Calpain Research Articles

Genetic polymorphism of the CAPN1 gene is associated with meat quality traits in Japanese quail

1. The objective of the study was to investigate the polymorphisms in two regions of the calpain 1 (CAPN1) gene and their association with breast and thigh meat quality in Japanese quail (ultimate pH (pHu), lightness, redness, yellowness, drip loss, thawing-cooking loss, water holding capacity and shear force, SF).2. Blood samples were collected randomly from 100 birds and DNA was extracted using a commercial kit. Genotypes were determined by PCR amplification followed by single-strand conformation polymorphism (SSCP) analysis. The effect of CAPN1 genotypes on meat quality traits were analysed using a general linear model (GLM) procedure.3. Genotypes of the CAPN1 gene in the first region (217-bp) analysed were significantly associated with yellowness and SF. The TT genotype showed significantly higher yellowness and lower shear force (more tenderness) than CT and CC genotypes. Genotypes of the second region of the gene (intron 4, 800-bp) were significantly associated with pHu, redness and SF of the breast meat. The BB genotype showed significantly lower pHu and redness and higher SF (lower tenderness) than other genotypes.4. Information on polymorphisms of the CAPN1 gene will eventually provide useful information for improving meat quality of Japanese quail through marker-assisted selection.

Limb-girdle Muscular Dystrophy Type 2A with Mutation in CAPN3: The First Report in Taiwan

The autosomal recessive limb-girdle muscular dystrophy type 2A (LGMD2A) is caused by mutations in the calpain 3 (CAPN3) gene, and it is characterized by selective atrophy and weakness of proximal limb and girdle muscles. We report a 33-year-old woman with initial presentations of exercise intolerance and running difficulty at age 15 years. At presentation, waddling gait, positive Gowers' sign, and marked muscle atrophy in pelvic and leg muscles were noted. Muscle computed tomography (CT) imaging demonstrated symmetric involvement of the posterior thigh muscles with relative sparing of vastus lateralis, sartorius, and gracilis. Muscle biopsy revealed a dystrophic change and many lobulated fibers on NADH-tetrazolium reductase staining. Genetic analysis of the CAPN3 gene identified a novel homozygous mutation of c2047_2050 del4, p.Lys683fs mutation, confirming the first LGMD2A patient in Taiwan.

Effect of N,N-dimethylformamide on calcium homeostasis and the calpain gene expression in human hepatocytes

To investigate the effect of N,N-dimethylformamide (DMF) on calcium homeostasis and calpain I and II gene expression in human hepatocytes (HL-7702). HL-7702 cells were exposed to different concentrations of DMF (10, 25, 50, 100, or 200 mmol/L); other HL-7702 cells, which were used as a control group, were exposed to the equal volume of DMEM; the intracellular Ca(2+) concentration was monitored using the calcium fluorescent probe (fluo-3/AM). After 24-h exposure to DMF (10, 25, 50, 100, 150, or 200 mmol/L), the morphology of hepatocytes was observed under an inverted phase contrast microscope, and the cell viability was measured by MTT assay. After 24-h exposure to DMF (10, 25, 50, 100, or 150 mmol/L), the mRNA expression levels of calpain I and II in hepatocytes were measured by real-time quantitative PCR. There were significant differences in cell viability among different exposure groups (P < 0.01); the 50, 100, 150, and 200 mmol/L DMF exposure groups had a significantly lower cell viability than the control group (P < 0.05). Under the inverted phase contrast microscope, HL-7702 cells gradually lost the original shape, with swelling and shrinking, as the dose of DMF increased, and those treated with 150 mmol/L DMF even became round and floated. The fluorescence density of fluo-3 in hepatocytes increased as the dose of DMF rose, demonstrating a dose-response relationship, and there were significant differences among these exposure groups (P < 0.05). There were significant differences in mRNA expression levels of calpain I and II among these exposure groups (P < 0.01), and the expression increased as the dose of DMF rose; but DMF did not promote the mRNA expression of calpain I at a concentration of 150 mmol/L. DMF can cause damage to hepatocytes, which is related to intracellular calcium increase and calpain mRNA increase.

A New Insight into the Role of Calpains in Post-mortem Meat Tenderization in Domestic Animals: A review.

Tenderness is the most important meat quality trait, which is determined by intracellular environment and extracellular matrix. Particularly, specific protein degradation and protein modification can disrupt the architecture and integrity of muscle cells so that improves the meat tenderness. Endogenous proteolytic systems are responsible for modifying proteinases as well as the meat tenderization. Abundant evidence has testified that calpains (CAPNs) including calpain I (CAPN1) and calpastatin (CAST) have the closest relationship with tenderness in livestock. They are involved in a wide range of physiological processes including muscle growth and differentiation, pathological conditions and post-mortem meat aging. Whereas, Calpain3 (CAPN3) has been established as an important activating enzyme specifically expressed in livestock’s skeletal muscle, but its role in domestic animals meat tenderization remains controversial. In this review, we summarize the role of CAPN1, calpain II (CAPN2) and CAST in post-mortem meat tenderization, and analyse the relationship between CAPN3 and tenderness in domestic animals. Besides, the possible mechanism affecting post-mortem meat aging and improving meat tenderization, and current possible causes responsible for divergence (whether CAPN3 contributes to animal meat tenderization or not) are inferred. Only the possible mechanism of CAPN3 in meat tenderization has been confirmed, while its exact role still needs to be studied further.

Calpastatin gene is not associated with late onset sporadic Parkinson’s disease in Chinese Han population

Objective To explore the association between late-onset sporadic Parkinson’s disease (PD) and single nucleotide polymorphisms (SNPs) of Ca2+ -dependent protease calpain inhibitor calpastatin (CAST) gene in a Chinese Han population. Methods 370 evaluable patients (221 male, 149 female) with PD (mean age 65.2±8.5 years) and 390 neurologically healthy controls (208 male, 182 female) matched for gender, ethnicity, and area of residence. PD cases were identified from the PD cohort of the Chinese National Consortium on Neurodegenerative Diseases (www.chinapd.cn). A total of 24 tag-SNPs were genotyped capturing 95% of the genetic variation across the CAST gene. Results There was no association found between any of the polymorphisms and PD in all models tested (co-dominant, dominant-effect and recessive-effect(P>0.05)). Similarly, none of the common haplotypes was associated with a risk for PD(P>0.05). Conclusion Results show no significant association between the CAST gene polymorphisms and late onset sporadic PD in the present population. Key words: Calpastatin; Parkinson’s disease; Single nucleotide polymorphisms

Identification and association of polymorphisms in CAPN1 and CAPN3 candidate genes related to performance and meat quality traits in chickens

Meat quality is an important feature for the poultry industry and is associated with consumer satisfaction. The calpain 1 (CAPN1) gene is related to the tenderness process of meat post- mortem, and the calpain 3 (CAPN3) gene plays an important role in myofibrillar organization and growth. The objective of the present study was to identify polymorphisms in these genes and to determine the association between these polymorphisms and traits of economic interest in poultry. Eleven animals (F₁) from an experimental poultry population at Embrapa Swine and Poultry were used to identify the polymorphisms. Four single nucleotide polymorphisms (SNPs) were found in the CAPN1 gene, and one SNP was found in the CAPN3 gene. A polymorphism from each gene was selected for genotyping in 152 chickens from the Embrapa F₂ experimental population and 311 chickens from a commercial population. Polymorphism g.2554T>C (CAPN1) was associated with body weight at 35 to 42 days, thigh weight, breast weight, carcass weight, and meat lightness content. SNP g.15486C>T (CAPN3) was associated with thigh yield, thawing-cooking loss, and shear force. Results suggest the possibility of using molecular markers in CAPN1 and CAPN3 genes as a tool for performance and meat quality traits in poultry breeding programs.

Associations of Polymorphisms in Four Candidate Genes with Carcass and/or Meat-Quality Traits in Two Meat-Type Chicken Lines

The associations between polymorphisms of five genes, calpain 1 ( CAPN1 ), follicle stimulating hormone beta (FSHB), follicle stimulating hormone receptor (FSHR), peroxisome proliferator-activated receptor gamma (PPARG), and retinol binding protein 7 (RBP7), and live weight, carcass composition, and meat-quality traits were estimated from two meat-type chickens lines (n = 311). Except for the variants of the FSHR gene, 11 SNPs of the other four genes and two diplotypes of PPARG were associated with one or more traits excluding shear factor (SF). SNP C31566680T of the CAPN1 gene was significantly associated with live weight (LW) carcass traits. The SNP A4580859C of FSHB gene was significantly associated with breast muscle weight (BrW) and LW. One of the PPARG SNPs, C5070948T, was associated with intramuscular fat content in breast (IMF br ). Diplotype P1 of the PPARG gene was significantly associated with LW and all carcass traits. P3 were significantly associated with abdominal fat weight (AbFW). SNPs in RBP7 were only associated with BrW. These results indicate that the four genes were associated with these traits and have promise as genetic markers for future marker-assisted selection. Supplementary materials for this paper are available online.

Calpain and calpastatin activity post mortem and meat tenderness

Calpain and calpastatin activity post mortem and meat tenderness

The effects of age, energy and protein intake on protein turnover and the expression of proteolysis-related genes in the broiler breeder hen

A study was conducted to determine the changes that occur to proteolysis and related genes due to age, protein, and energy intake in high-yield broiler breeder hens (Gallus gallus). Cobb 700 broiler breeders were randomly assigned to one of six diets in a 2×3 factorial fashion. Two levels of energy (390 and 450 kcal/day) and three levels of protein (22, 24, and 26 g CP/day) were utilized. Protein turnover was determined in the left pectoralis at 22, 26, 31 and 44 weeks. Relative mRNA expression of calpain 2 (CAPN2), proteasome C2 subunit (PSMA1), and F box protein 32 (FBXO32) were determined via RT-PCR at 20, 25, and 44 weeks. Contrasts indicate fractional synthesis rate (FSR) and FBXO32 increase to a maximum at 25-26 weeks and a decrease thereafter. A significant drop in PSMA1 and FBXO32 was observed between 25 and 44 weeks and matched the decrease observed in FBR. No differences were detected in the levels of fractional synthesis and degradation, or the expression of CAPN2, PSMA1, and FBXO32, due to protein or energy intake. In summary, protein turnover was upregulated during the transition into sexual maturity and decreased thereafter. The observed changes in degradation appeared to be mediated by the ubiquitin-proteasome pathway.

Erratum to: RVG-Mediated Calpain2 Gene Silencing in the Brain Impairs Learning and Memory

Kumar, P., Wu, H., McBride, J. L., Jung, K. E., Kim, M. H.,Davidson, B. L., Lee, S. K., Shankar, P., & Manjunath N. (2007).Transvascular delivery of small interfering RNA to the centralnervous system. Nature, 448, 39–43.The online version of the original article can be found underdoi:10.1007/s12017-012-8196-8.S. Zadran (&)Division of Biology, California Institute of Technology,1200 East California Blvd, Pasadena, CA 91125, USAe-mail: szadran@mednet.ucla.eduG. Akopian H. Zadran J. Walsh M. BaudryDepartment of Biological Sciences, University of SouthernCalifornia, 3641 Watt Way, HNB 514, Los Angeles, CA 90089,USA

Associations between candidate SNPs in the calpain 1, calpastatin and leptin genes and meat tenderness among Swedish beef populations

The contribution of three candidate genes to the variation in meat tenderness was tested in muscle samples from 243 pure-bred, young, beef bulls of Angus, Charolais, Hereford, Limousin and Simmental breeds, raised in Swedish commercial herds. The animals were genotyped for single nucleotide polymorphisms (SNPs) in the calpain 1 (CAPN1), calpastatin (CAST) and leptin genes. The frequent calpain 1 CAPN1:c.947G>C G allele showed an unfavourable association with tenderness. The calpastatin CAST:c.155C>T T allele, which was the most common allele, showed a favourable association with Warner–Bratzler shear force (WBSF) and compression tests. An association was observed between the leptin UASMS2C>T SNP and compression tests.

Genetic variability and clinical spectrum of Chinese patients with limb‐girdle muscular dystrophy type 2A

Previous studies of limb-girdle muscular dystrophy type 2A (LGMD2A) patients in many countries have suggested a heterogeneous genetic and clinical spectrum, but the genotypes and phenotypes of Chinese LGMD2A patients remain unclear. We directly screened calpain-3 (CAPN3) in 18 Chinese Han subjects who exhibited severely reduced or completely absent calpain-3 expression, as determined by Western blot analysis. We subsequently analyzed genotype/phenotype correlations. Seventeen patients (94.4%) were identified who had at least 1 causative mutation. All 18 mutations were distributed along the entire gene, and 11 of the mutations were novel, including 4 missense mutations, 5 deletions, and 2 splicing mutations. The phenotypes of these Chinese LGMD2A patients varied from severe LGMD to distal myopathy, and even asymptomatic hyper-CK-emia. No evidential correlation was found between the genotypes and phenotypes of the patients assessed in this study. Western blot analysis is still a useful diagnostic method when genetic analysis is unavailable.

Targeted suppression of μ-calpain and caspase 9 expression and its effect on caspase 3 and caspase 7 in satellite cells of Korean Hanwoo cattle

The calpains play an important role in cell death and cell signalling. Caspases catalyse wholesale destruction of cellular proteins which is a major cause of cellular death. The current study looks at the function of μ-calpain and caspase 9, using RNAi (RNA interference)-mediated silencing, and to observe the mRNA expression level of caspase genes during satellite cell growth. The satellite cells were treated with siRNA (small interfering RNA) of μ-calpain and caspase 9 separately. There was reduction of 16 and 24% in CAPN1 (calpain1)-siRNA2 and CAPN1-siRNA3 transfected cells respectively, whereas it was 60 and 56% in CAPN1-siRNA1 and CAPN1-siRNA4 transfected cells respectively. CAPN1-siRNA4 and CAPN1-siRNA1 treated cells showed more reduction in caspase 3 and 7 gene expression. CARD9 (caspase recruitment domain 9)-siRNA1 and CARD9-siRNA2-treated cells showed reduction of 40 and 49% respectively. CARD9-siRNA1 and CARD9-siRNA2 showed an increase in caspase 3 gene expression, whereas CARD9-siRNA2 showed reduction in caspase 7 gene expression. These results suggest a strong cross-talk between μ-calpain and the caspase enzyme systems. Suppression of target genes, such as μ-calpain and caspase 9, might have genuine potential in the treatment of skeletal muscle atrophy.

In ovoleptin administration accelerates post‐hatch muscle growth and changes myofibre characteristics, gene expression and enzymes activity in broiler chickens

To evaluate the effect of maternal leptin on muscle growth, we injected 0 μg (control, CON), 0.5 μg (low leptin dose, LL) or 5.0 μg (high leptin dose, HL) of recombinant murine leptin dissolved in 100 μl of PBS into the albumen of broiler eggs prior to incubation. The newly hatched chicks were all raised under the same conditions until 21 days of age (D21), when body weight was measured and samples of gastrocnemius muscle were collected and weighed. Myosin ATPase staining was applied to identify myofibre types and measure the cross-sectional area (CSA) of myofibres. Real-time PCR was performed to quantify leptin receptor (LEPR), insulin-like growth factor 1 (IGF-1), IGF-1 receptor (IGF-1R), growth hormone receptor (GHR) and myostatin (MSTN) mRNA expression in the gastrocnemius muscle. The activity of calpains (CAPNs) in the gastrocnemius muscle was measured using a quantitative fluorescence detection kit. Male chickens treated with both high and low doses of leptin had significantly higher (p < 0.05) body weight on D21. The high leptin significantly increased the CSA (p < 0.05) of gastrocnemius muscle in male chickens, which coincided with a 93% increase (p < 0.05) in IGF-1 mRNA expression. Likewise, the LL dose increased the weight of gastrocnemius muscle in male chickens (p < 0.05), which was accompanied by a 41% down-regulation (p < 0.05) of MSTN mRNA expression and a decreased activity of CAPNs. However, all these changes were not observed in female chickens. The proportion of myofibre types did not altered. No significant change was detected for LEPR and GHR mRNA expression. These results indicate that in ovo leptin treatment affects skeletal muscle growth in chickens in a dose-dependent and sex-specific manner. The altered expression of IGF-1, MSTN mRNA and activity of CAPNs in skeletal muscle may be responsible for such effects.

The effect of troglitazone on lipid accumulation and related gene expression in Hanwoo muscle satellite cell

The current study was undertaken to determine the effect of the troglitazone (TGZ) on the expression of peroxisome proliferator-activating receptor (PPARγ), CCAAT/enhancer-binding protein, fatty acid binding protein 4, calpain 1 (CAPN1), and lipid accumulation in the myotube of Hanwoo muscle satellite cells. The satellite cells were treated with 5, 10, and 50μM of TGZ for indicated time intervals. TGZ promoted the trans-differentiation with significant increase in glycerol accumulation. Polymerase chain reaction (PCR) and microarray results indicated that the TGZ treatment significantly increased the expression of adipogenic transcription factors. TGZ (10 and 50μM) increased the CAPN1 gene expression 2.2- and 2.6-fold in real-time polymerase chain reaction analysis and 0.52- and 0.25-fold in microarray analysis, respectively, when compared with their respective controls. This result suggests that CAPN1 gene might be involved in the adipogenic differentiation programs. In addition, 13 genes were upregulated and 12 genes were downregulated in microarray analysis. Most of the up/downregulated genes were directly linked with adipogenesis.

Calpains mediate epithelial-cell death during mammary gland involution: mitochondria and lysosomal destabilization

Our aim was to elucidate the physiological role of calpains (CAPN) in mammary gland involution. Both CAPN-1 and -2 were induced after weaning and its activity increased in isolated mitochondria and lysosomes. CAPN activation within the mitochondria could trigger the release of cytochrome c and other pro-apoptotic factors, whereas in lysosomes it might be essential for tissue remodeling by releasing cathepsins into the cytosol. Immunohistochemical analysis localized CAPNs mainly at the luminal side of alveoli. During weaning, CAPNs translocate to the lysosomes processing membrane proteins. To identify these substrates, lysosomal fractions were treated with recombinant CAPN and cleaved products were identified by 2D-DIGE. The subunit b(2) of the v-type H(+) ATPase is proteolyzed and so is the lysosomal-associated membrane protein 2a (LAMP2a). Both proteins are also cleaved in vivo. Furthermore, LAMP2a cleavage was confirmed in vitro by addition of CAPNs to isolated lysosomes and several CAPN inhibitors prevented it. Finally, in vivo inhibition of CAPN1 in 72-h-weaned mice decreased LAMP2a cleavage. Indeed, calpeptin-treated mice showed a substantial delay in tissue remodeling and involution of the mammary gland. These results suggest that CAPNs are responsible for mitochondrial and lysosomal membrane permeabilization, supporting the idea that lysosomal-mediated cell death is a new hallmark of mammary gland involution.

Abstract 4803: Quantitative proteome profiling of CD4+CD25+CCR4+ T-cells to identify potential therapeutic targets for adult T-cell leukemia (ATL) and Human T-lymphotropic virus type-1 associated myelopathy (HAM)

Abstract Human T-cell leukemia virus type 1 (HTLV-1) is a retro-viruse known as the one of the causative factors of adult T-cell leukemia/lymphoma (ATL) and HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP), both of which are developed after a long latency period over 30 years. ATL is developed in HTLV-1-infected individuals at the prevalence of 5% with poor prognosis. Meanwhile, HAM/TSP has a prevalence of approximately 0.25% of HTLV-1-infected individuals and is characterized by its neurological symptoms, such as sensory disturbance of lower extremities. Regardless of the importance of early diagnosis and appropriate therapeutic intervention for the patients, the diagnostic markers or the effective molecular treatment targets have not been discovered yet. The objective of the present study is to identify determinants responsible for the development of ATL or HAM/TSP, which could be useful as the potential therapeutic targets or prognostic biomarkers. Here we performed a quantitative proteomic profiling of CD4+CD25+CCR4+ T-cells, which are known as the predominant viral reservoir, by means of label-free quantification analysis on the Expressionist proteome server. CD4+CD25+CCR4+ T cells were isolated from the peripheral blood mononuclear cells derived from 6 uninfected volunteers, 5 asymptomatic carriers, 9 HAM/TSP patients, and 9 ATL patients. The isolated T cells from 29 individuals were lysed, digested, and subjected to LC/MS/MS analyses. Subsequently, we integrated the 29 LC/MS/MS data on the Expressionist proteome server and conducted label-free quantification analysis, obtaining 14,064 non-redundant peptides. Kruskal-Wallis test and leave-one-out cross validation test finally extracted 20 peptides derived from 17 distinct proteins, which allowed statistical classification of four pathological groups. The 17 candidates included a couple of proteins which are well known as an apoptotic enzyme-substrate pair; Calpain-2 (CAN2) and Spectrin alpha chain (SPTA2). Our mass spectrometric quantification data demonstrated that CAN2 expression was significantly down-regulated in CD4+CD25+CCR4+ cells from ATL patients, while SPTA2 was abnormally accumulated. Indeed, exogenous overexpression of CAN2 remarkably induced apoptosis to ATL cell line SO-4 (expressing high level of SPTA2) but not in KOB cells (SPTA2 negative). These observations indicate that CAN2 and SPTA2 coordinately regulate the apoptosis and the suppression of CAN2 in HTLV-1 infected T-cells would be one of the plausible causes of ATL onset. Our label-free quantification-based proteome profiling of CD4+CD25+CCR4+ T cells enabled us to identify 17 pathological determinants for HAM/TSP and ATL, which could contribute to developing new strategies for treatment and diagnosis in the future. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4803. doi:1538-7445.AM2012-4803

Impaired calcium calmodulin kinase signaling and muscle adaptation response in the absence of calpain 3

Mutations in the non-lysosomal, cysteine protease calpain 3 (CAPN3) result in the disease limb girdle muscular dystrophy type 2A (LGMD2A). CAPN3 is localized to several subcellular compartments, including triads, where it plays a structural, rather than a proteolytic, role. In the absence of CAPN3, several triad components are reduced, including the major Ca(2+) release channel, ryanodine receptor (RyR). Furthermore, Ca(2+) release upon excitation is impaired in the absence of CAPN3. In the present study, we show that Ca-calmodulin protein kinase II (CaMKII) signaling is compromised in CAPN3 knockout (C3KO) mice. The CaMK pathway has been previously implicated in promoting the slow skeletal muscle phenotype. As expected, the decrease in CaMKII signaling that was observed in the absence of CAPN3 is associated with a reduction in the slow versus fast muscle fiber phenotype. We show that muscles of WT mice subjected to exercise training activate the CaMKII signaling pathway and increase expression of the slow form of myosin; however, muscles of C3KO mice do not exhibit these adaptive changes to exercise. These data strongly suggest that skeletal muscle's adaptive response to functional demand is compromised in the absence of CAPN3. In agreement with our mouse studies, RyR levels were also decreased in biopsies from LGMD2A patients. Moreover, we observed a preferential pathological involvement of slow fibers in LGMD2A biopsies. Thus, impaired CaMKII signaling and, as a result, a weakened muscle adaptation response identify a novel mechanism that may underlie LGMD2A and suggest a pharmacological target that should be explored for therapy.

Functional cleavage of selenoprotein K is regulated by calpain/calpastation system in toll‐like receptor activated macrophages

Calpains (CAPN) are proteolytic enzymes that modulate cellular function through cleavage of targets. Calpastatin (CAST) is a key endogenous inhibitor of CAPN, thereby modifying its actions. An important role is emerging for CAPN/CAST system in regulating immune responses. Herein, we identify a novel target of CAPN, selenoprotein K (SelK), which is important for Ca2+ flux in immune cells. CAPN‐mediated cleavage of SelK was detected in myeloid cells but not in lymphoid cells. Both m‐ and μ‐CAPN were capable of cleaving SelK, but m‐CAPN was the predominant isoform. Specific inhibitors of CAPN were used to show that m‐CAPN cleaved SelK in macrophages. The cleavage site in SelK was identified between Arg81 and Gly82 and the resulting truncated SelK was shown to lack selenocysteine, the amino acid that defines selenoproteins. Resting macrophages predominantly expressed cleaved SelK and, when activated through Toll‐like receptors (TLRs), SelK cleavage was inhibited. We found that decreased CAPN cleavage was due to TLR‐induced up‐regulation of CAST. CAST knock down resulted in higher CAPN activity and sustained SelK cleavage in TLR activated macrophage. Moreover, the expression of the CAPN and CAST in macrophages was different from T and B cells. Overall, our findings identify SelK as a novel CAPN target and reveal dynamic changes in functional cleavage of SelK is regulated by CAPN/CAST system in TLR activated macrophages.

Production and processing studies on calpain-system gene markers for beef tenderness: Consumer assessments of eating quality1

We investigated the effects of calpain-system genetic markers on consumer beef quality ratings, including interactions of marker effects with hormonal growth promotant (HGP) use and tenderstretch hanging. Brahman cattle in New South Wales (NSW; n = 164) and Western Australia (WA; n = 141) were selected at weaning from commercial and research herds to achieve balance and divergence in calpastatin (CAST) and calpain 3 (CAPN3) gene marker status. Genotypes for μ-calpain (CAPN1-4751 and CAPN1-316) were also determined. Angus cattle (49 in NSW, 17 in WA) with favorable CAST and CAPN3 alleles, balanced for CAPN1-316 status, were also studied. Half the cattle at each site had HGP (Revalor-H, containing 200 mg trenbolone acetate and 20 mg 17β-estradiol) implants during grain finishing. One side of each carcass was suspended from the Achilles tendon (AT) and the other from the pelvis [tenderstretch (TS)]. Meat Standards Australia consumer panels scored 7-d aged striploin steaks from both AT and TS sides, and 7-d aged rump and oyster blade steaks from the AT side of each carcass. Two favorable CAST alleles increased tenderness ratings of AT-striploin, TS-striploin, rump, and oyster blade steaks by, respectively, 6.1, 4.2, 4.2, and 3.1 units, and overall liking by 4.7, 2.8, 2.9, 3.7 (all P < 0.04). Two favorable CAPN1-4751 alleles increased tenderness of AT-striploin, TS-striploin, and rump steaks by 6.5, 4.3, and 3.9 units, and overall liking by 5.6, 3.1, and 4.1 units. Two favorable CAPN3 alleles improved rump steaks by 3.7, 3.3, 3.7, and 3.5 units, for tenderness, juiciness, liking the flavor, and overall liking. There were no significant CAPN1-316 effects. The effect of HGP was greatest for the AT-striploin (reducing tenderness and overall liking by 8.2 units, P < 0.001), then TS-striploin (-5.6 for tenderness, -5.0 for overall liking, P < 0.001), and then rump (-4.4 for tenderness, -3.3 for overall liking, P < 0.007). Processing conditions differed considerably between NSW and WA. Rump steaks from NSW scored about 10 units greater than those from WA, but Angus and Brahman steaks from the same location with the same marker alleles had similar scores. In contrast, NSW Angus striploin steaks scored about 15 units greater for tenderness and overall liking (P < 0.001) than cattle with the same marker alleles at the other 3 location × breed combinations, which had generally similar scores. Therefore, calpain-system gene markers have beneficial effects on eating quality, consistent with our previous findings for objective meat quality.