Long lasting stickleback sperm; is ovarian fluid a key to success in fresh water?

Although the sperm of externally fertilizing fishes usually has a brief life span of up to a few minutes, this study showed that the sperm of the three-spined stickleback Gasterosteus aculeatus moved for several hours in brackish water and up to at least 10 h in the presence of ovarian fluid. Three-spined sticklebacks were able to spawn in waters ranging from full-strength sea water to fresh water, an ability unusual among fishes. The influence of salinity on sperm motility was examined, using three-spined sticklebacks from sea (salinity 30), brackish (5·5) and freshwater (0) populations. All three populations were found to have sperm with long motility periods in brackish water, lasting 165–270 min. Seawater three-spined sticklebacks had sperm motile for up to 65 min in sea water, whereas sperm from fresh- and brackish-water fish were quiescent in this medium. In fresh water, sperm from all three populations showed a very brief motility period, lasting <60 s. The presence of ovarian fluid, however, prolonged the motility period of sperm from both fresh- and brackish-water three-spined sticklebacks, for up to 7 and 10 h in fresh and brackish water, respectively, with some sperm found to be motile for up to 24 h. The results indicated that ovarian fluid created a favourable environment for the sperm and might have facilitated the three-spined sticklebacks' successful penetration of fresh water.     

Male traits influence odour‐based mate choice in the three‐spined stickleback

Female three‐spined sticklebacks Gasterosteus aculeatus preferred odour stemming from males with a nest over 'neutral' water and odour from males without any nest in an experimental flow channel. Males without any nest were preferred over females when either having the same major histocompatibility complex (MHC) class lI genotype or being unattractive in respect to MHC class II alleles.     

Relationships between sperm morphometry and sperm motility in the Atlantic salmon

Relationships between spermatozoal design and swimming behaviour were investigated using the significant natural variance in sperm traits in Atlantic salmon Salmo salar. In vitro motility and fertilization experiments were conducted with 86 Atlantic salmon to measure sperm form and function under natural fertilization conditions. Spermatozoal traits of Atlantic salmon showed narrow variance within individuals but differed extensively between samples: mean sperm length varied from 32·3 to 39·5 μm, mean velocity ranged from 18 to 127 μm s⁻¹, and ejaculate longevity varied from 18 to 78 s. In addition to variation in sperm morphometry between fish, a negative relationship was also found between sperm head length and flagellum length. This natural variation in sperm form and function between males is counter-intuitive since measures are from a single Atlantic salmon population where all males are adapted to a common fertilization environment. No evidence was found that longer sperm, or sperm with longer flagella, achieved faster swimming velocities. Also no evidence was found for a trade-off between mean sperm velocity and ejaculate longevity. There were significant negative associations, however, between sperm total and flagellum length and ejaculate longevity, so that males with longer sperm had shorter-lived gametes. This finding has previously been reported in a study across fish species, supporting the theory that increased hydrostatic forces generated by longer flagella may trade against sperm cell longevity.     

Preventive effect of Desferal on sperm motility and morphology

Transition metal ions, mainly iron, are involved in the generation of highly reactive hydroxyl radicals, which are the most powerful inducers of oxidative damage to all biomolecules. The lipids in sperm membranes are highly susceptible to oxidation. Sperm lipid peroxidation (LPO) leads to decrease of motility and reduction of likelihood for sperm‐oocyte fusion. The excess radical production may affect also the spermatozoa morphology. The aim of the present study was to investigate the effect of Desferal on the LPO, motility, and morphology of boar sperm subjected to oxidative stress. After collection, the ejaculates were equally separated and diluted in a commercial semen extender (experiment 1) or in physiological saline (experiment 2). The ejaculates of the 2 experiments were divided into aliquots, which were incubated with one of the following agents: FeSO₄ (0.1mM), H₂O₂ (0.5mM), or FeSO₄ + H₂O₂ (Fenton system), in the presence or absence of Desferal. The application of Desferal in the incubation medium had a protective effect against FeSO₄ + H₂O₂‐induced sperm damage, namely, decrease of LPO; decrease the quantity of immotile spermatozoa and decrease the number of morphological abnormalities, regardless of the used medium. In experiment 2, the presence of FeSO₄ in the incubation medium induced LPO in the same range as the combination FeSO₄ + H₂O₂, in which the effect was reduced by Desferal. Thus, the supplement of Desferal to media used for sperm storage and processing could be a useful tool for diminishing oxidative injury and improving the quality of the semen.     

Objective analysis of stallion sperm motility

An image-analysis system utilizing a microcomputer and CellSoft computer-assisted semen analysis software package was evaluated to assess stallion sperm motility characteristics. Analyses were performed at 37°C on a 6 μl drop of diluted semen placed on a glass slide and covered with an 18 mm² coverslip. Four groups of 25 cells each per slide, four slides per ejaculate and four ejaculates from each of three stallions were analyzed in a nested model. The percentage of motile sperm cells, mean velocity (μm/sec), mean linearity, and mean angular head displacement (μm) were measured. Statistical analysis of variance components showed that within ejaculates, more variation was accounted for in the differences among groups of 25 cells than among slides. Predicted standard deviations calculated for combinations of slides and groups of cells showed that a combination of two slides from which a total of 400 cells were analyzed resulted in a mean intra-assay coefficient of variation (CV) of 5.7% for the four measured variables. The following are individual coefficients of variation: percentage of motile cells (7.8%), mean velocity (6.4%), mean linearity (1.9%) and mean angular head displacement (6.6%). When ejaculate differences were included in the model and predicted standard deviations were calculated for a single ejaculate, the mean inter-assay CV was 9.2%. Mean velocity (6.4%) and mean linearity (4.7%) were more repeatable among ejaculates than either the percentage of motile sperm (14.4%) or angular head displacement (11.2%). It was concluded that this system is precise enough to determine differences in motility characteristics of stallion semen samples.     

Sperm motility in the horseshoe crab. III. Isolation and characterization of a sperm motility initiating peptide

Sperm motility in Limulus is initiated by a sperm motility initiating factor (SMI) that emanates from Limulus eggs. This report describes the partial purification of SMI (greater than 230-fold purification with respect to protein content) with 40% recovery. SMI appears to be a hydrophobic peptide of 500–2,000 MW. Although probably not purified to homogeneity, SMI is estimated to be active at a concentration of less than 0.2 μM.     

Relationship between sperm density, spermatocrit, sperm motility and spawning date in wild and cultured haddock

Semen was collected repeatedly from captive haddock Melanogrammus aeglefinus and the effect of seasonality on various sperm parameters was investigated. No differences in sperm traits were observed for wild and cultured haddock. A highly significant positive relationship existed between spermatocrit and spermatozoa density. A significant increase in mean spermatocrit occurred throughout the spawning season but the amount of variability explained by collection date was low (35·1%) due to variability between males. Each of 10 males sampled repeatedly throughout the spawning season demonstrated an increase in spermatocrit. No relationship existed between spermatocrit and proportion of motile spermatozoa when spermatocrit was ≤70%. Motility was reduced in semen samples with spermatocrits >70%. The proportion of spermatozoa that were motile decreased with time since activation. Some motility was still observed after 60 min in sea water (0·1–15·2%) for sperm collected at all times within the spawning season. Of those spermatozoa that were motile, the proportion that exhibited forward swimming motion decreased and the proportion that had only vibratory movement increased with time post‐activation. The speed of forward swimming spermatozoa showed no significant relationship with spermatocrit at any time between 0 and 60 min after activation. Swimming speed was negatively related to time since activation, decreasing from 174–240 μm s⁻¹ at 0 min to 80–128 μm s⁻¹ at 60 min after activation.     

Clutch characteristics of two populations of nine‐spined stickleback from south‐central Alaska

Clutch size of nine‐spined stickleback Pungitius pungitius from Airolo and Dog Bone Lakes, Alaska, showed small but significant differences between lakes and across 2 years. The variation in clutch size was independent of variation in egg mass, which did not differ appreciably between lakes or across years.     

Cholesterol and phospholipid levels of washed and percoll gradient centrifuged mouse sperm: Presence of lipids possessing inhibitory effects on sperm motility

Levels of DNA, cholesterol, and phospholipids of mouse caudal epididymal and vas deferens sperm that were processed through simple washing and Percoll gradient centrifugation were measured. The DNA and cholesterol contents of washed sperm and Percoll gradient centrifuged (PGC) sperm (DNA = 3.6 ± 0.3 pg/sperm and 3.4 ± 0.3 pg/sperm, respectively; cholesterol = 0.219 ± 0.057 nmole/μg DNA and 0.224 ± 0.030 nmole/μg DNA, respectively, for washed and PGC sperm) were not significantly different from each other; however, the phospholipid level of PGC sperm was only one half of that of washed sperm (0.315 ± 0.071 nmole/μg DNA versus 0.720 ± 0.075 nmole/μg DNA, respectively). The presence of 0.3% bovine serum albumin (BSA) in the culture medium used in sperm washing did not change the cholesterol and phospholipid contents of washed sperm. Similarly, the cholesterol and phospholipid levels of washed sperm and PGC sperm that were further incubated in BSA-containing medium for 30 min remained the same. Interestingly, substantial amounts of lipids, as determined by the cholesterol and phospholipid levels, were released into the supernatants of the sperm washes, and sperm needed to be washed at least twice to ensure their stable levels of cholesterol and phospholipids. The lipid mixture in the first sperm wash supernatant was shown to have inhibitory effects on PGC sperm motility. © 1996 Wiley-Liss, Inc.     

The effects of habitat‐ and diet‐based cues on association preferences in the three‐spined stickleback (Gasterosteus aculeatus)

A number of recent papers have investigated the potential of familiarity to organize the distributions of free‐ranging animals. It is not clear, however, to which extent individual recognition or a more general recognition of a group odour is responsible for familiarity preferences. First, we tested the sensory basis of the recognition of familiars in three‐spined stickleback ( Gasterosteus aculeatus ). When allowed to choose between a familiar and an unfamiliar stimulus shoal on the basis of both visual and chemical communication, visual communication only and chemical communication only, the preference of focal fish for familiars was shown to be dependent on the presence of chemical cues. We subsequently investigated the mechanisms underlying such association preferences, specifically the effects of recent habitat and diet on preferences. Experimental fish were divided into four treatment groups consisting of two environment treatments (saline and freshwater) and two diet treatments ( Daphnia spp. and chironimid larvae). Focal fish subsequently showed significant association preferences for groups of unfamiliar fish that had undergone the same environment or diet treatment as themselves, suggesting self‐referent matching. These data indicate that individual recognition is not a pre‐requisite for the expression of familiarity preferences.     

Monthly variation in sperm motility in common carp assessed using computer-assisted sperm analysis (CASA)

Sperm motility variables from the milt of the common carp Cyprinus carpio were assessed using a computer-assisted sperm analysis (CASA) system across several months (March-August 1992) known to encompass the natural spawning period. Two-year-old pond-raised males obtained each month were primed with an exogeneous hormone injection of carp pituitary extract and stripped of milt 18–24h later. The milt was diluted, activated and videotaped using a high-speed (200 Hz) videocamera and recorder. Videotaped samples were subsequently analysed using the CellTrak/S CASA system (Motion Analysis Corp.) for percent motility, curvilinear and straight-line velocity. In addition to assessing changes in motility parameters across several months, a comparison was made between two predilution/ activation media combinations (homologous seminal plasma/NaCl+HEPES v. 2-h incubation in 200 mM KCl+Tris/Tris). The percentage of motile cells assessed immediately after activation was significantly greater in the summer months (May, June, July) when compared to a spring sampling point (March); when assessed 1 min after activation, cells prediluted in seminal plasma maintained a higher percentage of motility than those prediluted in KC1. Curvilinear and straight-line velocities exhibited a slight seasonal trend; variations in response to the predilution treatments were observed with these measurements. Sperm count gradually increased through April and May (9–63 × 10⁹ to 2–38 × lO¹⁰ml^{– 1} milt), declined in June and July (to 1–83 × lO¹⁰ml^{– 1} milt), and was followed by a steep increase in August (2–74 × 10¹⁰ ml^{– 1} milt). Mean seminal plasma osmolality remained relatively constant (250–265 mOsmol kg ^{– 1}) throughout the sampling period.     

Ovarian fluid enhances sperm movement in Arctic charr

Like the spermatozoa of most other fish species spawning in fresh water, Arctic charr Sahelinus alpinus sperm were short-lived (mean 42 s) after activation and their swimming speed declined rapidly during this period, e.g. from a mean speed of 106 um s⁻¹ at 10 s after activation in fresh water to 21 μm s⁻¹ only 20 s later. Ovarian fluid significantly influenced sperm longevity (duration of forward mobility), per cent motility, swimming speed and the linearity of sperm movement. All of these variables generally increased as the concentration of ovarian fluid increased from 0 to 50%, even though ovarian fluid is more than three times as viscous as fresh water. It is concluded that ovarian fluid enhances sperm movement in this species and thus has the potential to influence both fertilization success and the outcome of sperm competition.     

Effect of overdose zinc on mouse testis and its relation with sperm count and motility

The purpose of this study was to investigate the effects of excessive zinc intake on the testes and on sperm count and motility in mice. Thirty Balb c mice were divided randomly into 3 groups of 10 animals in each. Group I acted as controls; group II was supplied with drinking water containing 1.5 g/100 mL Zn, and group III was supplied with drinking water containing 2.5 g/100 mL Zn. The animals were sacrificed after 3 wk supplementation and the epididymis and testis were quickly excised. A negative correlation between Zn dose and sperm count and motility was found. The sperm count in group III was significantly lower than in groups II and I (p<0.05). The sperm motility in group III was significantly lower than in the controls (p<0.05). Degenerative changes, including spermatic arrest, degeneration of seminiferous tubules, and fibrosis in interstitial tissue, were observed in group III animals. These results show that high doses of zinc significantly alter sperm motility.     

The response of rhesus monkey sperm motility to cervical mucus and solid surfaces

Movement characteristics of rhesus monkey spermatozoa were analyzed using high-speed cinemicrography. In the first experiment, spermatozoa were studied at 100 frames/sec in diluted semen near a surface, and after entering ovulatory cervical mucus from a bonnet monkey. In mucus, the spermatozoa swam more slowly, with reduced flagellar beat frequencies. The beat shape was altered, and there was less lateral yawing of the sperm head. In the second experiment, spermatozoa in diluted semen were studied at 500 frames/sec in deep preparations, while swimming near a surface or when in the midplane of these preparations. Those sperm in the midplane swam faster, but with lower beat frequencies than those near the surface, and exhibited much more pronounced yawing motions. Such distinctions in sperm motion are probably hydromechanical in origin and may be significant during transport in the female.     

紫外线辐射对西伯利亚鲟精子活力和寿命的影响

研究了不同剂量紫外线辐射(254nm,UVC)对西伯利亚鲟精子活力和寿命的影响.结果表明:紫外线辐射对精子的活力、快速运动时间和寿命均具有显著性影响.其中,精子活力随辐射剂量的增加而呈先迅速下降,后迅速上升,再迅速下降的趋势;精子快速运动时间的变化趋势与活力相似;精子寿命随辐射剂量的增加呈缓慢下降的趋势.当辐射剂量达288mJ.cm-2时,精子无快速运动,当辐射剂量达324mJ.cm-2时,精子活力和寿命均降为0.根据Hertwig效应判断,辐射剂量216mJ.cm-2为西伯利亚鲟精子灭活的最适剂量.     

The effect of Schistocephalus solidus infection on meal size of three‐spined stickleback

The effect of infection with the pseudophyllidean cestode Schistocephalus solidus on the meal size of individually housed three‐spined stickleback Gasterosteus aculeatus was quantified. Infected fish harboured plerocercoid loads that contributed from 1·1 to 33·9% of their total mass. Across this range of infection levels, the presence of S. solidus infection had no significant directional effect on standard length ( L _S) corrected meal size of host three‐spined sticklebacks. Amongst multiply infected fish there was a significant negative relationship between L _S‐corrected meal size and the proportion of host mass contributed by S. solidus parasites. This relationship, however, did not hold for singly‐infected fish. Furthermore, the data suggest that multiply‐infected fish that harbour a combined mass of parasites contributing < c . 15% to host body mass might exhibit meal sizes that exceed those of length‐matched uninfected fish. The results suggest that although heavy infections can significantly reduce the meal size of heavily infected three‐spined sticklebacks, in the early stages of multiple S. solidus infections host food intake may increase. The probable causes of these differential effects on meal size and their consequences for the host‐parasite system are discussed.     

Transglutaminase inhibitors,calmodulin antagonists,and calcium channel blockers: Influence on Arbacia sperm motility

Sea urchin spermatozoa react to transglutaminase inhibitors, which may antagonize the function of calmodulin, in dose-dependent fashion. The optimum concentration of diallyl-amino propionyl benzothiophenc (DAPBT), a potent, noncompetitive inhibitor of transglutaminase, 0.01 mM, causes a 3 1/2-fold increase in the forward swimming speed of Arbacia sperm. This effect apparently involves calcium-dependent enzymes since manipulation of both extracellular and intracellular calcium by means of chelating agents, calcium-channel blockers, and calmodulin antagonists depresses the stimulatory effect of DAPBT. These results suggest that in the spermatozoa the interaction of flagellar sliding filaments may be mediated by reversible cross-linking of the contractile proteins catalyzed by a calcium- dependent transglutaminase.     

Combined effects of physicochemical variables (pH and salinity) on sperm motility: characterization of sperm motility in European sea bass Dicentrarchus labrax

Gamete activation in fish is an important step in terms of artificial fertilization of oocytes, cryopreservation studies and other experimental manipulations. Salinity and pH differences in activation media affect to sperm motility and fertilizing ability. These experiments were therefore designed to investigate the combined effects of pH (range 5.0–9.0) and salinity (20, 30, 37, and 45‰) of activation media on sperm motility of European sea bass Dicentrarchus labrax. The best results were obtained at salinity 37‰ and a pH of 9.0. Our results also demonstrated that non-progressive motility at salinity 45‰ was observed in the range of 5.0–9.0 pH. In conclusion, spermatozoa can be motile at a wide range of pH and salinity values although the percent of motile spermatozoa and motility duration are negatively affected by low pH values.     

NMDA‐type glutamate receptors mediate the acrosome reaction and motility initiation in newt sperm

The N‐methyl d ‐aspartate type glutamate receptor (NMDAR) is a ligand‐gated cation channel that causes Ca²⁺ influx in nerve cells. An NMDAR agonist is effective to the sperm motility in fowls, although the actual role of NMDAR in sperm function is unknown. In the present study, RNA‐seq of the spermatogenic testes suggested the presence of NMDAR in the sperm of the newt Cynops pyrrhogaster. Glutamate of at least 0.7 ± 0.5 mM was detected in the egg‐jelly substances along with acrosome reaction‐inducing substance (ARIS) and sperm motility‐initiating substance (SMIS). In the egg‐jelly extract (JE) that included the ARIS and SMIS, the acrosome reaction was inhibited by a NMDAR antagonists, memantine and MK801. MK801 also inhibited the spontaneous acrosome reaction in Steinberg's salt solution (ST). Furthermore, memantine and MK801 suppressed the progressive motility of the sperm in JE and spontaneous waving of the undulating membrane, which is the tail structure giving thrust for forward motility, in ST. The spontaneous waving of the undulating membrane was promoted when Mg²⁺, which blocks Ca²⁺ influx through gated NMDARs, was removed from the ST. In addition, the ARIS‐induced acrosome reaction was inhibited by a selective antagonist of the transient receptor potential vanilloid 4, whose activation might result in the membrane depolarization to release Mg²⁺ from the NMDAR. These results suggest that NMDAR acts together with other cation channels in the induction of the acrosome reaction and motility of the sperm during the fertilization process of C. pyrrhogaster.