Sperm-derived sperm motility-initiating substance from amphioxus Branchiostoma belcheri

The sperm of amphioxus, Branchiostoma belcheri, were immotile when excised from the testis and suspended in seawater. The sperm became motile upon spawning in natural seawater, suggesting mechanisms that triggered sperm motility during spawning. When a male amphioxus that underwent spawning was transferred to a cup containing a small amount of natural seawater, and then the seawater containing the spawned sperm was centrifuged, the supernatant caused motility initiation in the immotile sperm from the testis. This sperm motility-initiating activity was also found in the supernatant of seawater in which immotile sperm from the testis were incubated overnight. These suggest that in the amphioxus, a sperm motility-initiating substance resides in the sperm, and upon spawning, the substance is transformed into a free and active form to activate the sperm. Partial purification of the substance revealed it as a small and heat-stable substance with maximum UV absorbance at 234 nm.     

Scallop Size does not Predict Amount or Rate of Induced Sperm Release

Within populations of broadcast spawning marine invertebrates such as scallops, larger animals typically have larger gonads. Presumably, this means those larger males have more sperm to release than small males. However, there has never been a direct test of whether larger males actually release more sperm, at a higher rate, during spawning. To address this, we compared the allometry of induced sperm release with that of reproductive investment (gonad weight) in ripe males of 2 species of scallops, Chlamys bifrons and Chlamys asperrima. We did not find that larger scallops released more sperm or released it faster than small scallops, and were able to reject the hypothesis that instantaneous sperm release was related to body size in the same way as gonad weight. Consequently, we speculate that if larger broadcast spawning males do release more sperm, they may do so by spawning on more occasions within a reproductive season.     

The ability of the amphipod Corophium volutator (Pallas) to follow chemical signals from con-specifics

The amphipod Corophium volutator (Pallas) is a tube-dwelling species that only occasionally leaves its burrow. Animals that do crawl on the sediment are mainly males, apparently in search of females. This study was designed to determine whether C. volutator males and females release chemical signals that attract con-specifics, and if so, whether these signals are gender specific. Laboratory experiments were conducted on the over-wintering generation during their reproductive season, using a Y-maze choice test apparatus. The results show that C. volutator females release chemical signals that attract males. In the field, the attractant may guide males in search of burrowed females. In addition, male odour tends to attract males, while females do not follow chemical signals from con-specifics. We thereby show on gender-specific chemical attraction of this species.     

Fertilization in an egg-brooding colonial ascidian does not vary with population density

The possibility that free-spawning marine organisms may be subject to fertilization failure at low population density (due to the effects of sperm dilution) has sparked much interest, but these effects have been demonstrated only in a few species that broadcast their eggs. Some egg-brooding species may overcome dilution effects by filtering low concentrations of sperm from seawater and fertilizing eggs throughout an extended period of time. We examined the effects of population density and size on fertilization in Botryllus schlosseri, a hermaphroditic colonial ascidian that free-spawns sperm, but broods eggs. We experimentally manipulated the size and density of mating groups and surveyed fertilization levels in natural populations that varied in density. Fertilization was not affected by variation in population size or density in either the experimental or natural populations. Near the end of the reproductive season, some eggs may have been fertilized too late to complete development, suggesting a temporal form of sperm limitation that has not been considered in other systems. We also detected greater variability in fertilization levels at lower population density. Nevertheless, these results suggest that caution must be used in extrapolating reported density effects on fertilization to all taxa of free-spawners; density effects may be reduced in brooders that have efficient sperm collection mechanisms.     

Effect of seawater temperature on reproductive seasonality and fecundity of Pseudoplexaura porosa (Cnidaria: Octocorallia): latitudinal variation in Caribbean gorgonian reproduction

Abstract. The majority of tagged colonies of Pseudoplexaura porosa in Bermuda were reproductive over 2 months in the summer. They spawned 5–8 d after the full moon, with a peak on the sixth day, similar to colonies in Panama. The months of spawning were August and September in 1998, but July and August in 1999 and 2000. This temporal difference between the months of spawning corresponded to inter-annual variations in seawater temperature profiles. Initial gamete development each year occurred only when the daily mean seawater temperature during the month before spawning exceeded 27°C. There was a significant positive relationship between reproductive effort (gamete volume) of colonies and rising seawater temperature in the month preceding spawning; this was true for both the initial and the second spawning months. The end of the reproductive season each year was triggered by the decline in seawater temperature past the summer maximum. The duration of the reproductive season of conspecifics at the central Caribbean reef of Panama is 2 months longer than in Bermuda. This can be explained by the smaller annual temperature range at the lower latitude and the earlier onset of temperatures favorable for gamete development. Fecundity estimates for members of P. porosa (mean oocyte and spermary densities) in Bermuda were lower than for conspecifics in Panama. The shorter reproductive season in Bermuda, in addition to the lower fecundity of colonies, indicates that reproduction in P. porosa is compromised at this high latitude reef.     

Ocean acidification during prefertilization chemical communication affects sperm success

Ocean acidification (OA) poses a major threat to marine organisms, particularly during reproduction when externally shed gametes are vulnerable to changes in seawater pH. Accordingly, several studies on OA have focused on how changes in seawater pH influence sperm behavior and/or rates of in vitro fertilization. By contrast, few studies have examined how pH influences prefertilization gamete interactions, which are crucial during natural spawning events in most externally fertilizing taxa. One mechanism of gamete interaction that forms an important component of fertilization in most taxa is communication between sperm and egg‐derived chemicals. These chemical signals, along with the physiological responses in sperm they elicit, are likely to be highly sensitive to changes in seawater chemistry. In this study, we experimentally tested this possibility using the blue mussel, Mytilus galloprovincialis, a species in which females have been shown to use egg‐derived chemicals to promote the success of sperm from genetically compatible males. We conducted trials in which sperm were allowed to swim in gradients of egg‐derived chemicals under different seawater CO₂ (and therefore pH) treatments. We found that sperm had elevated fertilization rates after swimming in the presence of egg‐derived chemicals in low pH (pH 7.6) compared with ambient (pH 8.0) seawater. This observed effect could have important implications for the reproductive fitness of external fertilizers, where gamete compatibility plays a critical role in modulating reproduction in many species. For example, elevated sperm fertilization rates might disrupt the eggs' capacity to avoid fertilizations by genetically incompatible sperm. Our findings highlight the need to understand how OA affects the multiple stages of sperm‐egg interactions and to develop approaches that disentangle the implications of OA for female, male, and population fitness.     

东海北部大管鞭虾的食性

根据对2009年6月至2010年5月在东海北部采集到的720尾大管鞭虾胃含物的分析,对不同季节、性别、个体大小以及发育条件下大管鞭虾的摄食习性进行了研究.结果表明: 东海大管鞭虾的饵料生物共有43种(包含未鉴定种),甲壳类、有孔虫类和多毛类为其主要的摄食群类,在食物中所占的数量百分比分别为34.7%、29.4%和12.4%.大管鞭虾的饵料组成具有明显的季节波动,春季以有孔虫、藻类和无脊椎卵为主要食物;夏、秋季以有孔虫、鱼类和多毛类为主要食物;冬季则以多毛类和有孔虫类为主要食物.其摄食强度在春季和夏季较高,秋季较低.不同甲长的个体食性变化显著,小于24 mm的个体主食有孔虫类和藻类,24 mm以上的个体主食有多毛类和十足类;随着性腺发育雌性个体摄食等级呈现减弱的趋势,表明大管鞭虾摄食活动受到繁殖周期的影响.不同性别和不同季节大管鞭虾的食物重叠指数均较高,与栖息环境饵料资源的波动有关.大管鞭虾的营养级为2.67,属低级肉食性动物.     

东海北部大管鞭虾的食性

根据对2009年6月至2010年5月在东海北部采集到的720尾大管鞭虾胃含物的分析,对不同季节、性别、个体大小以及发育条件下大管鞭虾的摄食习性进行了研究.结果表明: 东海大管鞭虾的饵料生物共有43种(包含未鉴定种),甲壳类、有孔虫类和多毛类为其主要的摄食群类,在食物中所占的数量百分比分别为34.7%、29.4%和12.4%.大管鞭虾的饵料组成具有明显的季节波动,春季以有孔虫、藻类和无脊椎卵为主要食物;夏、秋季以有孔虫、鱼类和多毛类为主要食物;冬季则以多毛类和有孔虫类为主要食物.其摄食强度在春季和夏季较高,秋季较低.不同甲长的个体食性变化显著,小于24 mm的个体主食有孔虫类和藻类,24 mm以上的个体主食有多毛类和十足类;随着性腺发育雌性个体摄食等级呈现减弱的趋势,表明大管鞭虾摄食活动受到繁殖周期的影响.不同性别和不同季节大管鞭虾的食物重叠指数均较高,与栖息环境饵料资源的波动有关.大管鞭虾的营养级为2.67,属低级肉食性动物.     

Cytoplasmic activation of starfish oocytes by sperm and divalent ionophore A-23187

The relationship between onset of the early cytoplasmic stages of oocyte activation (vitelline membrane separation and elevation) and nuclear meiotic maturation was investigated in starfish oocytes after their exposure to divalent ionophore (A-23187) or sperm. Meiotically mature oocytes, isolated in calcium-free seawater, underwent activation in response to sperm or ionophore as previously reported. Large, immature starfish oocytes, arrested in prophase I of meiosis (germinal vesicle stage), underwent vitelline membrane elevation when treated with divalent ionophore A-23187 or starfish sperm. Histological studies demonstrated that cortical granule breakdown in the oocyte cortex was associated with vitelline membrane elevation after these treatments. Activation of oocytes by sperm occurred only in response to starfish sperm. Sea urchin, sand dollar, surf clam, or marine worm sperm did not induce vitelline membrane elevation of either immature or mature starfish oocytes. Sperm- or ionophore-activated immature oocytes underwent nuclear maturation after addition of the meiosis-inducing hormone, l-methyladenine; however, parthenogenetic development did not occur and embryonic development was markedly inhibited. In contrast to previous studies, the present results indicate that cytoplasmic activation can be initiated before and without hormone induction of the nuclear maturation process. Differentiation of the oocyte cell surface or cortex reactivity therefore appears to occur during oogenesis rather than as a consequence of maturation. The data further support the view that divalent ions mediate certain of the early activation responses initiated by sperm at the time of fertilization and that synchronization of fertilization to the meiotic process in the oocyte is important for the occurrence of normal development.     

Ca2+ spikes in the flagellum control chemotactic behavior of sperm

The events that occur during chemotaxis of sperm are only partly known. As an essential step toward determining the underlying mechanism, we have recorded Ca2+ dynamics in swimming sperm of marine invertebrates. Stimulation of the sea urchin Arbacia punctulata by the chemoattractant or by intracellular cGMP evokes Ca2+ spikes in the flagellum. A Ca2+ spike elicits a turn in the trajectory followed by a period of straight swimming ('turn-and-run'). The train of Ca2+ spikes gives rise to repetitive loop-like movements. When sperm swim in a concentration gradient of the attractant, the Ca2+ spikes and the stimulus function are synchronized, suggesting that precise timing of Ca2+ spikes controls navigation. We identified the peptide asterosap as a chemotactic factor of the starfish Asterias amurensis. The Ca2+ spikes and swimming behavior of sperm from starfish and sea urchin are similar, implying that the signaling pathway of chemotaxis has been conserved for almost 500 million years.     

Comparative analysis of reproductive traits in black-chinned tilapia females from various coastal marine, estuarine and freshwater ecosystems

The black-chinned tilapia Sarotherodon melanotheron is a marine teleost characterised by an extreme euryhalinity. However, beyond a certain threshold at very high salinity, the species exhibits impaired growth and precocious reproduction. In this study, the relationships between reproductive parameters, environmental salinity and condition factor were investigated in wild populations of this species that were sampled in two consecutive years (2003 and 2004) from three locations in Senegal with different salinities: Guiers lake (freshwater, 0 psu), Hann bay (seawater, 37 psu) and Saloum estuary (hypersaline water, 66-127 psu). The highest absolute fecundity and spawning weight were recorded in seawater by comparison to either freshwater or hypersaline water whereas the poorest condition factors were observed in the most saline sampling site. These results reflect higher resource allocation to the reproduction due to the lowest costs of adaptation to salinity in seawater (the natural environment of this species) rather than differences in food resources at sites and/or efficiency at foraging and prey availability. Fecundities, oocyte size as well as spawning weight were consistent from year to year. However, the relative fecundity in the Saloum estuary varied significantly between the dry and rainy raisons with higher values in the wet season, which seems to reflect seasonal variations in environmental salinity. Such a reproductive tactic of producing large amounts of eggs in the rainy season when the salinity in the estuary was lower, would give the fry a better chance at survival and therefore assures a high larval recruitment. An inverse correlation was found between relative fecundity and oocyte size at the two extreme salinity locations, indicating that S. melanotheron has different reproductive strategies in these ecosystems. The adaptive significance of these two reproductive modes is discussed in regard to the heavy osmotic constraint imposed by extreme salinities and high inter-specific competition.     

Evaluation of DNA damage as a quality marker for rainbow trout sperm cryopreservation and use of LDL as cryoprotectant

Defining reliable and objective biomarkers of sperm quality is a complex matter, because it does not rely on a particular characteristic of the milt. Susceptibility to cryopreservation varies between ejaculations and throughout the year, and the evaluation of fresh sperm does not always provide accurate information about their fertilization ability after freezing and thawing. DNA is one of the cell components prone to suffering cryodamage and several studies have pointed out the importance of the maintenance of its integrity during sperm cryostorage. The authors analysed sperm from rainbow trout for four weeks during the natural reproductive season. Viability, DNA integrity, and fertilization ability were evaluated. Furthermore, in order to increase membrane and DNA protection during sperm cryopreservation, the authors optimized the use of LDL fraction from egg yolk as a cryoprotectant during the analysed period. Results revealed that the evaluation of DNA damage in fresh sperm reveals subtle cell damage, not evidenced in fresh sperm by the other parameters. DNA fragmentation increased from 8 to 31% during the reproductive season, indicating pre-freezing differences that render the cells more susceptible to cryodamage. Also, the use of 12% LDL (low density lipoprotein) fraction, instead of the commonly used pure egg yolk, improved sperm quality after freezing. When LDL was used, post-thaw quality remained constant throughout the analysed period, providing around 60% of eyed embryos. In contrast, when egg yolk was used, post-thaw quality decreased significantly at the end of the season and the percentage of eyed embryos dropped from 60% to 27%. Results demonstrated that reduction in DNA integrity takes place during the reproductive season affecting susceptibility to cryodamage and that the protective effect of egg yolk is very much improved when only their LDL fraction is added to the cryopreservation extender.     

闽江口-平潭海域有机解磷菌多样性及群落特征

有机磷的微生物矿化是海洋磷循环的重要环节,开展有机解磷菌研究有助于揭示富营养化海域有机磷矿化的微生物驱动机制。以phoX为标志基因,采用Illumina高通量测序技术分析了闽江口-平潭海域2019年4月(春季)及7月(夏季)有机解磷菌的多样性及群落特征。结果表明: 表层海水样品中有机解磷菌的Shannon指数介于3.21～7.91,各站位多样性春季均大于夏季;沉积物样品中有机解磷菌的Shannon指数介于2.04～8.70,各站位多样性夏季大于春季。春季各站位表层海水有机解磷菌的Shannon指数均高于沉积物,而夏季则相反。表层海水中检测到9个门类的有机解磷菌,主要为变形菌门和蓝细菌门;沉积物中则检测到12个门类,主要为变形菌门和拟杆菌门。在属水平上,有机解磷菌的群落组成呈现时空变异特征。表层海水样品中,春季以雷辛格氏菌属、褐杆菌属、深海球菌属和假单胞菌属等属为主,夏季则以聚球藻属、Halioglobus属、玫瑰变色菌属、褐杆菌属、亚硫杆菌属和生丝单胞菌属等属为主。在沉积物样品中,春季主要菌属包括雷辛格氏菌属、褐杆菌属、弧菌属和亚硫杆菌属;夏季主要菌属包括固氮螺丝菌属、氨基杆菌属、Sulfurifustis属、伯克氏菌属和Thiohalobacter属。此外,在表层海水和沉积物样品中均检测到大量未分类的有机解磷菌。冗余分析显示,溶解氧、水温、pH、可溶性无机氮、亚硝态氮、硝态氮等对闽江口-平潭海域表层海水有机解磷菌群落分布影响较大。表层海水和沉积物中存在的丰富有机解磷菌可能在该海域的磷循环中发挥重要作用。     

Biochemical Studies on the Acrosome Reaction of the Starfish, Asterias Amurensis I. Factors Participating in the Acrosome Reaction

In contrast with the case in sea urchin sperm, in starfish the acrosome reaction is not spontaneously induced by simply increasing the extracellular Ca²⁺ concentration or pH. At higher pHs, starfish sperm undergo morphological changes accompanied by exocytosis of the acrosomal vacuole, but they do not form acrosomal filaments. Nomarski-microscopic observation confirmed that spermatozoa undergo the acrosome reaction within the jelly coat. Acrosome reaction-inducing substance, a glycoprotein from the egg jelly, required a diffusible cofactor(s) present in the egg jelly for full activity. Several lines of evidence showed that this diffusible factor(s) is not merely Ca²⁺.     

Changes in Solea senegalensis sperm quality throughout the year

Some of the Senegalese sole (Solea senegalensis) broodstock reproductive constraints are related to sperm quality. Although they present two defined spawning season (spring and autumn), males gave semen during all the year thus an exhaustive annual sperm analysis is important to determine the seasonal changes in semen quality. Sampling was performed monthly during one year, analyzing different cellular parameters to better understand sperm quality limitations obstructing sole mass production. The percentage of progressive motile cells and their linear velocity showed a decrease from March (beginning of the first spawning season) to July (when the highest temperatures were observed), followed by a slight increase in August and October (second spawning season). DNA fragmentation values showed highest values between the two spawning seasons and decreased to the end of the year. The percentage of apoptotic cells was lowest in March (beginning of the first spawning season) and the highest in November. The percentage of cells resistant to seawater exposure presented two peaks related with both spawning seasons. There was a tendency for the semen to attain a quality peak between the beginning and the middle of the first spawning season (March-May), followed by a pronounced decrease, achieving the lowest values during the months with the highest temperature. Also, the different males present in the broodstocks reach their sperm quality peak at different times, which will result in an unequal contribution for the next generation.     

The annual reproductive cycle of the Smooth newt (Triturus vulgaris) in England

Newts were collected throughout the year from both breeding ponds and terrestrial sites and were weighed, measured and dissected; in males, the testes were examined histologically. Smooth newts show post-nuptial gametogenesis such that, during late summer and autumn they are producing mature gametes for the following year's breeding season. In males, the testes are at their smallest size during the spring, when they consist mostly of immature sperm cysts and evacuated tissue, mature sperm having been evacuated into the vasa deferentia during the newts' migration to water. Evacuated testicular tissue is glandular in function and there is evidence that secretions from this tissue control the development of secondary sexual characters: the dorsal crest, fringes of skin on the toes and the dorsal cloacal gland. In both sexes, fat body and liver weights are lowest in the spring and increase in the autumn. In females, oocytes vary in size, depending on the amount of yolk they contain. Only the larger oocytes are laid in a current breeding season, the smaller ones being retained and yolked in late summer and autumn. In both sexes, measures of fecundity (testis size in males, oocyte number in females) are strongly correlated with body size. The finding that male newts have a finite supply of sperm during the breeding season leads to an interpretation of various aspects of male courtship behaviour. These are adaptations for conserving sperm and allocating it to courtship encounters in a way likely to promote male reproductive success.     

Peptide-induced hyperactivation-like vigorous flagellar movement in starfish sperm

Asterosap, a sperm-activating peptide (SAP) from the starfish egg jelly coat, is diffusible and controls a cGMP-signalling pathway in starfish sperm in the same manner as resact, a potent chemoattracting SAP in sea urchins. This fact suggests that asterosap may serve as a chemoattractant like resact at concentrations with appropriate gradients. Since asterosap is one of three egg jelly components, which in concert induce the acrosome reaction, it is still worthwhile to evaluate how asterosap modulates sperm motility prior to this reaction. We analysed the flagellar movement of sperm of the starfish Aphelasterias japonica in artificial seawater (ASW) containing the asterosap isoform P15 at 1 micromol l(-1). We found that sperm swim straighter with more symmetrical flagellar movement in P15 than in ASW, but without any significant difference in the flagellar beat frequency and the swimming velocity. The flagellar movement is, however, dramatically different between sperm firmly attached to the solid surface by the head in P15 and those attached in ASW: in P15 the flagellum bends to a greater extent, with higher curvature and with higher shear angle up to a right angle to the flagellar wave axis, and beats at an increased frequency. The vigorous flagellar movement of sperm, which can be activated when sperm are placed in high-load circumstances just as entering into a jelly layer, may increase propulsive forces and hydrodynamic resistances, allowing sperm to undergo the acrosome reaction as effectively as possible.     

Anion Channel Blockers Inhibit the Acrosome Reaction of Echinoderm Sperm

Two types of anion channel blockers, SITS (4-acetamide-4'-isothiocyanostilbene-2,2'-disulfonic acid) and DIDS (4,4'-diisothiocyanostilbene-2, 2'-disulfonic acid), inhibited jelly-induced acrosorne reaction in starfish and sea urchin. In starfish sperm, both of the blockers reversibly inhibited the formation of acrosomal process but they had no effect on either the acrosomal exocytosis or acid release from the sperm. Complete acrosome reaction occurred even in Cl⁻- and SO₄²⁻-free artificial seawater whereas HCO₃⁻was required for the acrosomal exocytosis. Importance of anion transport in acrosome reaction is discussed.     

Morphology and evolutionary implications of the annual cycle of secretion and sperm storage in spermathecae of the salamander Ambystoma opacum (Amphibia: Ambystomatidae)

Females of the marbled salamander, Ambystoma opacum, store sperm in exocrine glands called spermathecae in the roof of the cloaca. Eggs are fertilized by sperm released from the spermathecae during oviposition. Some sperm remain in the spermathecae following oviposition, but these sperm degenerate within a month and none persists more than 6 mo after oviposition. Thus, sperm storage between successive breeding seasons does not occur. Apical secretory vaculoes are abundant during the fall mating season and contain a substance that is alcian blue+ at pH 2.5. Production of secretory vacuoles decreases markedly after oviposition, and the glands are inactive by the summer months. Ambystoma opacum is a terrestrial breeder, and some mating occurs prior to arrival at pond basins where oviposition occurs. Mating prior to arrival at the ovipository site may prolong the breeding season, leading to fitness implications for both males and females. Females have opportunities for more matings, and the possibilities for sperm competition in the spermathecae are enhanced. © 1995 Wiley-Liss, Inc.