CAPN1 Research Articles

Missense mutation of c.635 T > C in CAPN3 impairs muscle injury repair in a Limb-Girdel Muscular Dystropy Model.

Limb-girdle muscular dystrophy recessive 1 (LGMDR1), previously known as LGMD2A, is a specific LGMD caused by a gene mutation encoding the calcium-dependent neutral cysteine protease calpain-3 (CAPN3). In our study, the compound heterozygosity with two missense variants c.635 T > C (p.Leu212Pro) and c.2120A > G (p.Asp707Gly) was identified in patients with LGMDR1. However, the pathogenicity of c.635 T > C has not been investigated. To evaluate the effects of this novel likely pathogenic variant to the motor system, the mouse model with c.635 T > C variant was prepared by CRISPR/Cas9 gene editing technique. The pathological results revealed that a limited number of inflammatory cells infiltrated the endomyocytes of certain c.635 T > C homozygous mice at 10 months of age. Compared with wild-type mice, motor function was not significantly impaired in Capn3 c. 635 T > C homozygous mice. Western blot and immunofluorescence assays further indicated that the expression levels of the Capn3 protein in muscle tissues of homozygous mice were similar to those of wild-type mice. However, the arrangement and ultrastructural alterations of the mitochondria in the muscular tissues of homozygous mice were confirmed by electron microscopy. Subsequently, muscle regeneration of LGMDR1 was simulated using cardiotoxin (CTX) to induce muscle necrosis and regeneration to trigger the injury modification process. The repair of the homozygous mice was significantly worse than that of the control mice at day 15 and day 21 following treatment, the c.635 T > C variant of Capn3 exhibited a significant effect on muscle regeneration of homozygous mice and induced mitochondrial damage. RNA-sequencing results demonstrated that the expression levels of the mitochondrial-related functional genes were significantly downregulated in the mutant mice. Taken together, the results of the present study strongly suggested that the LGMDR1 mouse model with a novel c.635 T > C variant in the Capn3 gene was significantly dysfunctional in muscle injury repair via impairment of the mitochondrial function.

Analysis of polymorphism of genotypes of the lebedyn cattle population by the DGAT1 and CAPN1 genes

Analysis of polymorphism of genotypes of the lebedyn cattle population by the DGAT1 and CAPN1 genes

Effect of CAPN1 gene polymorphism on some physical and sensory traits in broiler meat

This study was conducted in the poultry field, Animal Production Department, College of Agriculture and Marshes, University of Thi-Qar, from 10/11/2021 to 3/2/2022, the period included field and laboratory stud. A total of 100, one day broilers Ross 308 were used. Laboratory analyzes were carried out in the laboratories of the Marshes Research Center, University of Thi-Qar and the laboratories of the College of Agriculture and Marshes, University of Thi-Qar. Three genotypes CC, CT and TT were identified whose frequency was 0.52, 0.34 and 0.14 respectively and the frequency of the C allele was 0.69 and the frequency of the T allele was 0.31. There were significant differences on Water Holding Capacity (WHC) among the genotypes, the CC genotype was superior to the CT and TT genotypes. As for the thawing and cooking loss, there were no significant differences between the genotypes. A significant differences on pH values among the genotypes, the CC genotype was superior to the CT and TT genotypes. Significant differences in flavor between genotypes, the CC genotype was superior to the CT genotype, and the CT genotype was superior to the TT genotype. Highly significant differences in tenderness, the CC genotype was superior to the TT genotype, while the CT genotype did not differ significantly from the CC and TT genotypes. Highly significant differences between the genotypes in juiciness, the CC genotype was superior to the CT and TT genotypes. No significant differences in color and general acceptable.

S3275 A Peculiar Case of Limb-Girdle Muscular Dystrophy Masquerading as Elevated Transaminases

Introduction: A disproportional elevation in aspartate transaminase (AST) and alanine transaminase (ALT) is considered a marker of hepatocellular injury. Guidance on the work up of chronically elevated liver chemistry suggests an algorithmic approach to testing of common causes of liver injury based on the degree of elevation and even mentions consideration of nonhepatic etiologies such as skeletal muscle damage. We present a unique case of asymptomatic elevation in liver chemistries as the herald finding in a patient with limb-girdle muscular dystrophy (LGMD). Case Description/Methods: Our patient is a 19-year-old adopted male who was referred to outpatient gastroenterology clinic for asymptomatic elevation in transaminases. AST was 133 IU/L and ALT was 223 IU/L. Total bilirubin and alkaline phosphatase were normal. He denied alcohol use, prescribed medications, herbal supplements, or weightlifting supplements. BMI was normal. Workup for viral hepatitis, hemochromatosis, autoimmune hepatitis, Wilson’s disease, alpha-1 antitrypsin, celiac disease, and thyroid disease were normal. Liver biopsy revealed reticulin collapse with complete regeneration without inflammation. Subsequent follow-up over years demonstrated persistently elevated but stable elevations. At the age of 24, he began to endorse proximal lower extremity muscle pain and weakness. Creatine kinase (CK) was elevated at 11,937 U/L; aldolase was elevated at 31.4 U/L; myositis workup was normal. Right thigh biopsy revealed a myopathic process. Genetic testing confirmed mutation in the CAPN3 gene consistent with calpainopathy, a subtype of LGMD. Discussion: LGMD is a rare, genetic disorder resulting in loss of ambulation within 20 years after disease onset. Concurrent liver disease is rare in LGMD, but elevations in transaminases can be found because of severe elevations in CK. Our patient was found to have calpainopathy, a subtype associated with a recessive mutation in the CAPN3 gene. Our case demonstrates that beyond the recommended algorithmic evaluation of abnormal liver chemistries, inconclusive evaluation of abnormal liver chemistries in children and young adults should include a CK level to further investigate nonhepatic causes, specifically a myopathic process. Although there is no cure for LGMD, interventions focused on muscle conditioning are more beneficial with early detection to mitigate the devastating effects of this disorder.

Exome sequencing reveals a homozygous frameshift variant in CAPN3 in a Tunisian patient with a neuromuscular disorder

Muscular dystrophy (MD) is a heterogeneous group of diseases that cause progressive weakness and loss of muscle mass. Specific signs and symptoms begin at different ages and in different muscle groups, depending on the type of muscular dystrophy. We report here a case of a Tunisian patient suffering from a neuromuscular disorder, highly suspicious of a Limb-Girdle Muscular Dystrophy (LGMD). Exome Sequencing revealed a one-base deletion variant in exon 4 of the CAPN3 gene. CAPN3 is associated with a recessive form of LGMD, also known as Muscular Dystrophy, Limb-Girdle, Type 2a (LGMD2A). The variant was shown to segregate with the disease in the family. The identification of the molecular defect in this family provided a rapid genotyping for the sister and an accurate diagnosis for the patient, allowing appropriate clinical management at an early stage of the disease.

Molecular and genetic features of calpainopathy

Calpainopathy is the most common form of limb-girdle muscular dystrophy, prevalence in the population is approximately 1 in 15,00042,700 individuals. In the Russian Federation, there is an insufficient number of studies, which researched prevalence of calpainopathy among patients with limb-girdle muscular dystrophy, but according to available data, approximately in 43% of cases the disease is associated with mutations of the CAPN3 gene. Molecular genetic analysis is the main method for diagnosing these patients. Studies indicate many pathogenic mutations that cause calpainopathy with corresponding phenotypes, however, it is quite difficult to establish clear correlations between genotype and phenotype due to the high variability of symptoms and severity, even among patients with the same CAPN3 gene mutations. Currently, there is no effective etiotropic treatment for limb-girdle muscular dystrophy, but new technologies are developing to improve patients condition and quality of life. This research collects data from various studies on the prevalence of calpainopathy in different countries and the main molecular genetic features of the CAPN3 gene and calpain-3 protein, which will further allow the development of possible treatment options for patients with limbgirdle muscular dystrophy.

HEMATOLOGICAL STATUS OF SIMMENTAL BREDINSKY MEAT TYPE HEIFERS WITH DIFFERENT ALLELIC SET OF CAPN1 AND TG5 GENES

The work carried out by geneticists in recent years connected with whole genome genotyping of cattle by SNP polymorphism with identification of associations with qualitative and quantitative parameters of productivity is used in innovative breeding programs. However, biochemical mechanisms of such determinations are not fully disclosed. Expression of polymorphic genes, which leads to a change of protein properties, is associated with biochemical processes which occur in the animal body at biomolecular level, which can manifest itself in neurohumoral and metabolic processes, which can possibly be traced by analyzing blood parameters. In this regard, blood was taken from the heifers of Bredinsky meat type of Simmental breed in Chelyabinsk region in order to isolate DNA to determine polymorphism of CAPN1 and TG5 genes associated with qualitative parameters of meat productivity and determine hematological parameters. The analysis of the genotyped population revealed a certain alignment of polymorphic state of the genes of the animals of the studied sample. The distribution of hematological parameters of heifers according to the genotypes of the studied genes did not show a clear significant difference, but outlined some tendencies. According to morphological composition of the blood, these tendencies were less expressed. However, the distribution of blood biochemical parameters by genotypes of CAPN1 gene showed superiority of GG genotype by blood glucose by 0.51 mmol / l, which is 21.9%, the content of ALT transamination enzymes by 11.9%, cholesterol by 18.8% . Difference tendencies were noted for genotypes of TG5 gene by creatinine (12.0%) and amylase (9.5%) content. The results of this study may be useful in terms of accumulating new research data and contribute to a better understanding of molecular mechanisms of relations among blood parameters, genes, and biochemical processes in animals.

Overview of Neuromuscular Disorder Molecular Diagnostic Experience for the Population of Latvia

Genetic testing has become an integral part of health care, allowing the confirmation of thousands of hereditary diseases, including neuromuscular disorders (NMDs). The reported average prevalence of individual inherited NMDs is 3.7-4.99 per 10,000. This number varies greatly in the selected populations after applying population-wide studies. The aim of this study was to evaluate the effect of genetic analysis as the first-tier test in patients with NMD and to calculate the disease prevalence and allelic frequencies for reoccurring genetic variants. Patients with NMD from Latvia with molecular tests confirming their diagnosis in 2008-2020 were included in this retrospective study. Diagnosis was confirmed in 153 unique cases of all persons tested. Next-generation sequencing resulted in a detection rate of 37%. Two of the most common childhood-onset NMDs in our population were spinal muscular atrophy and dystrophinopathies, with a birth prevalence of 1.01 per 10,000 newborns and 2.08 per 10,000 (male newborn population), respectively. The calculated point prevalence was 0.079 per 10,000 for facioscapulohumeral muscular dystrophy type 1, 0.078 per 10,000 for limb-girdle muscular dystrophy, 0.073 per 10,000 for nondystrophic congenital myotonia, 0.052 per 10,000 for spinobulbar muscular atrophy, and 0.047 per 10,000 for type 1 myotonic dystrophy. DNA diagnostics is a successful approach. The carrier frequencies of the common CAPN3, FKRP, SPG11, and HINT1 gene variants as well as that of the SMN1 gene exon 7 deletion in the population of Latvia are comparable with data from Europe. The carrier frequency of the CLCN1 gene variant c.2680C>T p.(Arg894Ter) is 2.11%, and consequently, congenital myotonia is the most frequent NMD in our population.

STUDY OF RS17872000 POLYMORPHISM IN CALPAIN (CAPN1) AND
 RS109221039 CALPASTATIN (CAST) GENES IN MEAT PRODUCTIVITY CATTLE

Allelic variants of single nucleotide CAPN1 and CAST polymorphisms in cattle are associated with “marbling” and “tenderness” of meat. SNPs in CAPN1 (rs17872000) and CAST (rs109221039) genes in beef cattle breeds being bred in the Republic of Belarus were studied using KASP and real-time PCR methods. Genetic structure analysis of studied samples from cattle populations by the CAPN1 gene showed that the frequency of occurrence of the preferential Callele associated with “marbling” in Aberdeen Angus cattle was 34.3%, in the Limousine breed ― 14.3%, and in the Hereford breed ― 13.9%. The frequency of occurrence of the preferential A allele (CAST), which determines increased “tenderness” of meat, was 87.1% in the Aberdeen Angus breed, 75.5% in the Limousin, and 77.8% in the Hereford. The use of genetic markers as an important additional criterion in meat cattle breeding in combination with main
 zootechnical methods will allow improving meat productivity.

Genetic diversity of Calpain 1 gene in Creole, Nellore and Brahman bovine breeds in Bolivia

In Bolivia, beef production is mainly based on two genotypes, Bos taurus (Creole cattle) and B. indicus (zebu), being weight gain the main selection criteria used by farmers. However, meat quality and especially tenderness must be incorporated in the selection process. Meat tenderness is partly determined by the calpain CAPN1)/ calpastatin (CAST) protein system. Thus, the objective of the present work was to determine and (compare the genetic variability of the CAPN1 gene in Creole (CreBo), Brahman (BraBo) and Nellore (NelBo) breeds in Bolivia. DNA was extracted from blood samples from 147 CreBo, 59 BraBo and 93 NelBo, and three polymorphisms were genotyped using ARMS­PCR (CAPN1­316 and CAPN1­4751) and PCR­RFLP (CAPN1­530). Furthermore, CAPN1­316 and CAPN1­4751 SNPs were analyzed with Axiom™ Bos 1 Genotyping Array r3 and the Axiom™ ArBos 1 Genotyping Array. Allele frequencies associated with higher tenderness in CreBo, BraBo and NelBo were 0.22, 0 and 0.09 (CAPN1­316 C; P < 0.001), 0.76, 0.16 and 0.08 (CAPN1­4751 C; P < 0.001), and 0.77, 0.92 and 0.94 (CAPN1­530 G; P < 0.001). Linkage disequilibrium (LD) analysis revealed the presence of two LD blocks. Our results evidence that CreBo has a higher frequency of alleles associated with higher meat tenderness than the cebuine breeds. These markers could be used in breeding programs to improve Bolivian cattle herd meat quality either by selection within Creole breeds or crosses with cebuine cattle

Sustained Effects of Muscle Calpain System Genotypes on Tenderness Phenotypes of South African Beef Bulls during Ageing up to 20 Days.

Simple SummaryWhen searching for genetic markers for the selection of more tender beef, it is important to maintain minimal environmental variation from pre-slaughter, right through to the ageing process, to ensure the accuracy of the obtained phenotypes. This is because beef quality traits have a large environmental component that can greatly alter the characteristics of the meat, which would not reflect a true genetic effect. We propose that variable ageing times are especially important in determining whether markers are associated with tenderization or not. Our analyses included candidate genes for the protein degrading enzyme system for calpains, because they contribute the most to tenderization. We were able to validate these markers in South African beef cattle, where they could be useful for selection. The timing of the collection of tenderness data was critical, as only a few (6/134) genetic markers sustained their association with tenderization over ageing to 20 days. A larger tenderization effect earlier in ageing, as shown here for the capn1_187 and capn1_4751 markers, would decrease the length of ageing. This would not only increase profits, but also decrease the energy needed during the storage and refrigeration of aged beef, decreasing the carbon footprint of beef production.The most important factor that determines beef tenderness is its proteolytic activity, and the balance between calpain-1 protease activity and calpastatin inhibition is especially important, while contributions can also arise from calpain-2 and, possibly, calpain-3. The meat ageing process itself affects these processes. To determine whether genotypes in the calpain–calpastatin system can enhance tenderness through a 20-day ageing period, South African purebred beef bulls (n = 166) were genotyped using the Illumina BovineHD SNP BeadChip through a gene-based association analysis targeting the cast, capn3, capn2 and capn1 genes. The Warner–Bratzler shear force (WBSF) and myofibril fragment length (MFL) of Longissimus thoracis et lumborum (LTL) steaks were evaluated between d 3 and d 20 of ageing, with protease enzyme activity in the first 20 h post-mortem. Although several of the 134 SNPs are associated with tenderness, only seven SNP in the cast, capn2 and capn1 genes sustained genetic associations, additive to the ageing-associated increases in tenderness for at least three of the four ageing periods. While most genomic associations were relatively stable over time, some genotypes within the SNP responded differently to ageing, resulting in altered genomic effects over time. The level of ageing at which genomic associations are performed is an important factor that determines whether SNPs affect tenderness phenotypes.

A New SNPs at 3’UTR Region of calpain 1 gene and its association with growth and meat quality traits in beef cattle

The objective of this study was to evaluate single nucleotide polymorphisms (SNP) in the 3'UTR region of CAPN1 gene by direct sequencing technique in beef cattle and its influence on growth and meat quality of the contrast mode ultrasound image in Bali cattle. The numbers of beef cattle used were Bali cattle (n=52 heads) which compared to purebred Belgian Blue (n=30 heads), Limousine (n=14 heads), Wagyu (n=7 heads), and PO (n=10 heads). Genetic diversity data were obtained from calculations by PopGen 1.32 software. Ultrasound imaging of the thoracic longissimus dorsi muscle in Bali cattle was carried out between the 12 − 13 ribs, and the sonogram was analyzed by Image-J NIH software. The result shown that 3’UTR of CAPN1 gene was found in six discovery SNPs that polymorphic in Bali cattle, they were g.15284 C>T, g.15347 T>G, g.15525 G>A, g.15853 G>A, g.15905 G>A, g.15915 G>A and Indel mutation was polymorphic in Bali cattle, Belgian Blue, Limousine, and PO. There was no association between these SNPs and growth traits. However, SNP g.15525 G>A was significantly associated (P<0.05) with a backfat thickness (BFT) in Bali cattle. In conclusion, the CAPN1 gene in Bali cattle is a candidate for Marker Assisted Selection (MAS) related to meat quality.

Experiences in the molecular genetic and histopathological evaluation of calpainopathies.

Calpainopathy is mainly characterized by symmetric and progressive weakness of proximal muscles. Several reports showed that the most common LGMD subtype is LGMDR1 or calpainopathy, which had previously been defined as LGMD2A. Until now, more than 500 likely pathogenic/pathogenic variants in the CAPN3 gene have been reported. However, a clear genotype-phenotype association had not yet been established and this causes major difficulties in predicting the prognosis in asymptomatic patients and in providing genetic counseling for prenatal diagnosis. In this report, we aimed to add new data to the literature by evaluating 37 patients with likely pathogenic/pathogenic variants for the detected variants' nature, patients' phenotypes, and histopathological features. As a result, the general clinical presentation of the 23 different variants was presented, the high frequency of NM_000070.3:c.550delA mutation in Exon 4 was discussed, and some novel genotype-phenotype associations were suggested. We have underlined that calpainopathy can be misdiagnosed with inflammatory myopathies histopathologically. We have also emphasized that, in young or adult patients with mild to moderate proximal muscle weakness and elevated CK levels, calpainopathy should be the first suspected diagnosis.

Neurodegeneration-related genes are differentially expressed in middle-aged rats compared to young-adult rats having equal performance on long-term memory and synaptic plasticity

The present study is concerned with assessing differences in plasticity-induced neurodegeneration-related gene expressions and tau phosphorylation between young-aged and middle-aged rats. The experiments were carried out in vivo under urethane anesthesia on adult male Wistar rats between the ages of 2-3 months and 11-12 months. Field potentials, composed of a field of excitatory-postsynaptic potential (fEPSP) and a population-spike (PS), were recorded from granule cells of the dentate gyrus. Plasticity was induced by high-frequency (HFS) or low frequency stimulation (LFS). mRNA of neurodegeneration-related genes and total-and phosphorylated-tau were measured in HFS-and LFS-induced hippocampus by using quantitative rt-PCR and Western blotting. In addition, naive rats (unstimulated) were tested for spatial learning and memory with a 5-day Morris water maze (MWM). HFS-induced LTP of PS had attenuated in middle-aged rats, but there were no gross differences in baseline synaptic function, HFS-induced fEPSP and LFS-induced fEPSP, and PS plasticity between young-aged and middle-aged rats. Relative to young-aged rats, in middle-aged rats, HFS-induced MAPT, CDK5, and AKT1 genes were more up regulated, while LFS-induced Bace1, PSEN2, CAPN1, ANXA, CDK5, and GSK-3β genes were more down-regulated. Tau and p-tauThr231 were increased by HFS/LFS in the hippocampus of middle-aged rats compared to those of young-aged rats. In MWM, despite the difference in searching strategy of both age groups of rats, memory was not affected by age. Impaired long-term potentiation (LTP) and accompanying changes in intracellular biological markers may underlie in neurodegenerative disease characterized by dementia that occurs gradually later ages. However, these changes were not reflected in behavioral spatial memory.

Limb Girdle Muskuler Distrofi Tip 2a: Vaka Sunumu

Limb Girdle Muscular Disease (LGMD) comprise a group of inherited muscular distrophy with chronic progressive weakness of hip and shoulder girdles. The inheritance pattern is either autosomal dominant (LGMD1) or autosomal recessive (LGMD2). LGMD 2A is known as calpainopathy in which there was a defect of gene encoding the protein named as calpain. There are three calpainopathy phenotypes according to distribution of muscle weakness and age at onset. In this report, we presented an asymptomatic child with persistant hyper CKemia diagnosed with muscle biopsy and genetic testing. Genetical examination results of the patient showed homozygote mutation of CAPN3 gene(c.2092C>A) and parents revealed that they were heterozygous unaffected carriers.

Effect of CAPN1 gene polymorphism on some physical and sensory traits in broiler meat

Effect of CAPN1 gene polymorphism on some physical and sensory traits in broiler meat

A Novel Mutation of CAPN1 Gene Causing Hereditary Spastic Paraplegia-76.

A Novel Mutation of CAPN1 Gene Causing Hereditary Spastic Paraplegia-76.

Effect of CAPN3 gene genotypes on productive traits and carcass traits of broiler

Determining the genotypes of the CAPN3 gene and studying its relationship to some economic characteristics of broiler chickens, which included weekly weights, final live weight, carcass weight and carcass parts weights, in addition to the correlation coefficient for some important Ross characteristics. 308 chickens, in which 100 chicks were bred for 35 days and taken. Primary birds were weighed and all birds were numbered. Genotypes were determined by analyzing the nitrogenous base sequences of the studied part of the CAPN3 gene the site of variation in the nitrogenous bases of the studied gene (G>C2426) was diagnosed and the resulting genotypes were calculated, which are three genotypes: GG, GC, and CC of the CAPN3 gene. As for the CAPN3 gene, the results showed statistically significant differences for some traits such as chest weight and wing weight, while the results did not indicate any statistically significant differences between the three models for other traits such as live body weight, carcass, and thigh weight, back weight, weekly weights, and superiority of genotype CC on the rest of the individuals in some traits. the study suggests the possibility of using molecular markers of the CAPN3 gene in poultry breeding and selection programs.

Effect of CAPN3 gene genotypes on productive traits and carcass traits of broiler

Effect of CAPN3 gene genotypes on productive traits and carcass traits of broiler

Molecular Diagnosis of Muscular Dystrophy Patients in Western Indian Population: A Comprehensive Mutation Analysis Using Amplicon Sequencing.

Muscular Dystrophies (MDs) are a group of inherited diseases and heterogeneous in nature. To date, 40 different genes have been reported for the occurrence and/or progression of MDs. This study was conducted to demonstrate the application of next-generation sequencing (NGS) in developing a time-saving and cost-effective diagnostic method to detect single nucleotide variants (SNVs) and copy number variants (CNVs) in a single test. A total of 123 cases clinically suspected of MD were enrolled in this study. Amplicon panel-based diagnosis was carried out for 102 (DMD/BMD) cases and the results were further screened using multiplex ligation-dependent probe amplification (MLPA). Whilst in the case of LGMD (N = 19) and UMD (N = 2), only NGS panel-based analysis was carried out. We identified the large deletions in 74.50% (76/102) of the cases screened with query DMD or BMD. Further, the large deletion in CAPN3 gene (N = 3) and known SNV mutations (N = 4) were identified in LGMD patients. Together, the total diagnosis rate for this amplicon panel was 70.73% (87/123) which demonstrated the utility of panel-based diagnosis for high throughput, affordable, and time-saving diagnostic strategy. Collectively, present study demonstrates that the panel based NGS sequencing could be superior over to MLPA.