Abstract The objective of this study was to identify in Angus and Brangus breed animals with extreme temperament, measured as exit velocity, genomic regions and candidate genes associated with bovine temperament. The population was genotyped with the Genomic Profiler HD 150K chip and after the genome-wide association analysis, the SNPs rs133956611 (P=2.65 E-06) and rs81144933 (P=9.58 E-06) were associated with temperament. The mapping analysis of the regions close to the SNP rs81144933 identified the SNCA (alpha-synuclein) and MMRN1 (multimerin-1) genes at 222.8 and 435.9 Kb downstream respectively, while for the rs133956611 loci the gene GPRIN3 (GPRIN family-member-3) was identified at 245.7 Kb upstream, all three genes are located on the BTA6 chromosome. The analysis of SNCA protein-protein interactions allowed the identification of the genes APP (β-amyloid precursor protein), PARK7 (parkinsonism-associated-deglycase), UCHL1 (ubiquitin-C-terminal-hydrolase-L1), PARK2 (parkin-RBR- E3-ubiquitin-protein-ligase), and genes of the SLC family as candidates to be associated with bovine temperament. All these candidate genes and their interacting were resequenced, which allowed the discovery of new SNPs in the SNCA and APP genes. Of these, the SNPs located in introns 5, 8 and 11 of the APP gene affect splicing site motifs. These results indicate that SNCA and its interacting genes are candidates to be related to bovine temperament.

Resumen El objetivo de este estudio fue identificar en animales de raza Angus y Brangus con temperamento extremo, medido como velocidad de salida, regiones genómicas y genes candidatos asociados con el temperamento bovino. La población fue genotipada con el chip Genomic Profiler HD 150K y después del análisis de asociación del genoma completo, los SNP rs133956611 (P= 2.65 E-06) y rs81144933 (P= 9.58 E-06) se asociaron con el temperamento. El análisis de mapeo de las regiones cercanas al SNP rs81144933 identificó los genes SNCA (alfa-sinucleína) y MMRN1 (multimerin-1) a 222.8 y 435.9 Kb corriente abajo, respectivamente, mientras que para los loci rs133956611 se identificó el gen GPRIN3 (familia GPRIN- miembro 3) a 245.7 Kb corriente arriba, los tres genes se encuentran en el cromosoma BTA6. El análisis de las interacciones proteína-proteína de SNCA permitió la identificación de los genes APP (proteína precursora de β-amiloide), PARK7 (deglicasa asociada al parkinsonismo), UCHL1 (ubiquitina C-terminal hidrolasa L1), PARK2 (parkina-RBR-E3-ubiquitina-proteína-ligasa), y genes de la familia SLC como candidatos a estar asociados con el temperamento bovino. Todos estos genes candidatos y su interacción fueron resecuenciados, lo que permitió el descubrimiento de nuevos SNP en los genes SNCA y APP. De estos, los SNP localizados en los intrones 5, 8 y 11 del gen APP afectan a los motivos del sitio de empalme. Estos resultados indican que el SNCA y sus genes interactuantes son candidatos para estar relacionados con el temperamento bovino.

ABSTRACT The objective of this study was to evaluate the effect of intronic single nucleotide polymorphisms (SNP) on temperament traits in a Brahman cattle population. The SNP located in CACNG4, EXOC4, NRXN3, and SLC9A4 candidate genes were genotyped in 250 animals with temperament records of exit velocity, pen score, and temperament score. Rs3423464051:G>A in the CACNG4 gene was associated with exit velocity and temperament score. An in silico analysis of the five intronic SNP showed that alternative alleles of CACNG4-rs3423464051, EXOC4-rs109393235, and SLC9A4-rs109722627 SNP could alter branch point sites during splicing, while a protein–protein interaction network analysis demonstrated a GRIA2 gene-mediated interaction between CACNG4 and NRXN3. The present results support previously reported evidence regarding bovine temperament-related candidate genes, particularly CACNG4, which is a confirmed candidate gene in need of more detailed analyses to reveal its role in temperament-related traits. (SNP population CACNG EXOC4 EXOC NRXN3 NRXN SLCA SLC A SLC9A 25 score Rs3423464051G>A Rs3423464051GA RsGA Rs3423464051 G>A G Rs CACNG4rs3423464051, CACNG4rs3423464051 CACNGrs rs3423464051, rs3423464051 rs CACNG4-rs3423464051 EXOC4rs109393235, EXOC4rs109393235 EXOCrs rs109393235, rs109393235 EXOC4-rs109393235 SLC9A4rs109722627 SLCArs rs109722627 SLC9A4-rs10972262 splicing proteinprotein protein GRIA genemediated mediated temperamentrelated related 2 Rs3423464051G GA Rs342346405 CACNG4rs342346405 rs342346405 CACNG4-rs342346405 EXOC4rs10939323 rs10939323 EXOC4-rs10939323 SLC9A4rs10972262 rs10972262 SLC9A4-rs1097226 RsG Rs34234640 CACNG4rs34234640 rs34234640 CACNG4-rs34234640 EXOC4rs1093932 rs1093932 EXOC4-rs1093932 SLC9A4rs1097226 rs1097226 SLC9A4-rs109722 Rs3423464 CACNG4rs3423464 rs3423464 CACNG4-rs3423464 EXOC4rs109393 rs109393 EXOC4-rs109393 SLC9A4rs109722 rs109722 SLC9A4-rs10972 Rs342346 CACNG4rs342346 rs342346 CACNG4-rs342346 EXOC4rs10939 rs10939 EXOC4-rs10939 SLC9A4rs10972 rs10972 SLC9A4-rs1097 Rs34234 CACNG4rs34234 rs34234 CACNG4-rs34234 EXOC4rs1093 rs1093 EXOC4-rs1093 SLC9A4rs1097 rs1097 SLC9A4-rs109 Rs3423 CACNG4rs3423 rs3423 CACNG4-rs3423 EXOC4rs109 rs109 EXOC4-rs109 SLC9A4rs109 SLC9A4-rs10 Rs342 CACNG4rs342 rs342 CACNG4-rs342 EXOC4rs10 rs10 EXOC4-rs10 SLC9A4rs10 SLC9A4-rs1 Rs34 CACNG4rs34 rs34 CACNG4-rs34 EXOC4rs1 rs1 EXOC4-rs1 SLC9A4rs1 SLC9A4-rs Rs3 CACNG4rs3 rs3 CACNG4-rs3 EXOC4rs EXOC4-rs SLC9A4rs CACNG4rs CACNG4-rs

Abstract Acute infections of bovine viral diarrhea virus (BVDV) lead to a range of clinical presentations. Laboratory tests for detection depend on collection of samples during a short viremia. Acutely infected animals remain largely undiagnosed. Transfer RNA halves (tsRNAs) are hypothesized to function like microRNAs to regulate gene expression during an immune response. The objective of this study was to identify tsRNAs in cattle that had been challenged with a non-cytopathic field strain of BVDV. Colostrum-deprived neonatal Holstein calves were either challenged with BVDV (n=5) or mock challenged (n=4). Sera was collected prior to challenge and days 4, 9, and 16 post challenge. RNA was extracted and read counts of small non-coding RNAs were assessed using next-generation sequencing. A total of 87,838,207 reads identified 41 different tsRNAs. Two 5’ tsRNAs, tsRNAProAGG and tsRNAValAAC, differed across time. Two 5’ tsRNAs, tsRNAGlyCCC and tsRNAGlyGCC, differed between treatment groups across time. Four days post challenge, 5’ tsRNAGlyCCC and tsRNAGlyGCC were significantly lower in the challenged group than the control group. Further studies are needed to identify the importance and function of 5’ tsRNAGlyCCC and tsRNAGlyGCC in serum samples of cattle challenged with BVDV.

Abstract Oocyte maturation involves several complex events which are partially regulated by the gonadotropins. However, in order to fully understand these events it is necessary to identify all those genes whith roles in maturation. One of these genes is the protein ATPase6, which is a component of ATP synthase and is upregulated during oocyte maturation, nevertheless, its participation has not been experimentally evaluated. Therefore, the aim of the present study was to determine if ATPase6 has a relevant role in pig oocyte maturation, by conforming two groups of oocytes: The experimental, oocytes microinjected with ATPase6-interfering RNA (RNAi) in order to silence its expression, and the control, oocytes microinjected with an invertebrate gene-RNAi. After microinjection, both groups of oocytes were cultured in maturation medium for 48 h. ATPase6 expression was reduced 71% in ATPase6-RNAi microinjected oocytes, while the percentage of matured oocytes was reduced 51% in this group (67.5% in control oocytes vs 33% in ATPase6-RNAi microinjected oocytes; p<0.05 with Student’s t test). These results allow us to conclude that ATPase6 has an important role in the maturation of pig oocytes.

Resumen La maduración del ovocito en los folículos preovulatorios comprende complejos eventos regulados en parte por gonadotropinas. Sin embargo para entender estos eventos, es necesario identificar los diferentes genes que participan en la maduración. Uno de estos genes corresponde a la proteína ATPasa6, componente de la ATP sintasa que se sobreexpresa durante la maduración de los ovocitos pero hasta ahora no ha sido comprobada su participación experimentalmente. Por lo tanto, el objetivo del presente estudio se encaminó a determinar si la ATPasa6 tiene un papel relevante en la maduración de los ovocitos del cerdo, a partir de la conformación de dos grupos de ovocitos: El experimental microinyectados con un RNA interferente para ATPasa6, silenciando su expresión, y el grupo control microinyectados con un RNAi para un gen de invertebrado. Después de la microinyección, ambos grupos se cultivaron in vitro durante 48 h en medio de maduración. El grupo microinyectado con siRNA para ATPasa6 mostró una disminución del 71% en la expresión de este gen a las 24 h de incubación, en tanto que el porcentaje de ovocitos maduros en este grupo disminuyó 51.1% (67.5% en el grupo control vs 33% en el grupo tratado con ATPasa6-RNAi; p<0.05 con t de Student). Estos resultados permiten concluir que la ATPasa6 juega un papel importante durante la maduración de los ovocitos del cerdo.

The objective of this work was to identify QTLs for liveweight in a candidate region of bovine chromosome 5. Half-sib families from two lines, one traditional and the other new, of Canchim beef cattle (5/8 Charolais + 3/8 Zebu) were genotyped for four microsatellite markers, including the microsatellite in the IGF-1 (insulin-like growth factor-1) promoter region. Significant differences in allele distribution between the two lines were found for three markers. Interval mapping analyses in this region indicated the presence of a QTL controlling birth weight (p < 0.05) and of a QTL influencing breeding value for yearling weight (p < 0.01) in the newer line of the breed. The previously identified interaction between the IGF-1 genotype and genetic group strengthens the hypothesis of a linked QTL rather than an IGF-1 effect on growth traits in the Canchim cattle.