Calpain Gene Research Articles

ANALYSIS OF THE GENETIC STRUCTURE OF THE POLISH BEEF BREED BY MOLECULAR MARKERS OF MEAT QUALITY INDICATORS PRODUCTIVITY

The results of determining the genotypes of Polish beef breed animals based on polymorphism of the thyroglobulin (TG 5) and calpain (CAPN1 530) genes are given. The PCR-PDRF method was used for the analysis. Based on the obtained data, the specifics of the genetic structure of the studied population were established based on the distribution of frequencies of alleles and genotypes. For the thyroglobulin gene, animals with the homozygous SS genotype had a quantitative advantage, which led to a high concentration of the C allele (0.775). For the calpain gene, a high frequency of the desired G allele was found in the studied group of animals (0.539). Based on the results of the analysis, the prospects of the researched group of animals of the Polish meat breed were determined for establishing an associative relationship of various allelic variants of the thyroglobulin and calpain genes with the parameters of the quality characteristics of the final product – marbling and tenderness of meat.

Calpain 1 gene expression in liver tissue and the association of novel calpain 1 single-nucleotide polymorphisms (SNPs) with meat quality in Bali cattle

Abstract. Calpain 1 (CAPN1) is an enzyme that influences meat tenderization, and it is involved in post mortem proteolysis. The bovine CAPN1 gene was chosen as a candidate gene for DNA sequencing to identify novel single-nucleotide polymorphisms (SNPs) in exons 8–10 and assess their associations with meat quality in Bali cattle. In an analysis of 95 Bali cattle, two novel SNPs (g.5327C>T and g.5959C>T) were identified in exons and four novel SNPs (g.5534C>T, g.5807A>C, g.5857G>A, and g.5869T>C) were discovered in introns. Allelic frequency was evaluated, and Hardy–Weinberg equilibrium was found for all SNPs in Bali cattle. The marbling score and intramuscular fat content as determined by ultrasound were associated with g.5869T>C and g.5959C>T. g.5327C>T was associated with pH and meat color in Bali cattle, whereas two other SNPs (g.5869T>C and g.5959C>T) were associated with Warner–Bratzler shear force and meat color. Furthermore, g.5869T>C was associated with linolenic acid content, and g.5959C>T with caprylic and linolenic acid levels. However, intronic SNPs (g.5534C>T, g.5807A>C, and g.5857G>A) did not significantly affect meat quality in Bali cattle. Quantitative real-time PCR of liver tissue revealed that the mRNA expression of CAPN1 significantly differed (P<0.05) among the CT, CC, and TT genotypes. The results suggest that genetic variability in loci within CAPN1 might be associated with meat quality in Bali cattle.

Identification of novel pathogenic variants of Calpain-3 gene in limb girdle muscular dystrophy R1

BackgroundLimb Girdle Muscular Dystrophy R1 (LGMDR1) is an autosomal recessive neuromuscular disease caused by mutations in the calpain-3 (CAPN3) gene. As clinical and pathological features may overlap with other types of LGMD, therefore definite molecular diagnosis is required to understand the progression of this debilitating disease. This study aims to identify novel variants of CAPN3 gene in LGMDR1 patients.ResultsThirty-four patients with clinical and histopathological features suggestive of LGMD were studied. The muscle biopsy samples were evaluated using Enzyme histochemistry, Immunohistochemistry, followed by Western Blotting and Sanger sequencing. Out of 34 LGMD cases, 13 patients were diagnosed as LGMDR1 by immunoblot analysis, demonstrating reduced or absent calpain-3 protein as compared to controls. Variants of CAPN3 gene were also found and pathogenicity was predicted using in-silico prediction tools. The CAPN3 gene variants found in this study, included, two missense variants [CAPN3: c.1189T > C, CAPN3: c.2338G > C], one insertion-deletion [c.1688delinsTC], one splice site variant [c.2051-1G > T], and one nonsense variant [c.1939G > T; p.Glu647Ter].ConclusionsWe confirmed 6 patients as LGMDR1 (with CAPN3 variants) from our cohort and calpain-3 protein expression was significantly reduced by immunoblot analysis as compared to control. Besides the previously known variants, our study found two novel variants in CAPN3 gene by Sanger sequencing-based approach indicating that genetic variants in LGMDR1 patients may help to understand the etiology of the disease and future prognostication.

Genetic polymorphisms of calpain1 and calpain3 genes and their effects on growth, carcass, and meat quality traits in Betong chicken (KU line).

Betong chicken (KU line) is a slow-growing Thai native chicken used for meat production. The objectives of this study were to identify polymorphisms of the calpain1 (CAPN1) and calpain3 (CAPN3) genes and to investigate their effects on growth, carcass, and meat quality traits in Betong chickens (KU line). A sample of 252 Betong chickens (KU line) was screened for CAPN1 and CAPN3 polymorphisms. The polymorphisms of CAPN1 were detected using gel electrophoresis and DNA sequencing, whereas the polymorphisms of CAPN3 were identified using restriction fragment length polymorphism. Polymorphisms were detected in both CAPN1 (AA, AB, and BB genotypes) and CAPN3 (CC, CT, and TT genotypes). The frequency of the B allele was higher than for the A allele (0.78 and 0.22, respectively) in CAPN1, while the C allelic frequency was higher than for the T allele (0.54 and 0.46, respectively) in CAPN3. The CAPN1 genotype and the combination of the CAPN1 and CAPN3 genotypes could be used as genetic markers for meat lightness. The CAPN3 could be useful for increasing body weight, live weight, and breast meat weight in Betong chickens (KU line).

Genotypic and allelic variability of the calpain CAPN1-316 gene in the Tropical Milking criollo breed and indication of its meat tenderness

Background: Genes and their variants related to milk and beef production in cattle can be identified through molecular markers. In particular, the Calpain gene CAPN1-316 has been linked to meat tenderness. The males of the Tropical Milking Criollo cattle not assigned as breeding animals are destined for commercial beef production. Objective: To estimate the genotypic and allelic frequencies in the calpain gene CAPN1-316 and its relation to meat tenderness in the Tropical Milking criollo breed. Methods: Molecular marker microsatellites of single sequence repeats (SSR) from DNA were extracted from blood samples of 423 purebred animals from three farms to identify the variants of the CAPN1-316 gene using the polymerase chain reaction (PCR). Genotypic and Allelic frequencies within populations, genetic distance, heterozygosity, and Xi squared (X2) test were performed using Pop Gene V.4. Results: The genotypic frequencies of the whole population were CC 0.31, GG 0.18, and CG 0.51. The allelic frequencies of variant C per farm were 0.62, 0.60, and 0.52. The genotypic and allelic frequencies of the CAPN1-316 polymorphisms were in Hardy-Weinberg equilibrium (X2, p>0.05). The CC genotype showed indication of greater tenderness. Conclusion: In the Tropical Milking criollo cattle, the C variant of the CAPN1-316, favorable to meat tenderness, had a bigger frequency than the G variant.

Polymorphisms of Calpain and Calpastatin Cascade Genes in the Beef Cattle Population

Polymorphisms of Calpain and Calpastatin Cascade Genes in the Beef Cattle Population

Baicalin attenuates pulmonary vascular remodeling by inhibiting calpain-1 mediated endothelial-to-mesenchymal transition

BackgroundPrevious studies have demonstrated the beneficial effect of baicalin on pulmonary arterial hypertension (PAH), but the mechanism is unclear. AimThe aim of the present study was to evaluate the effect of baicalin on pulmonary vascular remodeling (PVR) with a focus on calpain-1-mediated endothelial-to-mesenchymal transition (EndMT). MethodsPAH was induced by intraperitoneal injection of monocrotaline (MCT) in rats and hypoxia in calpain-1 gene knockout (Capn1−/−) and wild-type C57BL/6 mice. An in vitro PVR model was established in PASMCs and HPAECs. ResultsThe data showed that baicalin treatment and calpain-1 inhibition alleviated MCT and hypoxia-induced increases in right ventricular systolic pressure (RVSP), prevented right ventricle hypertrophy and PVR, and attenuated cardiopulmonary fibrosis. Moreover, baicalin ameliorated PAH-induced EndMT, as evidenced by the suppressed expression of mesenchymal markers vimentin, and α-SMA and restored expression of endothelial markers CD31, and VE-cadherin. In vitro studies showed that baicalin treatment blocked TGF-β1-induced EndMT in HPAECs and abolished hypoxia-induced PASMC proliferation and migration. All the beneficial effects of baicalin on PVR in vitro and in vivo were accompanied by suppressed calpain-1 expression. Further study demonstrated that baicalin treatment and calpain-1 inhibition inhibited the enhanced expression of PI3K and p-AKT both in vitro and in vivo. ConclusionsIn conclusion, baicalin treatment attenuates PVR by inhibiting calpain-1 and PI3K/Akt-mediated EndMT.

Family and literature analysis demonstrates phenotypic effect of two variants in the calpain-3 gene

Both, recessive (LGMD R1) and dominant (LGMD D4) inheritance occur in calpain 3-related muscular dystrophy. We report a family with calpain-related muscular dystrophy caused by two known variants in the calpain 3 gene (CAPN3, NM_000070.3; (I) c.700G>A, p.Gly234Arg and (II) c.1746-20C>G, p.?). Three family members are compound heterozygous and exhibit a relatively homogeneous phenotype characterized by progressive proximal weakness starting in the third to fourth decade of life in the shoulder girdle and spreading to the legs. Two family members affected only by the p.Gly234Arg heterozygous missense variants show a different phenotype characterized by severe exertional myalgia without overt pareses. We conclude that in our family, the missense variant causes a severe myalgic phenotype without pareses that is aggravated by the second intronic variant and put these findings in the context of previous studies of the same variants.

Analisis Perbandingan Sekuens Gen Calpain 1 (CAPN1) pada Sapi, Kerbau, dan Kambing Berdasarkan Database Genbank

Livestock genetic improvement is the main purpose of animal breeding program. Biotechnology brings novel method to improve the animal breeding program qualitatively and quantitatively. Identification of gene sequence of polymorphism are used widely to analyze gene sequence to understand genetic variation. Calpain 1 (CAPN1) gene is the gene that involve in the process of improving meat quality such as marbling and tenderness of the livestock including the cattle, buffalo, and goat species. CAPN1 is responsible in encoding the µ-calpain that regulate postmortem proteolysis. This study collect the gene database including calpain 1 [Bos taurus (cattle)] NC_037356.1, calpain 1 [Bos indicus (zebu cattle)] NC_032678.1, calpain 1 [Bos indicus x Bos taurus (hybrid cattle)] NC_040104.1, calpain 1 [Bubalus bubalis (water buffalo)] NC_059161.1 calpain 1 [Capra hircus (goat)] NC_030836.1 through NCBI. Those collected gene were analyze using the Clustal W alignment, MEGA 11, and BLAST program. The results indicate that the cattle, buffalo, and goat species CAPN1 gene of the 5’ UTR, Intron, Exon, dan 3’ UTR region show 9 SNPs, 1691 SNPs, 153 SNPs, and 197 SNPs, respectively. Total of the identified SNPs within the the cattle, buffalo, and goat species CAPN1 gene comparison are 2050. This study can be used for the initial report in order to develop further research related to CAPN1 gene for the livestock ruminant.

Selection signature analysis and genome-wide divergence of South African Merino breeds from their founders.

Merino sheep are a breed of choice across the world, popularly kept for their wool and mutton value. They are often reared as a pure breed or used in crossbreeding and are a common component in synthetic breed development. This study evaluated genetic diversity, population structure, and breed divergence in 279 animals of Merino and Merino-based sheep breeds in South Africa using the Illumina Ovine SNP 50K BeadChip. The sheep breeds analysed included the three Merino-derived breeds of Dohne Merino (n = 50); Meatmaster (n = 47); and Afrino (n = 52) and five presumed ancestral populations of Merinos (Merino (n = 46); South African Merino (n = 10); and South African Mutton Merino (n = 8)); and the non-Merino founding breeds of Damara (n = 20); Ronderib Afrikaner (n = 17); and Nguni (n = 29). Highest genetic diversity values were observed in the Dohne Merino (DM), with H o = 0.39 ± 0.01, followed by the Meatmaster and South African Merino (SAM), with H o = 0.37 ± 0.03. The level of inbreeding ranged from 0.0 ± 0.02 (DM) to 0.27 ± 0.05 (Nguni). Analysis of molecular variance (AMOVA) showed high within-population variance (>80%) across all population categories. The first principal component (PC1) separated the Merino, South African Mutton Merino (SAMM), DM, and Afrino (AFR) from the Meatmaster, Damara, Nguni, and Ronderib Afrikaner (RDA). PC2 aligned each Merino-derived breed with its presumed ancestors and separated the SAMM from the Merino and SAM. The iHS analysis yielded selection sweeps across the AFR (12 sweeps), Meatmaster (four sweeps), and DM (29 sweeps). Hair/wool trait genes such as FGF12; metabolic genes of ICA1, NXPH1, and GPR171; and immune response genes of IL22, IL26, IFNAR1, and IL10RB were reported. Other genes include HMGA, which was observed as selection signatures in other populations; WNT5A, important in the development of the skeleton and mammary glands; ANTXR2, associated with adaptation to variation in climatic conditions; and BMP2, which has been reported as strongly selected in both fat-tailed and thin-tailed sheep. The DM vs. SAMM shared all six sweep regions on chromosomes 1, 10, and 11 with AFR vs. SAMM. Genes such as FGF12 on OAR 1:191.3-194.7Mb and MAP2K4 on OAR 11:28.6-31.3Mb were observed. The selection sweep on chromosome 10 region 28.6-30.3Mb harbouring the RXFP2 for polledness was shared between the DM vs. Merino, the Meatmaster vs. Merino, and the Meatmaster vs. Nguni. The DM vs. Merino and the Meatmaster vs. Merino also shared an Rsb-based selection sweep on chromosome 1 region 268.5-269.9Mb associated with the Calpain gene, CAPN7. The study demonstrated some genetic similarities between the Merino and Merino-derived breeds emanating from common founding populations and some divergence driven by breed-specific selection goals. Overall, information regarding the evolution of these composite breeds from their founding population will guide future breed improvement programs and management and conservation efforts.

P.160 Clinical outcome assessments in limb girdle muscular dystrophy R1/2A

LGMDR1, an autosomal recessive LGMD due to mutations in calpain-3 gene (<i>CAPN3</i>), results in progressive muscle weakness. Advances in gene replacement technologies require planning for future clinical trials and better understanding of disease progression measured by COAs. The purpose of our study was to evaluate the relationship between clinical outcome assessments (COAs) and disease severity in a sample of patients with limb girdle muscular dystrophy R1 (LGMDR1). We report baseline data from our prospective observational study of clinically-affected, genetically-defined LGMDR1 participants. A battery of COAs were performed across two-day baseline visits. Test–retest reliability was evaluated using the intraclass-correlation coefficient (ICC). Linear regression was calculated to assess impact of disease duration on COAs based on genetic variant categorization. Thirty-two (75 target) participants have enrolled. The mean age 38.7 years (11-62 years) with mean symptom onset at 22.7 years (4-47 years). Participants were stratified into variant subcategories based on genetic mutations. The North Star Assessment for Limb Girdle-Type Dystrophies (NSAD) and Performance of Upper Limb (PUL) had excellent test-retest reliability (ICC 0.998 (95% confidence limit [CL] 0.994, 0.999) and 0.976 (95% CL 0.945, 0.990) respectively). Combination of COAs allows for continual evaluation across disease progression, while reducing floor/ceiling effects. Preliminary findings of COA performance in relation to disease duration suggest variants resulting in LoF may result in more rapid disease progression. Additional baseline data on the full cohort enrolled will be presented. Available COAs quantify function across the spectrum of abilities with potential to measure anticipated rate of progression based on genetic mutation and age of onset. Further research is needed to understand COA responsiveness to change in function over time in patients with LGMDR1.

Calpains in cyanobacteria and the origin of calpains

Calpains are cysteine proteases involved in many cellular processes. They are an ancient and large superfamily of enzymes responsible for the cleavage and irreversible modification of a large variety of substrates. They have been intensively studied in humans and other mammals, but information about calpains in bacteria is scarce. Calpains have not been found among Archaea to date. In this study, we have investigated the presence of calpains in selected cyanobacterial species using in silico analyses. We show that calpains defined by possessing CysPC core domain are present in cyanobacterial genera Anabaena, Aphanizomenon, Calothrix, Chamaesiphon, Fischerella, Microcystis, Scytonema and Trichormus. Based on in silico protein interaction analysis, we have predicted putative interaction partners for identified cyanobacterial calpains. The phylogenetic analysis including cyanobacterial, other bacterial and eukaryotic calpains divided bacterial and eukaryotic calpains into two separate monophyletic clusters. We propose two possible evolutionary scenarios to explain this tree topology: (1) the eukaryotic ancestor or an archaeal ancestor of eukaryotes obtained calpain gene from an unknown bacterial donor, or alternatively (2) calpain gene had been already present in the last common universal ancestor and subsequently lost by the ancestor of Archaea, but retained by the ancestor of Bacteria and by the ancestor of Eukarya. Both scenarios would require multiple independent losses of calpain genes in various bacteria and eukaryotes.

Targeting the Ubiquitin-Proteasome System in Limb-Girdle Muscular Dystrophy With CAPN3 Mutations.

LGMDR1 is caused by mutations in the CAPN3 gene that encodes calpain 3 (CAPN3), a non-lysosomal cysteine protease necessary for proper muscle function. Our previous findings show that CAPN3 deficiency leads to reduced SERCA levels through increased protein degradation. This work investigates the potential contribution of the ubiquitin-proteasome pathway to increased SERCA degradation in LGMDR1. Consistent with our previous results, we observed that CAPN3-deficient human myotubes exhibit reduced SERCA protein levels and high cytosolic calcium concentration. Treatment with the proteasome inhibitor bortezomib (Velcade) increased SERCA2 protein levels and normalized intracellular calcium levels in CAPN3-deficient myotubes. Moreover, bortezomib was able to recover mutated CAPN3 protein in a patient carrying R289W and R546L missense mutations. We found that CAPN3 knockout mice (C3KO) presented SERCA deficits in skeletal muscle in the early stages of the disease, prior to the manifestation of muscle deficits. However, treatment with bortezomib (0.8 mg/kg every 72 h) for 3 weeks did not rescue SERCA levels. No change in muscle proteasome activity was observed in bortezomib-treated animals, suggesting that higher bortezomib doses are needed to rescue SERCA levels in this model. Overall, our results lay the foundation for exploring inhibition of the ubiquitin-proteasome as a new therapeutic target to treat LGMDR1 patients. Moreover, patients carrying missense mutations in CAPN3 and presumably other genes may benefit from proteasome inhibition by rescuing mutant protein levels. Further studies in suitable models will be necessary to demonstrate the therapeutic efficacy of proteasome inhibition for different missense mutations.

Meat quality - Genetic background and methods of its analysis

Growing consumer awareness is forcing food producers to supply raw material and products of increasingly high quality and health-promoting properties. Knowledge of the genetic background of quality characteristics is taking on great importance, enabling selection based on molecular markers. The increasing throughput of molecular techniques, in combination with an expanding bioinformatics infrastructure, is leading to continual improvement in understanding of the molecular mechanisms influencing meat quality. This has resulted in the identification of polymorphic nucleotides [single nucleotide polymorphisms (SNPs)] showing a relationship with meat characteristics such as tenderness [polymorphism in the calpain (CAPN) and calpastatin (CAST) genes], marbling [diacylglycerol o-acyltransferase 1 (DGAT1)], colour, pH and water-holding capacity (WHC) [CAST, stearoyl-CoA desaturase (SCD) and others], and fatty acid profile (SCD1). An increasingly wide range of methods is used for analysis, from techniques based on amplification of nucleic acids [polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and amplification refractory mutation system-PCR (ARMS-PCR)] through Sanger sequencing to high-throughput next-generation sequencing (NGS) techniques. This paper is a review of the literature on polymorphism of genes determining the quality characteristics of meat and molecular methods used to detect them.

Association of single nucleotide polymorphisms in the CAPN, CAST, LEP, GH, and IGF-1 genes with growth parameters and ultrasound characteristics of the Longissimus dorsi muscle in Colombian hair sheep.

Recognition of the genes that influence livestock production characteristics has allowed researchers to identify single nucleotide polymorphisms (SNPs) associated with phenotypic traits that contribute to higher productivity. The objective of this research was to associate SNPs in the genes calpain (CAPN), calpastatin (CAST), leptin (LEP), growth hormone (GH), and insulin-like growth factor 1 (IGF-1) with the growth characteristics birth weight (BW), weaning weight adjusted at 120days (WW), daily pre-weaning gain (PRADG), adjusted weight at 210days (AW210), and daily post-weaning gain (POADG), and the measures of the Longissimus dorsi muscle based on ultrasound, namely loin eye area (LEA), loin depth (LD), and back fat thickness (BFT), in Colombian hair sheep (OPC). The association between phenotypic and genotypic characteristics was made using the PLINK v.1.9 program using linear regression analysis. There was a statistically significant association (p < 0.05) between the CAST polymorphism (M/N) and BW, a tendency (p = 0.07) for an association between the T → C SNP of the CAPN gene and AW210, and a trend (p = 0.07) for an association between the A → G SNP of the IGF-1 locus and POADG. The LEA and BFT characteristics were not associated with a SNP, while PL was significantly affected by SNPs in the GH and IGF-1 genes. In conclusion, all the genes evaluated were polymorphic, the CAST gene significantly influenced BW, and the GH and IGF-1 genes were associated with LD characteristics. These results could be used to identify individuals with favorable genotypes to implement a marker-assisted selection method.

A Discussion of Inbreeding Coefficients in the Herds of Turkish Grey Cattle by using Calpain and Calpastatin Gene Markers

The Turkish grey cattle (TGC) is the only local indigenous breed in Thrace and Western Anatolia. They can survive, feed, and breed without human intervention in nature. They spend the whole year, including winter, as free herds. TGC breed is under threat of extinction. Therefore they are protected by the Republic of Turkey Ministry of Agriculture and Forestry. Our investigation was supplied that purebred 51 TGC from the ex-situ protection program and 79 purebred TGC from smallholders into various villages. The research was conducted by Trakya University. Was studied that three different Single Nucleotide Polymorphisms (SNP) in the genes of calpain and calpastatin in purebred TGC. Variants of (CAPN1 316 and CAPN1 4751 SNPs) Calpain and (UoG-CAST SNP) Calpastatin genes were searched to monitor whether maintained the genetic diversity of the breed. Genetic analyses conducted on 130 cattle that have no consanguinity indicate that inbreeding coefficients calculated in each locus differed from each other. Average FIS, FIT, and FST values were determined as -0.0689, -0.0405, and 0.0266 respectively. The average gene flow was estimated to be 9.1590. The genetic variability was found to be mainly between families in subgroups (97.345%). The genetic diversity of the breed is maintained according to the results of the purebred TGC samples.

Targeted Next-Generation Sequencing Reveals Mutations in Non-coding Regions and Potential Regulatory Sequences of Calpain-3 Gene in Polish Limb-Girdle Muscular Dystrophy Patients.

Limb–girdle muscular dystrophy type R1 (LGMDR1) is caused by mutations in CAPN3 and is the most common type of recessive LGMD. Even with the use of whole-exome sequencing (WES), only one mutant allele of CAPN3 is found in a significant number of LGMDR patients. This points to a role of non-coding, intronic or regulatory, sequence variants in the disease pathogenesis. Targeted sequencing of the whole CAPN3 gene including not only intronic, 3′ and 5′ UTRs but also potential regulatory regions was performed in 27 patients suspected with LGMDR1. This group included 13 patients with only one mutated CAPN3 allele detected previously with exome sequencing. A second rare variant in the non-coding part of CAPN3 was found in 11 of 13 patients with previously identified single mutation. Intronic mutations were found in 10 cases, with c.1746-20C>G variant present in seven patients. In addition, a large deletion of exons 2–8 was found in one patient. In the patients with no causative mutation previously found, we detected rare CAPN3 variants in 5 out of 10 patients and in two of them in a compound heterozygous state. Rare variants within putative regulatory sequences distant from the CAPN3 gene were found in 15 patients, although in 11 of these cases, other variants are deemed causative. The results indicate that intronic mutations are common in Polish LGMDR patients, and testing for non-coding mutations in CAPN3 should be performed in apparently single heterozygous patients.

Clinical and genetic features of Calpainopathies in Saudi Arabia - a descriptive cross-sectional study.

Limb-girdle muscular dystrophies (LGMD) is a heterogeneous group of genetic disorders characterized by progressive weakness of pelvic and shoulder girdle muscles. The objective is to characterize the phenotypic, pathological, radiological, and genetic findings in LGMD2A phenotype (Calpainopathies). The National Saudi Arabian LGMD cohort database was screened for LGMD from January 2000 to January 2021. A descriptive cross-sectional study was done on a total of 112 families with LGMD. Screening for mutation in Calpain (CAPN3) gene was done. Clinical and genetic features of LGMD2A phenotype were the main outcome variables. Epi-info was used for statistical analysis. 34 subjects from 22 families (19.64%) had the specific LMGD2A phenotype. The mean age of onset was 9.9 ± 4.5 years (Range 4 to 19 years). The major initial symptoms were lower limb weakness, inability to climb stairs, and gait disturbance. Gower's sign occurred on an average of 3.75 to 7.25 years after onset. Loss of ambulation was observed in 55.8%. Two novel mutations in the CAPN3 gene were identified. The prevalence of LGMD2A was 19.64% among the national Saudi Arabian LGMD cohort. The clinical presentation was varied and was consistent with other reports from different ethnic groups.

Variants in Genes of Calpain System as Modifiers of Spinocerebellar Ataxia Type 3 Phenotype

Calpain-mediated proteolysis has been proposed to modulate the pathogenesis of spinocerebellar ataxia type 3, also known as Machado-Joseph disease (SCA3/MJD), a disorder due to a CAG repeat expansion (CAGexp) at ATXN3. We aimedto investigate if single-nucleotide polymorphisms (SNPs) at calpain gene CAPN2 and at calpastatin gene CAST modulate the age at onset (AO) and disease progression in SCA3/MJD. A total of 287 SCA3/MJD symptomatic subjects (151 families) were included. AO was analyzed and controlled by the CAG repeat length of expanded allele and family. Candidate polymorphisms were chosen based on the literature and on a priori criteria. The CAG repeat length and SNPs were genotyped according to standard methods. AO of carriers of AA and AG + GGrs1559085 genotypes in CAST and with the median value of 75 repeats at the expanded allele were 34.23 (33.07-35.38) and 36.42years (34.50-38.34), respectively (p = 0.049, mixed model treating the expanded CAG repeat size as fixed effect and family as random effect). Carriers of haplotype Crs27852/Grs1559085 had mean AO of 37.23 (12.76) and 33.42years (12.20) (p = 0.047, Student's t test). Our data suggest an association between allele Grs1559085 and haplotype Crs27852/Grs1559085 at CAST and variations in the AO of SCA3/MJD subjects, independent from the effects of the CAGexp and family. The present results support the potential role of calpain cleavage pathway over modulation of SCA3/MJD phenotype.

Association of SNP g.232 G&gt;T Calpain Gene with Growth and Live Meat Quality Prediction using Ultrasound Images in Bali Cattle

Bali cattle (&lt;em&gt;Bos javanicus&lt;/em&gt;) are native Indonesian cattle, domesticated from banteng (&lt;em&gt;Bibos banteng&lt;/em&gt;). Genes that have an important role in meat quality are calcium-activated neutral protease genes, known as calpains (CAPN). The objective of this study was to evaluate the polymophisms of calpain gene SNP g.232 G&amp;gt;T by PCR-RFLP technique and its influence on growth trait and meat quality of Bali cattle detected by ultrasound imaging of &lt;em&gt;longissimus dorsi&lt;/em&gt; thickness (LDT), back fat thickness (BFT), marbling score (MS), and intramuscular fat percentage (PIMF). The polymorphisms of CAPN1 gene were analyzed by PCR-RFLP using BglII restriction enzyme (n=52 cattle). The ultrasound images of longissimus dorsi muscle were carried out transversally and longitudinal between 12&lt;sup&gt;th&lt;/sup&gt; -13&lt;sup&gt;th &lt;/sup&gt;thoracic vertebrae then analyzed by Image-J NIH software. Result showed that SNP g.232 G&amp;gt;T of CAPN1 gene was polymorphic in Bali cattle. SNP g.232 G&amp;gt;T of CAPN1 gene in Bali cattle has higher diversity which was showed of 0.48 heterozygosity value and was in Hardy-Weinberg equilibrium. The polymorphisms of SNP g.232 G&amp;gt;T was associated significantly (P&amp;lt;0.05) with bodyweight at 730 days, marbling score (MS), and intramuscular fat percentage (PIMF). It suggests that the CAPN1 gene in Bali cattle is a candidate for Marker Assisted Selection (MAS), which influences body weight at 730 days, marbling score, and percentage of intramuscular fat.