Genetics of fresh and frozen-thawed semen traits and their relationship with growth rate in rabbits

Se utilizarán eyaculados procedentes de machos de la línea R (línea de conejos seleccionada por velocidad de crecimiento durante el periodo de engorde)alojados en diferentes centros de inseminación artificial. Una vez recuperados los eyaculados se procederá a su valoración y una muestra de todos ellos será crioconservada. La calidad seminal será de nuevo valorada tras el proceso de congelación. Junto con los anàlisis seminales se utilizarán los datos de crecimiento y pedigree de los machos y de todos los animales de la línea R desde su fundación para estimar por un lado los parámetros genéticos de las variables relacionadas con la producción y calidad de dosis seminales en fresco y tras un proceso de crioconservación y la correlación genética existente entre las variables seminales anteriormente citadas y la velocidad de crecimiento. A su vez se estimará mediante un modelo recursivo la relación entre las variables seminales en fresco y tras la descongelación.The general aim of this thesis was to study the genetic determinism for some traits related to artificial insemination (AI) dose production of fresh and frozen-thawed semen, in order to explore the interest and limitation of different strategies for their genetic improvement in a paternal line of rabbits selected for growth rate during the fattening period (28-63 days). In chapter 1, genetic parameters of sperm production traits are estimated as well as the genetic relationship with daily gain (DG). The heritabilities (h2) of the semen traits were 0.13±0.05, 0.08±0.04 and 0.07±0.03 for ejaculate volume (V), sperm concentration (CN) and sperm production (PROD) per ejaculate, respectively. A favourable and moderate genetic correlation was observed between V and DG (0.36±0.34). From this chapter it may be concluded that if a seminal trait is to be included as a selection objective, a useful one could be sperm production, as it is a trait in which both volume and concentration are included. Moreover, there is currently no evidence to suggest that selection for DG in rabbits will affect sperm production adversely. The aim of chapter 2 was to explore the genetic determinism of some sperm quality traits and their genetic relation with the selection criteria of the paternal rabbit line. The heritabilities (h2) of semen quality traits commonly evaluated in a classic spermiogram were 0.18, 0.19 and 0.12 for NAR (%, percentage of sperm with intact acrosome), ANR (%, percentage of sperm abnormalities) and MOT (%, percentage of total motile sperm cells) respectively. We also estimated the h2 of some motion CASA parameters 0.09, 0.11, 0.10, 0.11, 0.11 and 0.11 for VAP (µm/s; average path velocity), VSL (µm/s; straight-line velocity), VCL (µm/s; curvilinear velocity), LIN (%, linearity index), ALH (µm; amplitude of the lateral head displacement), STR (%, straightness). Genetic correlations between DG and semen traits showed a high HPD95% (interval of highest density of 95%). However there is some consistent evidence of the negativity of the genetic correlations of DG with NAR and MOT (-0.40 and -0.53, respectively). Chapter 3 aims to determine the repeatability and heritability of sperm head characteristics: width (W, ¿m), area (A, ¿m2),length (L, ¿m) and perimeter (P, ¿m), and explore the relationships between them and with the selection objective (DG). The results obtained showed that sperm head dimensions are heritable (ranged between 0.2 and 0.29). The genetic correlations between sperm traits were always high and positive (between 0.72 and 0.90), with the exception of L-W genetic correlation, which was moderate. Regarding the genetic correlations between DG and sperm head characteristics, the resulting means ranged from -0.09 for L-DG to -0.43 for W-DG, showing consistent evidence of the negativity of the genetic correlations. The environmental and male effects that could have an influence on sperm freezability are studied in Chapter 4. Six different traits were evaluated: sperm concentration (CONC, 106spermatozoa/mL), acrosome integrity in fresh (NAR, %) and frozen-thawed semen (Nar-FT, %), sperm motility in fresh (MOT, %) and frozen-thawed semen (Mot-FT, %) and the percentage of viable sperm in frozen-thawed semen (Live-FT, %). In addition, two synthetic traits were computed: the relative reduction of acrosome integrity (Rnar, %) and relative reduction of motility (Rmot, %) after the freezing-thawing process. A multiple-trait recursive model was used to analyse the relationships between the semen traits considered. For the fixed effects studied, the season had the highest impact on post-thaw semen characteristics. Results of the analysis of recursive coefficients showed that fresh semen concentration and motility influence the future freezability of the semen. All traits studied presented moderate repeatabilities, ranging from 0.11 to 0.38. These results provide conclusive evidence that sperm freezability in rabbits could be heritable. Regarding male correlations, there were large positive male correlations between fresh traits (rm=0.77-0.57), as well as between direct frozen-thawed traits (rm=0.72-1). Male effects on fresh and direct frozen-thawed traits were generally positively correlated. This correlation was moderate to high for MOT with all frozen-thawed traits (rm=0.41-0.74) and for Mot-FT and all fresh traits (rm=0.5-0.74); these results suggest that these traits could be genetically related. The final chapter of this thesis focused on estimating the heritability of semen freezability traits and estimating the genetic correlation between frozen-thawed sperm traits and the growth rate in a paternal rabbit line. Estimated heritabilities showed that frozen-thawed semen traits are heritable (ranged between 0.08 and 0.15). In the case of Live-FT, the estimated heritability is the highest and suggests the possibility of effective selection. After the study of genetic correlations, it seems that DG was negatively correlated with sperm freezability, but due to the high HPD95% no further conclusions could be drawn. More data should be included in order to obtain better accuracy for the estimates of these genetic correlations. If the results obtained in the present study were confirmed, it would imply that selection for DG could alter sperm cell membranes or seminal plasma composition, both components related to sperm cryoresistance.Lavara García, R. (2013). Genetics of fresh and frozen-thawed semen traits and their relationship with growth rate in rabbits [Tesis doctoral no publicada]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/31657TESI

Freezability genetics in rabbit semen

[EN] The aim of this study was to estimate the heritability of semen freezability and to estimate the genetic correlation between frozen-thawed sperm traits and the growth rate in a paternal rabbit line. Estimated heritabilities showed that frozen-thawed semen traits are heritable (ranged between 0.08 and 0.15). In the case of Live-FT (percentage of viable sperm after freezing), the estimated heritability is the highest one, and suggests the possibility of effective selection. After the study of genetic correlations it seems that daily weight gain (DG) was negatively correlated with sperm freezability, but no further conclusions could be drawn due to the high HPD95%. More data should be included in order to obtain better accuracy for the estimates of these genetic correlations. If the results obtained at present study were confirmed, it would imply that selection for DG could alter sperm cell membranes or seminal plasma composition, both components related to sperm cryoresistance. (C) 2017 Elsevier Inc. All rights reserved.This work was supported by the Generalitat Valenciana research program (Prometeo II 2014/036). Lavara R. acknowledges the partial support received from Generalitat Valenciana under VALid program (APOST/2014/034) and from Ministry of Economy and Competitiveness under subprogramme "Formation posdoctoral" (FPDI-2013-16707).Lavara García, R.; Moce Cervera, ET.; Baselga Izquierdo, M.; Vicente Antón, JS. (2017). Freezability genetics in rabbit semen. Theriogenology. 102:54-58. https://doi.org/10.1016/j.theriogenology.2017.07.013S545810

Semen evaluation of two selected lines of rabbit bucks

[EN] Twenty rabbit bucks of 9 months of age were used to evaluate semen quality of two lines of New Zealand rabbit bucks selected for litter size at weaning (A line) and growth rate from weaning to slaughter (R line). The morphological semen characteristics indicated that the A line spermatozoa had greater acrosome integrity (+3.6 percentage units; P<0.01) and smaller sperm head size (for example, ¿1.46 ¿m2 for sperm head area) than in the R line. Seminal functional traits were also significantly higher for the A line (+13.4 percentage units for viability, +10.6 percentage units for hypo-osmotic swelling test (HOST) and +3.3 g/L for seminal plasma protein. However, no differences were detected between lines for motility parameters and seminal plasma protein electrophoretic profiles. Both lines had the same twelve bands with the following molecular weights to the nearest 1 kD: 124, 117, 99, 86, 75, 62, 40, 32, 21, 19, 10 and 6 kD. A relationship (r=0.308 for A line and 0.359 for R line; P<0.01) was found between the integrity of the plasmatic membrane (viability rate) and tail membrane (HOST) of the spermatozoa in the A line, but not in the R line, which had greater sperm head size. There was also a significant positive correlation coefficient between sperm concentration and either viability or some kinetic traits (r=0.567 and 0.575 for VCL, r=0.584 and 0.561 for VSL and r=0.588 and 0.588 for VAP, for A and R lines, respectively; P<0.001). We concluded that the A line seems to have better semen characteristics than the R line. We also found an interesting correlation among the seminal morphological, functional and kinetic traits, which could possibly be used to facilitate semen evaluation.Hosam M. Safaa was supported by a grant from ICSC-World Laboratory, Switzerland. This work has been supported by the Spanish Research Project (CICYT AGL2004-02710/GAN). The authors are indebted to F. Marco-Jiménez, Universidad Politécnica de Valencia, Valencia, Spain, and G.M.K. Mehaisen, Department of Animal Production, Faculty of Agricultural, Cairo University, Giza, Egypt, for their excellent cooperation throughout this study and to D.A.A. El-Sayed, Faculty of Agricultural, Cairo University, Giza, Egypt, for her help in preparing the manuscript.Safaa, H.; Vicente, J.; Lavara, R.; Viudes De Castro, M. (2008). Semen evaluation of two selected lines of rabbit bucks. World Rabbit Science. 16(3). doi:10.4995/wrs.2008.62216

Embryo vitrification in rabbits: Consequences for progeny growth

[EN] The objective of this research is to examine if there are any effects of the rederivation procedures on rabbit growth pattern and on weight of different organ in adults. For this purpose, three experiments were conducted on two different groups of animals (control group and vitrified transferred group) to evaluate the possible effect of embryo manipulation (vitrification and transfer procedures) on future growth traits. The first experiment studies body weight from 1 to 9 weeks of age from the two groups. The second experiment describes the growth curve of progeny from experimental groups and analyzes their Gompertz curve parameters, including the estimation of adult body weight. The third experiment has been developed to study if there are any differences in different organ weight in adult males from the two experimental groups. In general, the results indicate that rederivation procedures had effect on the phenotypic expression of growth traits. The results showed that rabbit produced by vitrification and embryo transfer had higher body weight in the first four weeks of age than control progeny. Results from body weight (a parameter) and b parameter estimated by fitting the Gompertz growth curve did not show any difference between experimental groups. However, differences related with growth velocity (k parameter of the Gompertz curve) were observed among them, showing that the control group had higher growth velocity than the vitrified transferred group. In addition, we found that liver weight at 40th week of age exhibits significant differences between the experimental groups. The liver weight was higher in the control males than in the VF males. Although the present results indicate that vitrification and transfer procedures might affect some traits related with growth in rabbits, further research is needed to assess the mechanisms involved in the appearance of these phenotypes and if these phenotypes could be transferred to the future progeny.This study was supported by the Generalitat Valenciana research program (Prometeo II 2014/036) and Spanish Research Projects (CICYT AGL2011-29831-C03-01; AGL2014-53405-C2-1-P). Lavara R. acknowledges the partial support received from Generalitat Valenciana under VALid+ program (APOST/2014/034) and from Ministry of Economy and Competitiveness under subprogramme "Formacion posdoctoral" (FPDI-2013-16707).Lavara García, R.; Baselga Izquierdo, M.; Marco Jiménez, F.; Vicente Antón, JS. (2015). Embryo vitrification in rabbits: Consequences for progeny growth. Theriogenology. 84(5):674-680. https://doi.org/10.1016/j.theriogenology.2015.04.025S67468084

Rederivation by Cryopreservation of a Paternal Line of Rabbits Suggests Exhaustion of Selection for Post-Weaning Daily Weight Gain after 37 Generations

[EN] This study was conducted to evaluate the effect of a long-term selection for post-weaning daily weight gain after 37 generations, using vitrified embryos with 18 generational intervals to rederive two coetaneous populations, reducing or avoiding genetic drift, environmental and cryopreservation effects. This study reports that the selection programme had improved average daily weight gain without variations in adult body weight but, after 37 generations of selection, this trait seems exhausted. Rabbit selection programmes have mainly been evaluated using unselected or divergently selected populations, or populations rederived from cryopreserved embryos after a reduced number of generations. Nevertheless, unselected and divergent populations do not avoid genetic drift, while rederived animals seem to influence phenotypic traits such as birth and adult weights or prolificacy. The study aimed to evaluate the effect of a long-term selection for post-weaning average daily weight gain (ADG) over 37 generations with two rederived populations. Specifically, two coetaneous populations were derived from vitrified embryos with 18 generational intervals (R19 and R37), reducing or avoiding genetic drift and environmental and cryopreservation effects. After two generations of both rederived populations (R21 vs. R39 generations), all evaluated traits showed some progress as a result of the selection, the response being 0.113 g/day by generation. This response does not seem to affect the estimated Gompertz growth curve parameters in terms of the day, the weight at the inflexion point or the adult weight. Moreover, a sexual dimorphism favouring females was observed in this paternal line. Results demonstrated that the selection programme had improved ADG without variations in adult body weight but, after 37 generations of selection, this trait seems exhausted. Given the reduction in the cumulative reproductive performance and as a consequence in the selection pressure, or possibly/perhaps due to an unexpected effect, rederivation could be the cause of this weak selection response observed from generation 18 onwards.Funding from the Ministry of Economy, Industry and Competitiveness (Research project: AGL2014-53405-C2-1-P) is acknowledged. English text version was revised by N. Macowan English Language Service.Juarez, JD.; Marco-Jiménez, F.; Lavara, R.; Vicente Antón, JS. (2020). Rederivation by Cryopreservation of a Paternal Line of Rabbits Suggests Exhaustion of Selection for Post-Weaning Daily Weight Gain after 37 Generations. Animals. 10(8):1-15. https://doi.org/10.3390/ani10081436S115108Estany, J., Baselga, M., Blasco, A., & Camacho, J. (1989). Mixed model methodology for the estimation of genetic response to selection in litter size of rabbits. Livestock Production Science, 21(1), 67-75. doi:10.1016/0301-6226(89)90021-3Lukefahr, S. 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Elliptical selection experiment for the estimation of genetic parameters of the growth rate and feed conversion ratio in rabbits1. Journal of Animal Science, 82(3), 654-660. doi:10.2527/2004.823654xDrouilhet, L., Gilbert, H., Balmisse, E., Ruesche, J., Tircazes, A., Larzul, C., & Garreau, H. (2013). Genetic parameters for two selection criteria for feed efficiency in rabbits1. Journal of Animal Science, 91(7), 3121-3128. doi:10.2527/jas.2012-6176Larzul, C., Gondret, F., Combes, S., & de Rochambeau, H. (2005). Divergent selection on 63-day body weight in the rabbit: response on growth, carcass and muscle traits. Genetics Selection Evolution, 37(1), 105. doi:10.1186/1297-9686-37-1-105Piles M., & Blasco A. (2010). RESPONSE TO SELECTION FOR GROWTH RATE IN RABBITS ESTIMATED BY USING A CONTROL CRYOPRESERVED POPULATION. World Rabbit Science, 11(2). doi:10.4995/wrs.2003.497Gondret, F., Combes, S., Larzul, C., & de Rochambeau, H. (2002). 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Theriogenology, 81(7), 988-992. doi:10.1016/j.theriogenology.2014.01.030Lavara, R., Baselga, M., Marco-Jiménez, F., & Vicente, J. S. (2015). Embryo vitrification in rabbits: Consequences for progeny growth. Theriogenology, 84(5), 674-680. doi:10.1016/j.theriogenology.2015.04.025Garcia-Dominguez, X., Marco-Jiménez, F., Peñaranda, D. S., & Vicente, J. S. (2020). Long-Term Phenotypic and Proteomic Changes Following Vitrified Embryo Transfer in the Rabbit Model. Animals, 10(6), 1043. doi:10.3390/ani10061043Garcia-Dominguez, X., Vicente, J. S., & Marco-Jiménez, F. (2020). Developmental Plasticity in Response to Embryo Cryopreservation: The Importance of the Vitrification Device in Rabbits. Animals, 10(5), 804. doi:10.3390/ani10050804Marco-Jiménez, F., Baselga, M., & Vicente, J. S. (2018). Successful re-establishment of a rabbit population from embryos vitrified 15 years ago: The importance of biobanks in livestock conservation. 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Effects of pharmacological agents on the lifespan phenotype of Drosophila DJ-1β mutants

Mutations in the DJ-1 gene cause autosomal recessive, early-onset Parkinsonism. The DJ-1 protein exerts a protective role against oxidative stress damage, working as a cellular oxidative stress sensor, and it seems to regulate gene expression at different levels. In Drosophila, two DJ-1 orthologs have been identified: DJ-1β and DJ-1β. Several studies have shown that loss of DJ-1β function causes Parkinson's disease (PD)-like phenotypes in flies such as age-dependent locomotor defects, reduced lifespan, and enhanced sensitivity to toxins that induce oxidative stress, like the herbicide paraquat. However, no dopaminergic neurodegeneration is observed. These results suggested that both locomotor and lifespan phenotypes could be either related to defects in oxidative stress response, or in dopaminergic physiology as proposed in mice models. In this study, we have employed pharmacological approaches to modify the lifespan phenotype of DJ-1β mutant flies. We have assessed the effects of chronic treatments with antiparkinsonian drugs as well as with antioxidant compounds on such phenotype finding that only antioxidants show statistically significant beneficial effects on DJ-1β mutants' lifespan. These results strongly suggest that oxidative stress plays a causal role in the lifespan phenotype of DJ-1β mutants. Consistent with this, we find that loss of DJ-1β function results in cellular accumulation of reactive oxygen species (ROS) in adult brains, elevated levels of lipid peroxidation and an increased catalase enzymatic activity, thus indicating the existence of high oxidative stress levels in DJ-1β mutants and confirming the essential function of the DJ-1β protein in protecting the organism against oxidative insults. Our study further shows that the lifespan phenotype of DJ-1β mutant flies is amenable to pharmacological intervention, and validates Drosophila as a valuable model for testing and identifying new drugs with therapeutic potential for PD. © 2010 Elsevier B.V.E. L.-C. was supported by a predoctoral fellowship from Consellería de Cultura, Educació i Ciència and R.G.-P. by a predoctoral fellowship of the I3P program from Consejo Superior de Investigaciones Científicas. V. M.-S. is a postdoctoral researcher funded by the Ministerio de Educación y Ciencia. This work has been supported by grants from Consellería de Cultura, Educació i Ciència and, in part, by grants from the Ministerio de Educación y Ciencia to N.P.Peer Reviewe

Long-term and transgenerational effects of cryopreservation on rabbit embryos

The short-term effects of cryopreservation and embryo transfer are well documented (reduced embryo viability, changes in pattern expression), but little is known about their long-term effects. We examined the possibility that embryo vitrification and transfer in rabbit could have an impact on the long-term reproductive physiology of the offspring and whether these phenotypes could be transferred to the progeny. Vitrified rabbit embryos were warmed and transferred to recipient females (F0). The offspring of the F0 generation were the F1 generation (cryopreserved animals). Females from F1 generation offspring were bred to F1 males to generate an F2 generation. In addition, two counterpart groups of noncryopreserved animals were bred and housed simultaneously to F1 and F2 generations (CF1 and CF2, respectively). The reproductive traits studied in all studied groups were litter size (LS), number born alive at birth (BA), and postnatal survival at Day 28 (number of weaned/number born alive expressed as percentage). The reproductive traits were analyzed using Bayesian methodology. Features of the estimated marginal posterior distributions of the differences between F1 and their counterparts (F1 - CF1) and between F2 and their counterparts (F2 - CF2) in reproductive characters found that vitrification and transfer procedures cause a consistent increase in LS and BA between F1 and CF1 females (more than 1.4 kits in LS and more than 1.3 BA) and also between F2 and CF2 females (0.96 kits in LS and 0.94 BA). We concluded that embryo cryopreservation and transfer procedures have long-term effects on derived female reproduction (F1 females) and transgenerational effects on female F1 offspring (F2 females).Lavara García, R.; Baselga Izquierdo, M.; Marco Jiménez, F.; Vicente Antón, JS. (2014). Long-term and transgenerational effects of cryopreservation on rabbit embryos. Theriogenology. 81(7):988-992. doi:10.1016/j.theriogenology.2014.01.030S98899281

Evaluation by re-derivation of a paternal line after 18 generations on seminal traits, proteome and fertility

[EN] Males from a paternal line selected for growth traits were used to produce semen doses at insemination centres and farms in a breeding scheme for rabbit meat production. The aim of this study was to assess whether a program of selection by daily gain in fattening period changed the seminal traits, plasma and sperm proteome and the fertility of semen when used in artificial insemination. Thirty-nine males from a paternal line were obtained by re-derivation from vitrified embryos with a difference of 18 generations (G21V and G39V). Sperm production parameters, morphological traits, sperm motility parameters and viability were evaluated from ejaculates. Seminal plasma and sperm proteome of three pool ejaculates from 10 mature males of each group were analysed and semen doses were used to inseminate 311 females. Only the percentage of abnormal sperm showed significant differences, with G21V presenting fewer abnormal sperm than G39V (10.5 +/- 2.63 vs 23.8 +/- 1.98). The discriminant analysis (DA-PLS) showed a clear effect of the generation for plasma and sperm proteome. In seminal plasma, 643 proteins were reported and 64 proteins were differentially expressed, of which 56 were overexpressed in G39V (87.5%). Sperm proteome reported 1360 proteins with 132 differentially abundant proteins. Of the total, 89 proteins were overexpressed in G39V (67.4%). From the 64 and 132 differentially abundant proteins of plasma and sperm, 19 and 26 had a FC >1.5, 12 and 13 of them belonging to the Oryctolagus cuniculus taxonomy, respectively. Despite observing differences in important proteins related to capacitation, sperm motility or immunoprotection and consequently to the fertilization process (TMPRSS2, Serpin family, Farn71f1, ATPase H+ transporting accessory protein 2, carbonic anhydrase 2, UDP-glucose glycoprotein glucosyltransferase 2), no differences in fertility and prolificacy were detected when commercial seminal doses were used for insemination from both male groups. However, overabundance of KIAA1324 protein can be related to the increase in abnormal sperm after selection by growth rate.This research was supported by AGL2017-85162-C2-1-R research project funded by Ministerio de Economia, Industria y Competitividad (MICINN, Spain). X Garcia-Dominguez was supported by a research grant from MICINN (BES-2015-072429). English text version was revised by N. Macowan English Language Service.Juárez, JD.; Marco-Jiménez, F.; Talaván, AM.; García-Domínguez, X.; Viudes-De-Castro, MP.; Lavara, R.; Vicente Antón, JS. (2020). Evaluation by re-derivation of a paternal line after 18 generations on seminal traits, proteome and fertility. Livestock Science. 232:1-13. https://doi.org/10.1016/j.livsci.2019.103894S113232Antalis, T. M., Bugge, T. H., & Wu, Q. (2011). Membrane-Anchored Serine Proteases in Health and Disease. Proteases in Health and Disease, 1-50. doi:10.1016/b978-0-12-385504-6.00001-4Bezerra, M. J. B., Arruda-Alencar, J. M., Martins, J. A. M., Viana, A. G. A., Viana Neto, A. M., Rêgo, J. P. A., … Moura, A. A. (2019). Major seminal plasma proteome of rabbits and associations with sperm quality. Theriogenology, 128, 156-166. doi:10.1016/j.theriogenology.2019.01.013Brun, J.-M., Theau-Clément, M., & Bolet, G. (2002). The relationship between rabbit semen characteristics and reproductive performance after artificial insemination. Animal Reproduction Science, 70(1-2), 139-149. doi:10.1016/s0378-4320(01)00197-xBrun, J.-M., Theau-Clément, M., Esparbié, J., Falières, J., Saleil, G., & Larzul, C. (2006). Semen production in two rabbit lines divergently selected for 63-d body weight. Theriogenology, 66(9), 2165-2172. doi:10.1016/j.theriogenology.2006.07.004Brun, J. M., Sanchez, A., Ailloud, E., Saleil, G., & Theau-Clément, M. (2016). Genetic parameters of rabbit semen traits and male fertilising ability. Animal Reproduction Science, 166, 15-21. doi:10.1016/j.anireprosci.2015.12.008Bünger, L., Lewis, R. M., Rothschild, M. F., Blasco, A., Renne, U., & Simm, G. (2005). Relationships between quantitative and reproductive fitness traits in animals. Philosophical Transactions of the Royal Society B: Biological Sciences, 360(1459), 1489-1502. doi:10.1098/rstb.2005.1679Casares-Crespo, L., Fernández-Serrano, P., Vicente, J. S., Marco-Jiménez, F., & Viudes-de-Castro, M. P. (2018). Rabbit seminal plasma proteome: The importance of the genetic origin. Animal Reproduction Science, 189, 30-42. doi:10.1016/j.anireprosci.2017.12.004Casares-Crespo, L., Fernández-Serrano, P., & Viudes-de-Castro, M. P. (2019). Proteomic characterization of rabbit (Oryctolagus cuniculus) sperm from two different genotypes. Theriogenology, 128, 140-148. doi:10.1016/j.theriogenology.2019.01.026Castellini, C., Lattaioli, P., Moroni, M., & Minelli, A. (2000). Effect of seminal plasma on the characteristics and fertility of rabbit spermatozoa. Animal Reproduction Science, 63(3-4), 275-282. doi:10.1016/s0378-4320(00)00181-0Castellini, C., Cardinali, R., Dal Bosco, A., Minelli, A., & Camici, O. (2006). Lipid composition of the main fractions of rabbit semen. Theriogenology, 65(4), 703-712. doi:10.1016/j.theriogenology.2005.05.053Castellini, C., Mourvaki, E., Cardinali, R., Collodel, G., Lasagna, E., Del Vecchio, M. T., & Dal Bosco, A. (2012). Secretion patterns and effect of prostate-derived granules on the sperm acrosome reaction of rabbit buck. Theriogenology, 78(4), 715-723. doi:10.1016/j.theriogenology.2012.02.012Courtens, J., Bolet, G., & Theau-Clément, M. (1994). Effect of acrosome defects and sperm chromatin decondensation on fertility and litter size in the rabbit. Preliminary electron-microscopic study. Reproduction Nutrition Development, 34(5), 427-437. doi:10.1051/rnd:19940504Choucair, F., 2018. Unraveling the sperm transcriptome by nextgeneration sequencing and the global epigenetic and landscape in infertile men. Molecular Biology.Université Côted’ Azur; Université libanaise, NNT:2018AZUR4058. https://tel.archives-ouvertes.fr/tel-01958881.Davis, B. K., & Davis, N. V. (1983). Binding by glycoproteins of seminal plasma membrane vesicles accelerates decapacitation in rabbit spermatozoa. Biochimica et Biophysica Acta (BBA) - Biomembranes, 727(1), 70-76. doi:10.1016/0005-2736(83)90370-xEllerman, D. A., Myles, D. G., & Primakoff, P. (2006). A Role for Sperm Surface Protein Disulfide Isomerase Activity in Gamete Fusion: Evidence for the Participation of ERp57. Developmental Cell, 10(6), 831-837. doi:10.1016/j.devcel.2006.03.011Estany, J., Camacho, J., Baselga, M., & Blasco, A. (1992). Selection response of growth rate in rabbits for meat production. Genetics Selection Evolution, 24(6), 527. doi:10.1186/1297-9686-24-6-527García-Tomás, M., Sánchez, J., Rafel, O., Ramon, J., & Piles, M. (2006). Variability, repeatability and phenotypic relationships of several characteristics of production and semen quality in rabbit. Animal Reproduction Science, 93(1-2), 88-100. doi:10.1016/j.anireprosci.2005.06.011García-Tomás, M., Sánchez, J., Rafel, O., Ramon, J., & Piles, M. (2006). Heterosis, direct and maternal genetic effects on semen quality traits of rabbits. Livestock Science, 100(2-3), 111-120. doi:10.1016/j.livprodsci.2005.08.004Garénaux, E., Kanagawa, M., Tsuchiyama, T., Hori, K., Kanazawa, T., Goshima, A., … Kitajima, K. (2015). Discovery, Primary, and Crystal Structures and Capacitation-related Properties of a Prostate-derived Heparin-binding Protein WGA16 from Boar Sperm. Journal of Biological Chemistry, 290(9), 5484-5501. doi:10.1074/jbc.m114.635268Gerena, R. L., Irikura, D., Urade, Y., Eguchi, N., Chapman, D. A., & Killian, G. J. (1998). Identification of a Fertility-Associated Protein in Bull Seminal Plasma As Lipocalin-Type Prostaglandin D Synthase1. Biology of Reproduction, 58(3), 826-833. doi:10.1095/biolreprod58.3.826Gervasi, M. G., & Visconti, P. E. (2017). Molecular changes and signaling events occurring in spermatozoa during epididymal maturation. Andrology, 5(2), 204-218. doi:10.1111/andr.12320Jeyendran, R. S., Van der Ven, H. H., Perez-Pelaez, M., Crabo, B. G., & Zaneveld, L. J. D. (1984). Development of an assay to assess the functional integrity of the human sperm membrane and its relationship to other semen characteristics. Reproduction, 70(1), 219-228. doi:10.1530/jrf.0.0700219Kim, T. S., Heinlein, C., Hackman, R. C., & Nelson, P. S. (2006). Phenotypic Analysis of Mice Lacking the Tmprss2 -Encoded Protease. Molecular and Cellular Biology, 26(3), 965-975. doi:10.1128/mcb.26.3.965-975.2006Kwon, J. T., Ham, S., Jeon, S., Kim, Y., Oh, S., & Cho, C. (2017). Expression of uncharacterized male germ cell-specific genes and discovery of novel sperm-tail proteins in mice. PLOS ONE, 12(7), e0182038. doi:10.1371/journal.pone.0182038Larzul, C., Gondret, F., Combes, S., & de Rochambeau, H. (2005). Divergent selection on 63-day body weight in the rabbit: response on growth, carcass and muscle traits. Genetics Selection Evolution, 37(1), 105. doi:10.1186/1297-9686-37-1-105Lavara, R., Mocé, E., Lavara, F., Viudes de Castro, M. P., & Vicente, J. S. (2005). Do parameters of seminal quality correlate with the results of on-farm inseminations in rabbits? Theriogenology, 64(5), 1130-1141. doi:10.1016/j.theriogenology.2005.01.009Lavara, R., Vicente, J. S., & Baselga, M. (2010). Genetic parameter estimates for semen production traits and growth rate of a paternal rabbit line. Journal of Animal Breeding and Genetics, 128(1), 44-51. doi:10.1111/j.1439-0388.2010.00889.xLavara, R., Vicente, J. S., & Baselga, M. (2012). Estimation of genetic parameters for semen quality traits and growth rate in a paternal rabbit line. Theriogenology, 78(3), 567-575. doi:10.1016/j.theriogenology.2012.03.002Lavara, R., Vicente, J. S., & Baselga, M. (2013). Genetic variation in head morphometry of rabbit sperm. Theriogenology, 80(4), 313-318. doi:10.1016/j.theriogenology.2013.04.015Law, R. H., Zhang, Q., McGowan, S., Buckle, A. M., Silverman, G. A., Wong, W., … Whisstock, J. C. (2006). Genome Biology, 7(5), 216. doi:10.1186/gb-2006-7-5-216Leone, M. G., Haq, H. A., & Saso, L. (2002). Lipocalin type prostaglandin D-synthase: which role in male fertility? Contraception, 65(4), 293-295. doi:10.1016/s0010-7824(02)00280-9Lestari, S. W., Miati, D. N., Seoharso, P., Sugiyanto, R., & Pujianto, D. A. (2017). Sperm Na+, K+-ATPase α4 and plasma membrane Ca2+-ATPase (PMCA) 4 regulation in asthenozoospermia. Systems Biology in Reproductive Medicine, 63(5), 294-302. doi:10.1080/19396368.2017.1348565Liao, T.-T., Xiang, Z., Zhu, W.-B., & Fan, L.-Q. (2009). Proteome analysis of round-headed and normal spermatozoa by 2-D fluorescence difference gel electrophoresis and mass spectrometry. Asian Journal of Andrology, 11(6), 683-693. doi:10.1038/aja.2009.59Llobat, L., Marco-Jiménez, F., Peñaranda, D., Thieme, R., Navarrete, A., & Vicente, J. (2011). mRNA Expression in Rabbit Blastocyst and Endometrial Tissue of Candidate Gene Involved in Gestational Losses. Reproduction in Domestic Animals, 47(2), 281-287. doi:10.1111/j.1439-0531.2011.01855.xLoveland, K., Major, A., Butler, R., Jans, D., Miyamoto, Y., & Young, J. (2015). Putting things in place for fertilization: discovering roles for importin proteins in cell fate and spermatogenesis. Asian Journal of Andrology, 17(4), 537. doi:10.4103/1008-682x.154310Lukefahr, S. D., Odi, H. B., & Atakora, J. K. (1996). Mass selection for 70-day body weight in rabbits. Journal of Animal Science, 74(7), 1481. doi:10.2527/1996.7471481xMa, Q., Li, Y., Luo, M., Guo, H., Lin, S., Chen, J., … Gui, Y. (2017). The expression characteristics of FAM71D and its association with sperm motility. Human Reproduction, 32(11), 2178-2187. doi:10.1093/humrep/dex290Marai, I. F. ., Habeeb, A. A. ., & Gad, A. . (2002). Rabbits’ productive, reproductive and physiological performance traits as affected by heat stress: a review. Livestock Production Science, 78(2), 71-90. doi:10.1016/s0301-6226(02)00091-xMocé, E., Vicente, J. S., & Lavara, R. (2003). Effect of freezing–thawing protocols on the performance of semen from three rabbit lines after artificial insemination. Theriogenology, 60(1), 115-123. doi:10.1016/s0093-691x(02)01329-8Naturil-Alfonso, C., Lavara, R., Millán, P., Rebollar, P. G., Vicente, J. S., & Marco-Jiménez, F. (2016). Study of failures in a rabbit line selected for growth rate. World Rabbit Science, 24(1), 47. doi:10.4995/wrs.2016.4016Nizza, A., Di Meo, C., & Taranto, S. (2003). Effect of Collection Rhythms and Season on Rabbit Semen Production. Reproduction in Domestic Animals, 38(6), 436-439. doi:10.1046/j.1439-0531.2003.00458.xOsada, T., Watanabe, G., Kondo, S., Toyoda, M., Sakaki, Y., & Takeuchi, T. (2001). Male Reproductive Defects Caused by Puromycin-Sensitive Aminopeptidase Deficiency in Mice. Molecular Endocrinology, 15(6), 960-971. doi:10.1210/mend.15.6.0643Pascual, J. J., García, C., Martínez, E., Mocé, E., & Vicente, J. S. (2004). Rearing management of rabbit males selected by high growth rate: the effect of diet and season on semen characteristics. Reproduction Nutrition Development, 44(1), 49-63. doi:10.1051/rnd:2004016Pascual, J. J., Marco-Jiménez, F., Martínez-Paredes, E., Ródenas, L., Fabre, C., Juvero, M. A., & Cano, J. L. (2016). Feeding programs promoting daily feed intake stability in rabbit males reduce sperm abnormalities and improve fertility. Theriogenology, 86(3), 730-737. doi:10.1016/j.theriogenology.2016.02.026Pérez-Patiño, C., Parrilla, I., Li, J., Barranco, I., Martínez, E. A., Rodriguez-Martínez, H., & Roca, J. (2019). The Proteome of Pig Spermatozoa Is Remodeled During Ejaculation. Molecular & Cellular Proteomics, 18(1), 41-50. doi:10.1074/mcp.ra118.000840Peralta-Arias, R. D., Vívenes, C. Y., Camejo, M. I., Piñero, S., Proverbio, T., Martínez, E., … Proverbio, F. (2015). ATPases, ion exchangers and human sperm motility. REPRODUCTION, 149(5), 475-484. doi:10.1530/rep-14-0471Piles, M., & Tusell, L. (2011). Genetic correlation between growth and female and male contributions to fertility in rabbit. Journal of Animal Breeding and Genetics, 129(4), 298-305. doi:10.1111/j.1439-0388.2011.00975.xPiles, M., Mocé, M. L., Laborda, P., & Santacreu, M. A. (2013). Feasibility of selection for male contribution to embryo survival as a way of improving male reproductive performance and semen quality in rabbits1. Journal of Animal Science, 91(10), 4654-4658. doi:10.2527/jas.2013-6446Rahman, M. S., Kwon, W.-S., & Pang, M.-G. (2017). Prediction of male fertility using capacitation-associated proteins in spermatozoa. Molecular Reproduction and Development, 84(9), 749-759. doi:10.1002/mrd.22810Roca, J., Martínez, S., Orengo, J., Parrilla, I., Vázquez, J. M., & Martínez, E. A. (2005). Influence of constant long days on ejaculate parameters of rabbits reared under natural environment conditions of Mediterranean area. Livestock Production Science, 94(3), 169-177. doi:10.1016/j.livprodsci.2004.10.011De Rochambeau, H., de la Fuente, L., Rouvier, R., & Ouhayoun, J. (1989). Sélection sur la vitesse de croissance post-sevrage chez le lapin. Genetics Selection Evolution, 21(4), 527. doi:10.1186/1297-9686-21-4-527Saeed, A. I., Sharov, V., White, J., Li, J., Liang, W., Bhagabati, N., … Quackenbush, J. (2003). TM4: A Free, Open-Source System for Microarray Data Management and Analysis. BioTechniques, 34(2), 374-378. doi:10.2144/03342mt01Samanta, L., Parida, R., Dias, T. R., & Agarwal, A. (2018). The enigmatic seminal plasma: a proteomics insight from ejaculation to fertilization. Reproductive Biology and Endocrinology, 16(1). doi:10.1186/s12958-018-0358-6Sabés-Alsina, M., Planell, N., Torres-Mejia, E., Taberner, E., Maya-Soriano, M. J., Tusell, L., … Lopez-Bejar, M. (2015). Daily exposure to summer circadian cycles affects spermatogenesis, but not fertility in an in vivo rabbit model. Theriogenology, 83(2), 246-252. doi:10.1016/j.theriogenology.2014.09.013Shevchenko, A., Jensen, O. N., Podtelejnikov, A. V., Sagliocco, F., Wilm, M., Vorm, O., … Mann, M. (1996). Linking genome and proteome by mass spectrometry: Large-scale identification of yeast proteins from two dimensional gels. Proceedings of the National Academy of Sciences, 93(25), 14440-14445. doi:10.1073/pnas.93.25.14440Shilov, I. V., Seymour, S. L., Patel, A. A., Loboda, A., Tang, W. H., Keating, S. P., … Schaeffer, D. A. (2007). The Paragon Algorithm, a Next Generation Search Engine That Uses Sequence Temperature Values and Feature Probabilities to Identify Peptides from Tandem Mass Spectra. Molecular & Cellular Proteomics, 6(9), 1638-1655. doi:10.1074/mcp.t600050-mcp200Theau-Clément, M., Bolet, G., Sanchez, A., Saleil, G., & Brun, J. M. (2015). Some factors that influence semen characteristics in rabbits. Animal Reproduction Science, 157, 33-38. doi:10.1016/j.anireprosci.2015.03.011Thundathil, J. C., Rajamanickam, G. D., & Kastelic, J. P. (2018). Na/K-ATPase and Regulation of Sperm Function. Animal Reproduction, 15(Suppl. 1), 711-720. doi:10.21451/1984-3143-ar2018-0024Tusell, L., Legarra, A., García-Tomás, M., Rafel, O., Ramon, J., & Piles, M. (2012). Genetic basis of semen traits and their relationship with growth rate in rabbits1. Journal of Animal Science, 90(5), 1385-1397. doi:10.2527/jas.2011-4165Vicente, J. (2004). Study of fertilising capacity of spermatozoa after heterospermic insemination in rabbit using DNA markers. Theriogenology, 61(7-8), 1357-1365. doi:10.1016/j.theriogenology.2003.08.009Vicente, J. S., Llobat, L., Viudes-de-Castro, M. P., Lavara, R., Baselga, M., & Marco-Jiménez, F. (2012). Gestational losses in a rabbit line selected for growth rate. Theriogenology, 77(1), 81-88. doi:10.1016/j.theriogenology.2011.07.019Viudes-de-Castro, M. P., & Vicente, J. S. (1997). Effect of sperm count on the fertility and prolificity rates of meat rabbits. Animal Reproduction Science, 46(3-4), 313-319. doi:10.1016/s0378-4320(96)01628-4Viudes-de-Castro, M. P., Mocé, E., Lavara, R., Marco-Jiménez, F., & Vicente, J. S. (2014). Aminopeptidase activity in seminal plasma and effect of dilution rate on rabbit reproductive performance after insemination with an extender supplemented with buserelin acetate. Theriogenology, 81(9), 1223-1228. doi:10.1016/j.theriogenology.2014.02.003Viudes de Castro, M. P., Casares-Crespo, L., Monserrat-Martínez, A., & Vicente, J. S. (2015). Determination of enzyme activity in rabbit seminal plasma and its relationship with quality semen parameters. World Rabbit Science, 23(4), 247. doi:10.4995/wrs.2015.4064Vizcaíno, J. A., Deutsch, E. W., Wang, R., Csordas, A., Reisinger, F., Ríos, D., … Hermjakob, H. (2014). ProteomeXchange provides globally coordinated proteomics data submission and dissemination. Nature Biotechnology, 32(3), 223-226. doi:10.1038/nbt.2839Wandernoth, P. M., Mannowetz, N., Szczyrba, J., Grannemann, L., Wolf, A., Becker, H. M., … Wennemuth, G. (2015). Normal Fertility Requires the Expression of Carbonic Anhydrases II and IV in Sperm. Journal of Biological Chemistry, 290(49), 29202-29216. doi:10.1074/jbc.m115.698597Weininger, R. B., Fisher, S., Rifkin, J., & Bedford, J. M. (1982). Experimental studies on the passage of specific IgG to the lumen of the rabbit epididymis. Reproduction, 66(1), 251-258. doi:10.1530/jrf.0.0660251Yan, M., Zhang, X., Pu, Q., Huang, T., Xie, Q., Wang, Y., … Gu, J. (2016). Immunoglobulin G Expression in Human Sperm and Possible Functional Significance. Scientific Reports, 6(1). doi:10.1038/srep2016

Study of failures in a rabbit line selected for growth rate

[EN] Selection for growth rate is negatively related with reproductive fitness. The aim of this work was to analyse the causes of fertility failure in rabbit does selected for growth rate and characterised for reproductive deficiencies (line R). In the experiment, 82 does were divided into 2 groups: naturally mated (NM) and artificially inseminated (AI), to relate luteinizing hormone (LH) concentration with ovulation induction and pregnancy rate by laparoscopic determination. Additionally, in 38 of these females ovulation rate and metabolites determination (leptin, NEFA, BOHB and glucose) were analysed and perirenal fat thickness measurement and live body weight (LBW) determined. The results showed that all ovulated does (both NM and AI) presented higher concentrations of LH than non-ovulated females. In addition, non-ovulated females showed high levels of leptin and BOHB, as well as LBW. Females from line R have an inherit reduced fertility due to ovulation failure as a consequence of a reduction in LH release, which could be explained by a heavier body weight and higher leptin concentrations.This work was supported by the Spanish Research Project AGL2011-30170-C02-01 (CICYT). Carmen Naturil-Alfonso was supported by a research grant from the Education Ministry of the Valencian Regional Government (programme VALi+d. ACIF/2013/296). English text version was revised by N. Macowan English Language Service.Naturil Alfonso, C.; Lavara García, R.; Millán, P.; Rebollar, P.; Vicente Antón, JS.; Marco Jiménez, F. (2016). Study of failures in a rabbit line selected for growth rate. World Rabbit Science. 24(1):47-53. https://doi.org/10.4995/wrs.2016.4016SWORD475324

Breeding programmes to improve male reproductive performance and efficiency of insemination dose production in paternal lines: feasibility and limitations

[EN] This paper aims at reviewing the current genetic knowledge of the issues related to the efficient use of bucks in artificial insemination (AI). Differences between lines have been found relevant in semen production and quality traits not necessarily related to their specialization as maternal or paternal lines. Accurate heritability estimates indicate that genetic selection for increasing semen production by improving male libido and reducing the number of rejected ejaculates may not be effective. However, total sperm produced per ejaculate appears to be as an interesting trait to select for, despite that genetic correlation between ejaculate volume and sperm concentration has not been yet accurately estimated. Semen pH has shown low to medium heritability estimates and a low coefficient of variation, therefore it is not advisable to attempt improvement by direct selection. In general, sperm motility traits have shown low heritabilities but, the rate of motile sperms per ejaculate has been considered as convenient to select for. Morphological characteristics of the spermatozoa have revealed as medium to highly heritable. There are evidences of high genetic correlations between sperm traits before and after freezing-thawing. There are few studies regarding the estimation of heterosis of seminal traits but results indicate important and favorable direct and maternal heterosis in crosses between maternal lines. However, this has not been confirmed in a cross between two paternal lines. Until now, attempts to find parametric or non-parametric functions to predict ejaculate fertility through seminal characteristics recorded in routinely evaluations have been very unsatisfactory. Hence, it may be necessary to find other semen quality markers, or to evaluate some of the currently used ones in a more precise manner or closer to the AI time in order to improve the ability to predict ejaculate fertility. Several seminal characteristics phenotypically correlated to male fertility, could be considered as potential traits to select for in order to genetically improving this trait. However, only the semen pH has been checked for this purpose, and a negative result has been obtained. Other traits can be studied in the future but bearing in mind that the required experiments will need large number of bucks for an accurate estimation of the genetic correlation of the trait with male fertility. This means that these experiments will be expensive and difficult to set up. The most common criterion to select paternal lines, average daily gain, seems not to be genetically correlated to male fertility and seminal traits. Thus, selection for average daily gain has no detrimental consequences on these traits, and a multi-trait selection, including growth rate and seminal traits directly related to an efficient AI semen dose production, is feasible in paternal lines. The male contribution to fertility after natural mating and after AI with semen doses with high concentration is negligible, but it has been found that, under more restrictive conditions of AI, male contributions to fertility and litter size are low but higher in magnitude than the ones obtained after natural mating. The genetic correlation between the female and male contributions to fertility has been found to be moderate to high and positive.This study was supported by the Generalitat Valenciana Research Programme (Prometeo 2009/125) and Spanish Research Projects (INIA RTA2005-00088-CO2; INIA RTA2008-00070-CO2; CICYT AGL2008-03274). Raquel Lavara was supported by a research grant from the Spanish Ministry of Education (MEC, FPU AP2007-03755) and Llibertat Tusell by a research grant from INIA.Piles, M.; Tusell, L.; Lavara García, R.; Baselga Izquierdo, M. (2013). Breeding programmes to improve male reproductive performance and efficiency of insemination dose production in paternal lines: feasibility and limitations. World Rabbit Science. 21(2):61-75. doi:10.4995/wrs.2013.1240SWORD617521