Sperm Mobility Phenotype Research Articles

Mobility of Japanese quail spermatozoa and its relationship to egg fertility.

Sperm mobility (SM) appears to be primary determinant of fertility in chicken and turkey. The aims of this study were to extend the concept to the Japanese quail by developing an assay to quantify SM, explaining the basis of SM using motility properties measured by CASA, and exploring the relationship between SM and egg fertility. The study was carried out in three stages: i) males (n=20) and females (n=20) were mated individually; ii) ejaculates were collected from 20 males, and SM was measured; iii) males (n=20) and females (n=20) were mated individually. In Stages I and III, data were collected for egg fertility, SpermOPVL and HolesIPVL . In Stage II, SM assay was developed and assay conditions were defined: effect of sperm numbers on absorbance in Accudenz solution; effect of Accudenz concentration on sperm motility and mobility; effect of quail proctodeal gland foam extract and incubation temperature on SM at 37 and 41°C. The recorded absorbance of sperm movement was dependent on sperm numbers in the sperm suspension overlaying the Accudenz (p<.001). At 41°C, SM, progressively motile sperm, VCL, VSL and VAP were negatively affected by Accudenz concentration (p<.05). The effect of foam on SM and motility depended on an interaction between the concentration of foam extract and incubating temperature. Males were categorized into low, average and high SM phenotypes. These categories differed significantly (p<.001), but sperm motility and SM were not related to egg fertility. In conclusion, SM assay can be used to identify mobility phenotypes, but the poor relationship between SM and egg fertility indicates a need for further studies on interaction between the concentration of foam extract, incubating temperature, and in vivo sperm movement and egg fertilization success.

Low-mobility sperm phenotype in the domestic turkey: Impact on sperm morphometry and early embryonic death.

The sperm mobility assay measures the ability of sperm to swim through a dense layer of Accudenz® , and the sperm mobility phenotype has been shown to predict fertility and other sperm performance traits in roosters and turkeys. In this study, we examined turkey sperm morphometry and rates of early embryonic death associated with high- and low-mobility semen. We also assessed whether the hypo-osmotic stress test, which evaluates the structural integrity of the sperm plasma membrane, may be used as a faster and simpler assay for sperm mobility and viability. We confirmed previous work that found that high-mobility sperm are faster and swim more linearly than low-mobility sperm, and that mobility traits were repeatable within males. In contrast to previous studies, we did not find higher rates of fertility, but low-mobility sperm was associated with higher rates of early embryonic death, though this trend was not significant. High-mobility sperm had longer sperm heads, explained by longer nuclei, despite shorter acrosomes. Although these sperm were faster, midpiece length and flagellum length did not differ between high- and low-mobility sperm. Finally, mobility was not found to be associated with sperm performance in the hypo-osmotic stress test.

Premises for fowl sperm preservation based on applied bioenergetics1

The primary goal of this work was to test whether the sperm mobility assay could be used to derive mathematical relationships from which predictions could be made about sperm cell function. A precondition was random sampling from a pool of sperm. This precondition was met by centrifuging mobile sperm through 12% (wt/vol) Accudenz containing the Ca(2+) chelator 1,2-bis-(o-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid (BAPTA) and then holding washed sperm at 20°C within buffered potassium chloride. These 2 conditions rendered washed sperm immobile at 20°C. Resumption of sperm mobility was independent of time (P > 0.8558) when sperm were reactivated at body temperature with 2 mM Ca(2+) in isotonic sodium chloride at pH 7.4. Reactivated sperm mobility was 93% of the prewash control. Subsequent experiments served to define a dose response, predict optimal conditions for in vitro sperm mobility, and show how sperm can recover from an imposed non-physiological condition. Thus, functions were derived from which predictions were made. Whereas the utility of BAPTA treatment was confirmed in a new context, such utility did not address the question of whole-cell Ca(2+) flux during sperm cell manipulation. This issue is pivotal for the application of bioenergetics to fowl sperm preservation. Therefore, the secondary goal of this research was to investigate sperm cell Ca(2+) flux using a simulation of conditions encountered by sperm during centrifugation through 12% (wt/vol) Accudenz. These conditions included a temperature of 30°C, a Ca(2+) sink, and no exogenous substrate. Sperm motion was measured with a Hobson SpermTracker. Data points conformed to parabolic functions when motile concentration and velocity were plotted as functions of time. In each case, maximums were observed, e.g., 26 min for motile concentration. The upswing was attributed to a redistribution of intracellular Ca(2+) whereas the downswing was attributed to sperm cell Ca(2+) depletion. A pronounced isothermal increase was observed for each variable when the Ca(2+) sink was overcome with exogenous Ca(2+). Experimental outcomes supported four testable premises applicable to fowl sperm preservation research: 1) the importance of sperm mobility phenotype, 2) the relationship between mitochondrial Ca(2+) cycling and sperm mobility, 3) the utility of the sperm mobility assay for predicting experimental outcomes, and 4) understanding mitochondrial Ca(2+) cycling in terms of whole-cell Ca(2+) flux.

BREEDING AND GENETICS SYMPOSIUM: A systems biology definition for chicken semen quality1,2

Rooster semen is an effluent from paired reproductive tracts. Each tract includes a testis, epididymis, and deferent duct. Upon ejaculation, efficacy of sperm propulsion varies among roosters. This phenotype is sperm mobility, that is, the movement of sperm against resistance at body temperature. The present work 1) compares reproductive tract throughput between lines of chickens selected for low and high sperm mobility, 2) demonstrates how semen quality can be defined in terms of an interaction between reproductive tract throughput and the proportion of mobile sperm ejaculated, 3) confirms that phenotype can be linked to genomewide differences in SNPlotype, and 4) shows how breeding can affect semen quality. Sperm mobility phenotype distributions were based on the average of duplicate observations per male (n = 241 and 262 roosters for low and high lines, respectively). Distributions were skewed and normal for low and high lines, respectively. Subsequent analyses used these base populations as sources for test subjects. In the first analysis, 10 males were selected from the mode of each distribution, and sperm mobility data were evaluated by nested ANOVA. Variation was observed between lines (P < 0.0001) but not among males within lines (P = 0.980). Sperm mobility data along with data from paired reproductive tracts were used to estimate combined reproductive tract throughput. Whereas testicular output was 1.2-fold greater in the low line (P = 0.037), the output of mobile sperm per day was 10.5-fold greater in the high line (P < 0.0001). Deferent duct transit differed between tails of the low line (P < 0.0001) but not between the tails of the high line (P = 0.514). Males from the mode and upper tail of the low line were SNPlotyped using a 60k chip by DNA Landmarks. These test subjects were used to associate phenotype with SNPlotype because founder effects and genetic drift could be discounted. Loci of interest were found on multiple chromosomes. Loci on chromosome Z were of particular interest because roosters are homozygous for this sex chromosome and a pronounced maternal effect was observed in a prior heritability study. Midrange phenotypes were produced by crossing low and high sperm mobility lines. Our experimental outcomes demonstrate that genes affect reproductive tract function as well as sperm cell attributes and thereby make semen quality subject to genetic selection.

PHYSIOLOGY AND ENDOCRINOLOGY SYMPOSIUM: A proteome-based model for sperm mobility phenotype1,2

Sperm mobility is defined as sperm movement against resistance at body temperature. Although all mobile sperm are motile, not all motile sperm are mobile. Sperm mobility is a primary determinant of male fertility in the chicken. Previous work explained phenotypic variation at the level of the sperm cell and the mitochondrion. The present work was conducted to determine if phenotypic variation could be explained at the level of the proteome using semen donors from lines of chickens selected for low or high sperm mobility. We began by testing the hypothesis that premature mitochondrial failure, and hence sperm immobility, arose from Ca(2+) overloading. The hypothesis was rejected because staining with a cell permeant Ca(2+)-specific dye was not enhanced in the case of low mobility sperm. The likelihood that sperm require little energy before ejaculation and the realization that the mitochondrial permeability transition can be induced by oxidative stress arising from inadequate NADH led to the hypothesis that glycolytic enzymes might differ between lines. This possibility was confirmed by 2-dimensional electrophoresis for aldolase and phosphoglycerate kinase 1. This outcome warranted evaluation of the whole cell proteome by differential detergent fractionation and mass spectrometry. Bioinformatics evaluation of proteins with different expression levels confirmed the likelihood that ATP metabolism and glycolysis differ between lines. This experimental outcome corroborated differences observed between lines in previous work, which include mitochondrial ultrastructure, sperm cell oxygen consumption, and straight line velocity. Although glycolytic proteins were more abundant within highly mobile sperm, quantitative PCR of representative testis RNA, which included mRNA for phosphoglycerate kinase 1, found no difference between lines. In summary, we propose a proteome-based model for sperm mobility phenotype in which a genetic predisposition puts sperm cells at risk of premature mitochondrial failure as they pass through the excurrent ducts of the testis. In other words, we attribute mitochondrial failure to sperm cell and reproductive tract attributes that interact to affect sperm in a stochastic manner before ejaculation. In conclusion, our work provides a starting point for understanding chicken semen quality in terms of gene networks.

Characterizing the Glycocalyx of Poultry Spermatozoa: II. In Vitro Storage of Turkey Semen and Mobility Phenotype Affects the Carbohydrate Component of Sperm Membrane Glycoconjugates

The turkey sperm glycocalyx is known to contain residues of sialic acid, alpha-mannose/alpha-glucose, alpha- and beta-galactose, alpha-fucose, alpha- and beta-N-acetyl-galactosamine, monomers and dimers of N-acetyl-glucosamine, and N-acetyl-lactosamine. Potential changes in these carbohydrates during in vitro semen storage at 4 degrees C were evaluated using males of both high- and low-sperm-mobility phenotypes. Changes in carbohydrate residues were quantified by flow cytometry analysis using a battery of 14 fluorescein isothiocyanate-labeled lectins in combination with control (sialylated) or neuraminidase-treated (nonsialylated) sperm. Sperm were evaluated at 0, 2, 4, 8, 12, and 24 hours of storage. For control sperm, 4 different patterns of lectin binding were observed over time: 1) increased mean fluorescence intensity (MnFI) at 2 hours (Griffonia simplicifolia lectin-I [GS-I]) and 8 hours (Ricinus communis lectin-I [RCA-I]) that remained elevated during storage; 2) increased MnFI at specific time points (Limax flavus lectin [LFA], 2 hours; Artocarpus integrifolia lectin [jacalin] and succinyl Triticum vulgare lectin [sWGA], 8 hours; Galanthus nivalis lectin [GNA], 12 hours) followed by decreasing MnFI during the remainder of the 24-hour storage period; 3) increased MnFI only at the 24-hour time point (Lotus tetragonolobus lectin [lotus] and Arachis hypogaea lectin [PNA]); and 4) no changes in MnFI during the 24-hour storage period (Erythrina cristagalli lectin [ECA], GS-II, Pisum sativum lectin [PSA], Glycine max lectin [SBA], and Wisteria floribunda lectin [WFA]). For nonsialylated sperm, increased binding of ECA, GS-II, SBA, and WFA was observed at variable time points; only Canavalia ensiformis lectin (Con A) and PSA remained unchanged during storage. Differences between mobility phenotypes existed for lectins Con A, GS-II, LFA, PSA, SBA, and sWGA, with sperm from low-mobility males exhibiting higher MnFI than high-mobility males throughout 24 hours of storage. We concluded that the observed increases in lectin binding during semen storage indicate an augmentation of nonsialylated terminal residues, which could alter sperm antigenicity and negatively impact fertility. Further, spermatozoa from low-mobility males may have higher antigenicity even before semen storage. Other possible functional implications are discussed.

Application of the Sperm Mobility Assay to Primary Broiler Breeder Stock

A new quantitative trait affecting male fertility was discovered in the 1990s. The term sperm mobility denotes the net movement of a sperm cell population against resistance at body temperature. Even though sperm cells are self-propelled DNA delivery vehicles, their self-propulsive nature is neither uniform among sperm within an ejaculate nor among males within flocks. Such variation is evident when semen quality is evaluated by sperm penetration of an Accudenz solution, hence the term sperm mobility assay. It is noteworthy that populations showing modest variation with respect to semen volume or sperm concentration often show considerable variation with respect to sperm mobility. Likewise, it is noteworthy that broiler breeder fertility is a function of sperm mobility phenotype when hens are inseminated artificially. This article outlines the following: 1) a series of experiments in which the sensitivity of the commercial sperm mobility assay was improved using semen donors from lines of chickens selected for either low or high sperm mobility and 2) the application of the improved technique to pedigree broiler breeders. Male fertility is subject to genetic selection when sperm mobility is the selection criterion. Therefore, sperm mobility may be a useful trait for improving broiler breeder reproductive efficiency.

Sperm Mobility: Deduction of a Model Explaining Phenotypic Variation in Roosters (Gallus domesticus)1

In previous work, variation in sperm mobility phenotype was attributed to the proportion of ejaculated fowl sperm containing dysfunctional mitochondria. In the present work, latent mitochondrial dysfunction was inferred from patterns of sperm egress from the oviduct's sperm-storage tubules. In addition, experiments were performed to help explain how mitochondrial function could be compromised in viable sperm cells. Confocal microscopy demonstrated that sperm Ca2+ content differed between low and high sperm-mobility phenotypes when sperm were stained with rhod-2 AM, a Ca2+ -specific dye. Fluorescence was associated with the nuclear envelope, a variant of the endoplasmic reticulum, and greater fluorescence was observed in sperm from low sperm-mobility males. Fluorescence was reduced by 50% when motile sperm were rendered immotile by incubation with a Ca2+ chelator. Thus, a relationship was established between a dynamic intracellular Ca2+ pool and sperm motility. Sperm N-methy-D-aspartic acid (NMDA) receptors were inferred by the action of D-homocysteinesulfinic acid, a potent NMDA receptor agonist. Seminal plasma from low sperm mobility males was characterized by an elevated glutamate concentration. Thapsigargin, which inhibits the smooth endoplasmic reticulum Ca2+ pump and thereby promotes Ca2+ efflux, rendered sperm immotile. This effect was blocked by cyclosporin A, which prevents the formation of the mitochondrial permeability transition pore (PTP) in response to elevated mitochondrial Ca2+ content. In summary, we propose that 1) glutamate enables Ca2+ uptake into sperm before ejaculation, 2) excessive Ca2+ uptake triggers formation of the PTP in a subpopulation of sperm, and 3) sperm mobility is decreased in proportion.

Sperm Mobility: Phenotype in Roosters (Gallus domesticus) Determined by Mitochondrial Function1

Previously, inheritance of sperm mobility entailed a maternal additive genetic effect, and sperm ATP content was correlated (r = 0.80) with phenotype. The present study was conducted to determine if mitochondrial function was critical to phenotypic expression. Whereas phenotype was independent of mitochondrial helix length, phenotype was correlated with sperm oxygen consumption (r = 0.83) using random-bred roosters. Aberrant mitochondria characterized immobile sperm, as evidenced by transmission-electron microscopy. Such mitochondria were swollen and contained disorganized cristae. Additional experiments were performed with roosters from lines selected for low or high sperm mobility. A threefold difference in sperm oxygen consumption was observed between lines. Single nucleotide polymorphisms were observed in mitochondrial DNA by sequencing replicate mitochondrial genomes from each line. An A-to-G substitution in the gene encoding tRNA(Arg) was inherited consistently, as evidenced by restriction fragment length polymorphism analysis using two male and two female progeny per family group and 14 family groups per line. Motile concentration in semen from low-line males was half that observed in semen from high-line males, as evidenced by computer-assisted sperm motion analysis. Likewise, 47% of sperm from low-line males contained aberrant mitochondria, compared to 4% for high-line males. In summary, sperm mobility phenotype was dependent on mitochondrial function, which in turn was altered by genetic selection. Fowl deferent duct fluid contains a high concentration of glutamate. We propose that variation in sperm mobility phenotype stems from the extent to which glutamate induces excessive mitochondrial Ca2+ uptake before ejaculation.

Attributes of broiler breeder males characterized by low and high sperm mobility

The attributes of broiler breeder males characterized by sperm mobility phenotype were studied in replicate experiments. Low and high phenotypes were observed in the first experiment over a 10-wk interval. Average straight line velocity differed between phenotypes (P < or = 0.05) as evidenced by computer-assisted sperm motion analysis. The second experiment compared low and high phenotypes from the same broiler breeder strain over a 29-wk interval. Body weight and fertility were measured in this experiment. Body weight data approximated parallel lines. The low phenotype averaged 227 g more than the high phenotype over the course of the experiment. Individual males were used as semen donors at 50, 54, 58, 62, and 65 wk of age. Eggs were collected for 2 wk after a single insemination at each age. Mean overall fertility (+/− SEM) was 44 +/− 4 and 69 +/− 2% for low and high sperm mobility phenotypes, respectively (P < or = 0.0001). As determined by transmission electron microscopy, the percentage of sperm with aberrant mitochondria was greater for the low sperm mobility phenotype (P < or = 0.0001). Neither blood plasma testosterone concentration nor testis weight differed between phenotypes. Sperm mobility is a biologically significant predictor of broiler breeder semen quality. The relationship between sperm mobility phenotype and body weight warrants further investigation.

Sperm mobility phenotype not determined by sperm quality index

We tested the capacity of the AviMate sperm quality analyzer to predict sperm mobility phenotype. A preliminary experiment was performed to determine a sperm concentration that afforded maximal sperm quality index (SQI) values with sperm from males with high sperm mobility. In order to facilitate comparison of sperm motility indexes, semen was diluted to aconstant concentration of 1.25 million sperm/mL rather than a constant ratio of 1:75 as recommended for the sperm quality analyzer. Thereafter, sperm mobility, motile concentration, and SQI were measured from a single ejaculate collected from each of 70 broiler breeders. Coefficients of variation were 60, 37, and 30%, respectively. Neither sperm mobility (r = 0.01) nor motile concentration (r = 0.14) was highly correlated with SQI. Neither motile concentration nor straight line velocity (VSL) differed (P > or = 0.05) among roosters whose SQI values were below or above one standard deviation from the population mean (n = 10 per group). In contrast, both motile concentration (P < or = 0.01) and VSL (P < or = 0.05) differed between roosters whose sperm mobility values were below or above a standard deviation from the population mean (n = 10 per group). Sperm mobility was a function of the size of a highly motile subpopulation of sperm. A replicate experiment was performed with a flock of New Hampshire roosters (n = 126). Neither the properties of motile sperm nor fertility differed among males characterized by extreme SQI scores. We concluded that variation in SQI scores was enigmatic because sperm concentration was controlled in our experiments, sperm viability was invariant, and motile concentration did not differ among roosters characterized by extreme SQI scores. In summary, the AviMate sperm quality analyzer did not predict sperm mobility phenotype.

Efficacy of sperm mobility assessment in commercial flocks and the relationships of sperm mobility and insemination dose with fertility in turkeys

Our objectives were to evaluate: 1) the efficacy of the Sperm Mobility Test on commercial turkey farms, and 2) the influence of sperm mobility phenotype on fertility when insemination parameters are varied. In research flocks, differences in sperm mobility among toms are predictive of fertility. We wanted to test the efficacy of this sire selection test in practical, real-world situations, evaluating its usefulness in terms of assessing large numbers of toms, different strains of turkeys, and variable management practices. Utilizing field study results, controlled studies were then conducted to improve test parameters. For the field trials, semen from each of 405 breeder toms (11 strains or lines) was evaluated either in duplicate (n = 285) or in triplicate (n = 120). Sperm mobility was normally distributed among all toms tested, except for one strain. Because the sperm mobility indices for toms evaluated in these field trials were higher than those observed in research flocks, the Sperm Mobility Test was modified to increase the separation between high and low sperm mobility phenotypes by increasing the concentration of Accudenz. To determine the effects of sperm mobility and insemination dose on sustained fertility through time, hens from a research flock were inseminated twice before the onset of lay with sperm from toms classified as high-, average-, or low-mobility in concentrations of 25 to 400 million sperm per artificial insemination dose, and egg fertility was evaluated over a 5-wk period. Toms with the high-mobility sperm phenotype maintained higher fertility (P < 0.05) over the 5-wk period at all insemination doses compared with toms with low-mobility sperm. Toms with high-mobility sperm sired equal numbers of poults in a sperm competition study in which numbers favored low-mobility toms by 3:1. These results demonstrate that the Sperm Mobility Test can be used for on-farm evaluation of semen quality of toms in commercial flocks and that sperm mobility influences fertility and sire fitness.

Sperm mobility: phenotype in roosters (Gallus domesticus) determinedby concentration of motile sperm and straight line velocity.

Previous research demonstrated that sperm mobility, i.e., the net movement of a sperm population, is a quantitative trait of the domestic fowl. However, the cellular basis for this trait was unknown. In the present work, individual motile sperm were evaluated with a Hobson SpermTracker in order to identify one or more properties of motile sperm that could account for variation in sperm mobility observed among males. A method was validated for assessing sperm motion over an erythrocyte monolayer at body temperature. A small-scale experiment with roosters from the tails and center of a normal distribution of sperm mobility phenotypes (n = 33 roosters) demonstrated that straight line velocity (VSL) and motile concentration were critical to expression of phenotype. The importance of these variables was confirmed with a large-scale experiment using a representative subpopulation (n = 100 roosters). VSL of individual sperm at 41 degrees C ranged between 5 and 100 microm/sec. VSL averaged 32, 39, and 40 microm/sec for low, average, and high sperm mobility phenotypes. Sperm were diluted to 1.2 x 10(6)/ml for motion analysis. Mean motile concentrations were 0.52, 0.84, and 0.95 x 10(6)/ml for low, average, and high sperm mobility phenotypes. Motile concentration was correlated with sperm mobility (r = 0.71). VSL appeared to have an additive effect as it was correlated with straightness of sperm cell trajectory (r = 0.79).

Sperm mobility: A primary determinant of fertility in the domestic fowl (Gallus domesticus).

Previous research demonstrated that sperm mobility is a quantitative trait of the domestic fowl. The trait is quantified by measuring the absorbance of an Accudenz solution after overlay with a sperm suspension and brief incubation at body temperature. In the present work, average and high sperm mobility phenotypes (n = 30 males per phenotype) were selected from a base population. Differences were found between sperm oxygen consumption (p < 0.0001), acylcarnitine content (p < 0.05), linear velocity (p < 0.001), and straightness (p < 0.001), a trajectory variable measured with the Hobson SpermTracker. Oxygen consumption and stearoylcarnitine content of sperm from the high-mobility phenotype were twice those observed with sperm from average males, implying a pivotal role for mitochondria. On the basis of these results, a graded relationship was predicted between fertility and sperm mobility. Males (n = 48) were chosen at random from another base population, sperm mobility was measured per male, and each ejaculate was used to inseminate 8-12 hens (8 x 10(7) viable sperm per hen). When fertility was plotted as a function of sperm mobility, data points approximated a skewed logistic function. The hypothesis that vaginal immunoglobulins constitute an immunological barrier to sperm transport was tested and rejected. Therefore, we concluded that sperm mobility is a primary determinant of fertility in the fowl.

Turkey sperm mobility influences paternity in the context of competitive fertilization.

We have devised a novel means of investigating competitive fertilization in turkeys, using microsatellite genotyping to identify male parentage. Our results demonstrate that sperm mobility is a mechanism responsible in part for paternity efficiency in turkeys. Sperm mobility is composed of several parameters in which sperm motility is a component. Differences between ejaculates in the number of sperm penetrating into a dense, insert, nontoxic solution were measured and used to classify males into high, average, or low sperm mobility phenotypes. Microsatellite genotyping was used to determine parentage of poults after equal numbers of sperm from 10 males (either high or average phenotype, n = 5, mixed with low phenotype, n = 5) were inseminated simultaneously. In a separate study, the numbers of sperm hydrolyzing the perivitelline layer of eggs were compared between hens inseminated with sperm from high-, average-, or low-phenotype males. Overall, heterospermic inseminations resulted in consistently fewer offspring produced by low-mobility phenotype males. This correlated with physiological data in which semen from the low-mobility males had reduced numbers of sperm at the fertilization site as determined by sperm hole counts in the perivitelline layer of eggs. This is the first illustration of a measurable sperm trait predictive of paternity success in a competitive fertilization trial in turkeys, a species that is predominately reproduced by artificial insemination of multiple-sire pools.

Assessment of ejaculate quality and sperm characteristics in turkeys: sperm mobility phenotype is independent of time

Given a pool of genetically superior male turkeys, the subsequent selection of toms as semen donors for artificial insemination should be based upon criteria that are predictive of the tom's fertility and fecundity over the course of a production cycle. Recently, sperm mobility phenotype has been shown to correlate highly with fertility. Therefore, the objectives of this study were to determine: 1) whether sperm mobility phenotypes of individual toms were independent of time, and 2) the extent to which traditional semen quality variables changed as a function of time during the study interval. Sperm mobility phenotype was determined by objectively measuring the ability of sperm to penetrate 2% Accudenz solution at body temperature. During the initial assessments of a flock (n = 94), sperm mobility indexes (SMI) were significantly higher for the High Mobility Phenotype toms (56.61 +/-1.03% SMI) compared to the Low Mobility Phenotype (30.46+/-1.27% SMI, P < or = 0.0001) toms. Over the 5 mo of this study, the High Mobility Phenotype toms consistently had higher (P < or = 0.05) SMI values than the Low Mobility Phenotype toms, with SMI values similar to those of the initial screen. Ejaculate volume, concentration, and plasma membrane integrity showed no significant differences between phenotypes (P > or = 0.05). Sperm viability remained significantly lower (P < or = 0.05) over the course of this study for the High Mobility Phenotype toms than for the Low Mobility Phenotype toms. Because sperm mobility phenotype remained consistent through time, the Sperm Mobility Test provides a potentially important tool for selecting semen donors in turkeys.

Semen donor selection by in vitro sperm mobility increases fertility and semen storage in the turkey hen.

Commercial turkey production relies on the artificial insemination (AI) of pooled semen. However, semen quality ultimately depends on the quality of individual ejaculates. The purpose of this study was to evaluate semen from individual toms by means of an objective sperm-mobility assay. Semen was then pooled by mobility phenotype and inseminated into hens, and the percentages of fertile and hatched eggs were determined after egg incubation. To indirectly evaluate hens' sperm storage, we determined the number of sperm holes in the perivitelline layer (PL) of freshly laid eggs. Semen from individual ejaculates (two trials, total of 169 toms) was evaluated by use of the sperm-mobility test (SMT). Semen was diluted to 1 x 10(9) sperm/ml, in prewarmed N-tris-[hydroxymethyl] methyl-2-amino-ethanesulfonic acid (TES)-buffered saline, and was placed over 3 ml of a 2% (w/v) Accudenz solution at 41degrees C. After a 5-minute incubation period, the cuvette was placed in a densimeter, and percentage transmission was recorded after 1 minute. Semen samples from toms ranked, according to sperm mobility, in the highest 10% and the lowest 10%, after three evaluations, were pooled by group and were used to inseminate hens weekly (trial 1: n = 20 hens/group, for 10 weeks, AI dose 150 x 10(6) spermatozoa inseminated fresh and after 24-hour in vitro storage at 5 degrees C; trial 2: n = 60 hens/group, for 16 weeks, AI dose = 75 x 10(6) spermatozoa inseminated fresh). Each week, eggs from day 6 post-AI were evaluated for holes in the PL, vestiges of acrosomal induced hydrolysis. Spermatozoa from toms of different mobility phenotypes were also evaluated individually, for sperm chromatin structure and motility variables, by use of the Hobson Sperm Tracker. Toms characterized by high and low in vitro sperm-mobility phenotype were categorized as "high mobility" and "low mobility," respectively. The percentage of fertile eggs from hens inseminated with semen from the high-mobility toms was higher than the percentage of fertile eggs from the low-mobility group, in each trial (95.8+/-1.3% vs. 90.4+/-2.2%, and 88.7+/-4.0% vs. 82.4+/-0.4%, trials 1 and 2, respectively; P < 0.05). More sperm holes were observed in the PL of eggs fertilized by the high-mobility toms than in the PL of eggs fertilized by the low-mobility toms (P < 0.05). No differences in susceptibility of sperm nuclear DNA to denature in situ, as measured by the flow-cytometric sperm chromatin-structure assay, were detected between toms of differing mobility phenotypes. Sperm-motility variables, straight-line velocity, and average-path velocity were significantly greater for high-mobility toms compared to low-mobility toms (P < 0.05). Sperm-mobility differences between toms (detected by means of the SMT) influenced sperm storage, as indicated by the number of sperm in the PL and by the percentage of fertile eggs produced.

Sperm mobility: a quantitative trait of the domestic fowl (Gallus domesticus).

Sperm from each rooster within a base population (n = 271) were evaluated with a mobility assay validated in previous work. Frequency analysis confirmed a normal distribution for the variable of sperm mobility. Repeated-measure analysis of males categorized by phenotype demonstrated that average and high sperm mobility phenotypes were distinct and independent of time. Sperm morphology, fertilizing ability, and ATP content were compared between phenotypes. Fertility and sperm ATP content differed (p < 0.001) between phenotypes, whereas sperm morphology did not (p > 0.05). Experimentation with washed sperm demonstrated that phenotype was fully expressed when mitochondrial respiration was the only source of ATP. Sperm mobility increased (p < 0.001) when sperm from average males were exposed to calyculin A, a protein phosphatase inhibitor. Correlation analyses were performed with data from a subpopulation (n = 46) whose range, mean, and variance were equivalent to those of the base population. Neither body weight nor the combined weight of the testes was correlated with sperm mobility (r = -0.02 and 0.01, respectively). In contrast, sperm ATP content was correlated with sperm mobility (r = 0.80). We attribute phenotypic differences in sperm mobility to differential rates of mitochondrial ATP synthesis.