Characterization of eight microsatellite markers in the white sea bream, Diplodus sargus (Teleostei, Sparidae)

The white sea bream, Diplodus sargus (Teleostei, Sparidae), is a species with a high commercial importance in Mediterranean aquaculture. There is currently little information available about the genetic characteristics of cultured populations. In this survey, we have developed eight polymorphic microsatellites for the white sea bream using an enriched genome library protocol. All of them were polymorphic in the 67 individuals tested, 32 of which were wild specimens, and 35 were individuals from a captive F(1) broodstock. These markers can potentially be useful tools for use in population genetic studies.     

Structures and movements of the buccal and pharyngeal jaws in relation to feeding in Diplodus sargus

The present paper studies the possibly different feeding strategies of Diplodus sargus to crustaceans, molluscs, worms, and small fish. The buccal jaws are built strongly and bound together by numerous ligaments. The dentition is heterodont: incisors in front and molars in the middle and hind parts. The principal originality of the musculature of this species is the forward insertion of the adductores mandibulae. These are very thick and insert on both the upper and lower jaws, so that contraction of any individual muscle acts on the buccal pieces as a whole, which thus constitute a remarkable crushing device. The pharyngeal jaws are frail as in primitive perciforms: the lower ones are well separated, being bound only anteriorly, while the upper ones consist of the second and third pharyngobranchials and a posterior toothed plate. When feeding on crabs, Diplodus sargus always sucks in the prey and seizes it with the buccal jaws. Mouth opening is accompanied by extensive protrusion of the mouth, with or without neurocranial elevation. Mouth sucking and seizing movements vary little. Once seized, the prey is usually moved to the molars and crushed. The crushing movements may be fast and ample or slow. In the latter case, deformation of the prey is observable. Crushing usually results in the crab being broken into pieces. The pharyngeal jaws grip one part of the prey and shift it to the oesophagus, then seize the second part. Diplodus sargus adapts its feeding behaviour to the type of prey. A snail, for instance, is crushed by the buccal or pharyngeal teeth, the pieces of shell are ejected, and the soft parts conveyed with difficulty to the oesophagus by the pharyngeal jaws. A fish on the other hand, is sucked tail first into the mouth cavity and quickly shifted to the digestive tract by the pharyngeal bones. Behaviour toward different prey differs by the presence or absence of parts of the sequence of feeding movements (for example crushing) or by the fact that certain movements or parts of the sequence are repeated. The variability of any movement in the sequence is the same whatever the sort of prey. Crushing occurs between the buccal incisors and molars and was observed twice between the pharyngeal teeth. Usually, it seems, the latter are involved in transport only. In transport, the left and right pharyngeal jaws may perform different functions: their movements, unlike the symmetrical movements of the buccal jaws, sometimes differ.     

Sperm structure and motility of the freshwater teleost Cottus gobio

When motility of spermatozoa of Cottos gobio was initiated with distilled water, the motility rate decreased to 0% within 1 min, and significant signs of osmotic alterations were observed at the end of the motility period. By contrast, in 50 mmol 1⁻¹ NaCl solution, the motility rate persisted for 120–140 min. In both distilled water and in 50 mmol 1⁻¹ NaCl solution, the main swimming type of spermatozoa was linear motion during the whole motility period. The initial swimming velocity (50.0 ± 2.1 μm s⁻¹) measured 10 s after motility initiation was similar in both distilled water and in 50 mmol 1⁻¹ NaCl solution. In distilled water, the velocity decreased to <20 μm s⁻¹ (locally motile) during the first minute of the motility phase. In 50 mmol 1⁻¹ NaCl solutions, it remained at a constant level during the first 60 min of the motility period, but then started to decrease to <20 μm s⁻¹ after 120 min. When 5 mmol 1⁻¹ potassium cyanide, antimycin or atractyloside was added to the 50 mmol 1⁻¹ NaCl solution, the motility period was reduced to ≤2min. Ten millimoles per litre 2-deoxy-D-glucose, malonate or a mixture of 5 mmol 1⁻¹ atractyloside and 5 mmol 1⁻¹ carnithine did not effect the duration of the motility period. This indicates that sperm energy metabolism depends mainly on respiration rate and fatty acid metabolism.     

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.     

Sperm motility in the horseshoe crab. IV. Extracellular ions and intracellular pH are not mediators of motility initiation

Upon dilution into sea water, Limulus spermatozoa undergo a brief flurry of motility (duration < 60 sec), after which they are nonmotile until encountering a sperm motility initiating peptide (SMI) that emanates from eggs. Utilizing highly purified SMI extracts and simplified seawater formulations (from which individual ions have been deleted), we found that no specific extracellular ion is required for either dilution-initiated or SMI-initiated motility. Indeed, deletion of one ion (Na⁺) produced dilution-initiated motility of very long duration (several hours). When motility is initiated by SMI (in normal seawater) there is an increase in intracellular pH (pH_i), as indicated by the fluorescent probe, 9-amino acridine; however, this pH, change is not a trigger for motility. As a more general method examining ion movements, the fluorescent probe diS-C₃-(5) was used to qualitatively measure changes in the membrane potential of spermatozoa. Although crude SMI extracts caused membrane depolarization, further purification resulted in an almost complete separation of this activity from SMI, thus showing that SMI activation is apparently an electroneutral event. (The membrane-depolarizing factor has a molecular weight > 30,000 and does not initiate acrosome reactions.) Experiments utilizing the ionophore A23187 and Ca⁺²-blocking agents (verapamil and TMB-8) provided tentative evidence that mobilization of intracellular Ca⁺² may be required for motility initiation. These results show that neither changes in pH_i nor the influx of specific extracellular ions are direct mediators of SMI-initiated motility; however, experiments with pharmacologic agents indicate a possible role for intracellular Ca⁺².     

Initiation,prolongation, and reactivation of the motility of salmonid spermatozoa

The motility of salmonid spermatozoa initiated by dilution of the milt with ovarian fluid or isotonic saline is brief duration; it was believed that it can be activated only once in the life of the spermatozoon. Dilution of the milt with an equal volume of isotonic saline (0.12 M-NaCl) containing 5 mM-3-isobutyl-1-methylxanthine (MIX) prolonged and intensified sperm motiliy. When motility had stopped after initial mobilization with saline or ovarian fluid, it could be reactivated by addition of MIX; reactivated spermatozoa fertilized eggs. Dilution with saline containing K⁺ (24 mEq/liter) did not initiate sperm motility even in the presence of MIX. The spermatozoa were mobilized by subsequent with 0.12 M-NaCl. The concentration of adenosine triphosphate (ATP) in sperm suspensions dropped on dilution with saline and rose as motility ceased, but declined without subsequent recovery following dilution with MIX-saline. The concentration of cyclic adenosine monophosphate (cAMP) rose and fell sharply on initiation of motility and rose again after motility had declined. While salmonid spermatozoa can be mobilized by dilition with saline alone, the effectiveness of MIX in reactivating “spent” spermatozoa supports the assumption that cAMP plays a role in the initiation of sperm motility.     

Sperm motility and fertilization time span in Atlantic salmon and brown trout—the effect of water temperature

The effect of water temperature on the duration of sperm motility, the time lapse after activation by fresh water and the fertility of eggs was studied in Atlantic salmon and brown trout. Eggs of both species were fully fertile in fresh water after 512 s. No interspecific differences were noted in egg fertility at the lower water temperatures, but the brown trout eggs showed a higher resistance to high temperatures, indicating a better physiological thermotolerance. A highly significant effect of temperature on the overall duration of sperm motility was found, with a marked peak at 3−4°C for salmon and a weaker one for trout. After freshwater activation the eggs of both species remained fertile for a longer time than the sperm were mobile.     

The spermatozoon of the African catfish: fine structure, motility, viability and its behaviour in seminal vesicle secretion

The spermatozoon of the African catfish Clarias gariepinus is a simple organized aquasperm although it reveals very unique characteristics: the cytoplasmic channel is lacking, the mitochondria form a complex structure and the arrangement of the centriolar complex is species specific. Semen has high initial motility rates ( c. 70–90%) and swimming velocities ( c. 120–140 μm s⁻¹), the main swimming type is linear. Motility duration in water is 30 s and is prolonged only to 40 s in NaCl solutions or more complex bu ered motility activating saline solutions. A pH between 7.0 and 9.0 has no e ect on the sperm motility parameters. Motility is completely and reversibly suppressed in electrolyte and non-electrolyte solutions with an osmolality of 200 mosmol kg⁻¹. During immotile storage the sperm viability is influenced by the osmolality and the potassium levels of the storage medium, by the temperature and by the dilution. At optimal conditions (bu ered sperm motility inhibiting saline solution: 150 mmol l⁻¹ NaCl, 2.5 mmol l⁻¹ KCl, 1 mmol l⁻¹ CaCl₂, 1 mmol l⁻¹ MgSO₄, 20 mmol l⁻¹ Tris solution, pH 8.5; dilution rate 1: 5; storage temperature, 4°C) sperm viability persists for >7 days. High viscosity of the pure seminal vesicle secretion completely inhibits the sperm motility. When the seminal vesicle secretion is diluted in water the viscosity decreases and the motility suppressing e ect is neutralized. When semen is mixed with seminal vesicle secretion the sperm viability decreases to zero within 10 min.     

Demographic and genetic structures of white sea bream populations (Diplodus sargus, Linnaeus, 1758) inside and outside a Mediterranean marine reserve

We studied the white sea bream (Diplodus sargus), a protandrous hermaphroditic fish, in two protected and unprotected areas in southwestern France. We observed a significant difference in the demographic structure between the two areas. Females were present in two different age distributions inside and outside the marine reserve with younger females outside. This suggests plasticity in the age of sexual inversion in the case of an exploited population. Genetic differentiation was weak and apparent at only one locus of 26 surveyed (FST = 0.007, p = 0.04). Our data suggest that gene flow between the two areas is important, or the separation between the two sites is recent. Our data on the white sea bream show that fishes inside and outside the marine reserve are very similar genetically, which means that the 'reserve effect' is truly a demographic one, not the result of genetic differences.     

Fine structure and motility of spermatozoa and composition of the seminal plasma in the perch

The fine structure and motility of spermatozoa and the composition of the seminal plasma of the perch Perca fluviatilis are investigated by electron microscopy, computer assisted cell motility analysis (CMA) and biochemical methods. The spermatozoon is asymmetrical as the flagellum inserts mediolateral on the nucleus. It lacks an acrosome, has an ovoid head and a small midpiece with one mitochondrion. Sperm motility–initiated in distilled water (10° C)–is characterized as follows: 85·0 ± 2·7% of the spermatozoa are motile, the main swimming type (10 ± 1 s after motility initiation) is the linear motion (61·4 ± 24·4%) and the average swimming velocity is 122·4 ± 21·9 μm s^–1. When motility is initiated with NaCl, glucose or sucrose solutions of 100 mosmol kg^–1 the percentage of motile spermatozoa and the swimming types are similar as in water, but the swimming velocity (174·0 ± 22·3 μm s^–1) is significantly higher. Motility is inhibited by high osmolality of the diluent: when increasing the osmolality of the saline solutions to 350 mosmol kg^–1 sperm motility is totally suppressed while potassium (10–40 mmol 1^–1) does not affect motility parameters. pH optimum for sperm motility is between pH 7·0 and 8·5. The seminal fluid contains 124·01 ± 21·68 mmol 1^–1 sodium, 10·22 ± 1·11 mmol 1^–1 potassium and 0·72 ± 0·26 mmol 1^–1 calcium. pH is 8·25 ± 0·09, and osmolality 283·90 ± 37·19 mosmol kg^–1. The following organic components were determined: monosaccharides (glucose 63 ± 19 μmol 1^–1, fructose 54 ± 28 μmol 1^–1, galactose 59 ± 25 μmol 1^–1), lipids (cholesterol 5·51 ± 6·42 μmol 1^–1, triglycerides 72 ± l00 μmol l^–1, cholesteryloleate 15–150 μmol 1^–1, phosphatidylcholine 26 · 31 μmol 1^–1, glycolipids 1–10 mg 100 m1^–1), lactate 108 ± 99 μmol 1^–1, hydroxybutyrate 102 ± 99 nmol 1^–1, choline 59 ± 159 μmol 1^–1, protein 344·75 ± 59·06 mg 100m1^–1, enzymes (β-d -glucuronidase l.4 ± 0.7 μmol h^–1 100 ml^–1, protease (caseolytic activity) 1·0 ± 0·6 μmol h^–1 100 ml^–1, alkaline phosphatase 2520·0 ± 861·0 μmol h^–1 100 ml^–1, acid phosphatase 44.0 ± 16.0 μmol h^–1 100 ml^–1, glucose-6-phosphate dehydrogenase 38·9 ± 86·9 μmol h^–1 100 ml^–1, lactate dehydrogenase 134·4 ± 69·6 μmol h^–1 100 ml^–1, butyrylcholine esterase 0·014 ± 0·010 μmol h^–1 100 ml^–1, adenosine triphosphatase 562·8 ± 665·4 μmol h ^–1 100 ml^–1).     

Induction of motility of apyrene spermatozoa and dissociation of Eupyrene sperm bundles of the silkmoth,Bombyx mori,by initiatorin and trypsin

Summary

The activator, or inducer of motility, of apyrene spermatozoa of the silkmoth, Bombyx mori, was shown to be present in the posterior glandula (g.) prostatica. The activity of this factor in this gland was lost by boiling at 100°C for 10 min. Bundles of eupyrene sperm were dissociated by treatment with either a g. prostatica homogenate or trypsin, and their dissociation was concentration-dependent. On the contrary, for activation of apyrene sperm, the optimal trypsin concentration was 0.45–1.80 μg/ml and activation decreased at higher and lower trypsin concentrations. Microscopic observation showed that the dissociation rate of eupyrene sperm bundles by the g. prostatica homogenate corresponded to that by 0.45–9.0 μg/ml of trypsin. An autolysate of the g. prostatica and digests of seminal fluid with the g. prostatica homogenate or trypsin did not activate apyrene sperm. Of 11 endopeptidase inhibitors tested, antipain, leupeptin, TLCK, TPCK and PMSF strongly inhibited sperm activation by the g. prostatica homogenate, suggesting that the activator is an endopeptidase of the wine protease type. The 6 exopeptidase inhibitors tested did not inhibit activation of apyrene spermatozoa and the dissociation of eupyrene sperm bundles are both caused by the same factor, initiatorin, an endopeptidase of the serine protease type present in the prostatic secretion.     

Temporal change in the genetic structure between and within cohorts of a marine fish,Diplodus sargus,induced by a large variance in individual reproductive success

Temporal changes at 16 allozyme loci in the Diplodus sargus population of Banyuls-sur-Mer (Mediterranean Sea, France) were monitored. Temporal genetic variation within a single population was examined over two temporal scales: (i) among three year-classes sampled at the same age, and (ii) within a single year-class sampled three times over a two-year period. We observed a significant change in the genotypic structure within the same cohort during the first two years following settlement and before recruitment into the adult population. In addition, comparison of year-classes showed that cohorts differed significantly one year after settlement, whereas they became similar later on before recruitment into the adult population. The observed changes in the genetic structure within and between year-classes may be the result of complex selective processes or genetic drift. Linkage disequilibrium and genetic relatedness data suggest that these changes are due to large variation in reproductive success, followed by homogenization through adult movement. Overall, these results demonstrated a rapid genetic change within a population.     

Roles of the Na,K-ATPase alpha4 isoform and the Na+/H+ exchanger in sperm motility

The Na,K-ATPase generates electrochemical gradients that are used to drive the coupled transport of many ions and nutrients across the plasma membrane. The functional enzyme is comprised of an alpha and beta subunit and families of isoforms for both subunits exist. Recent studies in this laboratory have identified a biological role for the Na,K-ATPase alpha4 isoform in sperm motility. Here we further investigate the role of the Na,K-ATPase carrying the alpha4 isoform, showing again that ouabain eliminates sperm motility, and in addition, that nigericin, a H+/K+ ionophore, and monensin, a H+/Na+ ionophore, reinitiate motility. These data, along with the observation that the K+ ionophore valinomycin has no effect on the motility of ouabain-inhibited sperm, suggest that ouabain may change intracellular H+ levels in a manner that is incompatible with sperm motility. We have also localized NHE1 and NHE5, known regulators of intracellular H+ content, to the same region of the sperm as the Na,K-ATPase alpha4 isoform. These data highlight the important role of the Na,K-ATPase alpha4 isoform in regulating intracellular H(+) levels, and provide evidence suggesting the involvement of the Na+/H+ exchanger, which is critical for maintaining normal sperm motility.