Myostatin Mutation Research Articles

Myostatin is a negative regulator of adult neurogenesis after spinal cord injury in zebrafish.

Intrinsic and extrinsic inhibition of neuronal regeneration obstruct spinal cord (SC) repair in mammals. In contrast, adult zebrafish achieve functional recovery after complete SC transection. While studies of innate SC regeneration have focused on axon regrowth as a primary repair mechanism, how local adult neurogenesis affects functional recovery is unknown. Here, we uncover dynamic expression of zebrafish myostatin b (mstnb) in a niche of dorsal SC progenitors after injury. mstnb mutants show impaired functional recovery, normal glial and axonal bridging across the lesion, and an increase in the profiles of newborn neurons. Molecularly, neuron differentiation genes are upregulated, while the neural stem cell maintenance gene fgf1b is downregulated in mstnb mutants. Finally, we show that human fibroblast growth factor 1 (FGF1) treatment rescues the molecular and cellular phenotypes of mstnb mutants. These studies uncover unanticipated neurogenic functions for mstnb and establish the importance of local adult neurogenesis for innate SC repair.

Research Note: Growth promoting potential in Mstn mutant quail dependent and independent of increased egg size

In avian species, positive relationships between egg weight (EW) and body weight (BW) have been reported. However, the correlation between the body growth rate and different weights of eggs from genetically mutated avian species was not studied yet. Myostatin (Mstn), an anti-myogenic factor, mutant quail were recently developed, and it was reported that EW produced from Mstn homozygous mutant quail (HO) was heavier compared to those from wild-type quail (WT). In the current study, distributions of pre-incubated EW and associations between EW and BW were compared between the Mstn mutant and WT quail lines. Average egg weight for the HO group was significantly heavier than the WT (P < 0.001) and the number of eggs having heavier EW (over 11 g) was higher in the HO compared to the WT (P < 0.01). BWs at wk (W) 0, 4, and 6 after hatch were also significantly greater in the HO (P < 0.001 in all groups). In addition, linear regression analyses revealed positive relationships between EW and BW from W0 to W6, regardless of sexes and genotypes. Furthermore, Mstn mutant quail were a heavier BW compared to the WT quail originated from eggs with similar weights. These data indicate that increased BW by Mstn mutation is contributed by increased EW and/or growth promoting activity of Mstn mutation independent of increasing egg sizes. These findings provide Mstn as a desirable genetic factor for selection of poultry breeds with superior growth. In addition, the knowledge gained from this study could inspire similar proof-of-concept studies involving standard and commercial lines of poultry.

Germline transmission of MSTN knockout cattle via CRISPR-Cas9

Although the production of several founder animals (F0) for gene editing in livestock has been reported in cattle, very few studies have assessed germline transmission to the next generation due to the long sexual maturation and gestation periods. The present study aimed to assess the germline transmission of MSTN mutations (-12bps deletion) in MSTN mutant F0 male and female cattle. For this purpose, oocytes and semen were collected after the sexual maturation of MSTN cattle, and embryos produced by in vitro fertilization were analyzed. In addition, the embryos were subjected to additional gene (PRNP) editing using electroporation. Embryos produced by in vitro fertilization with MSTN male and female cattle were transferred to a surrogate, and one calf was successfully born. MSTN heterozygous mutation was shown by sequencing of the F1 calf, which had no health issues. As a further experiment, using electroporation, additional gene-edited embryos fertilized with the MSTN male sperm showed a high mutation rate of PRNP (86.2 ± 3.4%). These data demonstrate that the cattle produced through gene editing matured without health issues and had transmitted MSTN mutation from the germ cells. Also, additional mutation of embryos fertilized with the MSTN male sperm could enable further mutagenesis using electroporation.

Positive growth of smooth muscle in uterine horns of myostatin homozygous mutant gilt

Current studies on myostatin (MSTN), a well-known negative regulator of skeletal muscle, studies mainly focus on the its effects on skeletal muscle.However, its effects on smooth muscle are less studied, especially in the uterine horns. To identify the role of MSTN in uterine horn smooth muscle, this study used 6–8-month-old homozygous MSTN mutant (MSTN−/−) gilts in anoestrum as animal models. Histochemical and immunofluorescence staining, western blotting, and RT-qPCR were performed. The results showed that the uteri of the MSTN−/− gilts were morphologically normal, and the uterine horn smooth muscle content was increased (MSTN−/−: 75.19%, Wild type: 51.52%, P < 0.01). In vivo immunofluorescence staining showed that the expression of the uterine horn smooth muscle-specific marker proteins, namely α-smooth muscle actin (ACTA2) and calponin, increased after MSTN knockout (1.41- and 1.21-fold, respectively, P < 0.05). Increased gene expression was also seen in MSTN−/− gilts in vivo for ACTA2 (approximately 2-fold), smooth muscle myosin heavy chain (7.14-fold), myocardin (9.32-fold), and serum response factor (2.17-fold). Protein expression of smooth muscle-specific markers was increased (1.51-fold for ACTA2, 1.57-fold for calponin, P<0.05). MSTN knockout promoted proliferation of the smooth muscle cell and the gene expression of c-kit, a peristaltic marker (2.43-fold, P < 0.05). The results of the in vitro experiments were consistent with those of the in vivo experiments. The present study indicates that MSTN knockout can increase the smooth muscle content of uterine horns, thus providing potential therapeutic targets for pregnancy disorders caused by increased smooth muscle content.

Generation of Heritable Prominent Double Muscle Buttock Rabbits via Novel Site Editing of Myostatin Gene Using CRISPR/Cas9 System

Rabbits have been domesticated for meat, wool, and fur production, and have also been cherished as a companion, artistic inspiration, and an experimental model to study many human diseases. In the present study, the muscle mass negative regulator gene myostatin (MSTN) was knocked out in rabbits at two novel sites in exon3, and the function of these mutations was determined in subsequent generations. The prominent double muscle phenotype with hyperplasia or hypertrophy of muscle fiber was observed in the MSTN-KO rabbits, and a similar phenotype was confirmed in the F1 generation. Moreover, the average weight of 80-day-old MSTN-KO rabbits (2,452 ± 63 g) was higher than that of wild-type rabbits (2,393.2 ± 106.88 g), and also the bodyweight of MSTN-KO rabbits (3,708 ± 43.06g) was significantly higher (P < 0.001) at the age of 180 days than wild-type (WT) rabbits (3,224 ± 48.64g). In MSTN-KO rabbits, fourteen rabbit pups from the F1 generation and thirteen from the F2 generation stably inherited the induced MSTN gene mutations. Totally, 194 pups were produced in the F1 generation of which 49 were MSTN-KO rabbits, while 47 pups were produced in the F2 generation of which 20 were edited rabbits, and the ratio of edited to wild-type rabbits in the F2 generation was approximately 1:1. Thus, we successfully generated a heritable double muscle buttocks rabbits via myostatin mutation with CRISPR/Cas9 system, which could be valuable in rabbit's meat production and also a useful animal model to study the development of muscles among livestock species and improve their important economic traits as well as the human muscle development-related diseases.

Esophageal striated muscle hypertrophy and muscle fiber type transformation in MSTN knockout pigs.

Myostatin (MSTN) is a member of the transforming growth factor-β superfamily that inhibits skeletal muscle growth and development. The esophagus is composed of skeletal muscle and smooth muscle, but the effect of MSTN on esophagus striated muscle (ESM) is unknown. The present study investigated the role of MSTN in ESM using MSTN mutant pigs through histological, gene and protein expression analysis in ESM of MSTN knockout (MSTN-/-) pigs and their wild type (WT) littermates. Hematoxylin-eosin staining showed that the fiber cross-sectional areas in ESM of MSTN-/- pigs were significantly larger than WT pigs (P < 0.05). Immunofluorescence staining showed that the percentage of type I muscle fibers in MSTN-/- pigs were significantly lower than WT pigs (P < 0.01) and type IIA muscle fibers in MSTN-/- pigs were significantly higher than WT pigs(21% higher, P < 0.01). However, type IIB muscle fibers were not detected in the ESM of MSTN-/- or WT pigs indicating that muscle fiber types in pig ESM was composed of type I and IIA. The mRNA levels of myogenic regulatory factors (MRFs) including myogenic differentiation (MyoD), myogenin (MyoG), myogenic factor 5 (Myf5) and myogenic regulatory factor 4 (MRF4) in ESM of MSTN-/- pigs showed a significant increase (P < 0.05 at least) when compared to WT pigs while mRNA level of myocyte enhancer factor 2C (MEF2C) displayed a decrease (P < 0.001). Protein expression of myosin heavy chain I (MHC-I) in MSTN-/- ESM was decreased and myosin heavy chain IIA (MHC-IIA) was increased (P < 0.01, P < 0.05). These findings indicate that MSTN plays an important role in esophageal striated muscle development and regulates muscle fiber types.

Analysis of the gut microbiota composition of myostatin mutant cattle prepared using CRISPR/Cas9

Myostatin (MSTN) negatively regulates muscle development and positively regulates metabolism through various pathways. Although MSTN function in cattle has been widely studied, the changes in the gut microbiota due to MSTN mutation, which contribute to host health by regulating its metabolism, remain unclear. Here, high-throughput sequencing of the 16S rRNA gene was conducted to analyze the gut microbiota of wild-type (WT) and MSTN mutant (MT) cattle. A total of 925 operational taxonomic units (OTUs) were obtained, which were classified into 11 phyla and 168 genera. Alpha diversity results showed no significant differences between MT and WT cattle. Beta diversity analyses suggested that the microbial composition of WT and MT cattle was different. Three dominant phyla and 21 dominant genera were identified. The most abundant bacterial genus had a significant relationship with the host metabolism. Moreover, various bacteria beneficial for health were found in the intestines of MT cattle. Analysis of the correlation between dominant gut bacteria and serum metabolic factors affected by MSTN mutation indicated that MSTN mutation affected the metabolism mainly by three metabolism-related bacteria, Ruminococcaceae_UCG-013, Clostridium_sensu_stricto_1, and Ruminococcaceae_UCG-010. This study provides further insight into MSTN mutation regulating the host metabolism by gut microbes and provides evidence for the safety of gene-edited animals.

Myostatin mutation causing double muscling could affect increased psoroptic mange sensitivity in dual purpose Belgian Blue cattle

Belgian Blue cattle are known for their high degree of muscling and good carcass qualities. This high degree of muscling is mainly caused by a mutation in the myostatin gene (MSTN). Although the MSTN mutation is considered as fixed in the Belgian Blue breed, segregation is occurring in a sub-population bred for dual purpose. In the latter population, we observed an association between the mutation in MSTN and susceptibility to psoroptic mange, a skin disease caused by Psoroptes ovis mites that heavily plagues Belgian Blue cattle. In total, 291 animals were sampled and screened for their susceptibility for mange lesions and their MSTN genotype. Via linear mixed modelling, we observed that homozygous mutant animals had a significant increase in the size of mange lesions (+2.51% lesion extent) compared to homozygous wild type. These findings were confirmed with zero-inflated modelling, an animal model and odds analysis. Risk ratios for developing severe mange lesions were 5.9 times as high for homozygous mutant animals. All analyses confirmed an association between the MSTN genotype and psoroptic mange lesion size.

Myostatin Mutation Promotes Glycolysis by Increasing Phosphorylation of Phosphofructokinase via Activation of PDE5A-cGMP-PKG in Cattle Heart.

Myostatin (MSTN) is a primary negative regulator of skeletal muscle mass and causes multiple metabolic changes. However, whether MSTN mutation affects heart morphology and physiology remains unclear. Myostatin mutation (MT) had no effect on cattle cardiac muscle in histological examination, but in biochemical assays, glycolysis increased in cattle hearts with MT. Compared with wild-type cattle, there were no differences in mRNA and protein levels of rate-limiting enzymes, but phosphofructokinase (PFK) phosphorylation increased in cattle hearts with MT. Transcriptome analysis showed that phosphodiesterase-5A (PDE5A), a target for inhibiting cGMP-PKG signaling, was downregulated. For the mechanism, chromatin immunoprecipitation qPCR showed that the SMAD2/SMAD3 complex in the canonical downstream pathway for MSTN combined with the promoter of PDE5A. The cGMP-PKG pathway was activated, and PKG increased phosphorylation of PFK in cattle hearts with MT. In addition, activation of PKG and the increase in PFK phosphorylation promoted glycolysis. Knockdown of PKG resulted in the opposite phenomena. The results indicated that MT potentiated PFK phosphorylation via the PDE5A-cGMP-PKG pathway and thereby promoted glycolysis in the heart.

Erythrocyte Osmotic Fragility and Hematology in Myostatin Mutant Pigs

Erythrocyte Osmotic Fragility and Hematology in Myostatin Mutant Pigs

Myostatin Mutation in Japanese Quail Increased Egg Size but Reduced Eggshell Thickness and Strength.

Simple SummaryAvian eggs provide huge benefits to both science and society by providing an important model for developmental studies and a high-quality protein source for the human diet. Especially, the hard-shell layer existing at the outer part of eggs is a unique characteristic, which is exclusive in avian species compared to other egg-laying species. Among various avian models developed to investigate genetic factors for potential industrial application, myostatin (MSTN) mutations in quail and chickens were recently generated, resulting in improved meat yield. In addition to previously reported growth and egg production traits in MSTN mutant quail, eggshell quality of mutants was further investigated in this study. Although eggshell height, width, and weight were increased by the MSTN mutation, eggshell breaking strength (EBS) and eggshell thickness were decreased in mutant eggs compared to wild-type eggs. Although these data indicated that decreased eggshell thickness contributed to decreased EBS in mutant eggs, the cellular mechanism of thinner eggshell formation in uterus by MSTN mutation needs to be further investigated using MSTN mutant quail.Recently developed myostatin (MSTN) mutant quail and chickens demonstrated similar effects of MSTN on muscle and fat developments between avian and mammalian species. However, the effect of MSTN mutation on the quality of eggshells, an important avian specific characteristic, has not yet been investigated although egg production traits of mutant quail have been studied. In this study, several parameters for eggshell quality, including eggshell size, eggshell weight, eggshell breaking strength (EBS), and eggshell thickness, were all compared between MSTN mutant and wild-type (WT) eggs. MSTN mutant eggs had greater height and width along with heavier eggshell weight compared to WT eggs, which shows proportional improvement in egg size as affected by the MSTN mutation. However, EBS and eggshell thickness were decreased in mutant eggs compared to WT eggs. In addition, the palisade layer, the thickest and most important layer for the strength of an eggshell, was also decreased without a change in the number of vesicular holes. These data indicated that decreases in the thickness of the eggshell and the palisade layer would be a main factor contributing to a lower EBS in mutant eggs. MSTN mutant quail provide a useful model to better understand the function of MSTN on avian uterine cell development and eggshell biomineralization.

Production of MSTN-mutated cattle without exogenous gene integration using CRISPR-Cas9.

Many genome-edited animals have been produced using clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9 technology to edit specific genes. However, there are few guidelines for the application of this technique to cattle. The goal of this study was to produce trait-improved cattle using the genome-editing technology CRISPR-Cas9. Myostatin (MSTN) was selected as a target locus, and synthetic mRNA of sgRNA and Cas9 were microinjected into fertilized bovine embryos in vitro. As a result, 17 healthy calves were born, and three of them showed MSTN mutation rates of 10.5%, 45.4%, and 99.9%, respectively. Importantly, the offspring with the 99.9% MSTN mutation rate had a biallelic mutation (-12bps) and a double-muscling phenotype. In conclusion, we demonstrate that the genome-editing technology CRISPR-Cas9 can produce genetically modified calves with improved traits.

Effects of Myostatin Mutation on Onset of Laying, Egg Production, Fertility, and Hatchability.

Simple SummaryPoultry can be classified as broilers for meat production and layers for egg production. Modern poultry farming improved economically important traits of broilers and layers by breeding and genetic selection. Myostatin (MSTN) has gained attention as a potential selection marker for higher meat production in the poultry industry, because MSTN mutant chickens and quail showed increased muscle mass. In this study, the effect of MSTN mutation on egg production was investigated to evaluate potential use of MSTN for higher egg production in the layer industry. MSTN homozygous mutant quail showed a significantly delayed onset of egg laying, a higher egg weight, and a lower number of eggs produced during the active laying period compared to wild-type quail. However, there were no significant differences in total egg production for 20 days, percentage proportion of egg white and yolk in egg weight, and egg fertility, and hatchability between MSTN mutant and WT quail. Although a clear benefit on egg production by MSTN mutation in quail was not revealed, this study provided useful information to understand the productive performance of MSTN mutant hens. Increased body weight and muscle mass, along with improved feed efficiency, by myostatin (MSTN) mutation in quail, supports the potential use of MSTN as a selection marker for higher meat yield in the poultry industry. Although economically important traits of broilers have been studied using recently generated MSTN mutant quail, the effect of MSTN mutation on egg production has not yet been investigated. In this study, several economically important traits of layers, including egg production, reproduction, and body composition of hens, were compared between MSTN homozygous mutant, heterozygous mutant, and wild-type (WT) quail. In terms of egg production, MSTN homozygous mutant quail, showing significantly delayed onset of egg laying, laid significantly heavier eggs, but a significantly lower number of eggs compared to WT quail for 20 days after 3 months of age, resulting in similar total egg production among groups. In addition, the percentage proportion of egg white and yolk in egg weight were similar among groups. Furthermore, similar fertility and hatchability of eggs from MSTN homozygous mutant breeding pairs and WT breeding pairs indicated normal reproductive function of MSTN mutant quail. These findings will provide scientific rationales for the consideration of MSTN as a potential selection marker for layers in the poultry industry.

CRISPR/Cas9 system-based myostatin-targeted disruption promotes somatic growth and adipogenesis in loach, Misgurnus anguillicaudatus

The mud loach (Misgurnus anguillicaudatus) is one of the most important aquaculture fishes in Asia. The application of the CRISPR/Cas9 system in the cultivation of loach strains with valuable economic traits presents great potential. Myostatin is a candidate factor that can be used for the cultivation of fast-growing loach strains. In this study, the appropriate microinjection dose of the Cas9 protein (600 pg) for genome editing was determined considering both the mutation rate and survival rate of loach embryos. With this microinjection dose of the Cas9 protein, both the exogenous GFP and endogenous myostatin genes were efficiently edited using the CRISPR/Cas9 system. Compared with that in wild-type siblings, the average body weight of the fish in 1-, 2- and 3-month-old myostatin mutant chimeric loaches was significantly increased by 34.9% (P < 0.001), 15.5% (P = 0.024) and 27.8% (P = 0.012), respectively. Mutation of mstn significantly increased the number of muscle fibers and total area of muscle fibers. Also, lipid accumulation was significantly higher in chimeric mutant loach. Additionally, the mRNA levels of myogenesis (myod and myog) and lipogenesis-related genes (scd, fabp1, fabp2 and dgat) were significantly upregulated in chimeric mutants. In particular, the mRNA level of the stearoyl-CoA desaturase gene, whose product catalyses the formation of monounsaturated fatty acids from saturated fatty acids, was increased by 23-fold (P = 0.034) and 736-fold (P < 0.05) in 3 dpf and 1-month-old chimeric loaches, respectively. The gene expression results indicated that the targeted disruption of myostatin activated myogenesis and adipogenesis in loach. Our study is highly valuable for cultivating fast-growing loach strains and understanding the specific role of myostatin in lipogenesis.

Research Note: Improved feed efficiency in quail with targeted genome editing in the myostatin gene

Increased growth rate and decreased cost of feed are main focuses to increase revenue of poultry farms. Myostatin (MSTN) is a negative regulator of muscle growth and mutation on MSTN results in increased muscle growth. Due to the antimyogenic function of MSTN, MSTN gains high attention as a potential target and genetic selection marker to increase meat yield in the livestock industry. In addition, MSTN can affect feed efficiencies and, thus decrease total feed requirement as shown in increased feed efficiencies in pigs and cattle with MSTN mutations. Although MSTN mutation in various animal species has been previously studied, MSTN mutation in avian species has only recently been generated to characterize its biological function. However, beneficial effects of MSTN mutation on poultry production need to be further investigated. In this study, using the MSTN mutant quail, feed efficiency related to interplay of changes in body weight gain (WG), feed intake (FI), and fat accretion were investigated. WG of mutant quail were significantly higher (P< 0.001) than those of wild-type (WT) from all time periods, 10-d interval from post-hatching day (D)10 to 40. Feed intake of mutant quail were significantly higher than those of WT from D 10 to 20 (P< 0.01) and D 20 to 30 (P< 0.001), but not from D 30 to 40, resulting in a significantly lower feed conversion ratio (FCR) of mutant quail compared to WT quail only from D 30 to 40 (P< 0.001). From those results, overall (D 10 to 40) FCR was significantly lower in mutant quail (P< 0.001) indicating improved feed efficiency by MSTN mutation. In addition, percentages of leg or abdominal fat compared to body weight in mutant quail at 8 wk were significantly lower than WT (P< 0.05). In combination to greater WG, less fat accretion might partially contribute to improved feed efficiency in MSTN mutant quail. As there is a current preference of meat with lower fat as a healthy food, MSTN can be used for the potential selection marker for not only bigger and leaner poultry, but also better feed efficiency that can satisfy both producers and consumers.

Prediction of Myostatin on Carcass Composition in Crossbred Lambs

Abstract In order to maintain consumer acceptance of lamb meat, producers are aiming to produce leaner lambs through breeding for certain carcass characteristics. The Texel breed is known for its superior muscling phenotype due to a myostatin mutation. Because of this mutation, Texel and Texel cross lambs have been shown to have improved carcass lean with less fat in various locations throughout the carcass. The objective of this study was to observe the impacts of different sire and dam breeds on carcass composition. Lambs (n = 41) were produced by mating two dam breeds (Southdown or Suffolk) and two sire breeds (Southdown or Texel). Lambs were harvested and a hot carcass weight was obtained. At 24 h postmortem, a chilled carcass weight was taken, and each carcass was split in half. The left side of the carcass was cut into the four primal cuts and scanned using a DXA. The right half was used for standard carcass variables. After DXA scans, each primal was weighed and the major muscles were dissected from the primal cuts and weighed. Subsamples of muscles were used for total lipid analysis and Warner-Bratzler Shear Force testing. DXA scans showed a difference in the primal cut mass (P = 0.0207) with the Suffolk-Texel cross having the highest average primal cut mass at 12.52 kg. Southdown-Southdown lambs had the highest fat percentage (dam P = 0.0398; sire P = 0.0116). Dam breed had a more significant effect on muscle toughness (P &amp;lt; 0.0035). Southdown sired lambs had a higher SFA (P = 0.0046) and MUFA (P &amp;lt; .0001) but Texel sired lambs had a higher overall fatty acid ratio (P &amp;lt; .0001). On average, the Suffolk-Texel cross was shown to have heavier average weight for the primal cuts in the rack and leg, which are where some of the more profitable cuts of meat are located.

The Blonde d'Aquitaine T3811>G3811 mutation in the myostatin gene: association with growth, carcass, and muscle phenotypes in veal calves.

The mutation T3811 → G3811 (TG3811) discovered in the myostatin gene of the Blonde d'Aquitaine breed is suspected of contributing to the outstanding muscularity of this breed. An experiment was designed to estimate the effect of this mutation in an F2 and back-cross Blonde d'Aquitaine × Holstein population. By genotyping all known mutations in the myostatin gene, it was ensured that the TG3811 mutation was indeed the only known mutation segregating in this population. Fifty-six calves (43 F2, 13 back-cross) were intensively fattened and slaughtered at 24.0 ± 1.4 wk of age. The effects of the mutation were estimated by comparing the calves with the [T/T] (n = 18), [T/G] (n = 30), and [G/G] (n = 8) genotypes. Highly significant substitution effects (P < 0.001), above + 1.2 phenotypic SD, were shown on carcass yield and muscularity scores. Birth weight (P < 0.001) was positively affected by the mutation (+0.8 SD) but not growth rate (P = 0.97), while carcass length (P = 0.03), and fatness (P ≤ 0.03) were negatively affected (-0.5 to -0.7 SD). The characteristics of the Triceps brachii muscle were affected by the mutation (P < 0.001), with lower ICDH activity (oxidative) and a higher proportion of myosin type 2X muscle fibers (fast twitch). The effects of the TG3811 mutation were similar to those of other known myostatin mutations, although the Blonde d'Aquitaine animals, which are predominantly [G/G] homozygous, do not exhibit extreme double muscling.

Sequencing and Analysis of the Myostatin Gene (GDF-8) in Bubalus bubalis Young Animals to Determine the Existence of Possible Mutations Expressed in Double Musculature Phenotype

Since the 19th century, the presentation of bovines with disproportionate muscle development have been associated with mutations that inhibit the action of the myostatin gene, it is referred to as double muscle mutation, which is common in some European Bos taurus breeds but it is not reported in buffaloes Bubalus bubalis. This study aims to evaluate if the phenotype observed in 6 young buffaloes with disproportionate muscle development has the same myostatin mutation reported in cattle. DNA was obtained from the blood of the animals of the Murrah breed. First, second and third exon was amplified end point PCR; the fragments were sequenced using capillary electrophoresis. Holstein cattle (Bos taurus) was used As control for normal phenotype. The results obtained from the comparison of the sequence of the myostatin gene show that the observed double-muscled phenotype did not show differences from normal controls. Interspecific variation was demonstrated by comparing exons two and three of the gene, finding 12 variations between the Bos taurus and Bubalus bubalis species in the evaluated fragments. It is necessary to study physiology, and the animals to explain the phenotype observed in buffaloes.

Selection for increased visual muscling increases carcass leanness without compromising predicted Meat Standards Australia eating-quality index

Context Selection using visual muscle score (MS) has been proposed to increase carcass leanness (i.e. meat yield), without compromising eating quality. Aims The aim of the present study was to examine the impact that selection for divergent MS has on live animal, commercial carcass and carcass tissue weights by using computed tomography (CT) including Meat Standards Australia (MSA) index-predicted eating quality. Methods Data from 67 steers originating from three muscling lines, namely, low, high and heterozygous high (HighHet – heterozygous for the 821 del11 myostatin mutation), were used. Visual MS was assessed on all steers. All steers were slaughtered and the left-hand side of each carcass was processed with fat trimming limited to only that required for hygiene purposes and kidney fat was not removed. All carcasses were MSA graded and then boned-out into untrimmed boneless primals (e.g. rump, cube roll). A CT scan of each beef primal was processed with image analysis software to estimate lean and fat tissue weights. The following traits were analysed: MS, weaning and slaughter weights; commercial carcass traits, including cold carcass weight, rump fat, MSA rib fat, MSA eye-muscle area, MSA marble score and MSA index; and CT-scanned compositional carcass traits, including lean, fat and bone tissues (%) and lean:bone ratio. All data were analysed with a linear mixed-effects model using REML. Least-squares means for the three muscling lines are reported. Linear trends between MS and seven carcass traits, with and without the myostatin mutation, are presented graphically. Key results Muscling line effects (P &amp;lt; 0.05) were found for visual MS and carcass traits. Linear trends between MS and carcass traits with and without the myostatin mutation demonstrate that increases in MS (P = 0.24) did not compromise predictions of MSA index even though MSA marble score decreased (P = 0.026), but myostatin decreased MSA marble score and tended to decrease MSA index (P = 0.097). Increases in the MSA eye-muscle area were associated with increases in MS (P &amp;lt; 0.01), with little effect of myostatin. Increases in MS and the myostatin mutation were both associated with increases (P &amp;lt; 0.01) in lean tissue (%) and the lean:bone ratio, and decreases (P = 0.02) in fat tissue (%). Conclusions The results indicate selection for high MS can be used to increase carcass yield, without negatively affecting MSA index predictions of eating quality. Implications Producers can use MS to identify animals with higher yields to increase carcass leanness and decrease carcass waste fat, without compromising MSA index predictions of eating quality, but should do so while considering all traits that affect profitability, in particular marble score and its association with eating quality.

Reproduction traits of heterozygous myostatin knockout sows crossbred with homozygous myostatin knockout boars

Few studies exist on homozygous myostatin gene mutant (MSTN-/- ) pigs, especially on their reproductive ability. We have previously shown that semen quality of homozygous MSTN-/- boars is comparable to that of wild type (WT). However, no data exist on the reproductive ability of heterozygous MSTN gene mutant (MSTN+/ - ) sows. The present study highlights showed that the heterozygous MSTN+/ - sows have delayed pubertal age than WT sows (255.80±6.79 versus 191.10±3.42, respectively). The number of services per pregnancy of heterozygous MSTN+/ - sows is significantly higher than that of WT sows (3.33±0.43 versus 1.60±0.25, respectively). Moreover, although heterozygous MSTN+/ - sows have natural reproduction ability, their litter size was significantly lower than that of WT sows (7.75±0.44 versus 14.25±0.60, respectively). Offsprings generated from heterozygous MSTN+/ - sow and homozygous MSTN-/- boar were genotyped with the PCR and sequencing method to detect myostatin mutation and to identify whether the piglets are homozygous MSTN-/- or heterozygous MSTN+/ - . The proportion of homozygous MSTN-/- piglets was significantly lower than that of heterozygous MSTN+/ - piglets (2.50±0.35 versus 5.25±0.60, respectively). Furthermore, none of the sows presented dystocia, and the phenotype of heterozygous MSTN+/ - piglets was normal. However, 10% homozygous MSTN-/- piglets died of dyspnoea within 2hr after birth, 60% of homozygous MSTN-/- piglets showed large tongues, and 50% had umbilical hernias. In summary, this study for the first time reports the reproduction traits of heterozygous MSTN+/ - sows crossbred with homozygous MSTN-/- boars. This study will pave the way in a new direction for the breeding and development of super lean meat varieties in the future.