Mmp2 Deficiency Leads to Defective Parturition and High Dystocia Rates in Mice.

Parturition is the final and essential step for mammalian reproduction. While the uterus is quiescent during pregnancy, fundamental changes arise in the myometrial contractility, inducing fetal expulsion. Extracellular matrix (ECM) remodeling is fundamental for these events. The gelatinases subgroup of matrix metalloproteinases (MMPs), MMP2 and MMP9, participate in uterine ECM remodeling throughout pregnancy and parturition. However, their loss-of-function effect is unknown. Here, we determined the result of eliminating Mmp2 and/or Mmp9 on parturition in vivo, using single- and double-knockout (dKO) mice. The dystocia rates were measured in each genotype, and uterine tissue was collected from nulliparous synchronized females at the ages of 2, 4, 9 and 12 months. Very high percentages of dystocia (40-55%) were found in the Mmp2-/- and dKO females, contrary to the Mmp9-/- and wild-type females. The histological analysis of the uterus and cervix revealed that Mmp2-/- tissues undergo marked structural alterations, including highly enlarged myometrial, endometrial and luminal cavity. Increased collagen deposition was also demonstrated, suggesting a mechanism of extensive fibrosis in the Mmp2-/- myometrium, which may result in dystocia. Overall, this study describes a new role for MMP2 in myometrium remodeling during mammalian parturition process, highlighting a novel cause for dystocia due to a loss in MMP2 activity in the uterine tissue.

Etiology behind canine uterine inertia: Role of uterine expression of MLCK4, MYH2, and PKC genes.

Dystocia is an obstetrical emergency, and primary uterine inertia (PUI) is the major etiological reason among the more prevalent maternal causes in dogs. The present study involved the relative expression analysis of genes associated with myometrial contraction in medium-sized dog breeds with uterine inertia. Dogs without any progress in the parturition process even after four hours of the onset of labor and the absence of uterine contractions were considered to have complete primary uterine inertia (CPUI, n = 9). Dogs that had expelled at least one fetus and made no further progress in parturition in the absence of active uterine contraction were considered to be experiencing partial primary uterine inertia (PPUI, n = 6). Dogs with the fetal cause of dystocia (FCD), i.e., obstructive dystocia, were taken as the third (n = 7) group. Uterine tissue samples were collected during cesarean section in each group, RNA was isolated, and the relative expression of myometrial ACTA2, ACTG2, MLCK4, MYH2, and PKC genes was analyzed. The MLCK4 gene expression was downregulated in CPUI (P ≤ 0.05) and PPUI (P ≤ 0.01) when compared to FCD. The MYH2 gene expression was downregulated in PPUI in comparison to CPUI (P ≤ 0.01) and FCD (P ≤ 0.05). The PKC gene expression was upregulated in PPUI in comparison to FCD and CPUI (P ≤ 0.05). The downregulation of MLCK4 and MYH2 gene expressions recorded in PPUI indicated the possibility of myometrial defects. The possibility of myometrial defects was also observed in CPUI, but to a lesser degree, suggesting other etiologies.

Genetic analysis of retained placenta and its association with reproductive disorder, production, and fertility traits of Iranian Holstein dairy cows.

To facilitate breeding for improved resistance to the reproductive disorder of retained placenta (RP), genetic parameters were estimated for RP and its genetic correlation with other reproductive disorders as well as with production and fertility traits of Iranian Holstein dairy cows. Data were 154,048 lactation records collected between 2011 and 2018 from 59,610 Holstein dairy cows in 9 Iranian herds. Other reproductive disorders included dystocia, stillbirth, and twinning. Fertility records were available for days from calving to first service (DFS), days open (DO), number of inseminations per conception (NIC), and success of first insemination (SFI). Genetic parameters for RP were estimated using univariate linear and logistic animal models with ASREML software. The univariate linear animal model was used to implement bivariate analysis to investigate potential genetic correlations of RP with other reproductive disorders and with production and fertility traits. Heritability estimates for RP were low from both linear (0.031) and logistic (0.092) animal models. Estimated genetic correlations with RP were -0.04 for twinning, 0.32 for stillbirth, and 0.34 for dystocia, which indicates that selection against RP could indirectly select against dystocia and stillbirth. Estimated genetic correlations between RP and production traits (milk, fat, and protein yields) at 100, 200, and 305 d in milk ranged from -0.12 to -0.29; the greatest correlation (-0.29) was for the first 100 d in milk. A moderate positive genetic correlation (0.25) was found for RP and DO, DFS, and NIC, whereas a low negative genetic correlation (-0.09) was found between RP and SFI. The pedigree-based genetic analysis of RP showed that this trait has a low heritability, is linked to other reproductive disorders, and generally has an unfavorable relationship with production and fertility traits. Selection against RP can reduce the incidence of reproductive disorders and improve fertility and production traits.

Differential response from selection for high calving ease vs. low birth weight in American Simmental beef cattle.

Data on calving ease (CE) and birth, weaning weight (WW), and yearling weight (YW) were obtained from the American Simmental Association (ASA) and included pedigree and performance information on 11,640,735 animals. Our objective was to quantify differential response from selection for high CE vs. low birth weight (LBW) in first-calf Simmental heifers. We hypothesized that direct selection for CE should be used as the primary approach to reduce dystocia and mitigate losses in growth-related traits. WW and YW were adjusted to 205 and 365 d of age, respectively. Sire and maternal grandsire (co)variance components for CE, birth weight (BW), and 205-d weaning weight (205-d WW), and sire covariance components for 160-d postweaning gain (160-d gain) were estimated using a sire-maternal grandsire model. Direct and maternal expected progeny differences (EPD) for CE, BW, and 205-d WW and direct EPD for 160-d gain and 365-d yearling weight (365-d YW) for first-calf Simmental heifers population (465,710 animals) were estimated using a threshold-linear multivariate maternal animal model. This population was used to estimate genetic trends and as a selection pool (control) for various selection scenarios. Selection scenarios were high CE (HCE), LBW, the all-purpose selection index (API = -1.8 BW + 1.3 CE + 0.10 WW + 0.20 YW) of the ASA and its two derived subindices: (API1 = 1.3 CE + 0.20 YW) and (API2 = -1.8 BW + 0.20 YW), and lastly Dickerson's selection index (DSI = -3.2 BW + YW). Data for each selection scenario were created by selecting sires with EPD greater than or equal to the average along with the top 75% of dams. Comparison between selection scenarios involved evaluating the direct and maternal genetic trends from these scenarios. Direct heritabilities for CE, BW, 205-d WW, 160-d gain, and 365-d YW of Simmental cattle were 0.23, 0.52, 0.28, 0.21, and 0.33, respectively. The single trait, HCE, selection scenario, as opposed to LBW, increased the intercept for CE by 57.7% and the slopes (P < 0.001) for BW, 205-d WW, 160-d gain, and 365-d YW by 27.9%, 37.5%, 16%, and 28%, respectively. Comparisons of various selection scenarios revealed that the CE-based selection scenarios (HCE, API, and API1) had a greater response for CE and growth traits.

Could consanguineous marriage provide a cultural alleviation for the obstetric dilemma?

In placental mammals, a poor fit between the physical dimensions of the fetus and maternal pelvis increases the likelihood of obstructed labour. This problem is especially relevant to humans, as our species demonstrates both unique adaptations in pelvic shape and structure associated with bipedalism, and fetal encephalization. Natural selection is expected to have favoured adaptations that reduce the chances of such mismatch within individual mother-offspring dyads. Here, I hypothesise that the cultural practice of consanguineous marriage may have been favoured, on account of increasing the genetic similarity between mothers and offspring and hence the correlation between maternal and fetal physical dimensions. These benefits could be amplified if consanguineous marriage was accompanied by assortative mating for height. An additional benefit of consanguineous marriage for childbirth is the slight reduction in birth size of such offspring compared to non-consanguineous unions. Although the offspring of consanguineous unions have elevated risks of morbidity and mortality, these risks are moderate and the practice could still have been favoured by selection if the reduction in maternal mortality was greater than the increased mortality among individual offspring. This hypothesis could be tested directly by investigating whether rates of obstructed labour are lower in individuals and populations practising consanguineous marriage. At a broader level, phylogenetic analysis could be conducted to test whether consanguineous marriage appears to have originated in the areas where intensive agriculture was first practiced, as adult height typically fell in such populations, potentially exacerbating the risk of obstructed labour.

Genetic and environmental analysis of diseases with major economic impact in Israeli Holsteins.

Incidences of ketosis, metritis, mastitis, and retained placenta were studied in Israeli Holstein cows calving between 2008 and 2017. These diseases were selected based on their economic impact. Ketosis, metritis, and retained placenta were scored dichotomously. Mastitis was scored as absent, a single occurrence during the lactation, or more than 1 occurrence. Ketosis and metritis were recorded during the first 21 d after calving, retained placenta during the first 5 d after calving, and mastitis up to 305 d in milk. The effects of herd-year-season, calving age, month of calving, gestation length, and occurrence of dystocia were included in the first-parity analysis models. All effects were significant for metritis and retained placenta. For ketosis, all effects were significant, except for gestation length. For mastitis, only the effects of herd-year-season and calving age were significant. Variance components were computed by the multitrait animal model. The 4 diseases were analyzed jointly based on first-parity records, and each disease was analyzed separately for parities 1 to 3 with the different parities considered separate traits. The 4 disease traits in first parity were also analyzed jointly with the 6 major traits included in the Israeli breeding index: milk, fat, and protein production; somatic cell score; female fertility; and longevity. Heritability was highest for metritis and lowest for mastitis, but all heritabilities were <0.07, similar to previous studies. For all 4 diseases, genetic correlations among the first 3 parities were >0.65, and all residual correlations were <0.07. Selection of herd-years assumed to have more accurate recording of mastitis did not result in higher heritability estimates. Genetic correlations between the disease traits and milk, fat, and protein production were economically unfavorable, while correlations between the disease traits and somatic cell score, female fertility, and longevity were economically favorable. Expected genetic changes in the disease traits after 10 yr of selection with the current Israeli breeding index were all <1%, except for ketosis, which was predicted to increase by 1.5%. Inclusion of these traits in a proposed index with the disease traits constituting 7% of the index would result in only marginal improvements for the disease traits and adversely affect genetic gain for fat and protein production. Thus, inclusion of these traits in the breeding index cannot be justified economically.

A functional role for AMPK in female fertility and endometrial regeneration.

Adenosine monophosphate-activated protein kinase (AMPK) is a highly conserved heterotrimeric complex that acts as an intracellular energy sensor. Based on recent observations of AMPK expression in all structures of the female reproductive system, we hypothesized that AMPK is functionally required for maintaining fertility in the female. This hypothesis was tested by conditionally ablating the two catalytic alpha subunits of AMPK, Prkaa1 and Prkaa2, using Pgr-cre mice. After confirming the presence of PRKAA1, PRKAA2 and the active phospho-PRKAA1/2 in the gravid uterus by immunohistochemistry, control (Prkaa1/2 fl/fl ) and double conditional knockout mice (Prkaa1/2 d/d ) were placed into a six-month breeding trial. While the first litter size was comparable between Prkaa1/2 fl/fl and Prkaa1/2 d/d female mice (P = 0.8619), the size of all subsequent litters was dramatically reduced in Prkaa1/2 d/d female mice (P = 0.0015). All Prkaa1/2 d/d female mice experienced premature reproductive senescence or dystocia by the fourth parity. This phenotype manifested despite no difference in estrous cycle length, ovarian histology in young and old nulliparous or multiparous animals, mid-gestation serum progesterone levels or uterine expression of Esr1 or Pgr between Prkaa1/2 fl/fl and Prkaa1/2 d/d female mice suggesting that the hypothalamic-pituitary-ovary axis remained unaffected by PRKAA1/2 deficiency. However, an evaluation of uterine histology from multiparous animals identified extensive endometrial fibrosis and disorganized stromal-glandular architecture indicative of endometritis, a condition that causes subfertility or infertility in most mammals. Interestingly, Prkaa1/2 d/d female mice failed to undergo artificial decidualization. Collectively, these findings suggest that AMPK plays an essential role in endometrial regeneration following parturition and tissue remodeling that accompanies decidualization.

In silico analysis of the Mus musculus uterine gene expression landscape during pregnancy identifies putative upstream regulators for labour.

BACKGROUND: The molecular pathways involved in the transition from uterine quiescence to overt labour are mapped and form the currently established pharmacological targets for both the induction and inhibition of human labour. However, both spontaneous premature labour and functional dystocia occur and are difficult to treat adequately. The identification of upstream regulators involved in the onset and orchestration of labour pathways is essential to develop additional therapies that will contribute to the regulation of the timing of birth. OBJECTIVES: To define uterine biological processes and their upstream activators involved in the transition from uterine quiescence to overt labour. STUDY DESIGN: The uterus of non-pregnant and pregnant FVB M. musculus is collected at embryonic days (E) 6.5, 8.5, 10.5, 12.5, 15.5 and 17.5 and the uterine transcriptome is determined using the Illumina mouse Ref8v2 micro-array platform. K-means clustering and Ingenuity Pathway Analysis are applied to further dissect the transcriptome data. RESULTS: From E6.5 to E17.5, 5405 genes are significantly differentially expressed and they segregate into 7 unique clusters. Five of the 7 clusters are enriched for genes involved in specific biological processes that include regulation of gene-expression, T-cell receptor activation, Toll-like receptor signalling and steroid metabolism. The identification of upstream activators for differentially expressed genes between consecutive time points highlights the E10.5 to E12.5 window during which the role from progesterone switches from an activated state to the inhibited state reflecting the process of functional progesterone withdrawal essential for the transgression from myometrial quiescence to synchronized contractions. For this time window in which 189 genes are differentially expressed we define 22 putative upstream activators of which NUPR1 and TBX2 are the most significant with respectively an activated and an inhibited status. CONCLUSIONS: Gene expression profiling of mice uterus from E6.5 to E17.5 results in 7 unique gene expression clusters from early to late pregnancy that define the landscape of molecular events in ongoing pregnancy. In the current dataset progesterone is predicted as an activated upstream regulator and maintainer of myometrial quiescence and is active till E10.5. Progesterone is predicted as an inhibited upstream regulator at E12.5. We identify 22 upstream regulators in the E10.5 to E12.5 time window where the switch to progesterone withdrawal occurs. They are putative relevant upstream activators of labour.

Including gene networks to predict calving difficulty in Holstein, Brown Swiss and Jersey cattle.

BACKGROUND: Calving difficulty or dystocia has a great economic impact in the US dairy industry. Reported risk factors associated with calving difficulty are feto-pelvic disproportion, gestation length and conformation. Different dairy cattle breeds have different incidence of calving difficulty, with Holstein having the highest dystocia rates and Jersey the lowest. Genomic selection becomes important especially for complex traits with low heritability, where the accuracy of conventional selection is lower. However, for complex traits where a large number of genes influence the phenotype, genome-wide association studies showed limitations. Biological networks could overcome some of these limitations and better capture the genetic architecture of complex traits. In this paper, we characterize Holstein, Brown Swiss and Jersey breed-specific dystocia networks and employ them in genomic predictions. RESULTS: Marker association analysis identified single nucleotide polymorphisms explaining the largest average proportion of genetic variance on BTA18 in Holstein, BTA25 in Brown Swiss, and BTA15 in Jersey. Gene networks derived from the genome-wide association included 1272 genes in Holstein, 1454 genes in Brown Swiss, and 1455 genes in Jersey. Furthermore, 256 genes in Holstein network, 275 genes in the Brown Swiss network, and 253 genes in the Jersey network were within previously reported dystocia quantitative trait loci. The across-breed network included 80 genes, with 9 genes being within previously reported dystocia quantitative trait loci. The gene-gene interactions in this network differed in the different breeds. Gene ontology enrichment analysis of genes in the networks showed Regulation of ARF GTPase was very significant (FDR ≤ 0.0098) on Holstein. Neuron morphogenesis and differentiation was the term most enriched (FDR ≤ 0.0539) on the across-breed network. Genomic prediction models enriched with network-derived relationship matrices did not outperform regular GBLUP models. CONCLUSIONS: Regions identified in the genome were in the proximity of previously described quantitative trait loci that would most likely affect calving difficulty by altering the feto-pelvic proportion. Inclusion of identified networks did not increase prediction accuracy. The approach used in this paper could be extended to any instance with asymmetric distribution of phenotypes, for example, resistance to disease data.

Genetic evaluation for birth and conformation traits in dual-purpose Belgian Blue cattle using a mixed inheritance model.

The segregation of the causal mutation () in the muscular hypertrophy gene in dual-purpose Belgian Blue (dpBB) cattle is considered to result in greater calving difficulty (dystocia). Establishing adapted genetic evaluations might overcome this situation through efficient selection. However, the heterogeneity of dpBB populations at the locus implies separating the major gene and other polygenic effects in complex modeling. The use of mixed inheritance models may be an interesting option because they simultaneously assume both influences. A genetic evaluation in dpBB based on a mixed inheritance model was developed for birth and conformation traits: gestation length (GL), calving difficulty (CD), birth weight (BiW), and body conformation score (BC). A total of 27,362 animals having records were used for analyses. The total number of animals in the pedigree used to build the numerator relationship matrix was 62,617. Genotypes at the locus were available for 2,671 animals. Missing records at this locus were replaced with genotype probabilities. A total of 13,221 (48.3%) were registered as dpBB, 1,287 (4.7%) as beef Belgian Blue, and 12,854 (47.0%) were unknown. From those 13,221 dpBB animals, 650, 849, and 534 had double or single copies or no copy, respectively, of the causal mutation () in the muscular hypertrophy gene, whereas 11,188 had missing genotypes. This heterogeneity at the locus may be the reason for high variability in the studied traits, that is, high heritability estimates of 0.33, 0.30, 0.38, and 0.43 for GL, CD, BiW, and BC, respectively. In general, additive ( < 0.05) and dominance ( < 0.001) allele substitution for calves and dams had significant impact for all traits. The moderate coefficient of genetic variation (27.80%) and high direct heritability (0.28) for CD suggested genetic variability in dpBB and possible genetic improvement through selection. This variability has allowed dpBB breeders to successfully apply mass selection in the past. Genetic trend means from 1988 to 2016 showed that sire selection for CD within genotype was progressively applied by breeders. The selection intensity was more important for CD in double-muscled lines than in segregated lines. Our study illustrated the possible confusion caused by the use of major genes in selection and the importance of fitting appropriate models such as mixed inheritance models that combine polygenic and gene content information.

Cliff-edge model predicts intergenerational predisposition to dystocia and Caesarean delivery.

Recently, we presented the cliff-edge model to explain the evolutionary persistence of relatively high incidences of fetopelvic disproportion (FPD) in human childbirth. According to this model, the regular application of Caesarean sections since the mid-20th century has triggered an evolutionary increase of fetal size relative to the dimensions of the maternal birth canal, which, in turn, has inflated incidences of FPD. While this prediction is difficult to test in epidemiological data on Caesarean sections, the model also implies that women born by Caesarean because of FPD are more likely to develop FPD in their own childbirth compared with women born vaginally. Multigenerational epidemiological studies indeed evidence such an intergenerational predisposition to surgical delivery. When confined to anatomical indications, these studies report risks for Caesarean up to twice as high for women born by Caesarean compared with women born vaginally. These findings provide independent support for our model, which we show here predicts that the risk of FPD for mothers born by Caesarean because of FPD is 2.8 times the risk for mothers born vaginally. The congruence between these data and our prediction lends support to the cliff-edge model of obstetric selection and its underlying assumptions, despite the genetic and anatomical idealizations involved.

The impact of cross-breeding Egyptian and Italian buffalo on reproductive and productive performance under a subtropical environment.

The aim of this study was to investigate the reproductive and productive performance of pure Egyptian (PE) buffaloes and their crosses with Italian buffaloes. In this study, 2969 dairy buffaloes were used (1599 PE; 615 F1 crosses, 50% PE and 50% Italian buffaloes; and 755 backcross [BC], 75% PE and 25% Italian buffaloes). When compared to PE, the BC and F1 had a significantly lower incidence of calving difficulty (odds ratio [OR] = 0.18, p < .0001 and .34, p < .0001, respectively) in conjunction with a lesser incidence of stillbirth (OR = 0.06, p < .0001 and 0.43, p < .0001, respectively). Backcross buffaloes were also noted to have a lower rate of not conceived after first insemination (OR = 0.74, p < .0001) in comparison with PE. The cross F1 buffaloes were superior in terms of productive traits, whereas PE buffaloes were best in most of the reproductive traits (days of non-pregnant p = .001, service per conception p < .0001 and calf weight p = .01). Although the study results appeared to indicate towards a contrary association between milk production and reproductive traits, BC was similar to F1 in terms of production and similar to PE in terms of reproductive traits. Thus, the farmers should be encouraged to breed BC animals in their herds for enhancing milk yield to meet with the market demands.

Breed effects and genetic parameter estimates for calving difficulty and birth weight in a multibreed population.

Birth weight (BWT) and calving difficulty (CD) were recorded on 4,579 first-parity females from the Germplasm Evaluation Program at the U.S. Meat Animal Research Center (USMARC). Both traits were analyzed using a bivariate animal model with direct and maternal effects. Calving difficulty was transformed from the USMARC scores to corresponding -scores from the standard normal distribution based on the incidence rate of the USMARC scores. Breed fraction covariates were included to estimate breed differences. Heritability estimates (SE) for BWT direct, CD direct, BWT maternal, and CD maternal were 0.34 (0.10), 0.29 (0.10), 0.15 (0.08), and 0.13 (0.08), respectively. Calving difficulty direct breed effects deviated from Angus ranged from -0.13 to 0.77 and maternal breed effects deviated from Angus ranged from -0.27 to 0.36. Hereford-, Angus-, Gelbvieh-, and Brangus-sired calves would be the least likely to require assistance at birth, whereas Chiangus-, Charolais-, and Limousin-sired calves would be the most likely to require assistance at birth. Maternal breed effects for CD were least for Simmental and Charolais and greatest for Red Angus and Chiangus. Results showed that the diverse biological types of cattle have different effects on both BWT and CD. Furthermore, results provide a mechanism whereby beef cattle producers can compare EBV for CD direct and maternal arising from disjoined and breed-specific genetic evaluations.

Genetic analysis of calving traits by the multi-trait individual animal model.

Five alternative models were applied for analysis of dystocia and stillbirth in first and second parities. Models 1 and 2 were included only to estimate the parameters required for model 4, and models 3 and 5 are included only as comparisons to the model 4 estimates. Variance components were estimated by multi-trait REML, including cows with valid calving records for both parities. For the effects of sire of calf on first and second parities, variance components were estimated including only calvings with the same sire of calf for both parities. All heritabilities for the cow effect were quite low, but higher for dystocia than for stillbirth and higher in first parity. The sire-of-calf heritabilities were higher than the cow effect heritabilities, except for stillbirth in parity 2. Unlike the effect of cow correlations, all sire of calf correlations were >0.6, and the correlations for the same trait in parities 1 and 2 were >0.9. Thus, a multi-trait analysis should yield a significant gain in accuracy with respect to the sire of calf effects for bulls not mated to virgin heifers. A multi-trait individual animal model algorithm was developed for joint analysis of dystocia and stillbirth in first and second parities. Relationships matrices were included both for the effects of cow and sire of calf. In addition, random herd-year-season and fixed sex of calf effects were included in the model. Records were preadjusted for calving month and age. A total of 899,223 Israeli Holstein cows with first calvings since 1985 were included in the complete analysis. Approximate reliabilities were computed for both sire of cow and sire of calf effects. Correlations between these reliabilities and reliabilities obtained by direct inversion of the coefficient matrix for a sire of cow-sire of calf model were all close to 0.99. Phenotypic trends for cows born from 1983 through 2007 were economically unfavorable for dystocia and favorable for stillbirth in both parities. Genetic trends were economically unfavorable for both dystocia and stillbirth in first parity. First-parity sire of calf trends were unfavorable for dystocia, but favorable for stillbirth. All environmental trends were nearly zero. Regressions of evaluations of the complete analysis on a model including only calvings before 2011 were all >0.8. All evaluations met the Interbull Method 3 criterion for unbiasedness. Model 4, which computed genetic evaluations for all animals for all 4 traits accounting for all known relationships and correlations among the traits, is recommended for routine genetic evaluation of calving traits.

Genome-wide association study for calving performance using high-density genotypes in dairy and beef cattle.

BACKGROUND: Calving difficulty and perinatal mortality are prevalent in modern-day cattle production systems. It is well-established that there is a genetic component to both traits, yet little is known about their underlying genomic architecture, particularly in beef breeds. Therefore, we performed a genome-wide association study using high-density genotypes to elucidate the genomic architecture of these traits and to identify regions of the bovine genome associated with them. RESULTS: Genomic regions associated with calving difficulty (direct and maternal) and perinatal mortality were detected using two statistical approaches: (1) single-SNP (single nucleotide polymorphism) regression and (2) a Bayesian approach. Data included high-density genotypes on 770 Holstein-Friesian, 927 Charolais and 963 Limousin bulls. Several novel or previously identified genomic regions were detected but associations differed by breed. For example, two genomic associations, one each on chromosomes 18 and 2 explained 2.49 % and 3.13 % of the genetic variance in direct calving difficulty in the Holstein-Friesian and Charolais populations, respectively. Imputed Holstein-Friesian sequence data was used to refine the genomic regions responsible for significant associations. Several candidate genes on chromosome 18 were identified and four highly significant missense variants were detected within three of these genes (SIGLEC12, CTU1, and ZNF615). Nevertheless, only CTU1 contained a missense variant with a putative impact on direct calving difficulty based on SIFT (0.06) and Polyphen (0.95) scores. Using imputed sequence data, we refined a genomic region on chromosome 4 associated with maternal calving difficulty in the Holstein-Friesian population and found the strongest association with an intronic variant in the PCLO gene. A meta-analysis was performed across the three breeds for each calving performance trait to identify common variants associated with these traits in the three breeds. Our results suggest that a portion of the genetic variation in calving performance is common to all three breeds. CONCLUSION: The genomic architecture of calving performance is complex and mainly influenced by many polymorphisms of small effect. We identified several associations of moderate effect size but the majority were breed-specific, indicating that breed-specific alleles exist for calving performance or that the linkage phase between genotyped allele and causal mutation varies between breeds.

Genetic and environmental analysis of dystocia and stillbirths in draft horses.

Genetic parameters and environmental factors were estimated for foaling ease (FE) and stillbirths (SBs) in four breeds of draft horses based on 11 229, 38 877, 35 764 and 13 274 FE and SB scores recorded between 1998 and 2010 for Ardennais (A), Breton (B), Comtois (C) and Percheron (P), respectively. Incidences for the three FE categories were: easy or without help 91.0% (A) to 95.4% (B), difficult 3.4% (B) to 7.1% (A) and intervention of a veterinarian 1.1% (B) to 1.9% (A). The frequency of SB ranged between 5.4% (B) and 9.4% (A). A multiple-trait threshold animal model was used that included the effects of sex of foal, region, month, year of foaling, combined maternal age and parity, direct genetic, maternal genetic and permanent environments. Estimates were obtained using Markov Chain Monte Carlo Gibbs sampling. The most unfavourable effect was first parity, which decreased the probability of easy foaling to 78.6% for A and 88.3% for B. Interaction with age showed that the risk for first foaling for mares aged 3 years was higher than at 4 or 5 to 9 years. This was also observed for SB with an increased probability of SB at first foaling of 17.9% (A) or 9.6% (B). The most unfavourable month was found to be the most frequent month for foaling (April) and not the most demanding months weather-wise (winter). For FE, direct heritabilities were A 0.27 (0.06), B 0.14 (0.03), C 0.18 (0.03) and P 0.18 (0.04), and maternal heritabilities were A 0.25 (0.06), B 0.19 (0.04), C 0.12 (0.03) and P 0.21 (0.06). Genetic correlations between direct and maternal genetic effects were A -0.29 (0.14), B -0.39 (0.12), C -0.09 (0.14) and P -0.54 (0.17). For SB, direct heritabilities were A 0.52 (0.09), B 0.42 (0.04), C 0.28 (0.04) and P 0.39 (0.05), and maternal heritabilities were A 0.25 (0.05), B 0.10 (0.02), C 0.07 (0.02) and P 0.14 (0.02). Genetic correlations between direct and maternal genetic effects were A -0.85 (0.06), B -0.63 (0.06), C -0.64 (0.11) and P -0.69 (0.06). Direct genetic correlations between FE and SB traits were A 0.60 (0.10), B 0.58 (0.10), C 0.36 (0.10) and P 0.29 (0.15). Maternal genetic correlations between FE and SB traits were A 0.67 (0.10), B 0.47 (0.13), C 0.28 (0.15) and P 0.39 (0.15). These estimates are posterior means of the Gibbs samples and are within the upper limits of comparable results reported in cattle.

Genome-wide association study for calving traits in Holstein-Friesian dairy cattle.

Dystocia and perinatal mortality are quantitative traits that significantly impact animal productivity and welfare. Their economic importance is reflected by their inclusion in the national breeding goals of many cattle populations. The genetic architecture that influences these traits, however, has still yet to be thoroughly defined. Regions of the bovine genome associated with calving difficulty (direct and maternal) and perinatal mortality were detected in this study using a Bayesian approach with 43 204 single nucleotide polymorphisms (SNPs) on up to 1970 Holstein-Friesian bulls. Several SNPs on chromosomes 5, 6, 11, 12, 17,18 and 28 were detected to be strongly associated with these calving performance traits. Novel genomic regions with previously reported associations with growth, stature, birth weight and bone morphology were identified in the present study as being associated with the three calving performance traits. Morphological abnormalities are a known contributor to perinatal mortality and the most significantly associated SNP for perinatal mortality in the present study was located in a region in linkage disequilibrium with the gene SLC26A7. This gene, SLC26A7, has similarities and colocalises with SLC4A2, which has previously been associated with osteoporosis and mortality in cattle populations. The HHIP gene that is known to be associated with stature in humans was strongly associated with direct calving difficulty in the present study; large calves are known to, on average, have a greater likelihood of dystocia. A stemloop microRNA, bta-mir-1256, on chromosome 12, involved in post-transcriptional regulation of gene expression was associated with maternal calving difficulty. Previously reported quantitative trait loci associated with calving performance traits in other populations were again identified in this study; with one genomic region on chromosome 18 supporting very strong evidence of an underlying causative mutation and accounting for 2.1% of the genetic variation in direct calving difficulty. Overlapping genomic regions associated with one or more of the calving traits were also detected substantiating the known genetic covariances existing between these traits. Moreover, some genomic regions were only associated with one of the calving traits implying the selective genomic breeding programs exploiting these regions could help resolve genetic antagonisms.

The molecular effects of a polymorphism in the 5'UTR of solute carrier family 44, member 5 that is associated with birth weight in Holsteins.

Dystocia is a major problem for the dairy cattle industry, and the observed high rates of this condition stem from genetic selection to increase subsequent milk production of the calving female. Because smaller birth size does not adversely affect subsequent milk production, selecting for cows with a smaller birth size would reduce dystocia rates and be beneficial for both the cattle and the farmers. To identify genes that regulate birth weight, we conducted a genome-wide association study using 1151 microsatellite markers and identified a single nucleotide polymorphism (SNP) associated with birth weight: A-326G in the 5' untranslated region (UTR) of solute carrier family 44, member 5 (SLC44A5). Cows with higher birth weights carried the A polymorphism in the SLC44A5 5' UTR, and the presence of the A polymorphism correlated with a high rate of dystocia. Luciferase assays and quantitative polymerase chain reaction (QPCR) assays revealed that SLC44A5 transcripts with the A polymorphism are expressed at lower levels than those carrying the G polymorphism. SLC44A5 encodes a choline transporter-like protein, and choline is a component of the major phospholipids of cell membranes. Uptake studies in HeLa cells demonstrated that SLC44A5 knockdown reduces choline efflux, whereas SLC44A5 overexpression resulted in the opposite effect. Furthermore, cell viability assays indicated that SLC44A5 knockdown increased cell proliferation, whereas SLC44A5 overexpression repressed proliferation. Taken together, our results suggest that calves with reduced SLC44A5 expression are larger due to enhanced cell proliferation. This study provides novel insights into the molecular mechanisms that control birth weight in Holsteins and suggests that SLC44A5 may serve as a potential target for preventing dystocia.

Identification of a myometrial molecular profile for dystocic labor.

BACKGROUND: The most common indication for cesarean section (CS) in nulliparous women is dystocia secondary to ineffective myometrial contractility. The aim of this study was to identify a molecular profile in myometrium associated with dystocic labor. METHODS: Myometrial biopsies were obtained from the upper incisional margins of nulliparous women undergoing lower segment CS for dystocia (n = 4) and control women undergoing CS in the second stage who had demonstrated efficient uterine action during the first stage of labor (n = 4). All patients were in spontaneous (non-induced) labor and had received intrapartum oxytocin to accelerate labor. RNA was extracted from biopsies and hybridized to Affymetrix HuGene U133A Plus 2 microarrays. Internal validation was performed using quantitative SYBR Green Real-Time PCR. RESULTS: Seventy genes were differentially expressed between the two groups. 58 genes were down-regulated in the dystocia group. Gene ontology analysis revealed 12 of the 58 down-regulated genes were involved in the immune response. These included (ERAP2, (8.67 fold change (FC)) HLA-DQB1 (7.88 FC) CD28 (2.60 FC), LILRA3 (2.87 FC) and TGFBR3 (2.1 FC)) Hierarchical clustering demonstrated a difference in global gene expression patterns between the samples from dystocic and non-dystocic labours. RT-PCR validation was performed on 4 genes ERAP2, CD28, LILRA3 and TGFBR3 CONCLUSION: These findings suggest an underlying molecular basis for dystocia in nulliparous women in spontaneous labor. Differentially expressed genes suggest an important role for the immune response in dystocic labor and may provide important indicators for new diagnostic assays and potential intrapartum therapeutic targets.

X-linked cholestasis in mouse due to mutations of the P4-ATPase ATP11C.

Transporters at the hepatic canalicular membrane are essential for the formation of bile and the prevention of cholestatic liver disease. One such example is ATP8B1, a P4-type ATPase disrupted in three inherited forms of intrahepatic cholestasis. Mutation of the X-linked mouse gene Atp11c, which encodes a paralogous P4-type ATPase, precludes B-cell development in the adult bone marrow, but also causes hyperbilirubinemia. Here we explore this hyperbilirubinemia in two independent Atp11c mutant mouse lines, and find that it originates from an effect on nonhematopoietic cells. Liver function tests and histology revealed only minor pathology, although cholic acid was elevated in the serum of mutant mice, and became toxic to mutant mice when given as a dietary supplement. The majority of homozygous mutant females also died of dystocia in a maternal genotype-specific manner. ATP11C therefore represents a multifunctional transporter, essential for adult B-cell development, the prevention of intrahepatic cholestasis, and parturition, and is a new candidate for genetically undiagnosed cases of cholestasis and dystocia in humans.