BMPR1B Research Articles

Molecular cloning of ESR1, BMPR1B, and FOXL2 and differential expressions depend on maternal age and size during breeding season in cultured Asian yellow pond turtle (Mauremys mutica).

The reproductive capacity (egg size and egg number) of most of oviparous animals, including the Asian yellow pond turtle (Mauremys mutica), is constrained by the maternal age and body size, but the mechanism determining the maternal reproductive ability remains unclear. To disclose how maternal age and size affect reproductive ability of M. mutica, we first identified the full-length cDNAs from estrogen receptor 1 (ESR1), bone morphogenetic protein receptor 1B (BMPR1B), and forkhead box L2 (FOXL2). The ESR1 open reading frame (ORF) was 1, 767 bp encoding 588 amino acids. For BMPR1B, the ORF was 1599 bp encoding 532 amino acids, and an ORF of 906 bp encoding 301 amino acids was identified in FOXL2. The effects of maternal age and size on the expression of ESR1, BMPR1B, and FOXL2 in the ovary, brain, and uterus showed that ESR1 expression in large females was significantly lower than that in small females in the brain, but body size did not affect ESR1 expression in the ovary. The expression of ESR1 was significantly different in the different age groups and size groups, and there was interaction detected between maternal age and body size. However, BMPR1B expression in the ovary, brain, and uterus was independent of maternal age and size. In addition, we found different FOXL2 expression patterns between the brain and uterus, while detected interaction of female age and size in the brain and ovary. Our results imply the complexity and diversity of maternal age and size in regulating the expression of genes related to reproduction. These results provide more information for the maternal effects on the reproduction-related gene expression.

Association of polymorphisms in bone morphogenetic protein receptor-1B gene exon-9 with litter size in Dorset, Mongolian, and Small Tail Han ewes.

ObjectiveThe present study was to investigate the association of polymorphisms in exon-9 of the bone morphogenetic protein receptor-1B (BMPR-1B) gene (C864T) with litter size in 240 Dorset, 232 Mongolian, and 124 Small Tail Han ewes.MethodsBlood samples were collected from 596 ewes and genomic DNA was extracted using the phenol: chloroform extraction method. The 304-bp amplified polymerase chain reaction product was analyzed for polymorphism by single-strand conformation polymorphism method. The genotypic frequency and allele frequency of BMPR-1B gene exon-9 were computed after sequence alignment. The χ2 independence test was used to analyze the association of genotypic frequency and litter size traits with in each ewe breed, where the phenotype was directly treated as category.ResultsThe results indicated two different banding patterns AA and AB for this fragment, with the most frequent genotype and allele of AA and A. Calculated Chi-square test for BMPR-1B gene exon-9 was found to be more than that of p value at the 5% level of significance, indicating that the population under study was in Hardy-Weinberg equilibrium for all ewes. The χ2 independence test analyses indicated litter size differences between genotypes was not the same for each breed. The 304-bp nucleotide sequence was subjected to BLAST analysis, and the C864T mutation significantly affected litter size in singletons, twins and multiples. The heterozygosity in exon-9 of BMPR-1B gene could increase litter size for all the studied ewes.ConclusionConsequently, it appears that the polymorphism BMPR-1B gene exon-9 detected in this study may have potential use in marker assisted selection for litter size in Dorset, Mongolian, and Small Tail Han ewes.

MiR-125b Contributes to Ovarian Granulosa Cell Apoptosis Through Targeting BMPR1B, a Major Gene for Sheep Prolificacy.

Bone morphogenetic protein receptor 1B (BMPR1B) is a major gene for ovine ovulation rate and litter size, which plays a crucial role in follicle development. However, its role and regulation in the ovine granulosa cells (GCs) are unclear. Here, we showed that silencing of BMPR1B enhanced cell apoptosis, whereas overexpression of BMPR1B inhibited cell apoptosis in the ovine GCs. Luciferase reporter assays revealed that BMPR1B is a target of miR-125b, and miR-125b reduced BMPR1B messenger RNA and protein levels in the ovine GCs by directly binding to the 3' untranslated region of the ovine BMPR1B gene. Furthermore, miR-125b enhanced cell apoptosis by attenuating BMPR1B in the ovine GCs. Our data demonstrated that BMPR1B is an important factor in the ovine GC function in vitro and targeted by a specific regulator miR-125b.

Influence of fecundity gene on oestrus synchronization and gonadal steroid levels in booroola carrier and Non-Carrier ewes

Prolificacy in Garole sheep is due to the effect of a single point mutation in the bone morphogenetic protein receptor 1B (BMPR-1B) and expressed in the granulosa cells and oocytes. The GMM (25% Garole-75% Malpura) sheep was developed by FecB introgression from Garole sheep. An experiment was conducted in the major breeding season (July-August, 2016) to study the oestrus synchronization response and plasma progesterone and oestradiol hormone concentrations in prolific GMM ewes (FecBBB), non-prolific GMM (FecB++) and Malpura ewes (FecB++). GMM strain was developed from Malpura (75%) and Garole (25%) breed so considered as control group for estrus response. A total of 28 ewes (carrier GMM (n=9), non-carrier GMM (n=10) and Malpura (n=9)) were synchronized with progesterone impregnated intravaginal sponges. It was observed that the proportion of ewes that exhibited oestrus did not differ significantly between the carrier and non-carrier GMM ewes; however, the oestrus response was early in carrier GMM compared to non-carrier GMM and Malpura ewes. The interval time for the onset of oestrus was non-significantly lower in carrier GMM (32.00±5.93) as compared to noncarrier GMM (36.00±3.70h) and Malpura (39.43±3.43h) ewes. The duration of oestrus did not differ significantly between the genotypes. The plasma progesterone concentration was significantly (P<0.05) higher in carrier GMM ewes (2.73±1.29ng/ml) as compared to non-carrier GMM ewes (1.28±0.62ng/ml). The oestradiol concentration was significantly higher (P<0.001) in non-carrier GMM (181.95±30.38pg/ml) as compared to carrier GMM ewes (124.43±15.87pg/ml). The results indicated that the FecB mutation alters the circulating concentration of progesterone and estradiol in Booroola carrier GMM (FecBBB) ewes.

Canonical BMP signaling in tubular cells mediates recovery after acute kidney injury

Bone morphogenetic protein (BMP) signaling has been shown to modulate the development of renal fibrosis in animal models of kidney injury, but the downstream mediators are incompletely understood. In wild-type mice, canonical BMP signaling mediated by SMAD1/5/8 transcription factors was constitutively active in healthy renal tubules, transiently down-regulated after ischemia reperfusion injury (IRI), and reactivated during successful tubular regeneration. We then induced IRI in mice with a tubular-specific BMP receptor 1A (BMPR1A) deletion. These mice failed to reactivate SMAD1/5/8 signaling in the post-ischemic phase and developed renal fibrosis after injury. Using unbiased genomic analyses, we identified three genes encoding inhibitor of DNA-binding (ID) proteins (Id1, Id2, and Id4) as key targets of BMPR1A-SMAD1/5/8 signaling. BMPR1A-deficient mice failed to re-induce these targets following IRI. Instead, BMPR1A-deficiency resulted in activation of pro-fibrotic signaling proteins that are normally repressed by ID proteins, namely, p38 mitogen-activated protein kinase and cell cycle inhibitor p27. These data indicate that the post-ischemic activation of canonical BMP signaling acts endogenously to repress pro-fibrotic signaling in tubular cells and may help to prevent the progression of acute kidney injury to chronic kidney disease.

Copper-Catalyzed Synthesis, Bio-Evaluation, and in Silico Studies of 2-Aryl-N-alkylbenzimidazoles as Neuroprotective Agents

2-aryl-N-alkylbenzimidazole derivatives synthesized by CuI/PPh3 promoted direct coupling of N-alkylbenzimidazoles with aryl bromides. In vitro neurotoxicities of 20 compounds were evaluated, and the neuroprotective abilities of low-neurotoxic compounds (3b, 3g, 3h, 3i, 3j, 3k, 3o, 3q, 3s and 3t) were investigated against toxicity induced by 1-methyl-4-phenylpyridinium ion (MPP+) in SH-SY5Y neuronal cells. In silico studies revealed that compound 3g could have molecule docking with the following proteins: the bone morphogenetic protein receptor type 1B (BMPR1B), human cytochrome P450 1B1(CYP1B1), Metabotropic glutamate receptor 7 (GRM7), histone deacetylase 6 (HDAC6), 5-hydroxytryptamine receptor 5A (HTR5A), human topoisomerase II beta (TOP2B). A molecular docking simulation of model compound 3g and model protein CYP1B1 has been shown.

Expression Analysis of the Prolific Candidate Genes, BMPR1B, BMP15, and GDF9 in Small Tail Han Ewes with Three Fecundity (FecB Gene) Genotypes.

Simple SummaryAs important prolific candidate genes, BMPR1B, BMP15, and GDF9 may affect the lambing performance of sheep. Therefore, regarding the three FecB genotypes of Small Tail Han (STH) sheep (FecB BB, FecB B+, and FecB ++), this study explored the gene expression characteristics of different tissues using reverse transcription PCR (RT-PCR) and real-time quantitative PCR (qPCR). The results showed that BMPR1B, BMP15, and GDF9 expression differed between the selected tissues, with all being highly expressed in the ovaries. Further analysis indicated that there was no significant difference in BMPR1B expression among the three FecB genotypes, but both GDF9 and BMP15 had the highest expression in FecB B+. As for other non-ovarian tissues, expression also varied. This study is relevant to understanding the high prolificacy of the STH breed.The expression characteristics of the prolific candidate genes, BMPR1B, BMP15, and GDF9, in the major visceral organs and hypothalamic–pituitary–gonadal (HPG) axis tissues of three FecB genotypes (FecB BB, FecB B+, and FecB ++) were explored in STH ewes using RT-PCR and qPCR. The results were as follows, BMPR1B was expressed in all FecB BB genotype (Han BB) tissues, and GDF9 was expressed in all selected tissues, but BMP15 was specifically expressed in the ovaries. Further study of ovarian expression indicated that there was no difference in BMPR1B expression between genotypes, but the FecB B+ genotype (Han B+) had greater expression of GDF9 and BMP15 than Han BB and FecB ++ genotype (Han ++) (p < 0.05, p < 0.01). BMP15 expression was lower in the ovaries of Han BB than in Han ++ sheep, but the reverse was shown for GDF9. The gene expression in non-ovarian tissues was also different between genotypes. Therefore, we consider that the three genes have an important function in ovine follicular development and maturation. This is the first systematic analysis of the tissue expression pattern of BMPR1B, BMP15, and GDF9 genes in STH sheep of the three FecB genotypes. These results contribute to the understanding of the molecular regulatory mechanism for ovine reproduction.

MicroRNA-125b Regulates Apoptosis by Targeting Bone Morphogenetic Protein Receptor 1B in Yak Granulosa Cells.

The intronic microRNA, miR-125b, plays a vital role in promyelocytic and hematopoietic stem cells, and in the development and apoptosis of cancer cells. In this study, we showed that miR-125b regulates granulosa cell (GC) apoptosis in the yak ovary. Bioinformatic analyses and luciferase reporter assays demonstrated that bone morphogenetic protein receptor type 1B (BMPR1B) is an miR-125b target. miR-125b overexpression induced apoptosis in yak GC, and affected the mRNA and protein expression of BMPR1B and the ratio of Bcl2/Bax. Silencing of miR-125b decreased the rate of yak GC apoptosis and increased the ratio of Bcl2/Bax. In addition, the effects of an miR-125b inhibitor were overturned by cotransfection with siRNA-BMPR1B2 (siRNA-299) in yak GC. Together, these results demonstrated that miR-125b regulates GC apoptosis in the yak ovary by targeting BMPR1B.

Detection of single nucleotide polymorphisms at major prolificacy genes in the Mehraban sheep and association with litter size

Abstract The present study aimed to investigate the presence of polymorphisms at four known genes controlling ovine prolificacy i.e. BMP15, GDF9, BMPR1B and B4GALNT2 in a sample of 115 Iranian Mehraban ewes and their association with litter size (LS) and lambs’ birth weight (BW) traits. Using Sanger sequencing of exons and polymorphism specific genotyping, ten SNPs (Single Nucleotide Polymorphisms) were observed in only two genes, GDF9 and BMPR1B. Seven SNPs were found in the GDF9 gene on the chromosome 5. Among them, six were already described in the coding sequence, and a new one (g.41840985C&gt;T) was found in the 3’UTR. In the BMPR1B gene on the chromosome 6, three novel SNPs were detected in the exon 7 (g.29382184G&gt;A; g.29382337G&gt;A and g.29382340G&gt;A). Allelic frequencies were established for six SNPs among the ten identified and they were in Hardy-Weinberg equilibrium. A significant association was found between the novel SNPs found in the exon 7 of BMPR1B and LS. Present results indicate the potential role of the BMPR1B locus in controlling prolificacy of Mehraban sheep and provide genetic markers for further exploitation in selection to improve reproductive efficiency.

Polymorphism of GDF9 and BMPR1B genes and their association with litter size in Markhoz goats.

The main objective of this study was to investigate the polymorphism of GDF9 and BMPR1B genes and their relationship with litter size in Markhoz goats. The polymorphism of GDF9 and BMPR1B genes as well-documented genes regarding fecundity in sheep and goat was investigated using RFLP-PCR and a tetra-primer amplification refractory mutation system-PCR (T-ARMS-PCR) in Markhoz goats. The 164 blood samples were collected from the raised goats in Sanandaj Markhoz goat Performance Testing Station. The DNA extraction was carried out by salting-out procedure, and then, PCR was performed using four and two pairs of primers to detect polymorphism in GDF9 and BMPR1B genes, respectively. To disclose GDF9 loci polymorphism, PCR products were digested with SspI (G3288A), PvuII (G423A), MvaI (A959C) and MspI (G1189A) restriction enzymes. The results showed that these mutations are available in tested animals. Parity had no significant effect on litter size. Also, the effects of different genotypes of GDF9 and BMPR1B had no significant effect on litter size. Further studies with a high number of animals with minimum relatedness for testing the association of these SNPs and others in the fecundity genes with reproductive traits may be worthwhile.

MP28-12 SUPPRESSION OF ONCOGENIC MICRORNA-1274A INDUCES CELL APOPTOSIS THROUGH BMPR1B ACCELERATION IN RENAL CELL CARCINOMA

You have accessJournal of UrologyKidney Cancer: Epidemiology & Evaluation/Staging I1 Apr 2018MP28-12 SUPPRESSION OF ONCOGENIC MICRORNA-1274A INDUCES CELL APOPTOSIS THROUGH BMPR1B ACCELERATION IN RENAL CELL CARCINOMA Hideki Enokida, Hirofumi Yoshino, Satoshi Sugita, Takashi Sakaguchi, Youichi Osako, Shuichi Tatarano, Takashi Yamane, and Masayuki Nakagawa Hideki EnokidaHideki Enokida More articles by this author , Hirofumi YoshinoHirofumi Yoshino More articles by this author , Satoshi SugitaSatoshi Sugita More articles by this author , Takashi SakaguchiTakashi Sakaguchi More articles by this author , Youichi OsakoYouichi Osako More articles by this author , Shuichi TataranoShuichi Tatarano More articles by this author , Takashi YamaneTakashi Yamane More articles by this author , and Masayuki NakagawaMasayuki Nakagawa More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2018.02.912AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES There is considerable evidence that normal microRNA (miRNA) regulatory mechanisms are disrupted in cancer cells. However, the regulatory mechanisms by which upregulated miRNAs exert their effects has not been often understood compared to downregulated miRNAs. Our previous studies of the miRNA expression signature in renal cell carcinoma (RCC) indicated that miR-1274a was significantly upregulated in clinical specimens, suggesting that miR-1274a might act as an oncogenic miRNA in RCC. The aim of this study was to investigate the functional roles of miR-1274a and identify downstream tumor suppressive targets regulated by miR-1274a in RCC cells. METHODS Expression levels of miR-1274a and its target gene were determined by qRT-PCR. Functional studies of miR-1274a were carried out by anti-miRNA to investigate cell proliferation and apoptosis by using A498, ACHN, and Caki1 RCC cell lines. Putative target genes were determined by in-silico analyses with Gene Expression Omnibus (GEO) database and the microarray study of miR-1274a-transfected ACHN. In addition, statistical analyses were performed based on the TCGA database of RCC patients. RESULTS The expression levels of miR-1274a were significantly upregulated in RCC in comparison with normal kidneys (P < 0.0001). Suppression of miR-1274a significantly inhibited cell proliferation and induced apoptosis in RCC cells (P < 0.0001 and P < 0.05, respectively). In-silico analysis of the gene expression data and luciferase reporter assays demonstrated that miR-1274a directly regulated bone morphogenetic protein receptor type 1B (BMPR1B), a member of BMP receptor family of transmembrane serine/threonine kinases. It is known that the ligands of this receptor BMPs, members of the TGF-beta superfamily, bind their receptor including BMPR1B, which lead to apoptosis thorough activation of the SMAD-dependent pathway subsequently through phosphorylation of SMAD1/5/8. Moreover, TCGA database as well as the immunohistochemistry showed that low expression of BMPR1B was observed in RCC clinical specimens compared to normal kidneys (P < 0.0001 and P = 0.0015, respectively). CONCLUSIONS We conclude that loss of oncogenic miR-1274a reduced cancer cell proliferation and induced apoptosis in RCC through targeting tumor suppressive BMPR1B. Our data demonstrating molecular pathways and a target gene regulated by oncogenic miR-1274a will provide new insight into the potential mechanisms of RCC oncogenesis. © 2018FiguresReferencesRelatedDetails Volume 199Issue 4SApril 2018Page: e361 Advertisement Copyright & Permissions© 2018MetricsAuthor Information Hideki Enokida More articles by this author Hirofumi Yoshino More articles by this author Satoshi Sugita More articles by this author Takashi Sakaguchi More articles by this author Youichi Osako More articles by this author Shuichi Tatarano More articles by this author Takashi Yamane More articles by this author Masayuki Nakagawa More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

Screening for Causative Mutations of Major Prolificacy Genes in Iranian Fat-Tailed Sheep.

BackgroundThe presence of different missense mutations in sheep breeds have shown that the bone morphogenetic protein receptor 1B (BMPR1B), bone morphogenetic protein 15 (BMP15) and growth differentiation factor 9 (GDF9) genes play a vital role in ovulation rate and prolificacy in ewes. Therefore, the present study aims to investigate BM- PR1B, BMP15 and GDF9 gene mutations in prolific ewes of Iranian fat-tailed Lori-Bakhtiari sheep.Materials and MethodsIn the present experimental study, genomic DNA was extracted from whole blood of 10 prolific Lori-Bakhtiari ewes with at least two twinning records in the first four parities to identify point mutations of the BMPR1B, BMP15 and GDF9 genes, using DNA sequencing.ResultsThe results obtained from DNA sequencing showed a new synonymous mutation (g.66496G>A) in exon 8 of the BMPR1B gene, without any amino acid change. Sequencing of the BMP15 gene revealed a deletion of 3 bp (g.656_658delTTC) in exon 1, leading to an amino acid deletion (p.Leu19del). Four single nucleotide polymorphisms (G1:g.2118G>A, G2:g.3451T>C, G3:g.3457A>G and G4:g.3701G>A), were detected in exons 1 and 2 of the GDF9 gene, two of which caused amino acid substitutions (G1: p.87Arg>His and G4: p.241Glu>Lys). These amino acid alterations are proposed to have a benign impact on structure and function of the GDF9 polypeptide sequence.ConclusionThree major prolificacy genes (BMPR1B, BMP15 and GDF9) were polymorphic in Lori-Bakhtiari sheep, although none of the major causative mutation was detected in this sheep type. Further studies using high throughput methods such as genome-wide association study (GWAS) and evaluation of other candidate genes are necessary in the future.

Generation of gene-edited sheep with a defined Booroola fecundity gene (FecBB) mutation in bone morphogenetic protein receptor type 1B (BMPR1B) via clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated (Cas) 9.

Since its emergence, the clustered regularly interspaced short palindromic repeat (CRISPR)-CRISPR-associated (Cas) 9 system has been increasingly used to generate animals for economically important traits. However, most CRISPR/Cas9 applications have been focused on non-homologous end joining, which results in base deletions and insertions, leading to a functional knockout of the targeted gene. The Booroola fecundity gene (FecBB) mutation (p.Q249R) in bone morphogenetic protein receptor type 1B (BMPR1B) has been demonstrated to exert a profound effect on fecundity in many breeds of sheep. In the present study, we successfully obtained lambs with defined point mutations resulting in a p.249Q>R substitution through the coinjection of Cas9 mRNA, a single guide RNA and single-stranded DNA oligonucleotides into Tan sheep zygotes. In the newborn lambs, the observed efficiency of the single nucleotide exchange was as high as 23.8%. We believe that our findings will contribute to improved reproduction traits in sheep, as well as to the generation of defined point mutations in other large animals.

Whole-genome resequencing reveals candidate mutations for pig prolificacy.

Changes in pig fertility have occurred as a result of domestication, but are not understood at the level of genetic variation. To identify variations potentially responsible for prolificacy, we sequenced the genomes of the highly prolific Taihu pig breed and four control breeds. Genes involved in embryogenesis and morphogenesis were targeted in the Taihu pig, consistent with the morphological differences observed between the Taihu pig and others during pregnancy. Additionally, excessive functional non-coding mutations have been specifically fixed or nearly fixed in the Taihu pig. We focused attention on an oestrogen response element (ERE) within the first intron of the bone morphogenetic protein receptor type-1B gene (BMPR1B) that overlaps with a known quantitative trait locus (QTL) for pig fecundity. Using 242 pigs from 30 different breeds, we confirmed that the genotype of the ERE was nearly fixed in the Taihu pig. ERE function was assessed by luciferase assays, examination of histological sections, chromatin immunoprecipitation, quantitative polymerase chain reactions, and western blots. The results suggest that the ERE may control pig prolificacy via the cis-regulation of BMPR1B expression. This study provides new insight into changes in reproductive performance and highlights the role of non-coding mutations in generating phenotypic diversity between breeds.

Rare copy number variants identified in prune belly syndrome

Prune belly syndrome (PBS), also known as Eagle-Barrett syndrome, is a rare congenital disorder characterized by absence or hypoplasia of the abdominal wall musculature, urinary tract anomalies, and cryptorchidism in males. The etiology of PBS is largely unresolved, but genetic factors are implicated given its recurrence in families. We examined cases of PBS to identify novel pathogenic copy number variants (CNVs). A total of 34 cases (30 males and 4 females) with PBS identified from all live births in New York State (1998–2005) were genotyped using Illumina HumanOmni2.5 microarrays. CNVs were prioritized if they were absent from in-house controls, encompassed ≥10 consecutive probes, were ≥20 Kb in size, had ≤20% overlap with common variants in population reference controls, and had ≤20% overlap with any variant previously detected in other birth defect phenotypes screened in our laboratory. We identified 17 candidate autosomal CNVs; 10 cases each had one CNV and four cases each had two CNVs. The CNVs included a 158 Kb duplication at 4q22 that overlaps the BMPR1B gene; duplications of different sizes carried by two cases in the intron of STIM1 gene; a 67 Kb duplication 202 Kb downstream of the NOG gene, and a 1.34 Mb deletion including the MYOCD gene. The identified rare CNVs spanned genes involved in mesodermal, muscle, and urinary tract development and differentiation, which might help in elucidating the genetic contribution to PBS. We did not have parental DNA and cannot identify whether these CNVs were de novo or inherited. Further research on these CNVs, particularly BMP signaling is warranted to elucidate the pathogenesis of PBS.

Downregulation of microRNA-1274a induces cell apoptosis through regulation of BMPR1B in clear cell renal cell carcinoma.

Our previous studies of the microRNA (miRNA) expression signature in clear cell renal cell carcinoma (ccRCC) indicated that miRNA-1274a (miR-1274a) was significantly upregulated in clinical specimens, suggesting that miR-1274a may act as an oncogenic miRNA in ccRCC. The aim of this study was to investigate the functional roles of miR-1274a and identify downstream tumor-suppressive targets regulated by miR‑1274a in ccRCC cells. Functional studies of miR-1274a were carried out by anti-miRNA to investigate cell proliferation and apoptosis using the A498, ACHN and Caki1 ccRCC cell lines. Suppression of miR-1274a significantly inhibited cancer cell proliferation and induced apoptosis in the ccRCC cells. Gene expression data combined with insilico analysis and luciferase reporter assays demonstrated that bone morphogenetic protein receptor type1B (BMPR1B) was directly regulated by miR-1274a. Moreover, TCGA database as well as immunohistochemistry demonstrated low expression of BMPR1B in ccRCC clinical specimens compared to that in normal kidney tissues. We conclude that loss of oncogenic miR-1274a reduced cancer cell proliferation and induced apoptosis in ccRCC through targeting BMPR1B. Our data revealing molecular pathways and a target gene regulated by oncogenic miR-1274a provide new insight into the potential mechanisms of ccRCC oncogenesis.

A novel duplication downstream of BMP2 in a Chinese family with Brachydactyly type A2 (BDA2)

BackgroundBrachydactyly type A2 (BDA2) is an autosomal dominant disease characterized by the deformation of the middle phalanx of the second fingers and toes. It has been reported to be associated with three genes regulating the osteogenesis, including BMPR1B, GDF5 and BMP2. Materials and methods10 BDA2 patients and 7 unaffected individuals in a Chinese family were identified through clinical signs and radiographs. The mutation analyses of BMPR1B, GDF5 and BMP2 gene was performed in all the available family members and 100 control subjects. The duplication analysis for the downstream of BMP2 was also performed in all the samples. ResultsA novel 4671bp duplication downstream the BMP2 gene was identified in all the patients undergoing molecular analysis but not in the unaffected individuals and healthy controls, with a 28bp microhomology flanking it. There was no mutation in all the exons of BMPR1B, GDF5 and BMP2 in all the tested family members. ConclusionThe novel duplication has different breakpoints compared with the previous ones but highly overlapped with them. The duplication narrows the range of the potential cis-regulatory sequence, and further supports the association between BDA2 and the duplication downstream BMP2.

Using sheep genomes from diverse U.S. breeds to identify missense variants in genes affecting fecundity

Background: Access to sheep genome sequences significantly improves the chances of identifying genes that may influence the health, welfare, and productivity of these animals. Methods: A public, searchable DNA sequence resource for U.S. sheep was created with whole genome sequence (WGS) of 96 rams. The animals shared minimal pedigree relationships and represent nine popular U.S. breeds and a composite line. The genomes are viewable online with the user-friendly Integrated Genome Viewer environment, and may be used to identify and decode gene variants present in U.S. sheep. Results: The genomes had a combined average read depth of 16, and an average WGS genotype scoring rate and accuracy exceeding 99%. The utility of this resource was illustrated by characterizing three genes with 14 known coding variants affecting litter size in global sheep populations: growth and differentiation factor 9 ( GDF9), bone morphogenetic protein 15 ( BMP15), and bone morphogenetic protein receptor 1B ( BMPR1B). In the 96 U.S. rams, nine missense variants encoding 11 protein variants were identified. However, only one was previously reported to affect litter size ( GDF9 V371M, Finnsheep). Two missense variants in BMP15 were identified that had not previously been reported: R67Q in Dorset, and L252P in Dorper and White Dorper breeds. Also, two novel missense variants were identified in BMPR1B: M64I in Katahdin, and T345N in Romanov and Finnsheep breeds. Based on the strict conservation of amino acid residues across placental mammals, the four variants encoded by BMP15 and BMPR1B are predicted to interfere with their function. However, preliminary analyses of litter sizes in small samples did not reveal a correlation with variants in BMP15 and BMPR1B with daughters of these rams. Conclusions: Collectively, this report describes a new resource for discovering protein variants in silico and identifies alleles for further testing of their effects on litter size in U.S. breeds.

Using sheep genomes from diverse U.S. breeds to identify missense variants in genes affecting fecundity.

Background: Access to sheep genome sequences significantly improves the chances of identifying genes that may influence the health, welfare, and productivity of these animals. Methods: A public, searchable DNA sequence resource for U.S. sheep was created with whole genome sequence (WGS) of 96 rams. The animals shared minimal pedigree relationships and represent nine popular U.S. breeds and a composite line. The genomes are viewable online with the user-friendly Integrated Genome Viewer environment, and may be used to identify and decode gene variants present in U.S. sheep. Results: The genomes had a combined average read depth of 16, and an average WGS genotype scoring rate and accuracy exceeding 99%. The utility of this resource was illustrated by characterizing three genes with 14 known coding variants affecting litter size in global sheep populations: growth and differentiation factor 9 ( GDF9), bone morphogenetic protein 15 ( BMP15), and bone morphogenetic protein receptor 1B ( BMPR1B). In the 96 U.S. rams, nine missense variants encoding 11 protein variants were identified. However, only one was previously reported to affect litter size ( GDF9 V371M, Finnsheep). Two missense variants in BMP15 were identified that had not previously been reported: R67Q in Dorset, and L252P in Dorper and White Dorper breeds. Also, two novel missense variants were identified in BMPR1B: M64I in Katahdin, and T345N in Romanov and Finnsheep breeds. Based on the strict conservation of amino acid residues across placental mammals, the four variants encoded by BMP15 and BMPR1B are predicted to interfere with their function. However, preliminary analyses of litter sizes in small samples did not reveal a correlation with variants in BMP15 and BMPR1B with daughters of these rams. Conclusions: Collectively, this report describes a new resource for discovering protein variants in silico and identifies alleles for further testing of their effects on litter size in U.S. breeds.

Survey of mutations in prolificacy genes in Santa Ines and Morada Nova sheep

ABSTRACT Polymorphisms in the BMP-15 gene related to Galway (FecXG) and Inverdale (FecXI) and in the BMPR-1B gene known as Booroola (FecB) mutations were investigated using the Polymerase Chain Reaction - Restriction Fragment Length Polymorphism (PCR-RFLP) method, on sheep from the breeds Santa Inês (n= 574) and Morada Nova (n=282). DNA was extracted and amplified through PCR with specific primers that introduced a restriction site in association with the mutation. The PCR products were submitted to endonucleases. The experiment found no FecXG and FecXI mutations. Six samples of animals with multiple offspring/birth history presented polymorphism for FecB similar to control samples, but this pattern was not confirmed by nucleotide sequencing. Although the absence of these mutations in the studied breeds, other factors related to prolificacy should be investigated to explain the inherent prolificity mechanisms.