Potential of in-plant intramuscular fat predictions to enable sheep breeders to incorporate consumer preferences in breeding programmes.

The inclusion of eating quality traits in sheep genetic improvement programmes is desirable. Intramuscular fat (IMF) plays a key role in ensuring consumer satisfaction when eating lamb, but genetic progress for IMF is constrained by a lack of routine data collection. This study investigated the potential for IMF predictor traits to substitute for measured IMF in genetic improvement programmes. Carcass and predicted IMF (near-infrared estimated IMF and marbling score) data were available on 10,113 New Zealand lambs, 1678 of which also had measured chemical IMF on a slice of M. longissimus lumborum on which the predictions of IMF had been made. Genetic antagonisms were observed between carcass lean traits and IMF. The genetic correlation between the predictors and measured IMF approached one, indicating that predictors of IMF can be used in genetic improvement programmes. Through using selection indexes, simultaneous increases in IMF and the existing terminal selection index are possible, provided all traits are measured. This study highlights the importance and potential of predicted IMF to achieve genetic improvement in traits of importance to consumers.

Can we have our steak and eat it: The impact of breeding for lowered environmental impact on yield and meat quality in sheep.

Global agreements in place to reduce methane emissions in livestock are a potential threat to food security. Successful but independent breeding strategies for improved production and lower methane are in place. The unanswered questions are whether these strategies can be combined and how they impact one another, physically and economically. The New Zealand economy is largely dependent on pastoral agriculture from grazing ruminants. The sheep industry produces ∼20 million lamb carcasses for export each year primarily from grass. Methane emitted from the fermentation of forage by grazing ruminants accounts for one-third of all New Zealand's greenhouse gas emissions. Here, we use sheep selection lines bred for divergent methane production and large numbers of their relatives to determine the genetic and phenotypic correlations between enteric methane emissions, carcass yield, and meat quality. The primary objectives were to determine whether previously shown physiological differences between methane selection lines (differing by ∼12% in methane) result in a negative impact on meat production and quality by measuring close relatives. The results show no negative effects of breeding for lowered methane on meat and carcass quality. Gross methane emissions were highly correlated with liveweight and measures of carcass weight and negatively correlated with dressing-out percentage and fat yield (GR). Trends were similar but not significant for methane yield (g CH4/kg DMI). Preliminary evidence, to date, shows that breeding for low methane may result in animals with higher lean yields that are economically favorable even before carbon costs and environmental benefits are taken into account. These benefits were seen in animals measured for methane on fixed intakes and require validation on intakes that are allowed to vary.

Relationships between intramuscular fat percentage and fatty acid composition in M. longissimus lumborum of pasture-finished lambs in New Zealand.

This paper reports relationships between fatty acids (FAs) and intramuscular fat (IMF)% in M. longissimus lumborum samples from 108 pasture-fed ewe lambs. Samples ranged in IMF from 1 to 6%. Relationships between %FA with total IMF% were mainly linear with percentages of saturated and monounsaturated FAs (MUFA), including trans-FAs, increasing and polyunsaturated FAs decreasing as IMF% increased. Normalized FA content data at 5.5% relative to 1.5% IMF, showed the highest relative increase for C14:0 as rates of endogenous synthesis increase with higher IMF deposition. This can be related to enhanced C12:0 elongation and lower rates of C14:0 desaturation, supported by a preferential desaturation of C18:1 trans-11 and C18:0 compared with C14:0 and C16:0 as IMF increased. The greatest normalized increase after C14:0 was anteisoC17:0 followed by other branched chain FAs and then trans-MUFA and C18:2 cis-9,trans-11. Finally, C22:6 and C22:5 showed higher relative increase than C20:5 indicating greater rates of elongation and desaturation past C20:5 at higher levels of fatness.

Consumer liking of M. longissimus lumborum from New Zealand pasture-finished lamb is influenced by intramuscular fat.

Palatability of meat is known to be affected by intramuscular fat (IMF), but the effect in relatively low-fat New Zealand lamb is unknown. This study evaluated the eating quality of 108 loins (M. longissimus lumborum) from a single flock of ewe-lambs. Loins ranged from 1.09-5.68% IMF and were stratified into 6 groups: 1.65, 2.12, 2.65, 3.20, 3.58 and 4.40%. Consumers' (n = 165) overall liking of lamb increased significantly at around 3% IMF, achieving maximum scores at 4% IMF. One consumer cluster (n = 111) showed a linear increase in overall liking with increasing IMF%, regarded as 'IMF lovers: the more the better', while a second cluster (n = 54) preferred 2.5-3.5% IMF, described as 'IMF optimizers: just the right amount'. IMF% was modestly correlated (~ + 0.25) with all sensory attributes except juiciness. Liking scores were modestly correlated with monounsaturated (~ + 0.25) and polyunsaturated (~ - 0.20) fatty acids. Results suggest aiming for IMF% levels in New Zealand lamb beyond 3% to maximize eating quality for premium markets in particular.

Panic results in unique molecular and network changes in the amygdala that facilitate fear responses.

Recurrent panic attacks (PAs) are a common feature of panic disorder (PD) and post-traumatic stress disorder (PTSD). Several distinct brain regions are involved in the regulation of panic responses, such as perifornical hypothalamus (PeF), periaqueductal gray, amygdala and frontal cortex. We have previously shown that inhibition of GABA synthesis in the PeF produces panic-vulnerable rats. Here, we investigate the mechanisms by which a panic-vulnerable state could lead to persistent fear. We first show that optogenetic activation of glutamatergic terminals from the PeF to the basolateral amygdala (BLA) enhanced the acquisition, delayed the extinction and induced the persistence of fear responses 3 weeks later, confirming a functional PeF-amygdala pathway involved in fear learning. Similar to optogenetic activation of PeF, panic-prone rats also exhibited delayed extinction. Next, we demonstrate that panic-prone rats had altered inhibitory and enhanced excitatory synaptic transmission of the principal neurons, and reduced protein levels of metabotropic glutamate type 2 receptor (mGluR2) in the BLA. Application of an mGluR2-positive allosteric modulator (PAM) reduced glutamate neurotransmission in the BLA slices from panic-prone rats. Treating panic-prone rats with mGluR2 PAM blocked sodium lactate (NaLac)-induced panic responses and normalized fear extinction deficits. Finally, in a subset of patients with comorbid PD, treatment with mGluR2 PAM resulted in complete remission of panic symptoms. These data demonstrate that a panic-prone state leads to specific reduction in mGluR2 function within the amygdala network and facilitates fear, and mGluR2 PAMs could be a targeted treatment for panic symptoms in PD and PTSD patients.

PSD95 and nNOS interaction as a novel molecular target to modulate conditioned fear: relevance to PTSD.

Stimulation of N-methyl-D-aspartic acid receptors (NMDARs) and the resulting increase of nitric oxide (NO) production are critical for fear memory formation. Following NMDAR activation, efficient production of NO requires linking the 95 kDa postsynaptic density protein (PSD95), a scaffolding protein to neuronal nitric oxide synthase (nNOS). A variety of previously studied NMDAR antagonists and NOS inhibitors can disrupt fear conditioning, but they also affect many other CNS functions such as motor activity, anxiety, and learning. We hypothesized that disrupting nNOS and PSD95 interaction in the amygdala, a critical site for fear memory formation, will reduce conditioned fear. Our results show that systemic treatment with ZL006, a compound that disrupts PSD95/nNOS binding, attenuates fear memory compared to its inactive isomer ZL007. Co-immunoprecipitation after fear conditioning showed a robust increase in the amygdala PSD95/nNOS binding, which was blocked by systemic pre-administration of ZL006. Treatment of amygdala slices with ZL006 also impaired long-term potentiation (LTP), a cellular signature of synaptic plasticity. Direct intra-amygdala infusion of ZL006 also attenuated conditioned fear. Finally, unlike NMDAR antagonist MK-801, ZL006 does not affect locomotion, social interaction, object recognition memory, and spatial memory. These findings support the hypothesis that disrupting the PSD95/nNOS interaction downstream of NMDARs selectively reduces fear memory, and highlights PSD95/nNOS interaction as a novel target for fear-related disorders, such as posttraumatic stress disorder.

Application of Hyperspectral imaging to predict the pH, intramuscular fatty acid content and composition of lamb M. longissimus lumborum at 24h post mortem.

Cost-effective, rapid and objective measurement of lamb quality on a routine basis is an important step for lamb value chains wishing to manage lamb product quality. Hyperspectral imaging (HSI) technology has shown promise as a solution for objective non-invasive prediction of meat quality. The performance of HSI applied 24h post mortem to lamb M. longissimus lumborum (LL) within a processing plant environment was assessed over two sampling years to evaluate its suitability for an objective lamb meat quality assurance tool. Calibration and validation steps were undertaken to evaluate HSI prediction performance for predicting fatty acid content and composition (n=1020 lambs) and pH (n=2406 lambs). Practical considerations of reference meat quality data quality and validation strategies are discussed. HSI can be used to predict meat quality parameters of lamb LL with varying accuracy levels, but ongoing calibration and validation across seasons is required to improve robustness of HSI for objective non-invasive assessment of lamb meat quality.

Estimates of genetic parameters for breech strike and potential indirect indicators in sheep.

AIM: To determine flystrike incidence, and estimate genetic parameters and potential indicator traits to reduce breech strike susceptibility in Romney sheep in New Zealand. METHODS: Seventeen Romney-based flocks, on 11 farms from throughout New Zealand, were enrolled in a case-control study in 2009/10 and 2010/11. Farmers observed lambs with flystrike and recorded dag score, breech bareness score and flystrike location for each lamb. Flystrike location was classed as breech, body, belly, shoulders, head or feet. Paternity of lambs was determined from tissue samples collected from lambs and all potential sires on farms. Control lambs without flystrike were selected by matching birth year, flock and sex. Due to the majority of strike occurring in the breech, genetic parameters for breech strike were explored. Dag score and breech bareness were investigated as indirect indicators of breech strike. Heritabilities and genetic and phenotypic correlations were estimated for breech strike, dag score and breech bareness using an animal model. RESULTS: For the 2009/10 season, 484 cases of flystrike were recorded with mean incidence rate per farm of 1.76 (min 0.47, max 2.95)%. For the 2010/11 season, 352 cases were recorded with a mean incidence rate per farm of 2.54 (min 0.43, max 8.18)%. Over both years 694/792 (88%) cases of flystrike occurred on the breech. Heritability on the observed scale for breech strike was 0.32 (SE 0.10). Heritabilities for dag score and breech bareness were 0.23 (SE 0.09) and 0.35 (SE 0.11), respectively. Breech strike had a high positive genetic correlation with dag score (0.71) and a low negative genetic correlation with breech bareness (-0.17). Breech strike had a high phenotypic correlation with dag score (0.62) and negative phenotypic correlation with breech bareness (-0.06). CONCLUSIONS: The high genetic and phenotypic correlations between breech strike and dag score makes dag score a viable option for indirect selection for breech strike resistance in Romney sheep in New Zealand. The heritability, genetic and phenotypic correlations require validation in other dual-purpose breeds, before breeding values for breech strike can be implemented, for use throughout the New Zealand sheep industry.

Effect and mode of action of the Texel muscling QTL (TM-QTL) on carcass traits in purebred Texel lambs.

TM-QTL is a quantitative trait locus (QTL) on ovine chromosome 18 (OAR18) known to affect loin muscling in Texel sheep. Previous work suggested that its mode of inheritance is consistent with paternal polar overdominance, but this has yet to be formally demonstrated. This study used purebred Texel sheep segregating for TM-QTL to confirm its presence in the chromosomal region in which it was first reported and to determine its pattern of inheritance. To do so, this study used the first available data from a Texel flock, which included homozygote TM-QTL carriers (TM/TM; n=34) in addition to homozygote non-carriers (+/+; n=40 and, heterozygote TM-QTL-carriers inheriting TM-QTL from their sire (TM/+; n=53) or their dam (+/TM; n=17). Phenotypes included a wide range of loin muscling, carcass composition and tissue distribution traits. The presence of a QTL affecting ultrasound muscle depth on OAR18 was confirmed with a paternal QTL effect ranging from +0.54 to +2.82 mm UMD (s.e. 0.37 to 0.57 mm) across the sires segregating for TM-QTL. Loin muscle width, depth and area, loin muscle volume and dissected M. longissimus lumborum weight were significantly greater for TM/+ than +/+ lambs (+2.9% to +7.9%; P<0.05). There was significant evidence that the effect of TM-QTL on the various loin muscling traits measured was paternally polar overdominant (P<0.05). In contrast, there was an additive effect of TM-QTL on both live weight at 20 weeks and carcass weight; TM/TM animals were significantly (P<0.05) heavier than +/+ (+11.1% and +7.3%, respectively) and +/TM animals (+11.9% and +11.7%, respectively), with TM/+ intermediate. Weights of the leg, saddle and shoulder region (corrected for carcass weight) were similar in the genotypic groups. There was a tendency for lambs inheriting TM-QTL from their sire to be less fat with slightly more muscle than non-carriers. For example, carcass muscle weight measured by live animal CT-scanning was 2.8% higher in TM/TM than +/+ lambs (P<0.05), carcass muscle weight measured by carcass CT-scanning was 1.36% higher in TM/+ than +/+ lambs (P<0.05), and weight of fat trimmed from the carcass cuts was significantly lower for TM/+ than +/+ lambs (-11.2%; P<0.05). No negative effects of TM-QTL on carcass traits were found. Optimal commercial use of TM-QTL within the sheep industry would require some consideration, due to the apparently different mode of action of the two main effects of TM-QTL (on growth and muscling).

Genetic relationships between dagginess, breech bareness, and wool traits in New Zealand dual-purpose sheep.

Genetic and phenotypic parameters were estimated for dagginess, breech, wool, and fiber traits from approximately 29,500 progeny born in 2009 and 2010 in New Zealand dual-purpose ram breeding sheep flocks. Dagginess is adherence of fecal matter to the wool, and this study investigates the genetic and phenotypic correlations between dagginess and breech and wool traits. Estimates for heritability were moderate (0.21 to 0.44) for the following traits: dag score at 3 and 8 mo (DAG3, DAG8), breech bareness, wool length, wool bulk (BULK), mean fiber diameter, mean fiber diameter SD, mean fiber diameter CV, curvature (CURV), weaning weight at 3 mo, and autumn BW. Heritability estimates for fleece weight at 12 mo and proportion of medullated fibers were high (0.49 and 0.53, respectively). Dag score at 3 mo and DAG8 had low genetic and phenotypic correlations with all traits. Breech bareness had positive genetic and phenotypic correlations with CURV and BULK and mostly negative genetic correlations with all other wool traits. In summary the quantity and attributes of wool were not primary causative factors in fecal accumulation, leaving fecal consistency and composition as the major factors.

Genetic parameters for production traits in New Zealand dual-purpose sheep, with an emphasis on dagginess.

Genetic and phenotypic parameters were estimated for production and disease traits (including dagginess) from about 2 million pedigree-recorded animals born between 1990 and 2008 in New Zealand dual-purpose ram breeding flocks. This is the most comprehensive study of genetic parameter estimates for the New Zealand sheep industry to date and includes estimates that have not previously been reported. Estimates of heritability were moderate for BW at 8 mo (LW8), fleece weight at 12 mo (FW12), dagginess score at 3 and 8 mo (DAG3, DAG8; 0.31 to 0.37), typical for weaning weight (WWT), fecal egg count in summer (FEC1) and autumn (FEC2), and analogous Nematodirus counts (NEM1, NEM2; 0.17 to 0.21), and low for number of lambs born to ewes (NLB; 0.09). The genetic correlations among production traits, WWT, LW8, and FW12, were positive and moderate to high. Correlations of DAG3 and DAG8 with production and disease traits were low and mostly negative. The NLB had low, but typically positive, correlations with other traits. Disease traits also had low, but positive, correlations with production traits (WWT, LW8, and FW12), and were highly correlated among themselves. In general, the heritability estimate for BW and dagginess were greater than what is currently used in the New Zealand genetic evaluation service (Sheep Improvement Limited), and the availability of accurate estimates for dagginess plus parasite resistance and their genetic correlations with production traits will enable more accurate breeding values to be estimated for New Zealand sheep.

Topographical distribution of corticotropin-releasing factor type 2 receptor-like immunoreactivity in the rat dorsal raphe nucleus: co-localization with tryptophan hydroxylase.

Corticotropin-releasing factor (CRF) and CRF-related neuropeptides are involved in the regulation of stress-related physiology and behavior. Members of the CRF family of neuropeptides bind to two known receptors, the CRF type 1 (CRF1) receptor, and the CRF type 2 (CRF2) receptor. Although the distribution of CRF2 receptor mRNA expression has been extensively studied, the distribution of CRF2 receptor protein has not been characterized. An area of the brain known to contain high levels of CRF2 receptor mRNA expression and CRF2 receptor binding is the dorsal raphe nucleus (DR). In the present study we investigated in detail the distribution of CRF2 receptor immunoreactivity throughout the rostrocaudal extent of the DR. CRF2 receptor-immunoreactive perikarya were observed throughout the DR, with the highest number and density in the mid-rostrocaudal DR. Dual immunofluorescence revealed that CRF2 receptor immunoreactivity was frequently co-localized with tryptophan hydroxylase, a marker of serotonergic neurons. This study provides evidence that CRF2 receptor protein is expressed in the DR, and that CRF2 receptors are expressed in topographically organized subpopulations of cells in the DR, including serotonergic neurons. Furthermore, these data are consistent with the hypothesis that CRF2 receptors play an important role in the regulation of stress-related physiology and behavior through actions on serotonergic and non-serotonergic neurons within the DR.

Anxiety-like behavior is modulated by a discrete subpopulation of interneurons in the basolateral amygdala.

The basolateral amygdala (BL) is a putative site for regulating anxiety, where inhibition and excitation respectively lead to decreases and increases in anxiety-like behaviors. The BL contains local networks of GABAergic interneurons that are subdivided into classes based on neurochemical content, and are hypothesized to regulate unique functional responses of local glutamatergic projection neurons. Recently it was demonstrated that lesioning a portion of the BL interneuronal population, those interneurons that express neurokinin1 receptors (NK(1r)), resulted in anxiety-like behavior. In the current study, these NK(1r) expressing cells of the BL are further phenotypically characterized, demonstrating approximately 80% co-expression with GABA thus confirming them as GABAergic interneurons. These NK(1r) interneurons also colocalize with two distinct populations of BL interneurons as defined by the neuropeptide content. Of the NK(1r) positive cells, 41.8% are also positive for neuropeptide Y (NPY) and 39.7% of the NK(1r) positive cells are also positive for cholecystokinin (CCK). In addition to enhancing the phenotypic characterization, the extent to which the NK(1r) cells of amygdala nuclei contribute to anxiety-like responses was also investigated. Lesioning the NK(1r) expressing interneurons, with a stable form of substance P (SSP; the natural ligand for NK(1r)) coupled to the targeted toxin saporin (SAP), in the anterior and posterior divisions of the BL was correlated to increased anxiety-like behaviors compared to baseline and control treated rats. Furthermore the phenotypic and regional selectivity of the lesions was also confirmed.

Investigations into the GDF8 g+6723G-A polymorphism in New Zealand Texel sheep.

This work investigated effects of carrying 0, 1, or 2 copies of the A allele resulting from the g+6723G-A transition in growth differentiation factor gene (GDF8) in New Zealand Texel-cross sheep at different lamb ages and carcass weights. Two Texel-cross sires carrying 1 copy of the A allele were mated to approximately 200 ewes carrying 0, 1, or 2 copies of the A allele. A total of 187 progeny were generated and genotyped to determine whether they were carrying 0, 1, or 2 copies of the A allele. The progeny were assigned to 1 of 4 slaughter groups balanced for the 3 genotypes, sex, and sire. The 4 groups were slaughtered commercially when their average BW (across all progeny in the slaughter group) reached 33, 40, 43, and 48 kg, respectively. Measurements of BW, and carcass dimensions and yield were made on all animals using Viascan (a commercial 2-dimensional imaging system that estimates lean content of the carcass as a percentage of total carcass weight). Additional measurements were made on the fourth slaughter group, which was computed tomography scanned at each slaughter time point to obtain 4 serial measures of lean and fat as estimated from the computed tomography images. The A allele did not have an effect on any BW traits. The A allele was associated with increased muscle and decreased fat across the variety of measures of muscling and fat, explaining between 0.2 and 1.1 of a residual SD unit. Estimates for an additive effect were significant and were positive for muscle and negative for fat traits. No dominance effect estimates (positive or negative) were significant. There was no significant interaction between A allele number and carcass weight or slaughter group for any trait. This is the first systematic study of the effect of the A allele copy number over a range of carcass weights (13 to 20 kg) and ages and results suggest the size of the effect across these endpoints is proportionately the same. Testing for the A allele therefore offers breeders the potential to improve rates of genetic gain for lean-meat yield across most production systems.

The bone-muscle ratio of fetal lambs is affected more by maternal nutrition during pregnancy than by maternal size.

Bone formation and loss are related to the strain imposed on bone by muscle forces. Bone mineral content (BMC) and lean mass (LM) of fetal lambs was determined at day 140 of pregnancy in 8 groups of ewes, which were of either large or small body size, on either high (ad libitum) or maintenance pasture intake from day 21 of pregnancy, or carrying either singletons or twins. BMC and LM (using DXA scanning) of fetal hindquarters/spine were corrected to leg length. BMC and LM were less in twin than singleton groups (P < 0.001). Large ewes on high intake produced single fetuses with a (group mean) BMC/LM ratio that was higher (P < 0.002) than that in fetuses of large ewes with singletons on maintenance intake or twins on either high or maintenance intakes, the ratios of which were not different. In single fetuses from small ewes on high intake, the BMC/LM ratio was higher than those from small ewes with singletons on maintenance intake or twins on either high or maintenance intakes, the ratios of which were not different. The ratio was not different in singleton fetuses of ewes on high intake, whether they were large or small. Different fetal environments resulted in a given amount of muscle being associated with a higher or lower bone mass. Dietary intake during pregnancy was more important than maternal size in affecting the ratio. We conclude that intrauterine environmental factors may be important in determining bone mass postnatally, and possibly later in life.

Meat quality traits were unaffected by a quantitative trait locus affecting leg composition traits in Texel sheep.

A QTL affecting leg muscle and fat traits has been identified within the New Zealand Texel population. The QTL maps to a region on OAR 2 with a two-marker haplotype test established at markers BULGE20 and BM81124. These markers encompass the likely position of Growth Differentiation Factor 8 (GDF8). The pleiotropic effects of this QTL on meat quality traits are tested. Objective measures of meat quality including pH, color (L*, a*, and b*), and tenderness (as assessed by Warner-Bratzler shear force measurements) were assessed on longissimus and semi-membranosus muscles of 540 progeny from six Texel sires. Four of these sires were subsequently identified as segregating for leg muscle and fat traits. For these segregating sires, comparison of progeny that had inherited the favorable haplotype from their sire with those that had received the alternate haplotype revealed no significant differences in the meat quality traits assessed. This finding suggests that the muscling QTL does not have pleiotropic effects on meat quality. A general scan for meat quality QTL was carried out using genotype data for eight markers from FCB128 to RM356 flanking 122cM of OAR 2 using Haley-Knott regression. This analysis revealed two QTL for a single sire. A QTL detected in the region of Marker INRA40 for color L* mapped to a site close to the muscling QTL, but there was evidence to suggest it is at a distinct locus. The QTL in the region of Marker RM356 might map distal to Marker RM356, as no peak was observed. This QTL, which seems to affect pH, color a*, color b*, and Warner-Bratzler shear measurements, requires further characterization.

A directed search in the region of GDF8 for quantitative trait loci affecting carcass traits in Texel sheep.

A directed search for QTL affecting carcass traits was carried out in the region of growth differentiation factor 8 (GDF8, also known as myostatin) on ovine chromosome 2 in seven Texel-sired half-sib families totaling 927 progeny. Weights were recorded at birth, weaning, ultrasound scanning, and slaughter. Ultrasonic measures of LM cross-sectional dimensions and s.c. fat above the LM were made, with the same measurements made on the LM after slaughter. Following slaughter, linear measurements of carcass length and width were made on all carcasses, and legs and loins from 540 lambs were dissected. Genotyping was carried out using eight microsatellite markers from FCB128 to RM356 on OAR 2 and analyzed using Haley-Knott regression. There was no evidence for QTL for growth rates or linear carcass traits. There was some evidence for QTL affecting LM dimensions segregating in some sire families, although it was not consistent between ultrasound and carcass measures of the same traits. There was strong and consistent evidence for a QTL affecting muscle and fat traits in the leg that mapped between markers BM81124 and BULGE20 for the four sires that were heterozygous in this region, but not for the three sires that were homozygous. The size of the effect varied across the four sires, ranging from 0.5 to 0.9 of an adjusted SD for weight-adjusted leg muscle traits, and ranging from 0.6 to 1.2 of an adjusted SD for weight-adjusted leg fat traits. The clearest effect shown was for multivariate analysis combining all leg muscle and fat traits analyzed across sires, where the -log(10) probability was 14. Animals carrying the favorable haplotype had 3.3% more muscle and 9.9% less fat in the leg relative to animals carrying other haplotypes. There was evidence for a second peak in the region of marker TEXAN2 for one sire group. It seems that a QTL affecting muscle and fat traits exists within the New Zealand Texel population, and it maps to the region of GDF8 on OAR2.

Serotonergic systems associated with arousal and vigilance behaviors following administration of anxiogenic drugs.

Serotonergic systems play important roles in modulating behavioral arousal, including behavioral arousal and vigilance associated with anxiety states. To further our understanding of the neural systems associated with increases in anxiety states, we investigated the effects of multiple anxiogenic drugs on topographically organized subpopulations of serotonergic neurons using double immunohistochemical staining for c-Fos and tryptophan hydroxylase combined with topographical analysis of the rat dorsal raphe nucleus (DR). Anxiogenic drugs with diverse pharmacological properties including the adenosine receptor antagonist caffeine, the serotonin 5-HT2A/2C receptor agonist m-chlorophenyl piperazine (mCPP), the alpha2-adrenoreceptor antagonist yohimbine, and the benzodiazepine receptor partial inverse agonist N-methyl-beta-carboline-3-carboxamide (FG-7142) induced increases in behavioral arousal and vigilance behaviors consistent with an increase in anxiety state. In addition, these anxiogenic drugs, excluding yohimbine, had convergent actions on an anatomically-defined subset of serotonergic neurons within the middle and caudal, dorsal subdivision of the DR. High resolution topographical analysis revealed that at the mid-rostrocaudal level, caffeine and FG-7142 had convergent effects on c-Fos expression in serotonergic neurons that were restricted to a previously undefined region, which we have named the shell region of the dorsal part of the dorsal raphe nucleus (DRDSh), that overlaps the anatomical border between the dorsal part of the dorsal raphe nucleus, the ventral part of the dorsal raphe nucleus (DRV), and the ventrolateral part of the dorsal raphe nucleus (DRVL). Retrograde tracing methods revealed that DRDSh contains large numbers of neurons projecting to the basolateral amygdaloid nucleus, a forebrain structure important for emotional appraisal and modulation of anxiety-related physiological and behavioral responses. Together these findings support the hypothesis that there is a functional topographical organization in the DR and are consistent with the hypothesis that anxiogenic drugs have selective actions on a subpopulation of serotonergic neurons projecting to a distributed central autonomic and emotional motor control system regulating anxiety states and anxiety-related physiological and behavioral responses.

Carcass composition and meat quality differences between pasture-reared ewe and ram lambs.

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.

Ratiometry of transmembrane voltage-sensitive fluorescent dye emission in hearts.

Transmembrane voltage-sensitive fluorescence measurements are limited by baseline drift that can obscure changes in resting membrane potential and by motion artifacts that can obscure repolarization. Voltage-dependent shift of emission wavelengths may allow reduction of drift and motion artifacts by emission ratiometry. We have tested this for action potentials and potassium-induced changes in resting membrane potential in rabbit hearts stained with di-4-ANEPPS [Pyridinium, 4-(2-(6-(dibutylamino)-2-naphthalenyl) ethenyl)-1-(3-sulfopropyl)-, hydroxide, inner salt] using laser excitation (488 nm) and a two-photomultiplier tube system or spectrofluorometer (resolution of 500-1,000 Hz and <1 mm). Green and red emissions produced upright and inverted action potentials, respectively. Ratios of green emission to red emission followed action potential contours and exhibited larger fractional changes than either emission alone (P < 0.001). The largest changes and signal-to-noise ratio (signal/noise) were obtained with numerator wavelengths of 525-550 nm and denominator wavelengths of 650-700 nm. Ratiometry lessened drift 56-66% (P < 0.015) and indicated decreases in resting membrane potential. Ratiometry lessened motion artifacts and increased magnitudes of deflections representing phase-zero depolarizations relative to total deflections by 123-188% in intact hearts (P < 0.02). Durations of action potentials at different pacing rates, temperatures, and potassium concentrations were independent of whether they were measured ratiometrically or with microelectrodes (P > or = 0.65). The ratiometric calibration slope was 0.017/100 mV and decreased with time. Thus emission ratiometry lessens the effects of motion and drift and indicates resting membrane potential changes and repolarization.