Erythropoietin Receptor Gene (EPOR) Polymorphisms are Associated with Sow Litter Sizes

The erythropoietin receptor （EPOR） has shown to play an important role in fetal survival by promoting the maturation of red blood cells in many studies of uterine capacity and litter size in swine. In this study, we screened the porcine EPOR gene for mutations and identified five single nucleotide polymorphisms （SNPs）： g.705G〉T in intron 1, g.2 373C〉T in intron 4, and g.2 882C〉T, g.3 035A〉G, and g.3 132A〉T in intron 6. We then genotyped 247 Beijing Black （BB） sows and compared the polymorphism data with the litter sizes of 1 375 parities among the sows. At first parity, there was no association of g.2 882C〉T and g.3 132A〉T with litter sizes. However, the CT sows in g.2 882C〉T had 2.13 higher total number born （TNB） （P〈0.01） and 1.81 higher number born alive （NBA） （P〈0.01） than the CC sows and the heterozygous sows in g.3 132A〉T had the highest litter size when compared to the two homozygotes for the later parities （P〈0.05）. In the g.3 035A〉G SNP, for the later parities, the TNB of the sows with the GG genotype was 3.81 higher （P〈0.01） and the NBA was 2.75 higher （P〈0.01） than that with the AA genotype but no difference at first parity. The G allele of the EPOR g.705G〉T SNP was associated with a greater litter size at both the first parity （P〈0.05） and later parities （P〈0.01）. Furthermore, we determined the allele frequencies for this SNP among five Chinese indigenous pig breeds （Erhualian, Laiwu Black, Meishan, Min, and Rongchang） and three western commercial pig breeds （Duroc, Landrace, and Large White）. The G allele of the EPOR g.705G〉T SNP was significantly more common in the more prolific Chinese breeds. These results indicated that the EPOR could be an important candidate gene for litter size and g.705G〉T can serve as a useful genetic marker for improving litter size in both first and later parities in swine.

Analysis of PRLR and BF Genotypes Associated with Litter Size in Beijing Black Pig Population

This study is aimed at using the DNA mutations in the prolactin receptor （PRLR） and properdin （BF） genes to determine associations between the genotype and litter size in the Beijing Black pig population. A total of 321 Beijing Black pig sows were genotyped using the polymerase chain reaction-restriction fragment length polymorphism （PCR-RFLP） method, with the Alu I and Sma I for PRLR and BF genes, respectively. Two different alleles of PRLR and BF genes were identified： allele A （0.25） and B （0.75） of the PRLR gene, allele A （0.13） and B （0.86） of the BF gene. The association analysis between the genotypes and the litter size were estimated with the method of the general linear model. The analysis results of PRLR showed that in first parity, sows with genotype AA had a larger litter size than sows with genotype AB and BB, but the difference was statistically not significant. In later parities, statistically significant （P 〈 0.05） differences were seen between sows with genotypes AA and AB, and BB of the PRLR gene. The associated analysis results between genotypes and litter size （total number born, TNB, and number born alive, NBA） showed that there were no significant differences in the first parity sows with different genotypes of the BF gene, but significant differences appeared in NBA between the sows of genotypes AB and BB, in later parity, for which significantly higher values were observed in the offspring of heterozygotes. Considering the consistent genotypic effect on the litter size of both sows in first parity and later parity, it was concluded that the locus of the PRLR gene, digested with Alu I, could be the gene maker for the litter size in Beijing Black pigs.

Heritabilities and genetic and phenotypic correlations of litter uniformity and litter size in Large White sows

Litter uniformity, which is usually represented by within-litter weight coefficient of variation at birth （CVB）, could influence litter performance of sows and the profitability of pig enterprises. The objective of this study was to characterize CVB and its effect on other reproductive traits in Large White sows. Genetic parameters and genetic correlation of the reproductive traits, including CVB, within-litter weight coefficient of variation at three weeks （CVT）, total number born （TNB）, number born alive （NBA）, number born dead （NBD）, gestation length （GL）, piglet mortality at birth （Mo）, piglet mortality at three weeks （M3）, total litter weight at birth （TLW0）, and total litter weight at three weeks （TLW3） were estimated for 2 032 Large White litters. The effects of parity and classified litter size on CVB, CVT, TNB, NBA, NBD, GL, M0, M3, TLW0, and TLW3 were also estimated. The heritabilities of these reproductive traits ranged from 0.06 to 0.17, with the lowest heritability for CVB and the highest heritability for TLW0. Phenotypic and genetic correlations between these reproductive traits were low to highly positive and negative （ranging from -0.03 to 0.93, and -0.53 to 0.93, respectively）. The genetic correlations between TNB and CVB, and between M0 and CVB were 0.32 and 0.29, respectively. In addition, CVB was significantly influenced by parity and litter size class （P〈0.05）. All the results suggest that piglet uniformity should be maintained in pig production practices and pig breeding programs.

Study on the Effect of ESR and FSH β Genes on Litter Size in Local Northeast Three Pig Breeds

The method of PCR RFLP was used to analyze the effects of ESR and FSH β genotypes on the litter size,especially the corresponding changes with the difference of genotype distributions in the Min,Sanjiang and Junmu I pigs.The results show that the ESR and FSH β genes are the major genes of litter size;the heterozygous genotype for ESR as well as for FSH β locus has a more litter size than others.However,the combined genotype ABDD means a more litter size than other genotypes,the frequencies of the heterozygous genotypes in Sanjiang and Junmu I are lower than in Min which maybe one of the most important reasons for the little litter sizes of those two breeds.

Characterization of Porcine Matrix Metalloproteinase 23 (pMMP-23) Gene and Its Association with Litter Size Traits

The matrix metalloproteinase 23 （MMP-23）, which might play a role in ovulation in mammals, was one of the promising candidate genes for litter size traits in pigs. In the present research, partial sequence of porcine MMP-23 （pMMP-23） gene, including exons 2-8 （GenBank： EU360790）, was obtained. Real-time PCR analysis revealed that pMMP-23 gene was highly expressed in ovary. PCR-Sau3A I-RFLP and PCR-Acc II-RFLP assay were established to detect a C/T mutation in exon 3 （EU360790： g. 269C〉T） and an A/G mutation in exon 4 （EU360790： g. 505A〉G）, respectively. Association study for these two SNPs with litter size was assessed in three independent populations （Minzhu, Landrace I and Landrace II）. Statistical analysis demonstrated that for second and subsequent litters, TT sows produced more TNB than CC pigs in Landrace breed （P〈0.05） at g. 269C〉T locus, and the additive effect was significant （P〈0.05）; GG sows produced more TNB and NBA than AA pigs in Minzhu （P〈0.01） and Landrace breeds （P〈0.05） at g. 505A〉G locus, and the additive effect was significant （P〈0.01 or P〈0.05）. Our study suggested that the pMMP-23 gene might be a novel candidate gene for litter size traits, and g. 505A〉G locus might be a useful molecular marker for marker assisted selection （MAS）.

Identification of Novel Polymorphisms in Porcine Ring Finger Protein 4 and Matrix Metalloproteinase 9 Genes and Association Analysis with Litter Size Traits

Reproduction trait plays an important role in pig production. Identification of molecular markers that are linked to litter size may contribute to the genetic development of porcine reproduction traits. In this study, porcine ring finger protein 4 （pRNF4） and Matrix metalloproteinase 9 （pMMP-9） were selected as candidate genes on the basis of their physiological roles in reproduction. Two single-nucleotide polymorphisms （416C〉T in pRNF4 and - 1257G〉A in pMMP-9） that could be detected by PCR restriction fragment length polymorphism （PCR-RFLP） were discovered and tested for statistical associations with litter size traits in three populations. For 416C〉T, TT genotype was associated with a significantly higher （p〈0.05） number of live births than those recorded for CC sows and the additive effect was significant （p〈0.05） in Qingping and Min Pigs populations in later parities. For -1257G〉A, inconsistent results were found in three populations. The results suggested that T allele in pRNF4 gene might confer a high prolificacy in breeding and further studies were needed to confLrm the results.

Polymorphism of BMPR-IB Gene and its Relationship with Litter Size in Small Tail Han Sheep

[ Objective] To open an exit for breeding high-fecundity Small Tail Han sheep according to genotypes. [ Method] By the PCR-RFLP, the polymorphism of ovine BMPR-IB gene was detected in Small Tail Han sheep of basic population and breeding population. And its relationship with litter size was analyzed. [ Result] In the basic population, the frequencies of B ＋ and BB genotypes were 77.78% and 14.81%, respectively, and their litter size per parity was 1.77 and 2.40, respectively. In the breeding population, the frequencies of B ＋ and BB genotypes were 51.79% and 40.18%, respectively, and their litter size per parity was 2.54 and 3.02, respectively. [ Conclusion] The ovine BMPR-IB gene can be used as a molecular Qenetic marker for fecundity traits to establish high-fecundity Small Tail Han sheep.

Expression of genes involved in progesterone receptor paracrine signaling and their effect on litter size in pigs

Background： Embryonic mortality during the period of implantation strongly affects litter size in pigs. Progesterone receptor（PGR） paracrine signaling has been recognized to play a significant role in embryonic implantation. IHH,NR2F2, BMP2, FKBP4 and HAND2 were proved to involve in PGR paracrine signaling. The objective of this study was to evaluate the expression of IHH, NR2F2, BMP2, FKBP4 and HAND2 in endometrium of pregnant sows and to further investigate these genes＇ effect on litter size in pigs. Real-time PCR, western blot and immunostaining were used to study target genes/proteins expression in endometrium in pigs. RFLP-PCR was used to detect single nucleotide polymorphisms（SNPs） of target genes.Results： The results showed that the m RNA and protein expression levels of IHH, NR2F2 and BMP2 were up-regulated during implantation period（P 〈 0.05 or P 〈 0.01）. All target proteins were mainly observed in luminal epithelium and glandular epithelium. Interestingly, the staining of NR2F2 and HAND2 was also strong in stroma. SNPs detection revealed that there was a-204 C 〉 A mutation in promoter region of NR2F2 gene. Three genotypes were found in Large White, Landrace and Duroc sows. A total of 1847 litter records from 625 sows genotyped at NR2F2 gene were used to analyze the total number born（TNB） and number born alive（NBA）. The study of the effect on litter size suggested that sows with genotype CC tend to have higher litter size.Conclusions： These results showed the expression patterns of genes/proteins involved in PGR paracrine signaling over implantation time. And the candidate gene for litter size was identified from genes involved in this signaling. This study could be a resource for further studies to identify the roles of these genes for embryonic implantation in pigs.

Model Optimization of Litter Size and Genetic Analyses of Model Parameters in Sows

According to two properties of the life cycle and to fluctuation with parities, four mathemati- cal models, the Poisson cycle model, the cubic polyno- mial model, the modified quadratic polynomial model- I artd the modified quadratic polynomial model-H, were used to fit the records of litter size in Jiangquhai sows. From the viewpoint of statistics and biological significance, the modified quadratic polynomial mod- el-I was found to be the optimum model. A single traitanimal model and DFREML procedures were further used to estimate the heritability values of optimum model parameters. The results show that the heritabili- ty values for the coefficients A and B and the herita- bility value for the acme of the model pure quadric curve are larger than the heritability value for the litter size. This suggests that selection for model parameters may be more effective than direct selection for litter size.

Association Analysis between a Polymorphism in the 5' Regulatory Region of the IL-6 Gene and Litter Size in Pigs

The aim of this study was to determine the effects of an IL-6 gene polymorphism, discovered in the 5＇ regulatory region, on porcine litter size. An association analysis was performed between the polymorphism and total number born （TNB） and number born alive （NBA） in 421 sows. The polymorphism was at Hpy188I within the 5＇ regulatory region of IL- 6 gene. Three genotypes of AA, AG, and GG were detected in Landrace, and two genotypes, AA and AG, were detected in Yorkshire and Duroc pigs. The A allele was the superior allele in all three breeds, with allele frequencies ranging from 0. 901 to 0.993. The IL-6 genotype was highly significantly associated with TNB and NBA in the third and following parities （ P 〈 0.01 ）, and with total parities （ P 〈 0.05）. In general, the TNB and NBA showed a tendency of GG 〉 AG 〉 AA, indicating that the common allele was the least favorable for litter size. Thus, there is an enormous opportunity to increase litter size if this effect is confirmed in other studies.

Relevance between BMPR-IB Genotypes and Litter Size in Small Tail Han Sheep

[ Objective] To detect the polymorphism of BMPR-IB gene and analyze the relevance between genotype frequency of FecB gene and litter size. [Method] Mutation sites in the BMPR-IB gene of Small Tail Hart sheep were analyzed by PCR-RFLP. [ Resluts] The percentages of individuals with BB, B ＋ and ＋ ＋ genotypes were 24.69%, 54.89% and 20.42%, respectively, and their average litter size per parity was 3.01,2.81 and 2.16, respectively. E Conclusion] The BMPR-IB gene can be used as a molecular genetic marker in fecundity traits. The FecB gene can effectively increase litter size of the Small Tail Han sheep and has additive action on litter size.

Polymorphism of Exon 2 of FSHβ Gene and Its Relationship with Reproduction Performance in Two Goat Breeds

The objectives of the present study were to detect the polymorphism in follicle stimulating hormone beta （FSHβ） gene and investigate the relationship between FSHβ gene and high prolificacy in goats.According to the sequence of ovine FSHβ gene,two pairs of primers were designed to detect single nucleotide polymorphism of exon 2 of FSHβ gene in two goat breeds,Xinong Saanen dairy goat and Boer goat,by PCR-SSCP.The least square mean and genetic variance of different genotypes at polymorphic locus were analyzed,and the selection reaction was forecasted by genetic correlation between marker locus and genetic variance of litter size traits.There was one polymorphic locus occurring in exon 2 （P2） with three genotypes （EE,EF and FF）.In Xinong Saanen dairy goat,EE genotype had significantly higher （P 〈 0.05） litter size than EF and FF genotype in the first to fourth parity and average parity;EF genotype had significantly higher （P 〈 0.05） litter size than FF genotype in the second and average parity.In Boer goat,EE genotype had significantly higher （P 〈 0.01） litter size than EF and FF genotype in the second to fourth parity and average parity;EF genotype had significantly higher （P 〈 0.05） litter size than FF genotype in average parity.In Xinong Saanen dairy and Boer goat,the heredity of litter size was mainly influenced by genetic additive effect and the EE genotype is a favorable marker genotype for litter size in these two breeds.

DNA Polymorphisms of 5′-Flanking Region of Insulin-Like Growth Factor 1 Gene and Their Association with Reproduction Traits in Goats

Research on the identity of genes and their relationship with traits of economic importance in farm animals could assist in the selection of livestock. In this study, the polymorphisms of insulin-like growth factor 1 （IGF1） gene in 561 goats of ten breeds were detected by polymerase chain reaction （PCR）-single strand conformation polymorphism （SSCP） and their association with litter size and birth weight in three breeds were investigated. The effects of IGF1 polymorphisms on the breeding value for litter size and birth weight were examined using least square methods. Two deletions （CA） were detected in the microsatellite and two mutations （A1637G, T1640C） were found in 5′-flanking regulatory region. No significant association between the polymorphisms in 5′-flanking region of IGF1 and birth weight was found in the three breeds of goats. In Gulin Ma goats, two polymorphisms were found to affect litter size traits. In Chuandong White goats and Guizhou White goats, no significant difference （P0.05） in litter size between goats carrying different genotypes was observed. Further evaluation and confirmation studies in more goat populations with larger sample sizes are necessary.

Reexamination of Sexual Dimorphism and Female Reproduction in the Many-Lined Sun Skink Eutropis multifasciata from China

We reexamined sexual dimorphism and female reproduction in the Many-Lined Sun Skink Eutropis multifasciata from Hainan,China. Our data confirm that adults are sexually dimorphic in body size and shape,with males being the larger sex and larger in head size but shorter in abdomen length than females of the same snoutvent length(SVL). The rate at which head width increased with SVL was greater in males as opposed to the previous conclusion that the rate does not differ between the sexes. Maternal size was the main determinant of reproductive investment,with larger females generally producing more,as well as larger,offspring. Females produced up to nine offspring per litter as opposed to the previously reported 2–7. Most females gave birth between March and August,a time period approximately four months longer than that(May–June) reported previously. Females with a higher fecundity tended to produce smaller offspring as opposed to the previous conclusion that females do not tradeoff offspring size against number. Litter size,neonate mass and litter mass remained remarkably constant among years,and litter mass was more tightly related to female body size than litter size or neonate mass. Smaller females could produce relatively heavier litters without a concomitant reduction in postpartum body condition.

Reproductive performance and gestational effort in relation to dietary fatty acids in guinea pigs

Background： Dietary saturated（SFAs） and polyunsaturated（PUFAs） fatty acids can highly affect reproductive functions by providing additional energy, modulating the biochemical properties of tissues, and hormone secretions. In precocial mammals such as domestic guinea pigs the offspring is born highly developed. Gestation might be the most critical reproductive period in this species and dietary fatty acids may profoundly influence the gestational effort. We therefore determined the hormonal status at conception, the reproductive success, and body mass changes during gestation in guinea pigs maintained on diets high in PUFAs or SFAs, or a control diet.Results： The diets significantly affected the females＇ plasma fatty acid status at conception, while cortisol and estrogen levels did not differ among groups. SFA females exhibited a significantly lower body mass and litter size, while the individual birth mass of pups did not differ among groups and a general higher pup mortality rate in larger litters was diminished by PUFAs and SFAs. The gestational effort, determined by a mother＇s body mass gain during gestation, increased with total litter mass, whereas this increase was lowest in SFA and highest in PUFA individuals. The mother＇s body mass after parturition did not differ among groups and was positively affected by the total litter mass in PUFA females.Conclusions： While SFAs reduce the litter size, but also the gestational effort as a consequence, PUFA supplementation may contribute to an adjustment of energy accumulations to the total litter mass, which may both favor a mother＇s body condition at parturition and perhaps increase the offspring survival at birth.

Effects of catechins on litter size,reproductive performance and antioxidative status in gestating sows

This study was conducted to investigate the effects of catechins on reproductive performance, antioxidative capacity and immune function of gestating sows. A total of 60 cross-bred（Landrace × Large White） multiparious sows were blocked by body weight, parity and backfact and randomly allocated to 1 of 5 treatments: 0.100,200,300, or 400 mg/kg catechins. Dietary treatments were imposed from mating to d 40 of gestation of sows. At farrowing, litter total born, born alive, dead, and normal-（healthy piglets,>0.85 kg） and low-birth weight piglets（<0.85 kg） were recorded. Within 3.00 ± 0.50 days after farrowing litter size was standardized to 8,00 ± 1.50 piglets within treatment. The piglets were weighed at birth（d 1） and weaning（d 28）. Sows serum samples were obtained from blood samples collected on d 40 of gestation for analyses of glutathione peroxidase（GSH-Px）, superoxide dismutase（SOD）, catalase（CAT）,malondialdehyde（MDA）, hydrogen peroxide（H2 O2）, nitric oxide synthetase（NOS） and nitrogen monoxide（NO）. Our results showed that supplementation of catechins at levels of 200 or 300 mg/kg led to improvements in litter born alive（P < 0.01） and piglet born healthy（P < 0.01） and a decrease in stillborn（P < 0.05） at farrowing when compared with the control. In comparison with the control, catechins at any supplemental levels all enhanced the serum SOD（P < 0.05） and CAT（P < 0.01） activities of sows at farrowing but no obvious differences in the serum GSH-Px and NOS activities were observed in this trial（P > 0.05）. Sows received 200 mg catechin per kg diets showed a reduction（P < 0.05） of the serum MDA level at farrowing compared with all other treatments. Sows received all the levels of catechin showed a reduction（P < 0.05） of serum H2 O2 level compared with sows received the control diet on both d 40 of gestation and farrowing. Our results demonstrated that the catechins may be a potential antioxidant to increase the reproductive performance and antioxidative capacity of sows when it was added into diets during the early gestation.

Cathepsin D knockdown regulates biological behaviors of granulosa cells and affects litter size traits in goats

Cathepsin D(CTSD),the major lysosomal aspartic protease that is widely expressed in different tissues,potentially regulates the biological behaviors of various cells.Follicular granulosa cells are responsive to the increase of ovulation number,hence indirectly influencing litter size.However,the mechanism underlying the effect of CTSD on the behaviors of goat granulosa cells has not been fully elucidated.This study used immunohistochemistry to analyze CTSD localization in goat ovarian tissues.Moreover,western blotting was applied to examine the differential expression of CTSD in the ovarian tissues of monotocous and polytocous goats.Subsequently,the effects of CTSD knockdown on cell proliferation,apoptosis,cell cycle,and the expression of candidate genes of the prolific traits,including bone morphogenetic protein receptor IB(BMPR-IB),follicle-stimulating hormone(FSHR),and inhibinα(INHA),were determined in granulosa cells.Results showed that CTSD was expressed in corpus luteum,follicle,and granulosa cells.Notably,CTSD expression in the monotocous group was significantly higher than that in the polytocous group.In addition,CTSD knockdown could improve granulosa cell proliferation,inhibit cell apoptosis,and significantly elevate the expression of proliferating cell nuclear antigen(PCNA)and B cell lymphoma 2(Bcl-2),but it lowered the expression of Bcl-2-associated X(Bax)and caspase-3.Furthermore,CTSD knockdown significantly reduced the ratios of cells in the G0/G1 and G2/M phases but substantially increased the ratio of cells in the S phase.The expression levels of cyclin D2 and cyclin E were elevated followed by the obvious decline of cyclin A1 expression.However,the expression levels of BMPR-IB,FSHR,and INHA clearly increased as a result of CTSD knockdown.Hence,our findings demonstrate that CTSD is an important factor affecting the litter size trait in goats by regulating the granulosa cell proliferation,apoptosis,cell cycle,and the expression of candidate genes of the prolific trait.

Insertion/deletion variants within the IGF2BP2 gene identified in reported genome-wide selective sweep analysis reveal a correlation with goat litter size

Insulin-like growth factor 2 m RNA-binding protein 2(IGF2 BP2,also called IMP2)plays an essential role in the development and maturation of germ cells and embryos and is a candidate gene for goat litter size,based on a previous genomewide selective sweep analysis.In this study,the m RNA expression level of IGF2 BP2 was found to be significantly higher in a single-lamb group than in a multi-lamb group.Insertions/deletions(indels)within the goat IGF2 BP2 gene,including P4-Ins-13 bp and P5-Del-12 bp,were verified in 918 Shaanbei White Cashmere(SBWC)female goats.The minor allelic frequencies(MAFs)of P4-Ins-13 bp and P5-Del-12 bp loci were 0.349 and 0.295,respectively.Analysis using the Chi-square(χ^(2))test showed that the genotype(χ^(2)=14.479,P=0.006)distribution of P4-Ins-13 bp was significantly different between the single-lamb and multi-lamb groups.Correlation analysis demonstrated that P4-Ins-13 bp was significantly associated with goat litter size(P=0.022),and individual goats with the homozygous deletion/deletion(DD)genotype produced more litters than other goats.Therefore,considered as a potential molecular marker significantly related to lambing traits,the P4-Ins-13 bp mutation of the goat IGF2 BP2 gene can be used in goat breeding with practical molecular marker-assisted selection(MAS)to optimize female reproduction and improve economic efficiency in the goat industry.

Multiple aspects of the maternal reproductive investmentin a polytocous species:What do mothers really control?

One of the factors facilitating the expansion and proliferation of wild boar Sus scrofa is the plasticity of its reproductive biology.Nevertheless,the real influence of maternal and environmental factors on number and sex of the offspring is stll controversial.While the litter size was shown to be related with the maternal condition,the strength of this relation remains to be understood,together with the possible role played by environmental conditions.Analogously,it is unclear whether wild boar females can adjust their offspring sex.We investigated multiple aspects of wild boar maternal investment by means of a 10-year dataset of female reproductive traits and a set of biologically meaningful environmental variables.The maternal condition slightly affected the litter size but not the offspring sex,and environment did not affect the litter size or the offspring sex.Moreover,mothers did not cope with the higher costs entailed by producing sons by placing them in the most advantageous intrauterine position,nor by allocating less resources on daughters.Our set of results showed that the female reproductive investment is quite rigid in comparison with other aspects of wild boar reproductive biology.Wild boar females seem to adopt a typical rstrategy,producing constantly large litters and allocating resources on both sexes regardless of internal and external conditions.Such strategy may be adaptive to cope with environmental unpredictability and an intense human harvest,contributing to explain the extreme success of wild boar within human-dominated landscapes.