Calpastatin Research Articles

Genetic diversity and polymorphism of genes associated with meat quality and growth traits of A Luoi Yellow cattle in Vietnam

Background and Aim: A Luoi Yellow cattle is an indigenous cattle breed that is raised in the A Luoi District, Vietnam, characterized by its small body size, high adaptability, and meat quality favored by domestic consumers. Marker-assisted selection is an effective approach for improving breeding genetics and sustainably developing livestock production. Therefore, this study aimed to investigate the genetic diversity and polymorphism of genes associated with meat quality and productivity in the A Luoi Yellow cattle population with the goal of future breeding selection and sustainable development of the A Luoi Yellow beef brand. Materials and Methods: In this study, we genotyped six functional genes, including Leptin (LEP), Calpastatin (CAST), Calpain 1, pleomorphic adenoma gene 1, Sirtuin 1, and Sirtuin 2 (SIRT2), involved in meat quality and growth traits using polymerase chain reaction (PCR) or PCR-restriction fragment length polymorphis. We also investigated mitochondrial DNA (mtDNA) and the Y chromosome-specific gene on the Y chromosome to elucidate the genetic diversity and paternal and maternal origin of the A Luoi Yellow cattle using Sanger sequencing. Results: The results showed that A Luoi yellow cattle have Bos indicus origin from both paternal and maternal lineages. By mtDNA analysis, we identified two new haplotypes of the I1 haplogroup that were not previously detected. The genotyping of the six functional genes indicated that A Luoi Yellow cattle carry favorable alleles that increase meat tenderness and body size, with frequencies of 0.02–0.40. In particular, the presence of desirable homozygous genotypes of the CAST, LEP, and SIRT2 genes will be important for the future selection of animals based on their potential performance in meat quality and productivity. Conclusion: The findings of this study is useful for the future breeding and sustainable development of A Luoi Yellow cattle. Keywords: A Luoi yellow cattle, growth trait, indels, meat tenderness, single nucleotide polymorphisms.

Gene‒Environment Interaction Between CAST Gene and Eye-Rubbing in the Chinese Keratoconus Cohort Study: A Case-Only Study.

Keratoconus (KC), characterized by progressive corneal protrusion and thinning, is a complex disease influenced by the combination of genetic and environmental factors. The purpose of this study was to explore potential gene‒environment interaction between the calpastatin (CAST) gene and eye-rubbing in KC. A case-only study including 930 patients (676 patients with eye-rubbing and 254 patients without eye-rubbing) from the Chinese Keratoconus (CKC) cohort study was performed in the present study. Genotyping of single nucleotide polymorphism (SNP) was conducted using the Illumina Infinium Human Asian Screening Array (ASA) Beadchip. The gene‒environment interactions between CAST gene and eye-rubbing were analyzed using PLINK version 1.90. The interactions between CAST genotypes and eye-rubbing were analyzed by logistic regression models. The SNP-SNP-environment interactions were analyzed using generalized multifactor dimensionality reduction (GMDR). Three SNPs in CAST gene, namely, rs26515, rs27991, and rs9314177, reached the significance threshold for interactions (defined as P < 2.272 × 10-3). Notably, the minor alleles of these three SNPs exhibited negative interactions with eye-rubbing in KC. The results of logistic regression models revealed that the minor allele homozygotes and heterozygotes of rs26515, rs27991, and rs9314177 also exhibited negative interactions with eye-rubbing. Furthermore, GMDR analysis revealed the significant SNP-SNP-environment interactions among rs26515, rs27991, rs9314177, and eye-rubbing in KC. This study identified rs26515, rs27991, and rs9314177 in CAST gene existed gene-environment interactions with eye-rubbing in KC, which is highly important for understanding the underlying biological mechanisms of KC and guiding precision prevention and proper management.

Analysis of the Calpastatin (CAST ) and Androgen Receptor (AR) Gene Polymorphisms as Biomarkers for Meat Quality Traits in Reindeer Rangifer tarandus

Analysis of the Calpastatin (CAST ) and Androgen Receptor (AR) Gene Polymorphisms as Biomarkers for Meat Quality Traits in Reindeer Rangifer tarandus

Investigation of calpastatin (CAST) gene polymorphism in Norduz sheep by PCR-RFLP method

Investigation of calpastatin (CAST) gene polymorphism in Norduz sheep by PCR-RFLP method

Abstract 4378: Identification of novel TXNIP-dependent antitumor pathways in breast cancer

Abstract Thioredoxin-interacting protein (TXNIP) plays a critical role in glucose metabolism and redox signaling. TXNIP has recently emerged as a potent tumor suppressor in various cancer models. In previous studies, we found TXNIP was activated by UNC0642, an inhibitor of histone methyltransferase, in MDA-MB-231 breast cancer cells. TXNIP activation was coupled with significant suppression of cell proliferation and tumor growth. In contrast, TXNIP knockdown increased MDA-MB-231 cell proliferation and in tumor growth and metastasis in vivo. TXNIP-reconstitution in TXNIP-deficient HCC1954 cells reduced cell proliferation and migration coupled with increased reactive oxygen species production. TXNIP-reconstitution also significantly decreased mitochondrial respiration, mitochondrial membrane potential and glycolysis in HCC1954 cells. To understand the mechanisms underlying TXNIP’s antitumor activity, we performed co-immunoprecipitation and proteomic analyses to uncover TXNIP-interacting proteins. We identified calpastatin (CAST) as a novel TXNIP-interacting protein in breast cancer cells. CAST is an endogenous inhibitor of calpains, a family of calcium-activated cysteine proteases. Intriguingly, CAST association with TXNIP was readily detectable in TXNIP-low, but not in TXNIP-high breast cancer cells. The disassociation of CAST from TXNIP in TXNIP-high cells might allow CAST to inhibit calpain activity to augment TXNIP’s antitumor function. Through RNAseq analysis, we identified a positive correlation between TXNIP expression and IL-24 activation in various breast cancer cells, which were also confirmed by western blotting. IL-24 has emerged as an important cancer therapy target because of its role in promoting cancer-specific apoptosis. Our data also demonstrated that TXNIP upregulated the level of IL-24 and downregulated STAT-3. Moreover, TXNIP exerts multifaceted effects on BC development through the IL24-STAT3 signaling axis, deepening our understanding of its complex role in the disease. Taken together, these studies identified novel TNXIP-dependent pathways and their potential new mechanisms in breast cancer pathogenesis with significant clinical implications. Citation Format: Jasvinder Singh, Bindeshwar Sah, Robert Clarke, Liang Liu. Identification of novel TXNIP-dependent antitumor pathways in breast cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 4378.

Genetic markers in sheep breeding

The priority direction in developing domestic sheep breeding is the creation of new and improved breeding forms that combine high meat and wool productivity rates with good adaptive qualities to breeding conditions. In Russia, 404.2 thousand heads of coarse-wool sheep breeds and 21.2 thousand heads of semi-coarse-wool sheep are concentrated on breeding farms. Compared to 2000, the number of breeding animals increased by 5.0 times, while the yield of lambs for slaughter per 100 breeding queens increased by 5.7 times. One way to increase the selection and breeding work with breeds is to search and use genetic markers of productivity and vitality. The work aimed to summarise and analyse information about the genetic polymorphism of genes associated with traits of productivity and disease resistance in different breeds of sheep. The results of studies of polymorphic variants of genes related to the growth energy of young animals, wool quality and meat productivity in sheep are presented. The calpastatin (CAST) gene is described, which is associated with meat productivity and tenderness of meat the KRT (keratins) gene - with wool productivity and wool quality, the growth hormone gene GH and the differential growth factor gene GDF9 - with the regulation of growth and development, reproductive qualities and meat production, sheep productivity. In most studies, the predominant genotype in all breeds is CASTMM. The frequency of genotypes for the KRT and GH genes in breeds of different directions of productivity varies significantly. The relationship of genotypes CAST, GН, and GDF9 with economically valuable traits is described, making it possible to consider their genetic markers.

Study of genetic variation of myostatin (MSTN) and calpastatin (CAST) genes in two native Iraqi sheep by PCR-RFLP technique

Study of genetic variation of myostatin (MSTN) and calpastatin (CAST) genes in two native Iraqi sheep by PCR-RFLP technique

Associations of CAST, GH gene polymorphism in young sheep with a blood type of ? Charolais + ? Kalmyk kurdyuchnaya with measurements of body articles and indices of physique

To increase the production of meat productivity, it is necessary to improve the genetic resources of sheep. The purpose of this work was to establish the allelic spectrum of the calpastatin (CAST) and somatotropin (GH) genes, and their associations with meat productivity in young animals of mixed stock with a bloodline of ? Charolais + ? Kalmyk chicken and to identify the effects of polymorphism of gene markers on the exterior parameters of young animals. The object of the study is the repair young sheep reared in the farm "Arl" of the Yashkul district of the Republic of Kalmykia. The PCR-PDRF method determined the allelic spectrum of genes: calpastatin (CAST), somatotropin (GH), in young hybrids with a blood type of ? Charolais + ? Kalmyk chicken. The superiority of the CASTM allele (0.78) over the frequency of occurrence of the selectively significant CASTN allele (0.22) was noted in the calpastatin gene in crossbreed livestock. According to the growth hormone gene, molecular genetic analysis showed that the frequency of occurrence of the GHA allele in young mixed sheep was 0.88, and the frequency of occurrence of the selectively significant GHB allele was 0.12. A comparative analysis of the live weight of animals with different genotypes showed that sheep and sheep carriers of heterozygous CASTMN and GHAB genotypes have higher live weight indicators than their peers with homozygous CASTMM and GHAA genotypes. Measurements of articles of exterior and indices of physique depending on genotypes are described, since their use allows breeders to determine the type of constitution, individual characteristics, degree and proportionality of the development of the organism, age variability, condition and ability to a particular productivity.

Ahf-Caltide, a Novel Polypeptide Derived from Calpastatin, Protects against Oxidative Stress Injury by Stabilizing the Expression of CaV1.2 Calcium Channel.

Reperfusion after ischemia would cause massive myocardial injury, which leads to oxidative stress (OS). Calcium homeostasis imbalance plays an essential role in myocardial OS injury. CaV1.2 calcium channel mediates calcium influx into cardiomyocytes, and its activity is modulated by a region of calpastatin (CAST) domain L, CSL54-64. In this study, the effect of Ahf-caltide, derived from CSL54-64, on myocardial OS injury was investigated. Ahf-caltide decreased the levels of LDH, MDA and ROS and increased heart rate, coronary flow, cell survival and SOD activity during OS. In addition, Ahf-caltide permeated into H9c2 cells and increased CaV1.2, CaVβ2 and CAST levels by inhibiting protein degradation. At different Ca2+ concentrations (25 nM, 10 μM, 1 mM), the binding of CSL to the IQ motif in the C terminus of the CaV1.2 channel was increased in a H2O2 concentration-dependent manner. CSL54-64 was predicted to be responsible for the binding of CSL to CaV1.2. In conclusion, Ahf-caltide exerted a cardioprotective effect on myocardial OS injury by stabilizing CaV1.2 protein expression. Our study, for the first time, proposed that restoring calcium homeostasis by targeting the CaV1.2 calcium channel and its regulating factor CAST could be a novel treatment for myocardial OS injury.

Effects of Short-Term Transportation and Lairage Time on Meat Quality, CAST, CAPN1, CAPN3, HSPA1A Gene Expression Levels and Myofibrillary Structure of M. longissimus dorsi of Kıvırcık Breed Sheep

The objective of this study was to examine the effect of short term transportation and lairage time on meat quality, myofibrillary structure and calpastatin (CAST), µ-calpain (CAPN1), calpain 3 (CAPN3) and heat shock protein 70 (HSPA1A) expression levels of longissimus dorsi of Kıvırcık breed sheep (n=40). Meat quality was evaluated by muscle glycogen amounts, pH, temperature, cooking loss, water holding capacity, shear force, instrumental color, sarcomere length and organoleptic properties. Cellular changes in the muscle during the ageing process was scanned by electron microscopy and CAST, CAPN1, CAPN3, HSPA1A expression levels were measured to reveal the association on meat tenderness. As a result, positive effect of long lairage time groups was observed in the pH, glycogen, SF values and organoleptic evaluation. CAST, CAPN1, CAPN3 expression showed no difference, however, HSPA1A showed significant difference in aging process. In conclusion, genotypic differences, their effect on gene expression and protein level on meat quality should be further investigated.

THE USE OF GENOTYPING TO SELECT DESIRED-TYPE ANIMALS

Modern sheep breeding should be based on the production of high quality sheep products. Knowing the limits of animal productivity potential allows to manage the breeding process in the most effective way. Applying PCR-RFLP method makes it possible to identify allelic variants of genes associated with quantitative and qualitative characteristics of animals. By identifying different polymorphic variations and their phenotypic manifestations, one can create and replenish a database for managing the genetic progression of breeding-significant traits.&#x0D; The paper determines the gene polymorphism of growth hormone (GH), calpastatin (CAST) and growth differentiation factor (GDF9) of experimental animals and reflects an associative connection between the dynamics of growth and that of development of crossbred one-shear sheep of different origin (Russian meat merino ewes and imported producers). The article concludes that in order to increase sheep meat productivity, farms should give preference to animals carrying genes with the identified desired genotype on the studied polymorphisms and take into account their presence in the breeding process.

Marker-assisted and Genomic Selection of Beef Cattle

This article provides an overview of modern genetic technologies for improving production traits and predicting breeding value in beef cattle. In particular, in marker-assisted selection the most promising is the selectionby desirable genotypes in the genes of myostatin (MSTN), calpain (CAPN), calpastatin (CAST), growth hormone (GH), leptin (LEP), thyroglobulin (TG), fatty acid binding protein (FABP), retinoic acid C-receptor (RORC), diacyl-glycerol acyltransferase (DGATI), sterol-Co desaturase (SCD). A modern and much more advanced approach is the Single Step Genomic Best Linear Unbiased Predictions (ssGBLUP) method, which calculates a Genomic Estimated Breeding Value (GEBV) using DNA chip genotyping, phenotype and pedigree data. Genome-wide association studies (GWAS), based on the use of genetic markers distributed throughout the genome and in non-equilibrium linkage with at least one of the quantitative traits, are currently recognised as more informative for finding new genes for beef cattle productivity. New genes associated with live weight at different stages of ontogenesis, average daily live weight gain, residual feed intake, carcass weight and flesh content have been identified. Most of the identified genes control cell division, lipid and carbohydrate metabolism. The accumulated data on full-genome association studies and exome sequencing led to new methods of genetic analysis – gene ontology and gene networks. The use of gene networks provided the first detailed understanding of the genetic basis for the formation of complex phenotypic traits based on the complex interaction of regulatory networks of «major» and «peripheral» genes controlling the development of a particular trait.

The allelic spectrum of the GDF9/BSTHHI, CAST/MSPI, GH/HAEIII genes in Kal-myk short-tailed sheep and crossbreeds with a blood type of ? dorper + ? Kalmyk short-tailed breed

To increase the production of meat productivity, it is necessary to improve the genetic resources of sheep. The purpose of this work was to establish the allelic spectrum of meat productivity genes in sheep of the Kalmyk Kalmyk short-tailed breed and crossbreeds with a blood type of ? dorper + ? Kalmyk short-tailed breed. The object of the study was sheep raised by the farm "Arl" of the Yashkul district of the Republic of Kalmykia. Blood samples (5 ml each) were taken from random sheep of the Kalmyk short-tailed breed (n = 14) and crossbreeds with a blood density of ? dorper + ? Kalmyk short-tailed breed (n = 12) from the jugular vein into a test tube with ethylenediaminetetraacetic acid (EDTA). The PCR-PDRF method determined the allelic spectrum of genes: differential growth factor (GDF9), calpastatin (CAST), somatotropin (growth hormone (GH)), in Kalmyk sheepdogs and crossbreeds with a blood type of ? dorper + Kalmyk short-tailed breed. The methods of genetic and statistical analysis in the studied samples of sheep revealed complex genotypes for the studied marker genes. By genetic and statistical methods of analysis, it was found that the proportion of animals that had the desired complex genotype according to the studied genes in sheep of the Kalmyk chicken breed, including – 1 marker allele in three genes (GDF9AG/CTMM/GHAA and GDF9GG/CASTMN/GHAA), was 28.6%, in turn, in crossbreeds with a blood type of ? dorper + A combination of 2 marker alleles of three genes (GDF9AG/CASTMN/GHAA) appeared in the Kalmyk short-tailed breed - 33.3%, a combination of the 1st marker allele in three genes - 50.0%. It is concluded that the crossing of Kalmyk sheep of the Kurdyuk breed with sheep-producers of the Dorper breed, carrying alleles desirable for breeding in their genome, can have a positive effect on increasing the productivity of crossbreed livestock.

Polymorphism of the BMP-15 locus in Romanov sheep in Western Siberia

Currently, selective selection takes into account gene polymorphisms associated not only with multiple pregnancies (growth differentiation factor 9 (GDF-9), bone morphogenetic protein receptor (BMPR-IB), etc.) but also with milk parameters (β-lactoglobulin (β-lg), αS1-casein) and meat (myostatin (MSTN), calpastatin (CAST), calpain (CAPN1)) productivity. Thus, genes associated with the transforming growth factor began to be monitored to improve reproductive performance in sheep breeding: BMP-15, GDF-9, and BMPR-IB. Genetic markers related to animal reproduction are exciting, with several productive indicators and other evaluation criteria that have not been previously studied. Work on the relationship of animal gene inheritance with biochemical, haematological, environmental and zootechnical indicators is particularly relevant. In this paper, we consider genotypic variability in Romanov sheep for the BMP-15 gene locus, which belongs to the genes of the β-growth factor family. The distribution of genotypes in sheep of the Romanov breed in the conditions of Western Siberia was as follows: for ewes, the frequencies of genotypes WW–25%, WM–75, and MM–0%; for sheep - 0, 80 and 20%, respectively. All three genotypes (WW, WM, and MM) were identified in the studied sheep. The results differ from the data obtained in several other works on foreign sheep breeds (short-tailed sheep Khan, Awassi, Barki, Ossimi, Rahmani, etc.). It is interesting to study the BMP-15 locus further to establish its associations with biochemical and haematological parameters and the hormonal status of sheep.

Polymorphism of Genes and Their Impact on Beef Quality.

The single-nucleotide polymorphism (SNP) form of genes is a valuable source of information regarding their suitability for use as specific markers of desirable traits in beef cattle breeding. For several decades, breeding work focused on improving production efficiency through optimizing the feed conversion ratio and improving daily gains and meat quality. Many research teams previously undertook research work on single-nucleotide polymorphism in myostatin (MSTN), thyroglobulin (TG), calpain (CAPN), and calpastatin (CAST) proteins. The literature review focuses on the most frequently addressed issues concerning these genes in beef cattle production and points to a number of relevant studies on the genes' polymorphic forms. The four genes presented are worth considering during breeding work as a set of genes that can positively influence productivity and production quality.

Study of population-genomic structure of Vyatka horses in interline aspect

Abstract. The population-genomic structure of the Vyatka horses was studied in the context of 10 male genealogical lines bred in the breed. Purpose. Evaluation of the genetic and breeding potential of the Vyatka breed, as well as the influence of factory breeds. Methods. Isolation of DNA from the hair follicles of Vyatka horses (n = 38) was performed using “ExtraGene DNA Prep 2000” by allele-specific PCR. Statistical calculations of allele frequencies and the types of studied loci were performed using MS Excel 10 software. Results. The polymorphism of MSTN, GYS1, DMTR3, CAST, and PRLR genes in Vyatka breed lines was studied for the first time. According to the frequency of occurrence CAST G/A (0.472) and PRLR G/C (0.417), as well as the genotype MSTN T/T (0.579) is somewhat dominated by the heterozygous genotypes characteristic of local breeds. They revealed polysaccharide accumulation mutation PSSM1 (0.189), characteristic of draft horses, and the DMRT3 mutation (0.087), which determines the ability for a non-standard gait - amble, noted among Oryol trotters, which indicates the presence of draft and trotting blood in the lines carrying these alleles. MSTN/C and DMRT3/A alleles, which are not typical for aborigines, are noted in Znatok line, whose representatives are distinguished by a lightweight body type and productive movements. The carriers of the mutant defective GYS1 allele (PSSM1) were identified in five lines: Bob, Gabizon, Buran, Dobrik, and Malakhit. Genotyping of all used stallions for genes associated with economically useful traits will allow more efficient selection in the Vyatka breed and prevent the spread of unwanted alleles, which is especially important for small breeds. Scientific novelty. For the first time, an intrapopulation genomic analysis of the domestic horse breed was studied in the interline aspect, and the influence of factory breeds on certain genealogical lines was shown. The polymorphism of the calpastatin (CAST) and prolactin receptor (PRLR) genes has not previously been studied in horse breeding.

Bibliometric Analysis of Research on the Main Genes Involved in Meat Tenderness.

Tenderness is one of the main characteristics of meat because it determines its price and acceptability. This is the first bibliometric study on the trend of research on the role of genes in meat tenderness. A total of 175 original and English-language articles published up to 2021 were retrieved from Scopus. The bibliometric analysis was carried out with VOSviewer (version 1.6.18, Eck and Waltman, Leiden, Netherlands) and complemented with the Analyze search results service from Scopus. Erroneous and duplicate data were eliminated, and incomplete information was added to standardize the results. Scientific production was evaluated by means of quantity, quality and structure indicators. As a first glance, 8.816% of authors have published more than 50% of papers mainly related to genes encoding the calpain (CAPN)-calpastatin (CAST) system and single nucleotide polymorphisms (SNPs). Among other findings, a strong link was found between the contribution of the main countries (led by the United States with) and their institutions (led by the USDA Agricultural Research Service with) to their gross domestic product. Most studies on the topic are published in the Journal of Animal Science, and other journals with high impact according to the number of citations and different metrics. Finally, when evaluating the most cited articles, the occurrence and association of the main keywords, it was confirmed that research is focused on the role of CAPN and CAST genes and of SNPs in beef tenderness. The change in science was emphasized; although marker-assisted selection is still used, genes have an infinitesimal effect on complex traits. Therefore, since about 2010, new research groups adopted genomic selection to evaluate dense panels of SNPs and better explain genetic variation in meat tenderness.

Role of CAST-Drp1 Pathway in Retinal Neuron-Regulated Necrosis in Experimental Glaucoma.

Numerous studies have indicated that excitatory amino acid toxicity, such as glutamate toxicity, is involved in glaucoma. In addition, excessive glutamate can lead to an intracellular calcium overload, resulting in regulated necrosis. Our previous studies have found that the calpastatin (CAST)-calpain pathway plays an important role in retinal neuron-regulated necrosis after glutamate injury. Although inhibition of the calpain pathway can decrease regulated necrosis, necrotic cells remain. It has been suggested that there are other molecules that participate in retinal neuron-regulated necrosis. CAST is an important regulator of dynamin-related protein 1 (Drp1)-mediated mitochondrial defects. Thus, the aim of this study was to determine whether the CAST-Drp1 pathway may be an underlying signaling axis in neuron-regulated necrosis. Using cultured retinal neurons and in an in-vivo glaucoma model induced by glutamate overload, members of the CAST-Drp1 pathway were assessed by immunofluorescence, Western blotting, Phos-tagTM SDS-PAGE, and co-immunoprecipitation assays. Moreover, the black and white box test was performed on the rats. We found that more retinal neuron-regulated necrosis and Drp1 activation as well as lower CAST levels were present in the glutamate-induced glaucoma model. Rats with glutamate-induced glaucoma exhibited impaired visual function. We also observed retinal neuron-regulated necrosis and Drp1 activity decreased, and impaired vision recovered after CAST active peptide application, indicating that the CAST-Drp1 pathway plays a critical role in retinal neuron-regulated necrosis and visual function. The results of this study indicate that the CAST-Drp1 pathway protects against retinal neuron-regulated necrosis, which may expand the therapeutic targets for the treatment of neurodegenerative disorders involving dysfunction of glutamate metabolism, such as glaucoma.

Akkaraman Koyun Irkında Kalpastatin (CAST) ve Büyüme Farklılaşma Faktörü 9 (GDF9) Genlerin Polimorfizmi

Objective: In this study, the gene polymorphisms of Calpastatin (CAST) and Growth Differentiation factor 9 (GDF9) were determined in Akkaraman sheep breed.Material and Methods: Genomic DNA was obtained from blood samples of 50 Akkaraman sheep. All samples for CAST and GDF9 were genotyped by the Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) method using the enzymes MspI and Hhal.Results: Three (MM, MN, and NN) and two (GG and GA) genotypes, respectively, were detected at the CAST and GDF9 genes. The frequencies of the genotypes MM, MN and NN were determined to be 44 %, 54 % and 2 %, respectively. The frequencies of the GG and GA genotypes were found to be 84 % and 16 %, respectively. The AA genotype was not found.Conclusion: The study showed for the first time that the Akkaraman breed carries the GDF9 G1 mutation and has sufficient variation in the CAST. Previous studies provided that both genes can be used as biomarkers for increasing lambing yield and growth rates. Therefore, in order for the studied genes to be used as biomarkers in the Akkaraman breed, association studies should first be performed.

Assessment of calpastatin and insulin-like growth factor 1 genotypes in Tsigai sheep

The aim of the present study was to evaluate the genotypic distribution of calpastatin (CAST) and insulin-like growth factor 1 (IGF1) gene polymorphisms in the Tsigai sheep breed. Genomic DNA was isolated from blood samples by the phenol-chloroform method. A total of 56 sheep were genotyped by the PCR-RFLP method. Allele and genotype frequencies were calculated by the standard procedures. To evaluate the population-genetic properties, the Hardy–Weinberg Equilibrium (HWE) testing was performed. Moreover, genetic diversity was evaluated through gene heterozygosity (He), effective allele numbers (Ne), polymorphism information content (PIC), the level of possible variability realization (V%), the fixation index (FIS), and the Shannon-Weaver diversity index (H´) were estimated. Results revealed that the frequency of the heterozygous genotype (0.64) was remarkably higher than the homozygotes in the CAST locus. HWE testing showed a deviation (P