproductive activity including: (1) rapid gonadal recrudescence in the fall (August–December); (2) continued slow gondadal growth in females, or maintenance of the well developed gonads in males, during the winter (December–February); (3) a prespawning phase of gonadal maintenance in the spring (March–April); (4) spawning early in the summer (May–June) after the female gonads reach peak weight; and (5) the summer postspawned period (June–August) when the gonads remain regressed. Female gonadal recrudescence in August is characterized by small increases in plasma estrogen levels and recruitment of small oocytes (150 m) into yolk accumulation. For the winter months, estradiol-17 levels in the plasma remain stable, approximately 15 ng ml^-1, until rising again together with testosterone to peak hormone levels just prior to spawning in conjunction with the highest seasonal values for the gonadosomatic index (GSI) and oocyte diameter. After spawning, the ovaries are regressed and sex steroids in the plasma fall to very low levels. Rapid seasonal recrudescence of the gonads in males is evident from rising GSI values, which reach a maximum in October, and from substantial early seasonal increases in plasma testosterone and 11-ketotestosterone. Afterwards, although GSI values subsequently decline presumably reflecting the process of spermiogenesis and/or onset of spermiation in some males, the testes remain relatively well developed with the presence of sperm throughout the spawning season. As the proportion of spermiating males increases, the plasma levels of the androgenic steroid hormones rise to peak circulating levels from April to June. At the end of the summer reproductive season, the testes of post-spawned males become regressed and plasma testosterone and 11-ketotestosterone levels fall, reaching the lowest seasonal values.     

Control and coordination of reproduction and molting in the spiny lobster Panulirus argus

Behavioral and physiological correlates of vernal reproduction and molting were investigated experimentally in laboratory-held spiny lobsters (Panulirus argus Latreille) from Florida to determine their control by photoperiod and temperature. Behavioral measures included courtship, copulation and aggression; physiological measures included the timing and frequency of ecdysis, spermatophore emplacement, oviposition, egg hatching, and setal (pleopod) and gonadal development. Behavior resembled that observed in nature, as evidenced in the expression of complete reproductive cycles. Long daylengths and warmer temperatures enhanced courtship, spawning frequencies, and female gonadal development, although gonadal recrudescence occurred in large females irrespective of photoperiod. A significant interaction effect between photoperiod and temperature indicated a need to examine joint, non-independent factor effects upon reproductive parameters. Photoperiod and temperature did not significantly influence aggression and male gonadal development. Photoperiod did not affect molting rates significantly. However, warm temperatures enhanced molting and growth such that sub-adult and young adult lobsters (both sexes) had highest molting rates, followed by large adult males, which in turn had higher rates than reproductively active females. Female setal development correlated positively with female size, and changed abruptly at ecdysis. Although setal lengths were highly variable, setae of 9 mm or more signified sexual maturity. Reproduction and molting were size-dependent: the largest adults mated or spawned, the smallest molted, and intermediate-sized ones either mated, or molted before mating. Spiny-lobster reproductive and molting patterns were influenced by photoperiod and temperature in a complex manner depending upon sex, season, size and developmental state.     

Plasma sex steroid and vitellogenin profiles during gonad development in wild Mediterranean amberjack (<Emphasis Type="Italic">Seriola dumerilii</Emphasis>)

Mediterranean amberjacks, Seriola dumerilii Risso, were caught off the Pelagie Islands, in the south Mediterranean Sea, between May 1997 and June 1999. Fish blood was sampled, and gonads were collected at 10-day intervals throughout the spawning period and at monthly intervals during the resting period. Concentrations of plasma estradiol-17ß (E₂), testosterone (T), 17,20ß-dihydroxy-4-pregnen-3-one (DHP) and vitellogenin (Vtg) in females; and plasma T and 11-ketoT (11-KT) in males were correlated with changes in gonadal development. The first females that had already ovulated (F5) were found in late May. Most mature females (F4) were caught in June. Post-spawned females (F6) were found from late July until September. Estradiol-17ß was at baseline levels (<0.02 ng ml^–1) during autumn/winter and rapidly peaked (6.29±0.68 ng ml^–1) from May to early July. Plasma T levels showed a similar profile and were positively correlated to E₂ (r²=0.668) during the spawning period. Continuously elevated levels of E₂ and T were observed during the spawning season in vitellogenic females (stages F3, F4 and F5). Resting females were found during the autumn/winter months. Vitellogenin levels increased during May and peaked in June, reverting to undetectable levels in August, in parallel with sex steroid changes. Plasma DHP levels peaked in June (283.45±97.3 pg ml^–1), falling to basal values (<5 pg ml^–1) in August. DHP levels were higher in mature females (F4) than in maturing (F3) and in partially ovulated (F5) females. DHP values increased during germinal vesicle migration, peaked during germinal vesicle breakdown and decreased again during complete oocyte hydration. In males, changes in T and 11-KT plasma levels were related to testis development. The highest levels of T (5.76±2.64 ng ml^–1) were measured during spermatogenesis and highest 11-KT (5.28±3.6 ng ml^–1) in males with milt, from May to June. This study provides information, for the first time, on the relation between plasma sex steroid profiles and gonad development in wild Mediterranean amberjack, a useful benchmark for broodstock monitoring under controlled conditions.Communicated by R. Cattaneo-Vietti, Genova     

Interactions among endocrinology,seasonal reproductive cycles and the nesting biology of the female green sea turtle

We collected data on plasma levels of testosterone+5a-dihydrotestosterone (T+DHT) and corticosterone (CORT) from adult female green sea turtles (Chelonia mydas) from southern Queensland during distinct stages of their reproductive cycle. Those females capable of breeding in a given year had elevated plasma steroid levels (T+DHT 0.91ǂ.08; CORT 1.05ǂ.29 ng/ml), associated with follicular development, until courtship began in October. At the beginning of the nesting season in November plasma levels of CORT were related to when the female first nested (r²=0.06; F=10.45; P=0.01). However, they were not correlated with the number of clutches a female laid in that season (F=3.65; P=0.08). We repeatedly sampled 23 turtles over the nesting season and profiled changes in steroids immediately following oviposition of each clutch. Levels of T+DHT (range 0.41-0.58 ng/ml) and CORT (range 2.13-2.81 ng/ml) were similar through the early stages of the nesting season and inter-nesting period, and declined to near basal levels (T+DHT 0.37ǂ.03 and CORT 1.85-ng/ml) following the last clutch for the season. Steroid hormone levels were also low (T+DHT 0.38ǂ.16; CORT 0.46ǂ.21 ng/ml) in four independent post-breeding (atretic) females; samples for these females were taken at a time when body condition was presumably at the lowest for the season. Subtle changes in the nesting environment, such as variation in nesting habitat or the time of night that nesting occurred, were associated with a small and slow CORT increase. We suggest CORT is increased in nesting females to assist in lipid transfer to prepare the ovarian follicles and/or the reproductive organs for ovulation.     

Sexual selection on mature male parr of masu salmon (Oncorhynchus masou): does sneaking behavior favor small body size and less-developed sexual characters?

In many salmonid species, males exhibit morphological dimorphism associated with alternative mating behaviors. ”Precocious males” have a small body size with little or no development of sexual characters and adopt sneaking to gain access to females, while ”migratory males” of large body size and well-developed secondary sexual characters fight. We quantified selection on precocious male parr of masu salmon (Oncorhynchus masou) under simulated natural conditions to examine the contribution of morphology to sneaking success. In contrast to the prediction that sneaking behavior favors small body size, we detected selection favoring relatively large body size for sneaking success. This selection pressure was caused by the dominance hierarchy within parr and may have been facilitated by indifference of dominant migratory males to parr. Unlike the secondary sexual characters exhibited by migratory male salmon, such as the hooked snout and humped back, no morphological characters other than body size contributed to the reproductive success of masu salmon parr. This non-contribution may have been responsible for the lack of development of sexual characters in precocious males. Received: 15 November 1999 / Accepted: 20 May 2000     

Changes in dopamine,norepinephrine and serotonin levels in the pituitary,telencephalon and hypothalamus during gonadal development of male <Emphasis Type="Italic">Lutjanus argentiventris</Emphasis> (Teleostei)

Biogenic amines are important in the regulation of certain aspects of teleost reproduction, but relatively little is known about variations in their levels in the nervous system during the reproductive cycle. The aim of this study was to investigate changes in the levels of dopamine (DA), norepinephrine (NE), serotonin (5-HT), the DA metabolite 3,4-dihydroxyphenylacetic acid (DOPAC) and the 5-HT metabolite 5-hydroxyindoleacetic acid (5-HIAA) in the pituitary and brain (hypothalamus and telencephalon) of the adult male of the tropical marine yellow snapper Lutjanus argentiventris during the reproductive cycle (quiescent, prespawning and spawning periods). Gonad weight, gonad/body weight ratio and plasma-testosterone levels increased steadily throughout gonadal development, but 17#-estradiol levels did not vary significantly. Pituitary DA and DOPAC levels and the DOPAC/DA ratio declined with gonadal development, whereas hypothalamic DOPAC level and DOPAC/DA ratio increased. In the telencephalon, DA and DOPAC levels and DOPAC/DA ratio all peaked in the prespawning period; however, DA level was lowest in the spawning period, whereas DOPAC level and DOPAC/DA ratio were both lowest in the quiescent period. Telencephalic NE, 5-HT and 5-HIAA levels likewise all peaked in the prespawning period, and were lowest in the spawning period. Telencephalic 5-HIAA/5-HT ratio peaked in the quiescent period and was lowest in the spawning period. No significant variation was observed either in pituitary NE level or in hypothalamic NE, 5-HT and 5-HIAA levels. The present results indicate that males of L. argentiventris show a decline in pituitary DA activity during gonadal development. We discuss the possible involvement of biogenic amines in the regulation of GTH II secretion, through direct action on the pituitary or indirect modulation of gonadotrophin-releasing hormone neuron activity.     

Is avian humoral immunocompetence suppressed by testosterone?

A key issue in sexual selection theory is how a correlation between male secondary sexual characters and male genetic quality can be maintained. The immunocompetence-handicap hypothesis proposes that testosterone-dependent male characters remain honest signals because of the immunosuppressive effect of elevated steroid hormone levels. The hypothesis requires that physiological levels of testosterone depress immune system function. We quantified testosterone titers and humoral immunocompetence of captive male and female red-winged blackbirds (Agelaius phoeniceus) at three points in the annual cycle (autumn, prebreeding, and breeding). We also conducted an implant experiment on the males to assess the effects of prolonged, above-normal testosterone titers on humoral immune responses. Humoral immunocompetence was measured as secondary antibody production to a non-pathogenic protein antigen, keyhole limpet hemocyanin, using an enzyme-linked immunosorbent assay we developed for A. phoeniceus. Secondary antibody responses of individuals were highly repeatable between sampling periods. Neither physiological nor above-normal levels of plasma testosterone suppressed secondary antibody production. In paired tests of the same individuals between prebreeding and breeding, and between breeding and implant, plasma testosterone increased significantly but secondary antibody responses were unaffected. We are confident in these results because with 80% power, an 11–14% difference in antibody titers would have been detected. There was no relationship between plasma testosterone levels and humoral immunocompetence in free-ranging males tested at the peak of breeding. These results cast doubt on a key assumption of the immunocompetence-handicap hypothesis. Received: 13 April 1998 / Accepted after revision: 7 October 1998     

Reproductive success of nesting male longear sunfish (Lepomis megalotis peltastes)

Summary Although a few male longear sunfish nest solitarily, most territorial males excavate their nest in dense aggregations. The importance of sexual selection in the evolution of this breeding system was evaluated by 1) examining the influence of certain male and nest characteristics on spawning success and 2) by comparing the success of social and solitary nesters. Among group nesters, females spawned preferentially with males nesting early within a spawning period and occupying central nests. Male size and nest diameter were negatively correlated with nesting day and hence spawning success. However, female discrimination using these traits directly could not be demonstrated. The reproductive success of social and solitary males was compared by counting the larvae in their nests several days after spawning. Successful males (with larvae) were more likely than males without larvae to be encountered later in the breeding season and to nest in small groups or solitarily. Spawning period (of which there were 5 or 6 in a season) was significantly correlated with larval abundance in the nest, while male size was not. Social and solitary males were equally successful, but aggregations may nonetheless result from sexual selection: males unlikely to attract females may nest around more attractive males to steal fertilizations from them. High spawning synchrony may prevent attractive males from renesting elsewhere once other nesters aggregate around them.     

Solitary nesting as an alternative breeding tactic in colonial nesting bluegill sunfish (Lepomis macrochirus)

Colonial breeding can evolve in response to benefits afforded by clumped individuals, such as reduced predation and increased ease of assessing potential mates. However, colonial breeding can also impose costs such as increased disease transmission or increased cuckoldry. Here, we investigate solitary nesting as a potential alternative breeding tactic in colonial breeding bluegill sunfish (Lepomis macrochirus). Most male bluegill, termed parentals, compete for nesting sites in colonies and then court and spawn with females and provide sole care of the eggs. Although nesting in a colony results in reduced predation and fungal infection of broods, it comes at a cost of increased parasitism by specialized cuckolder males that do not nest. We found that 4.5% of parentals forgo spawning in a colony and instead construct nests solitarily. Solitary males were of similar size and age to colonial males, but were in significantly better condition. Solitary males obtained as many eggs as males nesting in the center of colonies, and significantly more than males nesting on the periphery of colonies. Thus, females do not appear to discriminate against solitary males. Solitary males had smaller ear tabs, a presumed sexually selected character used by parental males in intrasexual competition, than colonial males. Tracking data revealed consistency in nesting tactic (but not nest position within the colony) between spawning attempts. We suggest that solitary nesting represents either a facultative decision made by parental males in top condition at the onset of breeding, or a life history decision to forgo spawning in colonies.Communicated by K. Lindström     

Changes in the content and fatty acid composition of lipid in an isolated population of the capelin Mallotus villosus during sexual maturation and spawning

Changes in the wet weights and lipid contents of muscle, liver and gonad were determined in male and female Mallotus villosus in Balsfjorden, northern Norway, in 1981, from January, when gonadal development becomes noticeable, until May when the fish are spawning. Fatty acid compositions of tissue lipids were also determined. Over 4 mo prior to spawning, the weight of muscle in female capelin decreased by 32% while the weight of the ovary increased exponentially by 830%. In males the weight of the muscle remained constant and that of the testis decreased slightly. The lipid contents of the muscle of both males and females decreased by 76% over the period and an inverse relationship existed between the water and lipid contents of muscle in both sexes. Male liver weight remained constant over the period of study whereas female liver weight increased transiently by 300% between January and March. 38% of the lipid lost from female muscle was accounted for by lipid deposited in ovary whereas negligible amounts of the lipid lost from male muscle was accounted for by lipid in the testis. Gonadal lipid was always richer in polyunsaturated fatty acids than muscle lipid and, immediately prior to spawning, 42% of the fatty acids in ovarian lipid were polyunsaturated. Muscle lipid of males and females showed a progressive increase in the percentage of the long-chain monoenes 20:1 and 22:1 between January and May. It is concluded that male capelin catabolise more of their muscle lipid reserves than females in the 4 mo prior to spawning and that most of the lipid catabolism in males is associated with physical activity. Conversely, females deposit much more of their muscle lipid in gonads than males, although considerable selectivity occurs in the mobilisation of fatty acids from muscle lipid into ovarian lipid. Additionally, biosynthesis of gonadal constituents accounts for a considerable proportion of the lipid catabolised in females.     

Sperm characteristics associated with different male reproductive tactics in bluegills (Lepomis macrochirus)

We examined the availability and motility of sperm from parental and sneaker male bluegills (Lepomis macrochirus), a colonially nesting sunfish (Family Centrarchidae) with male parental care and a high incidence of cuckoldry by both sneaker and satellite males. We found no differences between sneakers and parentals in length and swimming speed of sperm, or percent and duration of sperm activity. In sneaker milt, however, sperm was almost 50% more concentrated than in parental milt (16.5×10⁶ vs 11.5×10⁶ sperm/μl of milt, respectively). Despite this difference in sperm concentration, stripped ejaculates from sneakers contained almost 400 million fewer sperm (only 32% as many sperm) than those from parentals due to their much smaller stripped ejaculate volumes (only about 19% that of parentals). Thus unless sneakers can compensate by releasing more sperm or gaining closer proximity to eggs at the time of spawning, they may be at a disadvantage with respect to sperm competition. We discuss these results in relation to models for the evolution of alternative reproductive behaviours in this species and suggest that the cuckolders may be making the best of a bad situation. Received: 18 February 2000 / Revised: 23 March 2000 / Accepted: 14 September 2000     

Why do males tolerate sneakers? Tests with the European bitterling, Rhodeus sericeus

In most species, males attack other males that attempt to gain fertilizations through sneak copulations. Here we report on a system where dominant males show a low level of aggression against sneakers at the initial stages of territory establishment. Females of the European bitterling, Rhodeus sericeus, lay their eggs in living mussels and males fertilize the eggs by releasing sperm over the mussels both before and after egg laying. When we allowed males to court females to a mussel containing no eggs at different male densities - one, two, four, or six males - the dominant male showed a low level of aggression against other males that released sperm. The dominant male became aggressive toward the other males only after eggs had been laid. This unusual pattern could be due to either some benefit of accepting sneakers or a high cost of aggression. We found support for both possibilities. The presence of several males decreased the time until a female spawned, whereas increased aggression by the dominant male against other males during a second female presentation, when the male was more territorial, interrupted courtship and increased the time until spawning. Females appeared to be attracted by both the presence of several males around a mussel and increased courtship under male competition. The bitterling mating system possibly differs from that of other species due to lack of investment in nest building and parental care, and high costs of defending the spawning site against sneakers.     

Does female aggression prevent polygyny? An experiment with pied flycatchers (Ficedula hypoleuca)

The female aggression hypothesis states that resident females may be able to prevent polygyny by behaving aggressively towards intruding females. A critical test of the hypothesis is to provide prospecting females with a choice between displaying mated males some of which have initial mates with artificially reduced levels of aggressiveness. Here we present a mate choice experiment on pied flycatchers Ficedula hypoleuca. The species is a cavity nester, and resident females were prevented from behaving aggressively by enclosing them within their own nestboxes: narrowing the entrance hole so that they could not escape but could still let their head out and have some contact with their mate. This treament had only a minor influence on male behaviour. We studied whether the experimental males were better able to attract a new female than a control group of mated males. Four predictions from the female aggression hypothesis were supported. (1) Mating success of control males was positively related to the distance between their primary and secondary territory. (2) For experimental males, mating success was unrelated to interterritorial distance. (3) Experimental males had higher mating success than control males when the interterritorial distance was short but (4) not when it was long. Experimental males had much lower mating success than unmated males, as would be expected if prospecting females are able to discover male mating status from cues other than visits by primary females to their mates' secondary nest sites. Received: 5 January 1998 / Accepted after revision: 30 December 1998     

Correlates between number of mates,shelter availability and reproductive behavior in the Tautog Tautoga onitis

Earlier studies on reproductive behavior of the tautog Tautoga onitis under laboratory conditions indicated that differences in courtship and spawning behavior were apparently related to the number of males attempting to spawn with a female. It had also been conjectured that certain ecological factors such as availability and distribution of food and shelter would play a role in the establishment of territoriality and defense of a female, indirectly influencing courtship activities and the resultant spawning mode. In the present work we have examined this question more closely by varying the number of males, females and amount of shelter available. Regardless of the social situation, in every instance single males, through aggression, attempted to limit access to a female and pair spawn. The degree to which this succeeded depended on the male's place in the dominance hierarchy, the number of males in proximity which were actively attempting to spawn and the availability of shelter. Courtship activity of the female was influenced by the amount of male attention. When attention was high, the nuptial shading of the female never developed maximally. On the other hand, when male attention was low, the female showed progressive and maximal shading development to attract males. The significance of the findings is discussed.     

Experimental induction of gonadal maturation and spawning in the giant jellyfish <Emphasis Type="Italic">Nemopilema nomurai</Emphasis> (Scyphozoa: Rhizostomeae)

We studied the processes of gonadal maturation, spawning, fertilization and embryonic development of the giant jellyfish Nemopilema nomurai, which has been blooming in recent years in the Sea of Japan. Healthy medusae actively swimming offshore had immature gonads, but damaged and ashore-drifted animals had relatively mature gonads. The animals maintained in a small net on the nearshore to mimic the drifted condition showed induced gonadal maturation by the increase in oocyte diameter and darkness in the matrix. A similar maturation process also occurred in isolated pieces of ovary incubated at the same temperature. Fully grown oocytes that probably stopped at the prophase of the first meiosis reinitiated maturation divisions in response to light exposure, and massive spawnings occurred 80–100 min after the exposure. The spawned oocytes were first released within the subgenital sinus, then transported peripherally and finally shed into the gastrovascular cavity. Maturation and spawning in male gonads were similar to those of females except that the male spawning occurred within 30 min after the light exposure and always preceded the female spawning. Hence, it was suggested that fertilization might take place in the female gastrovascular cavity by the sperm that came in from surrounding seawater by animals’ pumping activities after dawn. Having mechanical damages might promote fertilization success of this species in the Sea of Japan.     

Plasticity in spawning visits of female damselfish, Stegastes nigricans : effect of distance to mates

Both sexes of the damselfish Stegastes nigricans hold individual territories in which they feed on filamentous algae. At dawn, females visit males' territories to spawn, and the males guard the eggs until hatching. We examined how females' spawning behavior varied according to the distances to their mates. Females usually mated with a single male per morning. The distance to the territory of a mate (0.7–12.8 m) did not affect a female's total spawning time per morning, but affected the number of her spawning visits with that male. Females made many repeated spawning visits when spawning with males at short distances, while they spawned the entire clutch in one visit when spawning with males at long distances. This plasticity in female behavior appears to be related to two costs during the spawning visits: (1) intrusions by other fish to feed on algae in the female's territory during her absence, which may cause the female to return repeatedly to her territory for defense, and (2) attacks on the female by other territorial fish, which increased with the distance to their mates' territories. To minimize the sum of both costs, females should change the number of spawning visits depending on the distance to the males' territories. Received: 30 September 1996 / Accepted after revision: 17 March 1997     

Reproductive investment reduces recuperation from exhaustive escape activity in the tropical scallop Euvola zizac

In scallops, mobilization of reserves from the adductor muscle to support maintenance and reproductive activity may impinge upon a major role of the adductor muscle, the movement of the valves during swimming and escape responses. The tropical scallop Euvola ziczac (Linnaeus 1758) invests energy reserves to different degrees during its two periods of reproduction each year. We evaluated the impact of reproductive investment on the escape responses (clapping capacity and recovery after exhaustive exercise) of E. ziczac sampled at different reproductive stages (immature, mature, spawned) during the two reproductive periods in 1997. We compared the escape response capacities with the levels of muscle energetic reserves (glycogen, proteins, and arginine phosphate) and muscle metabolic capacities [activity of the glycolytic enzymes: glycogen phosphorylase (GP), pyruvate kinase (PK), phosphofructokinase (PFK), octopine dehydrogenase (ODH), arginine kinase (AK), and the mitochondrial enzyme, citrate synthase (CS)]. Gonad size, gamete volume fraction, and levels of gonadal protein and lipid were greater for mature scallops during the first than during the second reproduction. Numbers of claps during escape responses (49–57) and levels of muscle arginine phosphate remained similar throughout the different reproductive stages in both reproductive periods. In contrast, recovery was slowed during gonadal maturation in both reproductive periods and during spawning in the first reproduction. Scallops generally took more time to regain their initial clapping capacity during the first (25–40 min) than during the second reproduction (20–30 min). Muscle glycogen decreased markedly during both gonadal maturation and spawning in both reproductions; whereas, muscle proteins decreased only in the first reproduction. The levels of most enzymes decreased during gonadal maturation in both reproductions, and also after spawning, particularly during the first reproduction. We concluded that gonadal maturation and spawning did not decrease clapping capacity of E. ziczac, but decreased its capacity to recover from exhaustive exercise most likely due to decreased levels of energetic reserves and a reduced metabolic capacity of the adductor muscle. Moreover, these effects were probably stronger during the first cycle because of the greater reproductive investment coincident with decreased food availability. Received: 28 April 2000 / Accepted: 21 September 2000     

Body color change and serum steroid hormone levels throughout the process of sex change in the adult wrasse, <Emphasis Type="Italic">Pseudolabrus sieboldi</Emphasis>

Gonadal sex steroid hormones are the principal factors that directly control the gonadal and morphological alterations during sex change in hermaphrodite fish; however, the physiological mechanism of action by which these hormones govern body coloration is poorly understood. The protogynous wrasse Pseudolabrus sieboldi is a good model for understanding the physiological mechanisms of gonadal and body color change during sex change in hermaphrodite fish. To obtain information on the relationship between sex steroids and body color change during the process of gonadal sex change, we analyzed body color, gonadal histology, and serum levels of sex steroids. Body color was analyzed using a quantitative analytical method based on the hue value. Compared to other body parts of the fish, the anal fin changed color the most, becoming increasingly redder in association with gonadal changes that converted ovaries to testes. Levels of serum 11-ketotestosterone (11KT) increased as the gonadal sex change proceeded, whereas no significant change was observed in estradiol-17β (E2) levels. Moreover, we found a significant correlation between the hue value of the anal fin and serum 11KT levels, but not E2 levels. These results suggest that androgen, but not estrogen, plays a principle role in the changes in both gonadal morphology and body color in the transformation from female to male in this species. To our knowledge, this is the first quantitative demonstration of the relationship between body color and serum steroid levels during sex change in fish.     

Vocalizations by Alaskan moose: female incitation of male aggression

Evidence of female fomentation of male–male aggression as a mechanism of mate choice is rare, especially in mammals. Female choice of mates in polygynous species may be masked by intense male competition or by males attempting to restrict female choice. We studied protest moans of female Alaskan moose Alces alces gigas in interior Alaska, USA, from 1987 to 1990, to determine if moans incited male–male aggression. Alaskan moose exhibit a mating system in which one dominant male (the harem master) herds, defends, courts, and attempts to mate with females in his harem. Protest moans were given by females only in response to courtship. We hypothesized that if protest moans were related to females reducing harassment and exercising mate choice, females should give protest moans more frequently when courted by small males and less often when courted by large males, and that rates of male–male aggression would be elevated following protest moans. Harems were composed of one large male, with a mean of 4.4 females (median = 3 females); 10% of 132 harems included ≥10 females. The temporal pattern of protest moans from late August through November was associated with, but tended to lag behind, mating behavior. The rate of protest moans given by females decreased with increasing size of males courting them. Male–male aggression was significantly less during periods without protest moans than during periods in which protest moans occurred. These results indicate that female moose gave protest moans to reduce harassment by smaller males, and assure a mating opportunity with the most dominant male. Such a subtle mechanism of indirect mate choice by females may occur in other vertebrates in which choice is limited by a mating system in which male–male combat and male dominance over females reduces opportunities for female choice. The importance of female choice may be undervalued in studies of sexual selection in mammals.