RESUMEN

The molecular mechanisms regulating the physiological adaptation of tissues important for nutrient partitioning and metabolism in lactating cows are still not completely understood. The aim of our study was to identify tissue-specific regulatory mechanisms necessary to accommodate metabolic changes associated with different genetic potential for milk performance. For this purpose, we analyzed mRNA expression of genes involved in energy metabolism of segregating F(2) beef type cows with a combined genetic dairy and beef background (Charolais × German Holstein cross, CH×GH) in contrast to purebred German Holstein (GH) dairy cows. Three groups of cows differing in milk performance were examined using quantitative real-time PCR in liver, mammary gland, and skeletal muscle. Our results describe substantial tissue-specific differences in mRNA transcription profiles between cow groups in relation to their genetic potential for milk performance and highlight genes exhibiting specific, partially yet-unknown functions in dairy and beef type cows, e.g., upregulation of PCK2 transcripts in the mammary gland and FBP2 transcripts in skeletal muscle of dairy cows. Noticeably, PCCA and PPARGC1A mRNA abundance varied significantly across experimental groups in all three tissues, pointing to potential key gene functions in the metabolic adaptation relative to divergent milk production performance. Correlations of mRNA expression levels to milk performance traits indicate that gene transcriptional processes may play a regulatory role in liver, mammary gland, and skeletal muscle to enable cows with different genetic potential for milk performance to cope with metabolic lactation-associated challenges.

Asunto(s)

Bovinos/fisiología , Metabolismo Energético/fisiología , Regulación de la Expresión Génica/fisiología , Lactancia/fisiología , Leche/fisiología , ARN Mensajero/metabolismo , Selección Genética/fisiología , Animales , Cruzamientos Genéticos , Cartilla de ADN/genética , Femenino , Lactancia/metabolismo , Modelos Lineales , Hígado/metabolismo , Glándulas Mamarias Animales/metabolismo , Músculo Esquelético/metabolismo , Especificidad de Órganos/fisiología , Reacción en Cadena en Tiempo Real de la Polimerasa

RESUMEN

Six genes that were known to exhibit expression levels that are correlated to drip loss BVES, SLC3A2, ZDHHC5, CS, COQ9, and EGFR have been for candidate gene analysis. Based on in silico analysis SNPs were detected, confirmed by sequencing, and used for genotyping. The SNPs were genotyped in about 1,800 animals from six pig populations including commercial herds of Pietrain (PI) and German Landrace (DL), different commercial herds of Pietrain×(German Large White×German Landrace) (PIF1(a/b/c)), and one experimental F2-population Duroc×Pietrain (DUPI). Comparative and genetic mapping established the location of BVES on SSC1, of SLC3A2 and ZDHHC5 on SSC2, of CS on SSC5, of COQ9 on SSC6 and of EGFR on SSC9, respectively, coinciding with QTL regions for carcass and meat quality traits. BVES, SLC3A2, and CS revealed association at least with drip loss and with several other measures of water holding capacity (WHC). Moreover, COQ9 and EGFR were associated with several meat quality traits such as meat color and/or thawing loss. This study reveals statistic evidence in addition to the functional relationship of these genes to WHC previously evidenced by expression analysis. This study reveals positional and genetic statistical evidence for a link of genetic variation at these loci or close to them and promotes those six candidate genes as functional and/or positional candidate genes for meat quality traits.

Asunto(s)

Agua Corporal/química , Genes/genética , Carne/análisis , Carne/normas , Fenotipo , Sitios de Carácter Cuantitativo/genética , Animales , Mapeo Cromosómico , Cartilla de ADN/genética , Estudios de Asociación Genética , Genotipo , Polimorfismo de Nucleótido Simple/genética , Análisis de Secuencia de ADN , Especificidad de la Especie , Porcinos

RESUMEN

The proximal half of Bos taurus chromosome 27 (BTA27prox) delimited by microsatellite markers BM3507 and CSSM043 reveals complex rearrangements compared to its corresponding Homo sapiens chromosome (HSA) fragments. A comparative mapping approach combining somatic and radiation hybrid cell mapping techniques and related cytogenetic data resulted in an improved physical map for BTA27prox, which provides candidate genes for several important economic traits. The generated comprehensive map includes anchor loci for 103 genes and microsatellite markers. Mapping of genes proximal to BM3507 matching a region from 0.60 to 2.78 megabase pairs (Mb) of HSA8 confirmed recent sequence annotations on BTA27. Assignments of loci predicted to be on BTA27 to BTA1, BTA8, and BTA17 narrowed down evolutionary chromosome break points compared with corresponding chromosome segments in human. New physical anchors obtained in this study confirm in more detail the described evolutionary conservation between the proximal half of BTA27 and homologous segments of HSA4 and HSA8 and will contribute to the completion of the cattle DNA genome sequence.

Asunto(s)

Bovinos/genética , Genoma Humano , Sintenía , Animales , Rotura Cromosómica , Mapeo Cromosómico , Evolución Molecular , Humanos , Repeticiones de Microsatélite , Mapeo de Híbrido por Radiación , Especificidad de la Especie

RESUMEN

A radiation hybrid (RH) map of sheep X chromosome (Ovisaries; OARX) containing 146 physically anchored loci was generated in this study, providing information for comparative X chromosome analysis between the maps of sheep, human, and cattle. Primers typed on the USUoRH5000 ovine whole-genome radiation hybrid panel were designed from sequences predicted to be on the ovine X chromosome, based on comparative mapping within the virtual sheep genome browser (v1.2). The resulting RH map for the ovine X chromosome consists of 4 linkage groups composed of 76 BAC end sequences (BES), 28 gene loci that were confirmed within ovine BAC clones in the CHORI-243 ovine BAC library, 28 additional gene loci from the ovine comparative map and 14 polymorphic sequence tagged sites (STS) from the OARX linkage map. This first-generation RH map of OARX contributes to the expansion of a comprehensive ovine genome map for sheep and provides evidence of rearrangements in loci order compared to the human and cattle orders.

Asunto(s)

Bovinos/genética , Cromosomas Humanos X/genética , Mapeo de Híbrido por Radiación/veterinaria , Ovinos/genética , Cromosoma X/genética , Animales , Cromosomas Artificiales Bacterianos/genética , Humanos , Repeticiones de Microsatélite , Mapeo de Híbrido por Radiación/métodos , Especificidad de la Especie

RESUMEN

A preliminary radiation hybrid (RH) map containing 50 loci on chromosome 7 of the domestic river buffalo Bubalus bubalis (BBU; 2n = 50) was constructed based on a comparative mapping approach. The RH map of BBU7 includes thirty-seven gene markers and thirteen microsatellites. All loci have been previously assigned to Bos taurus (BTA) chromosome BTA6, which is known for its association with several economically important milk production traits in cattle. The map consists of two linkage groups spanning a total length of 627.9 cR(5,000). Comparative analysis of the BBU7 RH(5,000) map with BTA6 in cattle gave new evidence for strong similarity between the two chromosomes over their entire length and exposed minor differences in locus order. Comparison of the BBU7 RH(5,000) map with the Homo sapiens (HSA) genome revealed similarity with a large chromosome segment of HSA4. Comparative analysis of loci in both species revealed more variability than previously known in gene order and several chromosome rearrangements including centromere relocation. The data obtained in our study define the evolutionarily conserved segment on BBU7 and HSA4 to be between 3.5 megabases (Mb) and 115.8 Mb in the HSA4 (genome build 36) DNA sequence.

Asunto(s)

Búfalos/genética , Bovinos/genética , Cromosomas de los Mamíferos/genética , Genoma/genética , Mapeo de Híbrido por Radiación , Animales , Secuencia de Bases , Marcadores Genéticos , Humanos

Asunto(s)

Enfermedades de los Bovinos/genética , Genes bcl-2 , Linfoma de Células B/genética , Linfoma de Células B/veterinaria , Proteína bcl-X/genética , Animales , Bovinos , Mapeo Cromosómico , Hibridación in Situ

Asunto(s)

Mapeo Cromosómico , Células Híbridas/fisiología , Proteínas Serina-Treonina Quinasas/genética , Péptidos y Proteínas Asociados a Receptores de Factores de Necrosis Tumoral/genética , Animales , Bovinos , Hibridación de Ácido Nucleico , Proteína Serina-Treonina Quinasa 2 de Interacción con Receptor

Asunto(s)

Bovinos/genética , Proteínas Proto-Oncogénicas c-bcl-2/genética , Animales , Mapeo Cromosómico

Asunto(s)

Proteínas de Neoplasias/genética , Mapeo de Híbrido por Radiación , Animales , Secuencia de Bases , Bovinos , Mapeo Cromosómico , Cromosomas de los Mamíferos , Cartilla de ADN , Etiquetas de Secuencia Expresada , Humanos , Reacción en Cadena de la Polimerasa

RESUMEN

Coordination of the primary defense mechanisms against pathogens relies on the appropriate expression of pathogen recognition receptors (PRRs) triggering the early release of effector molecules of the innate immune system. To analyze the impact of this system on the counteraction of infections of the mammary gland (mastitis), we characterized the bovine gene encoding the key PRR Toll-like receptor 9 (TLR9) and mapped its precise position on chromosome BTA22. The sequence information was used to establish real-time PCR quantification assays to measure the mRNA abundances of TLR9, TLR2, and TLR4 together with those of beta-defensin 5 (BNBD5), an early bactericidal effector molecule of the innate system, in healthy and infected mammary glands. Mastitis strongly increased (4- to 13-fold) the mRNA abundances of all of these genes except TLR9. Slight subclinical infections already caused a substantial increase in the copy numbers, though they did so the least for TLR9. Induction was not systemic, since mRNA abundance was low in uninfected control quarters of the udder but high in the severely infected quarters of the same animal. The number of TLR2 copies correlated well with those of TLR4, indicating coordinated regulation of these two PRRs during infection of the udder. Their coordinated regulation explains our unexpected observation that pure Staphylococcus aureus infections caused a strong increase also in TLR4 mRNA abundance. In situ hybridizations revealed that BNBD5 is expressed predominantly in the mammary epithelial cells (MEC) of the infected gland. Our data therefore suggest a significant contribution of the innate immune system to counteract mastitis and attribute a prominent effector function to the MEC.

Asunto(s)

Proteínas de Unión al ADN/genética , Mastitis Bovina/genética , Mastitis Bovina/inmunología , Glicoproteínas de Membrana/genética , Receptores de Superficie Celular/genética , beta-Defensinas/genética , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Bovinos , Mapeo Cromosómico , ADN Complementario/genética , Femenino , Expresión Génica , Inmunidad Innata , Glándulas Mamarias Animales/inmunología , Glándulas Mamarias Animales/patología , Mastitis Bovina/patología , Datos de Secuencia Molecular , ARN Mensajero/genética , ARN Mensajero/metabolismo , Homología de Secuencia de Aminoácido , Staphylococcus aureus/patogenicidad , Receptor Toll-Like 2 , Receptor Toll-Like 4 , Receptor Toll-Like 9 , Receptores Toll-Like

RESUMEN

The coagulation factor IX gene (F9), the hypoxanthine phosphoribosyl transferase 1 gene (HPRT1), and the X-inactive specific transcript gene (XIST) were physically assigned in cattle to analyze chromosomal breakpoints on BTAX recently identified by radiation hybrid (RH) mapping experiments. Whereas the FISH assignment of XIST indicates a similar location on the q-arm of the human and cattle X chromosomes, the locus of HPRT1 supported the assumption of a chromosome rearrangement between the distal half of the q-arm of HSAX and the p-arm of BTAX identified by RH mapping. F9 previously located on the q-arm of BTAX was assigned to the p-arm of BTAX using RH mapping and FISH. The suggested new position of F9 close to HPRT1 supports the homology between HSAXq and BTAXp. The F9 locus corresponds with the gene order found in the homologous human chromosome segment. XIST was assigned on BTAXq23, HPRT1 and F9 were mapped to BTAXp22, and the verification of the location of F9 in a 5000 rad cattle-hamster whole genome radiation hybrid panel linked the gene to markers URB10 and HPRT1.

Asunto(s)

Bovinos/genética , Cromosomas Humanos X/genética , Factor IX/genética , Hipoxantina Fosforribosiltransferasa/genética , ARN no Traducido/genética , Cromosoma X/genética , Animales , Mapeo Cromosómico , Cricetinae , Humanos , Hibridación Fluorescente in Situ , ARN Largo no Codificante , Mapeo de Híbrido por Radiación , Sintenía

Asunto(s)

Bovinos/genética , Proteínas del Tejido Nervioso/genética , Proteínas Tirosina Fosfatasas/genética , Animales , Cromosomas de los Mamíferos/ultraestructura , Cricetinae , Células Híbridas , Hibridación Fluorescente in Situ , Mapeo Físico de Cromosoma , Proteínas Tirosina Fosfatasas Clase 5 Similares a Receptores

Asunto(s)

Bovinos/genética , Proteínas HMGB/genética , Glicoproteínas de Membrana/genética , Proteoglicanos/genética , Animales , Bandeo Cromosómico , Cromosomas de los Mamíferos , Hibridación Fluorescente in Situ , Sindecano-2 , Timo

RESUMEN

A PCR-based method for sex determination of bovine DNA samples and embryo biopsies is presented. Using only one primer pair both the male-specific sequence FBNY (127 bp) and a sex-independent control PCR-fragment, the microsatellite marker FBN17 (136-140 bp) are generated in the same PCR reaction. Synteny mapping assigned the male-specific sequence to bovine chromosome Y (BTA Y), whereas FBN17 was mapped to bovine chromosome 2. Localisation of FBNY on BTA Y was confirmed by fluorescence in hybridisation of two BAC clones containing the male-specific sequence. There was no amplification of the male-specific target sequence FBNY in sheep, pig, goat, mice, man, and several wild species of the tribe Bovini. The bovine male-specific fragment was detected in dilutions containing as little as 10 pg genomic DNA and in blastomeres from embryo biopsies. The PCR assay presented here does require neither restriction endonuclease digestion of the PCR product nor additional nested PCR steps. Owing to the advantage of parallel amplification of the autosomal locus FBN17 no additional control fragment is necessary to detect PCR failure. The results of sex determination in embryo biopsies using FBNY were in agreement with the outcome from a reference assay used in commercial breeding programs.

Asunto(s)

Bovinos/genética , Reacción en Cadena de la Polimerasa/métodos , Caracteres Sexuales , Análisis para Determinación del Sexo/métodos , Cromosoma Y/genética , Animales , Cruzamiento , Cartilla de ADN/genética , Embrión de Mamíferos/citología , Embrión de Mamíferos/metabolismo , Femenino , Cabras/genética , Humanos , Hibridación Fluorescente in Situ , Masculino , Ratones , Mapeo Físico de Cromosoma , Sensibilidad y Especificidad , Ovinos/genética , Especificidad de la Especie , Porcinos/genética

RESUMEN

OBJECTIVE: To determine if orally delivered tizanidine will control spastic hypertonia due to acquired brain injury. DESIGN: Randomized, double-blind, placebo-controlled, crossover design, with 2 8-week treatment arms separated by a 1-week washout period at baseline. Patients were randomly assigned to receive tizanidine or a matching placebo. SETTING: Tertiary care outpatient and inpatient rehabilitation center attached to a university hospital. PARTICIPANTS: Seventeen persons recruited in a consecutive manner, 9 of whom had suffered a stroke and 8 a traumatic brain injury, and had more than 6 months of intractable spastic hypertonia. INTERVENTION: Over a 6-week period, subjects were slowly titrated up to their maximum tolerated dose (up to 36 mg/d). Following a 1-week drug taper and 1-week period in which no study drug was administered, patients were then crossed over to the other study medication following an identical titration regime. MAIN OUTCOME MEASURES: Subjects were evaluated for dose and effect throughout the trial as well as for side effects. Data for Ashworth rigidity scores, spasm scores, deep tendon reflex scores, and motor strength were collected on the affected upper extremity (UE) and lower extremity (LE). Differences over time were assessed via descriptive statistics, Friedman's analysis, and Wilcoxon's signed-rank. Data are reported as the mean +/- 1 standard deviation. RESULTS: Following 4 weeks of treatment when subjects reached their maximal tolerated dosage, the average LE Ashworth score on the affected side decreased from 2.3 +/- 1.4 to 1.7 +/- 1.1 (p <.0001). The spasm score decreased from 1.0 +/- 0.9 to 0.5 +/- 0.8 (p =.0464), while the reflex score was not statistically significant decreasing from 2.2 +/- 1.0 to 2.0 +/- 1.1 (p =.0883). The average UE Ashworth score on the affected side decreased from 1.9 +/- 1.1 to 1.5 +/- 0.9 (p <.0001). There was no significant change in the UE spasm and reflex scores. While there were positive placebo effects on motor tone, the active drug was still significantly better than placebo for decreasing LE tone (p =.0006) and UE tone (p =.0007). With a reduction in motor tone, there was an increase in motor strength (p =.0089). The average dosage at 4 weeks was 25.2mg/d. CONCLUSION: Tizanidine is effective in decreasing the spastic hypertonia associated with acquired brain injury, which is dose-dependent. There are limitations on its use due to side effects related to drowsiness.

Asunto(s)

Lesiones Encefálicas/complicaciones , Clonidina/uso terapéutico , Hipertonía Muscular/tratamiento farmacológico , Hipertonía Muscular/etiología , Relajantes Musculares Centrales/uso terapéutico , Espasticidad Muscular/tratamiento farmacológico , Espasticidad Muscular/etiología , Administración Oral , Adulto , Anciano , Lesiones Encefálicas/fisiopatología , Clonidina/análogos & derivados , Clonidina/farmacología , Estudios Cruzados , Relación Dosis-Respuesta a Droga , Método Doble Ciego , Femenino , Humanos , Masculino , Persona de Mediana Edad , Hipertonía Muscular/clasificación , Hipertonía Muscular/diagnóstico , Hipertonía Muscular/fisiopatología , Relajantes Musculares Centrales/farmacología , Espasticidad Muscular/clasificación , Espasticidad Muscular/diagnóstico , Espasticidad Muscular/fisiopatología , Estudios Prospectivos , Reflejo Anormal/efectos de los fármacos , Índice de Severidad de la Enfermedad , Fases del Sueño/efectos de los fármacos , Resultado del Tratamiento

RESUMEN

mRNA differential display was applied to identify hepatic and intestinal expressed sequence tags (ESTs) in lactating cows of different metabolic types (milk type, meat/milk type, meat type) that are potentially associated with energy turnover and involved in the regulation of these processes. Altogether, 277 ESTs (liver: 161, intestine: 116) were identified. For 150 transcripts (liver: 99, intestine: 51), the sequences showed similarity to previously described genes and ESTs. Many of these homologous sequences are reported to be involved in hepatic metabolism. Ninety-four ESTs (liver: 43, intestine: 51) did not match with any database entries. Semi-quantitative RT-PCR revealed quantitative differences in transcript represented by randomly chosen ESTs in liver samples of animals of the Holstein and Charolais breeds. One hundred twenty-two ESTs were mapped physically by using a bovine-hamster somatic cell hybrid panel (SCP) and a 5000-rad bovine whole genome radiation hybrid panel (WGRH). These ESTs were assigned to the bovine syntenic groups and positioned in the recently established RH-based ordered comparative map of the cattle and human genomes. The mapped, differentially expressed sequence tags are a useful prerequisite for cloning of genetic variation underlying economic traits.

Asunto(s)

Bovinos/genética , Mapeo Cromosómico/veterinaria , Etiquetas de Secuencia Expresada , Mucosa Intestinal/metabolismo , Hígado/metabolismo , Carácter Cuantitativo Heredable , Animales , ADN/química , ADN/genética , Cartilla de ADN , Femenino , Marcadores Genéticos , Células Híbridas/efectos de la radiación , Datos de Secuencia Molecular , ARN Mensajero/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Análisis de Secuencia de ADN

RESUMEN

Comparative mapping of four genes and one unknown coding DNA sequence in breakpoint positions of bovine chromosomes (BTA) 7 and 25 are presented. Performing a genome data base search five bovine expressed sequence tags from the MARC library matched with human genes coding for the general transcription factor IIIC polypeptide 1 (GTF3C1), the hypothetical protein KIAA0556, the interleukin 4 receptor (IL4R), the regulatory factor X-associated ankyrin-containing protein (RFXANK), and with an unknown human coding sequence partially homologous to the genomic cosmid clone R30923. Loci for these sequences were COMPASS predicted on BTA7 or BTA18 and to BTA18 or BTA25. Mapping was performed in a cattle-hamster somatic hybrid cell panel and a cattle-hamster 5000 rad whole genome radiation hybrid panel. GTF3C1, KIAA0556 and IL4R were assigned to the centromere region of BTA25 and RFXANK and R30923 close to the centromere of BTA7. The assignments contribute to the identification of evolutionary chromosome break points between human chromosomes 16 and 19 and BTA7, BTA18, and BTA25.

Asunto(s)

Mapeo Cromosómico , Receptores de Interleucina-4/genética , Factores de Transcripción TFIII/genética , Factores de Transcripción/genética , Animales , Bovinos , Cromosomas , Cósmidos , Cricetinae , Proteínas de Unión al ADN , Marcadores Genéticos , Humanos , Células Híbridas

RESUMEN

The STAT transcription factors form a family of signal transducers and activators of transcription. We sequenced the bovine STAT5B cDNA and both STAT5-encoding genes, STAT5A and STAT5B, representing the first complete description of any STAT5-encoding gene. DNA fiber FISH hybridization revealed that the genes reside only 40 kbp apart on BTA19. Both genes are segmented into 19 exons and all but two of the homologous exons are of equal size. The genes harbor a central block of nearly identical DNA sequence (97.5% sequence identity over 3373 bp), spanning from intron 5 to intron 9. Isolation and sequencing of the homologous segments from mouse revealed the same unusually high degree of intronic sequence conservation in these segments of the murine STAT5-encoding genes. However, the respective sequences are completely divergent between the two species. A comparison of the inter- and intragenic cDNA sequence preservation at nonsynonymous sites reveals that the DNA-binding domain is under the strongest selection pressure for both intergenic and factor-specific intragenic sequence preservation. The so-called "SH3" segment of the linker domain, in contrast, shows species-specific sequence identity in all but one amino acid residues in both factors, in cattle, human, and mouse. This indicates that the same species-specific selection pressure occurs on the linker domain from both factors, STAT5A and STAT5B. Thus, the comparison of evolutionary selection pressures resting on various domains suggests that the DNA-binding domain might contribute to differential DNA binding of STAT5A and STAT5B factors, while both might interact equally well with other cellular factors through a segment of the linker domain.

Asunto(s)

Proteínas de Unión al ADN/genética , Evolución Molecular , Proteínas de la Leche , Transactivadores/genética , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Bovinos , Clonación Molecular , Secuencia Conservada , ADN/genética , ADN Complementario/genética , Proteínas de Unión al ADN/química , Proteínas de Unión al ADN/metabolismo , Exones/genética , Humanos , Hibridación Fluorescente in Situ , Intrones/genética , Ratones , Datos de Secuencia Molecular , Filogenia , Ratas , Mapeo Restrictivo , Factor de Transcripción STAT5 , Análisis de Secuencia de ADN , Homología de Secuencia de Ácido Nucleico , Transactivadores/química , Transactivadores/metabolismo , Proteínas Supresoras de Tumor

Asunto(s)

Aromatasa/genética , Cabras/genética , Ovinos/genética , Animales , Bovinos , Mapeo Cromosómico , Cromosomas/genética , Hibridación Fluorescente in Situ

RESUMEN

An alternative approach for the direct analysis of chromosome regions corresponding to economical traits on the basis of chromosome microdissection is described. Large fragment clones isolated with primer pairs designed from chromosome fragment-specific DNA sequences were localized by FISH to the scraped chromosome region of interest. The chromosome fragment-specific clones are a useful tool for the generation of region specific high density marker and gene maps and represent the source material for the development of a DNA contig including the economical trait.

Asunto(s)

Bovinos/genética , Mapeo Cromosómico/métodos , Cromosomas/ultraestructura , Animales , Clonación Molecular , Cartilla de ADN , Hibridación Fluorescente in Situ