RESUMEN
The mammary gland is a complex tissue consisting of multiple cell types which, over the lifetime of an animal, go through repeated cycles of development associated with pregnancy, lactation and involution. The mammary gland is also known to be sensitive to maternal programming by environmental stimuli such as nutrition. The molecular basis of these adaptations is of significant interest, but requires robust methods to measure gene expression. Reverse-transcription quantitative PCR (RT-qPCR) is commonly used to measure gene expression, and is currently the method of choice for validating genome-wide expression studies. RT-qPCR requires the selection of reference genes that are stably expressed over physiological states and treatments. In this study we identify suitable reference genes to normalize RT-qPCR data for the ovine mammary gland in two physiological states; late pregnancy and lactation. Biopsies were collected from offspring of ewes that had been subjected to different nutritional paradigms during pregnancy to examine effects of maternal programming on the mammary gland of the offspring. We evaluated eight candidate reference genes and found that two reference genes (PRPF3 and CUL1) are required for normalising RT-qPCR data from pooled RNA samples, but five reference genes are required for analyzing gene expression in individual animals (SENP2, EIF6, MRPL39, ATP1A1, CUL1). Using these stable reference genes, we showed that TET1, a key regulator of DNA methylation, is responsive to maternal programming and physiological state. The identification of these novel reference genes will be of utility to future studies of gene expression in the ovine mammary gland.
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Perfilación de la Expresión Génica , Lactancia/genética , Glándulas Mamarias Animales/metabolismo , Fenómenos Fisiologicos Nutricionales Maternos , Preñez , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Animales , Metilación de ADN , Femenino , Expresión Génica , Regulación de la Expresión Génica , Modelos Genéticos , Embarazo , Ovinos , Oveja Doméstica/genéticaRESUMEN
A type of lower motor neuron (LMN) disease inherited as autosomal recessive in Romney sheep was characterized with normal appearance at birth, but with progressive weakness and tetraparesis after the first week of life. Here, we carried out genome-wide homozygosity mapping using Illumina Ovine SNP50 BeadChips on lambs descended from one carrier ram, including 19 sheep diagnosed as affected and 11 of their parents that were therefore known carriers. A homozygous region of 136 consecutive single-nucleotide polymorphism (SNP) loci on chromosome 2 was common to all affected sheep and it was the basis for searching for the positional candidate genes. Other homozygous regions shared by all affected sheep spanned eight or fewer SNP loci. The 136-SNP region contained the sheep ATP/GTP-binding protein 1 (AGTPBP1) gene. Mutations in this gene have been shown to be related to Purkinje cell degeneration (pcd) phenotypes including ataxia in mice. One missense mutation c.2909G>C on exon 21 of AGTPBP1 was discovered, which induces an Arg to Pro substitution (p.Arg970Pro) at amino-acid 970, a conserved residue for the catalytic activity of AGTPBP1. Genotyping of this mutation showed 100% concordant rate with the recessive pattern of inheritance in affected, carrier, phenotypically normal and unrelated normal individuals. This is the first report showing a mutant AGTPBP1 is associated with a LMN disease in a large mammal animal model. Our finding raises the possibility of human patients with the same etiology caused by this gene or other genes in the same pathway of neuronal development.
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Proteínas de Unión al GTP/genética , Enfermedad de la Neurona Motora/genética , Enfermedad de la Neurona Motora/veterinaria , Mutación Missense , Enfermedades de las Ovejas/genética , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Mapeo Cromosómico , Modelos Animales de Enfermedad , Proteínas de Unión al GTP/química , Humanos , Datos de Secuencia Molecular , Polimorfismo de Nucleótido Simple , Alineación de Secuencia , OvinosRESUMEN
OBJECTIVES: The objectives of this study were to examine the effects of voluntary exercise during pregnancy on maternal post-lactation bone parameters and offspring growth. METHODS: Pregnant Wistar rats were housed in conventional cages (control), or were housed in raised cages requiring them to rise to an erect, bipedal stance to obtain food/water, throughout pregnancy. Dual energy X-ray absorptiometry and peripheral quantitative computed tomography scans were performed pre-mating and post-weaning. Maternal stress was assessed by fecal corticosterone measurement. Offspring weights were assessed at postnatal days 1 and 25 (weaning). RESULTS: Changes in bone mineral over the pregnancy/lactation period were site-specific. Exercise did not affect loss of bone mineral from the lumbar spine, but did attenuate the loss of trabecular bone mineral from the tibial metaphysis and enhance the strength strain index and cross-sectional moment of inertia at the tibial diaphysis (P≤0.05) in dams in the exercised group. Fecal corticosterone did not differ between dam groups. There were no significant differences in offspring weight between the exercised and control group at either time point. CONCLUSIONS: Voluntary exercise in the pregnant rat can improve some post-lactation bone parameters and does not adversely affect early postnatal outcomes of the offspring.
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Densidad Ósea/fisiología , Huesos/fisiología , Condicionamiento Físico Animal/fisiología , Estrés Fisiológico/fisiología , Absorciometría de Fotón , Animales , Huesos/diagnóstico por imagen , Corticosterona/análisis , Heces/química , Femenino , Embarazo , Ratas , Ratas WistarRESUMEN
Chondrodysplasia in Texel sheep is a recessively inherited disorder characterized by dwarfism and angular deformities of the forelimbs. A genome-wide association study using the Illumina OvineSNP50 BeadChip on 15 sheep diagnosed as affected and eight carriers descended from three affected rams was conducted to uncover the genetic cause. A homozygous region of 25 consecutive single nucleotide polymorphism (SNP) loci was identified in all affected sheep, covering a region of 1 Mbp on ovine chromosome 4. Seven positional candidate genes - including the solute carrier family 13 (sodium/sulphate symporters), member 1 (SLC13A1) - were identified and used to search for new SNPs for fine mapping of the causal locus. The SLC13A1 gene, encoding a sodium/sulphate transporter, was the primary candidate gene attributable to similar phenotypes observed in the Slc13a1 knockout mouse model. We discovered a 1-bp deletion of T (g.25513delT) at the 107 bp position of exon 3 in the SLC13A1 gene. Genotyping by direct sequencing and restriction fragment length polymorphism analysis for this mutation showed that all 15 affected sheep were g.25513delT/g.25513delT; the eight carriers were g.25513delT/T and 54 normal controls were T/T. The mutation g.25513delT shifts the open reading frame of SLC13A1 to introduce a stop codon and truncate C-terminal amino acids. It was concluded that the g.25513delT mutation in the SLC13A1 gene was responsible for the chondrodysplasia seen in these Texel sheep. This knowledge can be used to identify carriers with the defective g.[25513delT] allele to avoid at-risk matings to improve animal welfare and decrease economic losses.
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Genómica , Osteocondrodisplasias/genética , Osteocondrodisplasias/patología , Osteocondrodisplasias/veterinaria , Oveja Doméstica/genética , Alelos , Secuencia de Aminoácidos , Animales , Proteínas de Transporte de Catión/genética , Mapeo Cromosómico , Cromosomas de los Mamíferos/genética , Femenino , Dosificación de Gen , Sitios Genéticos , Estudio de Asociación del Genoma Completo , Homocigoto , Masculino , Datos de Secuencia Molecular , Mutación , Fenotipo , Polimorfismo de Longitud del Fragmento de Restricción , Polimorfismo de Nucleótido Simple , Cotransportador de Sodio-Sulfato , Simportadores/genéticaRESUMEN
[This corrects the article DOI: 10.3168/jdsc.2020-0043.].
RESUMEN
A deterministic model was developed in 1998 to evaluate the concurrent effects of selection and crossbreeding on the rate of genetic gain and increases in productivity of New Zealand dairy cattle over the ensuing 25-yr period. The predictions of today's breed composition of the national dairy herd and genetic trends for body weight and lactation yields of milk, fat, and protein are compared with today's actual values. Selection was assumed to use an index that included live weight and lactation yields of milk, fat, and protein. Mating strategies involving the Holstein-Friesian (F), Jersey (J), and Ayrshire (A) breeds were evaluated. Effects of heterosis and age were included to calculate phenotypic live weight and yields of milk, fat, and protein per cow. At the time the model was developed, New Zealand had an across-breed evaluation system, but only straightbred bulls were used after progeny testing based on records of straightbred and crossbred daughters. The model predicted that if crossbred cows and bulls could be considered as bull parents, faster rates of genetic gain may result because of increased selection intensities in the cow to breed bull selection pathway. This scenario transpired, and the best bulls and cows for farm profit were used regardless of breed. Under that mating strategy for the 2018 birthyear, the model predicted the national breed composition would be 11% F, 34% J, 52% F×J, and 2% A; the actual breed composition was 36% F, 9% J, 53% F×J, and 1% A. The model-predicted annual genetic gains would be 16.7 L of milk, 1.2 kg of fat, 1.5 kg of protein, and -0.7 kg of body weight; the realized annual improvements were 13.6 L of milk, 1.31 kg of fat, 1.17 kg of protein, and -0.36 kg of body weight. Predicted long-term responses to selection can closely mirror realized improvements, confirming the value of modeling to inform animal breeding decision-making.
RESUMEN
Greater rates of genetic gain can be achieved by selecting animals born to younger parents. However, little is known about the lifetime performance of dual purpose ewes (Ovis aries) that are born to primiparous ewe lambs (8 to 9 months old at breeding). This experiment investigated the effect of being born from either a ewe lamb or mixed age dam as either a single or twin on the lifetime performance of ewe progeny. Lifetime performance was measured in terms of the life time live weights of the ewes, the weight and number of lambs born and weaned, the efficiency of production (kilograms of lamb weaned / predicted pasture intake (kgDM) of the ewes), and ewe survival. The study followed the lifetime production of 17 single and 41 twin female lambs born to mature ewes (M1 and M2, respectively), and 28 single and 29 twin lambs born to ewe lambs (L1 and L2, respectively). Over their lifetime L2 ewes were lighter (P<0.05) but had similar body condition scores to the other three ewe groups. There was no difference in average progeny weaning weight or total progeny litter weaning weights between groups. The M1 ewes had the greatest longevity (P<0.05) of the four groups. Even though L2 ewes were lighter than the other three groups, this was insufficient to increase their lifetime efficiency of production (kg lamb weaned/predicted pasture consumption), relative to the other groups. These results suggest farmers could select replacements born to ewe lambs without sacrificing animal production.
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Selección Artificial , Oveja Doméstica/fisiología , Animales , Animales Recién Nacidos , Peso al Nacer , Peso Corporal , Femenino , Tamaño de la Camada , Longevidad , Edad Materna , Nueva Zelanda , Paridad , Embarazo , Oveja Doméstica/anatomía & histología , Oveja Doméstica/crecimiento & desarrolloRESUMEN
We recorded head direction (HD) cells from the lateral mammillary nucleus (LMN) and anterior thalamus (ATN) of freely behaving rats and also made bilateral lesions of LMN while recording HD cells from ATN. We discovered that the tuning functions of LMN HD cells become narrower during contraversive head turns, but not ipsiversive head turns, compared to when the head is not turning. This narrowing effect does not occur for ATN HD cells. We also found that the HD signal in LMN leads that in ATN by about 15-20 ms. When LMN was lesioned bilaterally, HD cells in ATN immediately lost their directional firing properties and never recovered them. Based on these findings, we argue that LMN may be an essential component of an attractor-integrator network that participates in generating the HD signal.
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Mapeo Encefálico , Movimientos de la Cabeza/fisiología , Tubérculos Mamilares/fisiología , Neuronas/fisiología , Tálamo/fisiología , Animales , Lateralidad Funcional , Masculino , Modelos Neurológicos , Postura , Ratas , Ratas Long-EvansRESUMEN
The foetal mammary gland is sensitive to maternal weight and nutrition during gestation, which could affect offspring milk production. It has previously been shown that ewes born to dams offered maintenance nutrition during pregnancy (day 21 to 140 of gestation) produced greater milk, lactose and CP yields in their first lactation when compared with ewes born to dams offered ad libitum nutrition. In addition, ewes born to heavier dams produced greater milk and lactose yields when compared with ewes born to lighter dams. The objective of this study was to analyse and compare the 5-year lactation performance of the previously mentioned ewes, born to heavy or light dams that were offered maintenance or ad libitum pregnancy nutrition. Ewes were milked once per week, for the first 6 weeks of their lactation, for 5 years. Using milk yield and composition data, accumulated yields were calculated over a 42-day period for each year for milk, milk fat, CP, true protein, casein and lactose using a Legendre orthogonal polynomial model. Over the 5-year period, ewes born to heavy dams produced greater average milk (P=0.04), lactose (P=0.01) and CP (P=0.04) yields than offspring born to light dams. In contrast, over the 5-year period dam nutrition during pregnancy did not affect average (P>0.05) offspring milk yields or composition, but did increase milk and lactose accumulated yield (P=0.03 and 0.01, respectively) in the first lactation. These results indicate that maternal gestational nutrition appears to only affect the first lactational performance of ewe offspring. Neither dam nutrition nor size affected grand-offspring live weight gain to, or live weight at weaning (P>0.05). Combined these data indicate that under the conditions of the present study, manipulating dam weight or nutrition in pregnancy can have some effects of offspring lactational performance, however, these effects are not large enough to alter grand-offspring growth to weaning. Therefore, such manipulations are not a viable management tool for farmers to influence lamb growth to weaning.
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Fenómenos Fisiológicos Nutricionales de los Animales , Leche/metabolismo , Fenómenos Fisiologicos de la Nutrición Prenatal , Ovinos/fisiología , Animales , Peso Corporal , Femenino , Lactancia , Glándulas Mamarias Animales/fisiología , Parto , Embarazo , Ovinos/crecimiento & desarrollo , Destete , Aumento de PesoRESUMEN
Familial episodic ataxia of lambs is a congenital transient autosomal dominant disorder of newborn lambs, with varying expressivity. Affected lambs show episodes of an asymmetric ataxic gait, base-wide extensor hypertonia of the thoracic limbs and flexor hypertonia of the pelvic limbs. The aim of the study was to determine the genetic variant causing familial episodic ataxia in lambs. Using whole genome sequencing of two half-sib affected lambs, their sire, and their two normal dams, a heterozygous C>T transition at OAR10:77593415 (Oar_v3.1) in exon 1 of the fibroblast growth factor 14 (FGF14) gene (c.46C>T) was identified. The c.46C>T transition resulted in a premature stop codon at position 16 of the 247 amino acid FGF14 protein (p.Q16*). PCR and Sanger sequencing was used to genotype an additional 20 clinically affected animals, demonstrating all lambs carried the c.46C>T variant but 1 clinically more severely affected inbred lamb was homozygous (TT). A further 11 unrelated normal ewes were positionally sequenced, none of which had the variant, while in 18 lambs of unknown status born over 2 years of breeding trials six lambs were found to have the c.46C>T variant, likely clinically unidentified heterozygotes due to the variable expressivity, while 12 did not. In conclusion, familial episodic ataxia of lambs is potentially associated with a c.46C>T variant in the FGF14 gene. Further research is required into the mechanism behind the apparent recovery of lambs.
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Ataxia/genética , Factores de Crecimiento de Fibroblastos/genética , Mutación , Oveja Doméstica/genética , Animales , Cruzamiento , Codón sin Sentido , Modelos Animales de Enfermedad , Exones , Femenino , Genotipo , Homocigoto , Masculino , Análisis de Secuencia de ADN , OvinosRESUMEN
As a rat navigates through space, neurons called head-direction (HD) cells provide a signal of the rat's momentary directional heading. Although partly guided by landmarks, the cells also show a remarkable ability to track directional heading based on angular head movement. Theoretical models suggest that the HD cells are linked together to form an attractor network, and that cells which signal angular velocity update the directional setting of the attractor. Recently, cell types similar to those required theoretically have been discovered in the lateral mammillary and dorsal tegmental nuclei. Lesion and anatomical data suggest these nuclei might constitute the postulated attractor-path integration mechanism, and that they provide the HD cell signal to cortical areas where it has been observed.
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Movimientos de la Cabeza/fisiología , Hipocampo/citología , Tubérculos Mamilares/citología , Neuronas/fisiología , Percepción Espacial/fisiología , Animales , Hipocampo/fisiología , Tubérculos Mamilares/fisiología , Vías Nerviosas , RatasRESUMEN
Pavlovian fear conditioning has emerged as a leading behavioral paradigm for studying the neurobiological basis of learning and memory. Although considerable progress has been made in understanding the neural substrates of fear conditioning at the systems level, until recently little has been learned about the underlying cellular and molecular mechanisms. The success of systems-level work aimed at defining the neuroanatomical pathways underlying fear conditioning, combined with the knowledge accumulated by studies of long-term potentiation (LTP), has recently given way to new insights into the cellular and molecular mechanisms that underlie acquisition and consolidation of fear memories. Collectively, these findings suggest that fear memory consolidation in the amygdala shares essential biochemical features with LTP, and hold promise for understanding the relationship between memory consolidation and synaptic plasticity in the mammalian brain.
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Condicionamiento Psicológico/fisiología , Miedo/fisiología , Potenciación a Largo Plazo/fisiología , Memoria/fisiología , Neuronas/fisiología , Animales , HumanosRESUMEN
The major histocompatibility complex (MHC) in sheep, Ovar-Mhc, is poorly characterised, when compared to other domestic animals. However, its basic structure is similar to that of other mammals, comprising class I, II and III regions. Currently, there is evidence for the existence of four class I loci. The class II region is better characterised, with evidence of one DRA, four DRB (one coding and three non-coding), one DQA1, two DQA2, and one each of the DQB1, DQB2, DNA, DOB, DYA, DYB, DMA, and DMB genes in the region. The class III region is the least characterised, with the known presence of complement cascade (C4, C2 and Bf), TNFalpha and CYP21 genes. Products of the class I and II genes, MHC molecules, play a pivotal role in antigen presentation required for eliciting immune responses against invading pathogens. Several studies have focused on polymorphisms of Ovar-Mhc genes and their association with disease resistance. However, more research emphasis is needed on characterising the remaining Ovar-Mhc genes and developing simplified and cost-effective methods to score gene polymorphisms. Haplotype screening, employing multiple markers rather than single genes, would be more meaningful in MHC-disease association studies, as it is well known that most of the MHC loci are tightly linked, exhibiting very little recombination. This review summarises the current knowledge of the structure of Ovar-Mhc and polymorphisms of genes located in the complex.
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Antígenos de Histocompatibilidad Clase II/genética , Antígenos de Histocompatibilidad Clase I/genética , Inmunidad Innata/genética , Polimorfismo Genético/genética , Enfermedades de las Ovejas/genética , Animales , Antígenos HLA/genética , Haplotipos/genética , Humanos , Ratones , Ovinos , Enfermedades de las Ovejas/inmunología , Relación Estructura-ActividadRESUMEN
This perspective considers genetic disorders of domestic animal populations, in particular their epidemiology and control. Inherited disorders of animals share the same basic molecular biology as those of human beings, but they differ in their epidemiology due largely to the breed structure of the various species, human control of breeding and a greater influence of the founder effect, particularly due to extensive use of a limited number of sires, and inbreeding. Control of genetic disorders in animals is also more practical through extensive screening for disease, or heterozygous animals within defined breed populations, followed by exclusion of affected or carrier animals from breeding. This is assisted by the fact that, within a breed, many inherited monogenic disorders are associated with a single mutation. However some of the more important disorders may be inherited in a non-Mendelian manner, being influenced by multiple genes as well as environmental factors. These aspects are discussed and contrasted with similar aspects in human medical genetics.
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Enfermedades Genéticas Congénitas/veterinaria , Variación Genética , Animales , Cruzamiento , EndogamiaRESUMEN
There is limited information on factors affecting twin lamb growth before weaning, which limits the options available to farmers to actively manage lamb growth. Data from 2 multiyear experiments involving 402 twin-bearing Romney ewes were used to evaluate the effects of prenatal ewe traits (live weight at mating and set stocking and BCS at mating and set stocking) and combined twin lamb birth weight on ewe milk production and lamb growth from birth to weaning as well as the proportion of variation in twin lamb growth that could be explained by these variables. Additionally, the effect of accumulated ewe milk yield over a 42-d period (MY; Days 0 to 42) and accumulated milk components (protein, fat, and lactose) on twin lamb growth were investigated. The effects of prenatal variables on MY, birth weight, and combined twin lamb live weight gain from Day 0 to 42 (LWG) were inconsistent across the 2 experiments. In addition, prenatal ewe traits ( < 0.05) explained less than 30% of the variation in MY and lamb growth from birth to weaning in both experiments. Combined twin lamb birth weight was positively ( < 0.001) correlated with MY ( = 0.34 and = 0.43 in Exp. 1 and 2, respectively). Combined twin lamb LWG was dependent on ewe MY ( = 0.43 for Exp. 1 and = 0.30 for Exp. 2). Lactose, fat, and milk CP yields explained 47 and 42% of the variation in lamb LWG in Exp. 1 and 2, respectively. Lactose and milk CP yield positively affected ( < 0.05) LWG in Exp. 1 and 2, respectively. Fat yield had a positive relationship with LWG in Exp. 1 and a negative relationship with LWG in Exp. 2. In conclusion, the measured prenatal ewe traits had a minimal effect on milk yield and twin lamb growth to weaning. Milk yield and composition explained the greatest proportion of variation in LWG. This suggests that farmers should select ewes with higher milk yields to maximize twin lamb growth to weaning. However, less than 50% of the variation in LWG and weaning live weight was explained by the measured ewe and lamb parameters. Therefore, further studies are required to determine additional ewe or lamb variables that control variation in twin lamb growth.
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Peso al Nacer/fisiología , Lactancia/fisiología , Tamaño de la Camada , Ovinos/fisiología , Aumento de Peso/fisiología , Animales , Femenino , Leche , Ovinos/crecimiento & desarrollo , DesteteRESUMEN
Estimation of metabolizable energy (ME) requirement for maintenance (ME) and growth (ME) in pre-weaned lambs have been limited to milk-only fed lambs. This study aimed to determine energy and nitrogen metabolisability of milk and pellets when fed together, compare the growth and chemical body composition of lambs fed varying levels of pellets in addition to milk, and to estimate ME, ME, and the CP:ME ratio requirements for growth. The study included 32 twin-born Romney-cross ram lambs. Four lambs were slaughtered at 24 h post-partum to estimate initial body composition and the remaining 28 were assigned to 1 of 4 treatment groups of 7. Group 1 was fed milk replacer (MR) only; group 2 was fed MR and allowed ad libitum access to pellets; groups 3 and 4 were offered 30% and 60%, respectively of the average pellet intake of the ad libitum group the previous day while being fed MR. Milk replacer was fed as a proportion of the lamb's live weight (LW). Lambs from each treatment were placed in metabolic cages at 17 kg LW for 4 d to allow for total fecal and urine collection. All lambs were slaughtered at 18 kg LW. The ADG, ADG:ME ratio, stomach and liver weight, and rumen papillae lengths increased ( < 0.05) with increasing pellet intake. Increasing daily ME intake increased ( < 0.05) both daily energy and protein deposition but had no effect ( > 0.05) on fat deposition. However, the total chemical body composition was unaffected ( > 0.05) by dietary treatment. Digestibility of energy and N decreased ( < 0.05) with increasing ME intake. Percent energy and N retained for growth were 96% vs. 71% and 72% vs. 30% for milk and pellets, respectively. The ME and ME values obtained were 0.40 MJ ME/kg LW·d and 13.8 MJ ME/kg ADG, respectively. The CP:ME ratio of MR and pellet was 11.1 and 15.7, respectively. However, a simulation model suggested that lambs require a CP:ME ratio of 13.1 at 5 kg and 10.9 at 18 kg LW, indicating that protein intake may be limiting to lamb growth in early life and in excess by 18 kg LW. In conclusion, increasing pellet intake was associated with decreased N retention. The inclusion of pellets, however, improved the efficiency of ME utilization for growth in pre-weaned lambs and was beneficial for rumen development. The ME was higher than previously recommended values and the CP:ME intake of lambs does not match their requirements which may warrant further studies.
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Alimentación Animal/análisis , Proteínas en la Dieta/metabolismo , Necesidades Nutricionales , Ovinos/fisiología , Fenómenos Fisiológicos Nutricionales de los Animales , Animales , Composición Corporal , Peso Corporal , Dieta/veterinaria , Heces , Tracto Gastrointestinal/anatomía & histología , Masculino , Leche/metabolismoRESUMEN
The objective of this study was to investigate the gene expression of progesterone and estrogen receptor α (PR, ERα), insulin-like growth factor (IGF) 1, IGF-2, their receptor (IGFR1), IGF-binding proteins (BP) 1 to 6, insulin receptor, adiponectin receptors (AdipoR1/2), cyclooxygenase 2 (PTGS2), mucin 1 and to localize PR, ERα, IGF-1, IGFR1, PTGS2, and proliferating cellular nuclear antigen (PCNA) in the endometrium of pregnant (Day 19) Suffolk and Cheviot ewes carrying Suffolk and Cheviot embryos transferred within and reciprocally between breeds. Gene expression was determined by real-time quantitative polymerase chain reaction (RT-qPCR), and antigen determination was measured by immunohistochemistry in the luminal epithelium (LE), superficial and deep glands (SG, DG, respectively) and superficial and deep stroma. Gene expression of PR, IGF-1, IGFBP2, and IGFBP5 was higher in Suffolk than that in Cheviot ewes (P < 0.05). Greater abundance of IGF-2 and IGBP3 expression was found in Cheviot ewes carrying Cheviot embryos than Cheviot ewes carrying Suffolk embryos (P < 0.05). No staining for PR and ERα was observed in the LE, very scarce staining in SG and DG, whereas positive staining was observed in both superficial and deep stroma. No differences were found for PR staining, but Cheviot ewes had higher ERα staining intensity than Suffolk ewes (P < 0.05). Positive staining for IGF-1 was observed in all cell types except DG, and staining of IGFR1 was observed in all cell types. No differences among groups in staining were found for IGF-1 or IGFR1 in any cell type. Positive staining of PTGS2 was observed in LE and SG in all groups. An interaction between ewe and embryo breed affected PTGS2 staining (P < 0.05), whereby Cheviot ewes carrying Suffolk embryos had a lower PTGS2 staining than Suffolk ewes carrying Suffolk embryos. Positive staining of PCNA was found in LE and SG. Suffolk ewes carrying Suffolk embryos showed lower PCNA immunostaining than Cheviot ewes carrying Suffolk embryos (P < 0.05), whereas no differences were observed in ewes carrying Cheviot embryos. This study showed that gestation-related protein expression in the endometrium of Suffolk and Cheviot ewes is affected by both ewe and embryo breed at Day 19 of pregnancy.
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Transferencia de Embrión/veterinaria , Endometrio/química , Endometrio/metabolismo , Expresión Génica , Ovinos/genética , Animales , Ciclooxigenasa 2/análisis , Ciclooxigenasa 2/genética , Desarrollo Embrionario/genética , Receptor alfa de Estrógeno/análisis , Receptor alfa de Estrógeno/genética , Femenino , Edad Gestacional , Inmunohistoquímica , Proteínas de Unión a Factor de Crecimiento Similar a la Insulina/genética , Factor I del Crecimiento Similar a la Insulina/análisis , Factor II del Crecimiento Similar a la Insulina/genética , Mucina-1/genética , Embarazo , Antígeno Nuclear de Célula en Proliferación/análisis , Receptor IGF Tipo 1/análisis , Receptor IGF Tipo 1/genética , Receptor de Insulina/genética , Receptores de Adiponectina/genética , Receptores de Progesterona/análisis , Receptores de Progesterona/genética , Especificidad de la EspecieRESUMEN
The amygdala is critical for acquiring and expressing conditioned fear responses elicited by sensory stimuli that predict future punishment, but there is conflicting evidence about whether the amygdala is necessary for perceiving the aversive qualities of painful or noxious stimuli that inflict primary punishment. To investigate this question, rats were fear conditioned by pairing a sequence of auditory pips (the conditioned stimulus, or CS) with a brief train of shocks to one eyelid (the unconditioned stimulus, or US). Conditioned responding to the CS was assessed by measuring freezing responses during a test session conducted 24 h after training, and unconditioned responding to the US was assessed by measuring head movements evoked by the eyelid shocks during training. We found that pre-training electrolytic lesions of the amygdala's lateral (LA) nucleus blocked acquisition of conditioned freezing to the CS, and also significantly attenuated unconditioned head movements evoked by the US. Similarly, bilateral inactivation of the amygdala with the GABA-A agonist muscimol impaired acquisition of CS-evoked freezing, and also attenuated US-evoked responses during training. However, when amygdala synaptic plasticity was blocked by infusion of the NR2B receptor antagonist ifenprodil, acquisition of conditioned freezing was impaired but shock reactivity was unaffected. These findings indicate that neural activity within the amygdala is important for both predicting and perceiving the aversive qualities of noxious stimuli, and that synaptic plasticity within LA is the mechanism by which the CS becomes associated with the US during fear conditioning.
Asunto(s)
Amígdala del Cerebelo/fisiología , Reacción de Prevención/fisiología , Condicionamiento Clásico/fisiología , Miedo/fisiología , Estimulación Acústica/métodos , Amígdala del Cerebelo/efectos de los fármacos , Animales , Reacción de Prevención/efectos de los fármacos , Reacción de Prevención/efectos de la radiación , Conducta Animal , Condicionamiento Clásico/efectos de los fármacos , Condicionamiento Clásico/efectos de la radiación , Relación Dosis-Respuesta a Droga , Electrochoque/efectos adversos , Lateralidad Funcional , Agonistas del GABA/farmacología , Inhibición Psicológica , Masculino , Muscimol/farmacología , Ratas , Ratas Sprague-Dawley , Reflejo de Sobresalto/efectos de los fármacos , Reflejo de Sobresalto/fisiología , Reflejo de Sobresalto/efectos de la radiaciónRESUMEN
Comparisons were made of carcass and meat quality characteristics of pasture-raised Texel-cross ewe (n=269) and ram (n=275) lambs between 5 and 8 months of age with an average carcass weight of 17.2kg. Carcass assessment was based on linear dimensions and dissection of a leg into muscle, fat and bone, and meat quality measurements were made on M. semimembranosus and M. longissimus. At a set carcass weight, ewe lambs had higher dressing percentages (2%), shorter carcasses (0.7cm), and heavier leg cuts (35g) (P<0.01) than males. At the same leg weight, legs of ewe lambs were fatter than males (subcutaneous plus intermuscular fat; 11.2% vs 9.6%; P<0.001), whereas legs of ram lambs contained significantly more muscle and bone than females (P<0.001). Leg muscle to bone ratio (4.7 vs 4.4) and muscularity were higher for females than males (P<0.001). However, the relationship between leg muscle to bone ratio and muscularity was not the same between the sexes, and for any given muscularity value the muscle to bone ratio of ram lambs was lower than females. As a result, if carcass lean meat yield is predicted from a measure of carcass shape, such as muscularity, lean meat yields will be overestimated for males and underestimated for females. Meat quality was lower in ram lambs than in females (P<0.001) as shown by higher Warner-Bratzler shear values (peak value 109.8 vs 97.0N for M. Semimembranosus), higher ultimate meat pH values, and lower redness (a(∗)) and lightness (L(∗), for the longissimus muscle only) values (P<0.001). It is concluded that significant differences between ewe and ram lambs do exist for many carcass and meat quality traits, but for most quality traits the differences are small.
RESUMEN
Embryo transfer of large sheep breed embryos (Suffolk) into small breed ewes (Cheviot) constrains birth size, but the maternal factors influencing fetal growth restriction are unknown. We hypothesized that reciprocal embryo transfer crosses between breeds of divergent size would affect pregnancy-related development of maternal insulin resistance in midgestation, thereby influencing fetal growth. Following superovulation, embryos were surgically collected 6 d postmating and transferred to recipients on the same day. Between- and within-breed transfers were performed. Between 60 and 70 d of pregnancy overnight-fasted ewes underwent hyperinsulinemic-euglycemic clamps for assessment of insulin sensitivity. Maternal insulin sensitivity did not vary with transferred lamb breed. Overall, Cheviot ewes tended to have higher fasting glucose (P = 0.068), fasting insulin (P = 0.052), and steady-state glucose (P = 0.065) concentrations than Suffolk ewes at the stage of pregnancy studied. As expected, transferred between-breed Suffolk lambs were born lighter (P = 0.014), and transferred between-breed Cheviot lambs tended to be heavier at birth (P = 0.056) than respective lambs transferred within breed. Midgestation insulin sensitivity does not appear to be a major factor constraining growth of large breed sheep fetus transferred into smaller breed or a factor in releasing constraint in growth of a small breed fetus within a larger breed ewe. However, as embryo size is already different between transferred groups by 19 d, factors other than maternal gestational insulin resistance may determine fetal growth in this embryo transfer paradigm.