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1.
Genet Mol Res ; 15(4)2016 Dec 19.
Article in English | MEDLINE | ID: mdl-28002603

ABSTRACT

The application of assisted reproductive technology in animal production benefits the economy and conservation of biological resources. Single nucleotide polymorphism (SNPs) was used as predictive markers for breeding and reproduction. In the present study, we examined the association between a SNP of the grb10 gene and superovulation traits in cattle. Sequencing results indicated a point mutation and statistical analysis showed a significant association of the mutation with superovulation traits. The high number of embryos collected from the heterozygotes suggested that the mutation in the grb10 gene exerted a significant effect on the number of embryos recovered although the quality was not affected. The grb10 gene may serve as a useful biomarker for donor selection.


Subject(s)
GRB10 Adaptor Protein/genetics , Polymorphism, Single Nucleotide , Quantitative Trait Loci , Superovulation/genetics , Animals , Cattle , Embryo Culture Techniques , Female , Gene Expression Regulation, Developmental , Point Mutation , Sequence Analysis, DNA
2.
Genet Mol Res ; 15(4)2016 Nov 03.
Article in English | MEDLINE | ID: mdl-27819741

ABSTRACT

Heroin dependence is a chronic relapsing brain disease. Researchers have reported that the dopamine D2 receptor (DRD2) is involved in the development of opiate dependence. To identify markers that contribute to the genetic susceptibility to heroin addiction, we examined the potential association between heroin dependence and six polymorphisms of the DRD2 gene using the MassARRAY system. Three hundred and thirty-four patients with heroin dependence and 299 healthy controls participated in the research. Compared with the healthy controls, heroin-dependent patients had a significantly lower frequency of the AA genotype of rs6275 (P = 0.038), and a significantly higher frequency of the C allele of rs1125394 (P = 0.030). Statistically significant differences were observed in the genotypic and allelic frequencies of rs17115583 (P = 0.005 and P = 0.001, respectively) and rs1079597 (P = 0.03 and P = 0.02, respectively). Haplotype analysis revealed that the T-G-A (block 1) haplotype of the DRD2 gene conferred a protective effect (P = 0.020). These findings point to a role for DRD2 polymorphism in heroin dependence in the Chinese Han population, and may be informative for future genetic or neurobiological studies on heroin dependence.


Subject(s)
Genetic Association Studies , Genetic Predisposition to Disease , Heroin Dependence/genetics , Polymorphism, Single Nucleotide/genetics , Receptors, Dopamine D2/genetics , Alleles , Case-Control Studies , Gene Frequency , Haplotypes , Humans , Linkage Disequilibrium , Male , Middle Aged , Risk Factors
3.
Genet Mol Res ; 15(4)2016 Oct 05.
Article in English | MEDLINE | ID: mdl-27808368

ABSTRACT

Hepatocellular carcinoma (HCC) is a major cause of cancer-related deaths worldwide. MicroRNA-34 (miR-34) gene plays a key role in altering the apoptotic cycle and pathways of downstream cells, and therefore influences carcinogenesis. In this case-control study, we assessed the role of the pri-miR-34b/c rs4938723 polymorphism in HCC risk. The pri-miR-34b/c polymorphic genotype was determined in 286 patients with HCC and 572 controls using polymerase chain reaction-restriction fragment length polymorphism. The male gender (X2 = 12.95, P < 0.001), regular alcohol consumption (X2 = 16.81, P < 0.001), and a family history of cancer (X2 = 11.88, P = 0.001) were associated with HCC risk. However, the age (t = 1.19, P = 0.12) and tobacco smoking habit (X2 = 0.64, P = 0.42) of HCC patients were comparable to those of the controls. The TC (adjusted OR = 1.46, 95%CI = 1.06-2.01) and CC (adjusted OR = 3.07, 95%CI = 1.77-5.34) genotypes of pri-miR-34b/c rs4938723 were correlated with a higher risk of HCC compared to the TT genotype. Moreover, the TC+CC genotype was correlated with an increased risk of HCC compared to the TT genotype (adjusted OR = 1.64, 95%CI = 1.21-2.22). In the recessive model, the CC genotype of pri-miR-34b/c rs4938723 was significantly correlated with an elevated risk of HCC compared to the TT+TC genotype (adjusted OR = 2.50, 95%CI = 1.49-4.22). Further large-scale and multi-center studies are required to confirm these results.


Subject(s)
Carcinoma, Hepatocellular/genetics , Genetic Association Studies , Genetic Predisposition to Disease , MicroRNAs/genetics , Polymorphism, Single Nucleotide/genetics , Promoter Regions, Genetic , Case-Control Studies , Demography , Female , Humans , Liver Neoplasms/genetics , Male , MicroRNAs/metabolism , Middle Aged , Risk Factors
4.
Genet Mol Res ; 15(2)2016 Apr 07.
Article in English | MEDLINE | ID: mdl-27173213

ABSTRACT

Our study clarifies the role of the autocrine motility factor receptor (AMFR) gene in porcine preadipocyte differentiation. AMFR-siRNA was transfected into porcine preadipocytes and the preadipocytes were induced to differentiation. Subsequently, qRT-PCR was conducted to examine changes in mRNA expression of a series of genes in porcine preadipocytes, including AMFR, sterol-regulatory element-binding protein-1a (SREBP1a), SREBP2, insulin-induced gene 1 (Insig1), and Insig2. Expression changes in the mRNA of genes regulating adipocyte differentiation were also analyzed using qRT-PCR, including peroxisome proliferator-activated receptor gamma (PPARγ), CCAAT/enhancer-binding protein alpha (C/EBPα), and Kruppel-like factor 2 (KLF2). Western blot analysis was conducted to examine the changes in AMFR protein expression in porcine preadipocytes. Additionally, morphological changes in differentiated porcine preadipocytes were examined by oil red O staining, and changes in optical density (OD) values were measured using an ultraviolet spectrophotometer. At 24 h after transfection with AMFR-siRNA, AMFR mRNA expression significantly reduced (P < 0.01), and AMFR protein expression markedly decreased (P < 0.05). The mRNA expression of SREBP1a, SREBP2, Insig1, and C/EBPα was significantly reduced (P < 0.01), whereas the expression of KLF2 mRNA was significantly elevated (P < 0.01). After induction of preadipocyte differentiation, the number of lipid droplets decreased in the AMFR-silenced group, and the OD value markedly reduced (P < 0.05). In addition, the expression of C/EBPα mRNA significantly decreased (P < 0.05), whereas the expression of KLF2 mRNA considerably increased (P < 0.05). Taken together, silencing of the AMFR gene inhibits the differentiation of porcine preadipocytes.


Subject(s)
Adipocytes/metabolism , Cell Differentiation , Receptors, Autocrine Motility Factor/metabolism , Adipocytes/cytology , Animals , CCAAT-Enhancer-Binding Protein-alpha/genetics , CCAAT-Enhancer-Binding Protein-alpha/metabolism , Cells, Cultured , Gene Silencing , Kruppel-Like Transcription Factors/genetics , Kruppel-Like Transcription Factors/metabolism , PPAR gamma/genetics , PPAR gamma/metabolism , Receptors, Autocrine Motility Factor/genetics , Sterol Regulatory Element Binding Protein 1/genetics , Sterol Regulatory Element Binding Protein 1/metabolism , Sterol Regulatory Element Binding Protein 2/genetics , Sterol Regulatory Element Binding Protein 2/metabolism , Swine
5.
Genet Mol Res ; 15(1)2016 Mar 22.
Article in English | MEDLINE | ID: mdl-27050969

ABSTRACT

Aeromonas hydrophila, a widespread bacterium in the aquatic environment, causes hemorrhagic septicemia in fish. In the last decade, the disease has caused mass mortalities and tremendous economic loss in cultured fish. The complement component C7 is a terminal component of complement that interacts in a sequence of polymerization reactions with other terminal complement components to form a membrane attack complex. The formation of the membrane attack complex creates a pore in the membranes of certain pathogen that can lead to their death. The objective of this study was to identify single nucleotide polymorphisms (SNPs) in the C7 gene and to assess their association with A. hydrophila resistance in grass carp. A resource population consisting of 186 susceptible and 191 resistant grass carp was constructed. We sequenced a total of 7826 bp of the C7 gene and identified 6 SNPs that were genotyped in the resource population. The SNP -1575 A>C was positioned in the promoter region of the gene. The SNP 425 C>T identified in the coding exon was a synonymous substitution in the fourth exon. Statistical analysis showed that SNP 425 C>T was associated with the incidence of hemorrhagic septicemia. The SNPs -1575 A>C, -688 T>C, and -266 A>C were highly linked together (r(2) > 0.85). No haplotypes generated with these 3 SNPs were associated with resistance to A. hydrophila in grass carp. These findings suggest that the 425 C>T polymorphism in C7 gene may be a significant molecular marker for resistance to A. hydrophila in grass carp.


Subject(s)
Aeromonas hydrophila/pathogenicity , Carps/genetics , Carps/microbiology , Complement C7/genetics , Polymorphism, Single Nucleotide/genetics , Animals , Carps/metabolism , Genotype , Haplotypes
6.
Genet Mol Res ; 14(4): 15062-72, 2015 Nov 25.
Article in English | MEDLINE | ID: mdl-26634468

ABSTRACT

In this study, the performance of 300 Changbaishan Black cattle treated for superovulation from June to September was evaluated to determine the optimal conditions and herds for bovine embryo production. Data analysis revealed that cattle treated in July and August had higher numbers of available embryos (NAE), M1 embryos (NM1), and total embryos (NTE), as well as a higher percentage of M1 embryos (PM1). The temperature and precipitation observed during July and August were greater than those seen in the other two months; strong correlations were observed between these traits and the choice of month of treatment. In addition, multiparous cattle showed a better performance, higher NTE, NAE, NM1, and PM1 values, higher percentages of available embryos, and a lower percentage of degenerated embryos. The co-efficient correlation analysis showed that the month chosen for the treatment did not affect the superovulation traits of nulliparous cattle; however, the choice of the month affected multiparous cattle. Multiparous and nulliparous cattle exhibited many significant differences when treated in July and in August. In addition, the superovulatory traits of multiparous cattle, and not the nulliparous cattle, were strongly correlated to the choice of month of treatment. The results suggested that superovulation is more effective during a period with appropriate environmental temperature and humidity, and that multiparous cattle are more suitable for morula production.


Subject(s)
Cattle/genetics , Superovulation/genetics , Animals , Cattle/physiology , Embryo Transfer , Female , Follicle Stimulating Hormone/administration & dosage , Parity , Phenotype , Pregnancy , Rain , Sunlight , Superovulation/drug effects , Temperature , Time Factors
7.
Genet Mol Res ; 14(4): 14539-47, 2015 Nov 19.
Article in English | MEDLINE | ID: mdl-26600513

ABSTRACT

This study was designed to examine a single nucleotide polymorphism (SNP) in the HIF-3α gene in three hundred Changbaishan black cattle using PCR-restriction fragment length polymorphism to determine whether there is an association between this SNP and superovulation. The cloning and sequencing results indicate that the polymorphism is due to a point mutation at the 278-bp position in the HIF-3α gene, resulting in 3 genotypes (AA, AB, and BB). Association analysis indicated that the polymorphism has a significant effect on the number of unfertilized embryos (NUE) (P < 0.05) in the cattle. Cattle with genotype BB had a higher NUE than those with genotype AA, but the difference in NUE between AB and AA or BB was not significant. The polymorphism also has a highly significant effect on the number of degenerative embryos (NDE) and the number of total embryos (NTE) (P < 0.01). Genotype BB was associated with a higher NDE than AA, but the difference in NDE between AB and AA or BB was not significant. Genotype BB showed a higher NTE than AA or AB, but the difference in NTE between AA and AB was not significant. No significant conclusions could be drawn with respect to susceptibility to other traits. HIF-3α could serve as a useful biomarker for donor selection, superovulation improvement, and assisted fertility.


Subject(s)
Basic Helix-Loop-Helix Transcription Factors/genetics , Genetic Association Studies , Reproduction/genetics , Superovulation/genetics , Animals , Cattle , Female , Fertility/genetics , Phenotype
8.
Genet Mol Res ; 14(4): 12505-19, 2015 Oct 16.
Article in English | MEDLINE | ID: mdl-26505401

ABSTRACT

Cells isolated from human first trimester umbilical cord perivascular layer (hFTM-PV) tissues display the pluripotent characteristics of stem cells. In this study, we examined whether hFTM-PV cells can differentiate into islet-like clusters (ILCs) in vitro, and whether transplantation of the hFTM-PV cells with and without differentiation in vitro can alleviate diabetes in nude mice. The hFTM-PV cells were differentiated into ILCs in vitro through a simple stepwise culture protocol. To examine the in vivo effects of the cells, the hFTM-PV cells with and without differentiation in vitro were transplanted into the abdominal cavity of nude mice with streptozotocin (STZ)-induced diabetes. Blood glucose levels, body weight, and the survival probability of the diabetic nude mice were then statistically analyzed. The hFTM-PV cells were successfully induced into ILCs that could release insulin in response to elevated concentrations of glucose in vitro. In transplantation experiments, we observed that mice transplanted with the undifferentiated hFTM-PV cells, embryonic body-like cell aggregations, or ILCs all demonstrated normalized hyperglycemia and showed improved survival rate compared with those without cell transplantation. The hFTM-PV cells have the ability to differentiate into ILCs in vitro and transplantations of undifferentiated and differentiated cells can alleviate STZ-induced diabetes in nude mice. This may offer a potential cell source for stem cell-based therapy for treating diabetes in the future.


Subject(s)
Cord Blood Stem Cell Transplantation/methods , Diabetes Mellitus, Experimental/therapy , Animals , Cell Differentiation/physiology , Diabetes Mellitus, Experimental/blood , Female , Humans , Islets of Langerhans/cytology , Mice , Mice, Nude , Pregnancy , Umbilical Cord/cytology
9.
Genet Mol Res ; 14(3): 8901-8, 2015 Aug 03.
Article in English | MEDLINE | ID: mdl-26345821

ABSTRACT

The aim of this study was to investigate the effect of montelukast on the expression of interleukin (IL)-18, telomerase reverse transcriptase (TERT), and Bcl-2 in the brain tissue of neonatal rats with hypox-ic-ischemic brain damage (HIBD). To establish the model of HIBD, 8% oxygen was applied to rats after the unilateral carotid artery was ligated. Twenty rats were randomly assigned to the control group, while another 40 were used to establish the HIBD model and were randomly divided equally into model group and treatment group. A 0.1 mg/kg dose of montelukast or an equal volume of saline was intraperitoneally injected to the rats in the treatment group and the model group, respectively. Brain tissue from 4 rats in each group was sampled at 0, 6, 12, 24, and 72 h after brain damage, and immunohistochemistry was used to measure IL-18, TERT and Bcl-2 expressions. IL-18, TERT, and Bcl-2 levels increased after 12 h in both the model group and treatment group, peaked after 48 h, and then decreased. Although not statistically significant, IL-18, TERT, and Bcl-2 expressions after 24, 48, and 96 h were all lower in the treatment group than those in the model group. In conclusion, montelukast has a protective effect on the cerebral tissue of neonatal rats with HIBD, and may mediate an increase of TERT and Bcl-2 levels but not of IL-18. Further study is required to elucidate the mechanism of the protective effect of montelukast on HIBD.


Subject(s)
Acetates/pharmacology , Hypoxia-Ischemia, Brain/drug therapy , Hypoxia-Ischemia, Brain/metabolism , Interleukin-18/biosynthesis , Proto-Oncogene Proteins c-bcl-2/biosynthesis , Quinolines/pharmacology , Telomerase/biosynthesis , Animals , Animals, Newborn , Case-Control Studies , Cyclopropanes , Disease Models, Animal , Immunohistochemistry , Rats , Sulfides
10.
Genet Mol Res ; 14(3): 9753-63, 2015 Aug 19.
Article in English | MEDLINE | ID: mdl-26345908

ABSTRACT

In this study, expression levels of miRNAs (miRNAs), miR-375 and miR-7, were detected in different tissues of cattle to determine whether adenohypophysis-prefer or exclusively expressed miRNAs, and target genes could be predicted by TargetScan, RNA22, and other software. Target genes related to pituitary function or reproductive traits were identified using a dual-luciferase assay. miR-375 and miR-7 were expressed differently in various tissues. miR-375 and miR-7 showed higher expression in the adenohypophysis, and there was a significant difference compared with expression in other tissues (P < 0.01). The binding sites for miR-7 were the mRNAs of bone morphogenetic protein receptor type II (BMPR2), prostaglandin F2 receptor negative regulator, gonadotropin-releasing hormone receptor, follicle-stimulating hormoneß, somatostatin receptor 1, and interleukin-1ß by bioinformatic analysis; similarly, the mRNAs of BMPR2 and leptin contained binding sites for miR-375, suggesting that these genes are affected by miR-7 or miR-375. Dual-luciferase reporter assays showed that miR-7 regulated prostaglandin F2 receptor negative regulator expression, while miR-375 regulated BMPR2 expression. The mutated plasmid and miRNA mimics were used to co-transfect NIH3T3 cells; luciferase reporter assays showed that the inhibition of luciferase activity in the wild-type cells dramatically decreased from 75 to 26% with a 3-5-nucleotide mismatch mutation into the seed region of miR-7. miR-375 had nearly lost the ability to inhibit luciferase activity, suggesting that GTCTTCC is the site of interaction between miR-7 and the prostaglandin F2 receptor negative regulator sequence and that GAACAAA is the site of interaction between miR-375 and the BMPR2 sequence.


Subject(s)
MicroRNAs/genetics , Pituitary Gland, Anterior/metabolism , RNA, Messenger/genetics , Animals , Base Sequence , Binding Sites , Cattle , Gene Expression , Gene Expression Profiling , Gene Expression Regulation , Genes, Reporter , MicroRNAs/chemistry , Nucleic Acid Conformation , Organ Specificity/genetics , RNA Interference , RNA, Messenger/chemistry
11.
Genet Mol Res ; 14(1): 457-63, 2015 Jan 23.
Article in English | MEDLINE | ID: mdl-25729979

ABSTRACT

Calpain-3 (CAPN3) is a member of the calpain family of Ca(2+)-regulated cysteine proteases, which play an important role in sarcomere remodeling and mitochondrial protein turnover, and thus, regulating beef tenderness in cattle. Currently, multiple CAPN3 transcripts have been detected in human, monkey, rat, and rabbit. However, whether this transcript is present in cattle remains unknown. In this study, we identified 2 CAPN3 transcripts in the skeletal muscle individuals of local black cattle from Jilin, China. One transcript corresponded to the known full-length protein and was referred to as CAPN3a, while the second transcript did not contain exons 2-19 and contained a single-nucleotide insert in the penultimate base of exon 1 compared to CAPN3a; this protein was referred to as CAPN3b. The expression level of CAPN3b was approximately 50-fold lower than that of CAPN3a. Moreover, CAPN3b mRNA was not translated into a functional protein because it had lost essential domains according to bioinformatic analysis. Our results not provide a foundation for understanding the function of CAPN3, but also are useful for further elucidating the effect of CAPN3 on meat quality in cattle.


Subject(s)
Alternative Splicing/genetics , Calpain/genetics , Cattle/genetics , Amino Acid Sequence , Animals , Base Sequence , Calpain/chemistry , Molecular Sequence Data , RNA, Messenger/genetics , RNA, Messenger/metabolism , Sequence Analysis, DNA
12.
Genet Mol Res ; 14(4): 18249-58, 2015 Dec 28.
Article in English | MEDLINE | ID: mdl-26782472

ABSTRACT

Cell reprogramming mediated by histone methylation and demethylation is crucial for the activation of the embryonic genome in early embryonic development. In this study, we employed quantitative real-time polymerase chain reaction (qRT-PCR) to detect mRNA levels and expression patterns of all known histone demethylases in early germinal vesicle stage and in vitro-matured metaphase II (MII) oocytes (which are commonly used as donor cells for nuclear transfer). On screening, the Jumonji domain containing 1C (JMJD1C) gene had the highest level of expression and hence was used for subsequent experiments. We also found that JMJD1C was primarily expressed in the nucleus and showed relatively high levels of expression at the 2-cell, 4-cell, 8-cell, 16-cell, morula, and blastocyst stages of embryos developed from MII oocytes fertilized in vitro. Further, we knocked down the JMJD1C gene in MII oocytes using siRNA and monitored the cleavage of zygotes and development of early embryos after in vitro fertilization. The results showed that the zygote cleavage and blastocyst rates of the transfection group were reduced by 57.1 ± 0.07 and 50 ± 0.01% respectively, which were significantly lower than those of the negative control group (P < 0.05). These data suggest that JMJD1C plays a key role in the normal development of early bovine embryos. Our results also provide a theoretical basis for the investigation of the role and molecular mechanism of histone demethylation in the early development of bovine embryos.


Subject(s)
Cell Nucleus/genetics , Embryo, Mammalian , Embryonic Development/genetics , Jumonji Domain-Containing Histone Demethylases/biosynthesis , Animals , Blastocyst/metabolism , Cattle , Cell Nucleus/metabolism , Female , Fertilization in Vitro , Jumonji Domain-Containing Histone Demethylases/genetics , Methylation , Morula/metabolism , Nuclear Transfer Techniques , Oocytes/enzymology , Oocytes/growth & development , Pregnancy , RNA, Messenger/genetics , Zygote/growth & development
13.
Genet Mol Res ; 12(1): 235-41, 2013 Jan 30.
Article in English | MEDLINE | ID: mdl-23408410

ABSTRACT

This study was designed to investigate a single nucleotide polymorphism in intron 1 of the liver fatty acid-binding protein (L-FABP) gene in 156 Junmu No. 1 white swine using PCR-single-strand conformational polymorphism. The association between the polymorphism and meat quality traits was also studied. The cloning and sequencing results indicated that the polymorphism in intron 1 was due to a T→C mutation at position 1740 of L-FABP, yielding three genotypes (TT, TC, and CC). Association analysis revealed that the polymorphism had a significant effect on marbling (P < 0.05): genotype CC had more marbling than TC, and TC had more marbling than TT. The polymorphism also had a highly significant effect on intramuscular fat content (P < 0.01). Genotypes CC and TC had higher intramuscular fat content than TT; there was no significant difference between CC and TC (P > 0.05). However, no significant conclusions concerning other traits could be drawn. We tentatively conclude that L-FABP is a candidate gene or a quantitative trait locus-linked gene associated with meat quality traits.


Subject(s)
Fatty Acid-Binding Proteins/genetics , Swine/genetics , Animals , Food Quality , Genotype , Introns , Meat , Polymerase Chain Reaction/methods , Polymorphism, Single Nucleotide , Quantitative Trait Loci , Sequence Analysis, DNA/methods
14.
Genet Mol Res ; 11(4): 3744-54, 2012 Oct 15.
Article in English | MEDLINE | ID: mdl-23096694

ABSTRACT

The mannan-binding lectin gene (MBL) participates as an opsonin in the innate immune system of mammals, and single nucleotide polymorphisms (SNPs) in MBL cause various immune dysfunctions. In this study, we detected SNPs in MBL2 at exon 1 using polymerase chain reaction single-strand conformation polymorphism analysis and DNA sequencing techniques in 825 Chinese Holstein cows. Four new SNPs with various allele frequencies were also found. The g.1164 G>A SNP was predicted to substitute arginine with glutamine at the N-terminus of the cysteine-rich domain. In the collagen-like domain, SNPs g.1197 C>A and g.1198 G>A changed proline to glutamine, whereas SNP g.1207 T>C was identified as a synonymous mutation. Correlation analysis showed that the g.1197 C>A marker was significantly correlated to somatic cell score (SCS), and the g.1164 G>A locus had significant effects on SCS, fat content, and protein content (P < 0.05), suggesting possible roles of these SNPs in the host response against mastitis. Nine haplotypes and nine haplotype pairs corresponding to the loci of the 4 novel SNPs were found in Chinese Holsteins. Haplotype pairs MM, MN, and BQ were correlated with the lowest SCS; MN with the highest protein yield; MM with the highest protein rate, and MN with the highest 305- day milk yield. Thus, MM, MN, and BQ are possible candidates for marker-assisted selection in dairy cattle breeding programs.


Subject(s)
Cattle/genetics , Genetic Association Studies , Mannose-Binding Lectin/genetics , Milk/metabolism , Polymorphism, Single Nucleotide/genetics , Quantitative Trait, Heritable , Alleles , Amino Acid Sequence , Animals , Base Sequence , China , Exons/genetics , Gene Frequency/genetics , Haplotypes/genetics , Heterozygote , Least-Squares Analysis , Mannose-Binding Lectin/chemistry , Molecular Sequence Data , Polymorphism, Single-Stranded Conformational/genetics , Sequence Analysis, DNA , Silver Staining
15.
Genet Mol Res ; 11(2): 1721-30, 2012 Jun 29.
Article in English | MEDLINE | ID: mdl-22843048

ABSTRACT

The luteinizing hormone receptor (LHR) plays a key role in testosterone production through its interaction with the gonadotropins, LH and chorionic gonadotropin. We examined the LHR splicing pattern in bovine Leydig cells; LH-induced expression of eight cloned splicing variants was detected by real-time PCR. Luteinizing hormone applied to cultured Leydig cells resulted in expression of full-length LHR and the A and B isoforms, as well as secretion of testosterone, which first increased, then declined, and then increased further, with increased LH levels. The secretion of testosterone progressively increased with increasing LH, but the expression levels of LHR (FL, A, and B) did not increase correspondingly. We conclude that the LHR splicing pattern is complex in bovine Leydig cells, and that expression of full-length LHR and isoforms A and B changes when induced with LH.


Subject(s)
Leydig Cells/metabolism , Receptors, LH/metabolism , Alternative Splicing , Amino Acid Sequence , Analysis of Variance , Animals , Cattle , Cells, Cultured , Exons , Gene Expression , Luteinizing Hormone/physiology , Male , Molecular Sequence Data , Protein Isoforms/genetics , Protein Isoforms/metabolism , Receptors, LH/genetics , Testosterone/metabolism
16.
Genet Mol Res ; 10(3): 1504-13, 2011.
Article in English | MEDLINE | ID: mdl-21823101

ABSTRACT

Follicle-stimulating hormone (FSH) plays an essential role in mammalian spermatogenesis and follicular development. In a previous study, we demonstrated that some bulls carry numerous linked mutations in the FSH beta-subunit (FSHB) gene, and that these bulls have poor-quality semen, low fertility, and slightly lower serum FSH concentration compared to those without such mutations. Here, we identified the different FSHB mRNA transcripts in such individuals and analyzed the evolutionary pattern of the FSHB open reading frame (ORF) in different species. Two different lengths of FSHB mRNA transcripts corresponding to two different polyadenylation sites in the 3'-UTR were detected in wild-type bull pituitary glands, and four different mRNA transcripts resulting from the different polyadenylation sites and linked mutations were identified in mutation-bearing bull pituitaries. All transcripts had almost the same putative FSHB precursor molecule. When the ORF sequences of wild-type and mutation-bearing genes were compared with those of other tetrapod species, the leopard frog had the lowest level of homology (57.8 and 58.1%) and the buffalo had the highest level (95.9 and 96.7%), respectively. These results indicated that the bovine FSHB gene transcribes at least two classes of mRNA in the wild-type and four classes of mRNA in the mutation-bearing individuals, which provides a new insight into the bovine FSHB evolutionary pattern. In addition, these findings lay a foundation for further study of gene expression regulation and the effects of mutations on male fertility traits in cattle.


Subject(s)
Cloning, Molecular/methods , Follicle Stimulating Hormone, beta Subunit/chemistry , Follicle Stimulating Hormone, beta Subunit/metabolism , Pituitary Gland/metabolism , Sequence Analysis, DNA/methods , Animals , Cattle , DNA, Complementary/genetics , Follicle Stimulating Hormone, beta Subunit/classification , Follicle Stimulating Hormone, beta Subunit/genetics , Phylogeny , RNA, Messenger/genetics
17.
Genet Mol Res ; 10(2): 779-91, 2011 May 03.
Article in English | MEDLINE | ID: mdl-21563072

ABSTRACT

Recent attention in pig breeding programs has focused on the improvement of pork quality in response to increasing consumer demands. Compared to the fatty-type Northeastern Indigenous (Chinese) breed of pigs, the lean-type Large White has lower intramuscular fat and inferior eating quality from the perspective of the Chinese consumer. In order to investigate the molecular basis of differences in pork quality in Chinese indigenous and Western breeds, longissimus dorsi samples were collected from three adult Northeastern Indigenous and three adult Large White pigs. The RNAs were extracted and hybridized to the porcine Affymetrix GeneChip. Microarray analysis demonstrated differential expression of 1134 genes of which 401 have a known function. One hundred and thirty-six genes were up-regulated and 998 down-regulated in Northeastern Indigenous breed compared to Large White pigs. We screened 10 genes as candidate genes associated with pork quality. We investigated a single nucleotide polymorphism in the 5' regulatory region of the gene FABP4 in 65 Songliao black swine, using PCR-single-strand conformational polymorphism. We found this polymorphism to be highly significantly associated with marbling and intra-muscular fat content (P ≤ 0.01). Genotype BB had higher marbling than AB and AA, but there was no significant difference between AB and AA. Genotype BB and AB had higher intra-muscular fat content than AA, but there was no significant difference between BB and AB. These results help to elucidate the genetic mechanisms behind differences in pork quality and provide a theoretical basis for selection and genetic improvement of meat quality traits in pigs.


Subject(s)
Meat Products/analysis , Swine/genetics , Animals , Body Fat Distribution , Fatty Acid-Binding Proteins/genetics , Food Preferences , Gene Frequency , Microarray Analysis , Muscle, Skeletal , Polymerase Chain Reaction , Polymorphism, Single-Stranded Conformational , Reverse Transcriptase Polymerase Chain Reaction
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