Polymorphism detection of <i>DGAT1</i> and <i>Lep</i> genes in Anatolian water buffalo (<i>Bubalus bubalis</i>) populations in Turkey

Acyl-CoA: diacylglycerol–acyltransferase 1 (DGAT1) enzyme plays a key role in controlling the synthesis rate triglyceride from diacylglycerol. Leptin (LP, OB, obese) is an important hormone that synthesizes mostly from adipose tissue and regulates glucose metabolism and homeostasis. DGAT1 and Lep genes are closely related to reproduction, growth, milk yield and composition in water buffalo breeds. This study aimed to identify genetic variation in the DGAT1 and Lep gene regions in 150 water buffalo individuals from five different provinces of Turkey using DNA sequencing. A total of 38 nucleotide variations and indels have identified 761 bp long partial intron 2 and exon 3 and 5′ UTR regions of the Lep gene in Anatolian water buffalo populations; 422 bp long partial exon 7–9 and exon 8 regions of DGAT1 gene were amplified and two mutations were defined in the point of 155 and 275 nucleotide that is three genotypes for S allele and Y allele of DGAT1 gene in intron 7 in Anatolian buffalo populations, respectively. These SNPs may have an effect on reproduction, growth, milk yield and composition in water buffalo populations and may prove to be useful for water buffalo breeding.

Associations of MC4R, LEP, and LEPR Polymorphisms with Obesity-Related Parameters in Childhood and Adulthood

MC4R, LEP, and LEPR genes are involved in the hypothalamic leptin-melanocortin regulation pathway, which is important for energy homeostasis. Our study aimed to evaluate the associations between the MC4R rs17782313, LEP rs7799039, and LEPR rs1137101 polymorphisms with obesity-related parameters in childhood and adulthood. The data were obtained from the Kaunas Cardiovascular Risk Cohort study, which started in 1977 with 1082 participants aged 12–13 years. In 2012–2014, the follow-up survey was carried out. Genotype analysis of all respondents (n = 509) aged 48–49 years was performed for the gene polymorphisms using Real-Time Polymerase Chain Reaction. Anthropometric measurements were performed in childhood and adulthood. In childhood, only skinfold thicknesses were associated with gene variants being the lowest in children with MC4R TT genotype and LEP AG genotype. In adulthood, odds of obesity and metabolic syndrome was higher in MC4R CT/CC genotype than TT genotype carriers (OR 1.8; 95% CI 1.2–2.8 and OR 1.6; 95% CI 1.1–2.4, respectively). In men, physical activity attenuated the effect of the MC4R rs17782313 on obesity. The LEP GG genotype was associated with higher BMI, waist circumference, and visceral fat level only in men. No associations of the LEPR rs1137101 polymorphisms with anthropometric measurements and leptin level were found. In conclusion, the associations of the MC4R and LEP gene polymorphisms with obesity-related parameters strengthened with age.

DNA Markers for Meat Quality of Pigs

This paper presents the results of determining genotypes of 3-breed pig hybrids (Landrace x Yorkshire x Duroc) by marker genes MC4R (melanocortin receptor-4 gene), IGF2 (insulinoid growth factor-2), POU1F1 (pituitary transcription factor), H-FABP (protein gene binding fatty acids), GH (growth hormone gene), LEP (leptin gene) and their interrelation with meat productivity. Allele frequencies for the studied genes were determined (MC4R: А = 0.58, G = 0.42; IGF2: Q = 0.96, q = 0.04; POU1F1: Е = 0.66, F = 0.34; H-FABP: D = 0.38, d = 0.62; GH: А = 0.26, G = 0.74; LEP: С = 0.27, Т = 0.73). The study showed that 3-breed hybrids did not have the highest level of heterozygosity in most allelic genes. MC4R (AG = 48%), POU1F1 (EF = 68%) and H-FABP (Dd = 52%) genes had the greatest number of heterozygotes; IGF2 (QQ = 92%), GH (GG = 58%) and LEP (TT = 56%) genes had the greatest number of homozygotes. The data showed that the breeds used to obtain three-breed hybrids were selected for analogous (meat quality) traits and that they had a higher frequency of the desired Q (IGF2), G (GH) and T (LEP) gene alleles, which were lost from hybrids in the homozygous condition. We identified the most desirable genotypes for the studied genes (GGMC4R, AGMC4R, QQIGF2, EFPOU1F1, DDH-FABP, AAGH, CTLEP) which are recommended for pig selection as well as for selection of parent pairs for producing commercial hybrids with high meat productivity. Keywords: gene-dependent selection, slaughter and meat qualities of pigs, marker genes, MC4R, IGF2, POU1F1, H-FABP, GH, LEP

INFLUENCE OF LEPTIN (LEP) GENE POLYMORPHISM ON DAIRY AND MEAT PRODUCTIVITY OF HOLSTEIN HEIFERS

The aim of this study was to research the association between leptin gene polymorphism in Holstein cows with their dairy and meat traits of productivity. To conduct DNA testing on the LEP gene blood samples were selected from 228 Holstein heifers. Animal genotyping according to the leptin gene was performed by AC-PCR. All possible genotypes of the LEP gene were identified in the course of DNA testing. Frequency of occurrence of genotypes and alleles was ranked as fol-lows: CC – 32.5 %, TC – 49.1 %, TT – 18.4 % and C – 0.57, T – 0.43. The results of this study show that animals with genotype TT of LEP gene can be useful in breeding for the improvement of population of Holstein cattle by economic traits associated to the dairy and meat productivity. This research was supported by the FASO Russia project АААА-А18-118031390148-1.

Leptin G-2548A Gene Polymorphism is Positively Associated with Increased Plasma Leptin and Glucose Levels in Obese Saudi Patients Irrespective Status of Blood Pressure

The association between LEP G-2548A gene polymorphism with increased plasma leptin and glucose levels and blood pressure in a sample of obese Saudi patients has been evaluated. This is a cross-sectional study involved 206 Saudi adult subjects (94 males and 112 females), randomly selected from the primary health care centers, Riyadh, Saudi Arabia. The study sample was categorized into three groups: 50 normotensive ND controls (age: 47.9±5.4 yr.; BMI 22.9±2.1 Kg/m2), 80 obese normotensive ND (age: 47.7±6.0 yr.; BMI 34.1±4.2 Kg/m2) and 76 obese hypertensive with T2D patients (age: 49.4±5.9 yr.; BMI: 35.1±4.7 Kg/m2). Analyses of LEP G-2548A gene polymorphism were determined using polymerase chain reaction (PCR), followed by restriction fragment length polymorphism (RFLP) with 2U of HhaI restriction enzyme. Plasma leptin and insulin levels were measured using the Luminex instrument. Fasting plasma glucose, total cholesterol, HDL-cholesterol, and triglycerides were measured using a chemical autoanalyzer Konelab machine. Also, blood pressure and anthropometric data were measured. The association analysis with metabolic parameters showed that homozygous AA of the LEP gene had significantly higher plasma glucose levels and HOMA-IR compared with homozygous GG (6.8±0.55 vs. 5.8±0.30; p< 0.04; 4.1±0.84 vs. 2.6±0.67; p=0.03) respectively. Besides, heterozygous GA had significantly higher plasma leptin levels compared with homozygous GG (40.0±2.6 vs. 29.6±2.6; P= 0.04). GA, AA, GA+AA genotypes of the LEP G-2548A gene polymorphism are more prevalent among individuals with hyperglycemia (OR= 3.7, 95% CI= 1.6 to 8.4, P= 0.001; OR= 3.2, 95% CI= 1.2 to 8.6, P= 0.03; OR= 3.5, 95%CI= 1.6 to 7.7, P= 0.001) respectively. A allele of the LEP gene is more prevalent among subjects with hyperglycemia (OR= 1.9, 95%CI= 1.2 to 3.0, P=0.006). G-2548A variant of the LEP gene may not be considered as a genetic risk factor for hypertension in Saudi obese patients. However, the genotypes (GA and AA) and -2548AA allele of this gene may represent important risk factors predisposing healthy subjects to develop T2DM irrespective of the status of blood pressure.

Leptin G-2548A Gene Polymorphism is Positively Associated with Increased Plasma Leptin and Glucose Levels in Obese Saudi Patients Irrespective Status of Blood Pressure

The association between LEP G-2548A gene polymorphism with increased plasma leptin and glucose levels and blood pressure in a sample of obese Saudi patients has been evaluated. This is a cross-sectional study involved 206 Saudi adult subjects (94 males and 112 females), randomly selected from the primary health care centers, Riyadh, Saudi Arabia. The study sample was categorized into three groups: 50 normotensive ND controls (age: 47.9±5.4 yr.; BMI 22.9±2.1 Kg/m2), 80 obese normotensive ND (age: 47.7±6.0 yr.; BMI 34.1±4.2 Kg/m2) and 76 obese hypertensive with T2D patients (age: 49.4±5.9 yr.; BMI: 35.1±4.7 Kg/m2). Analyses of LEP G-2548A gene polymorphism were determined using polymerase chain reaction (PCR), followed by restriction fragment length polymorphism (RFLP) with 2U of HhaI restriction enzyme. Plasma leptin and insulin levels were measured using the Luminex instrument. Fasting plasma glucose, total cholesterol, HDL-cholesterol, and triglycerides were measured using a chemical autoanalyzer Konelab machine. Also, blood pressure and anthropometric data were measured. The association analysis with metabolic parameters showed that homozygous AA of the LEP gene had significantly higher plasma glucose levels and HOMA-IR compared with homozygous GG (6.8±0.55 vs. 5.8±0.30; p< 0.04; 4.1±0.84 vs. 2.6±0.67; p=0.03) respectively. Besides, heterozygous GA had significantly higher plasma leptin levels compared with homozygous GG (40.0±2.6 vs. 29.6±2.6; P= 0.04). GA, AA, GA+AA genotypes of the LEP G-2548A gene polymorphism are more prevalent among individuals with hyperglycemia (OR= 3.7, 95% CI= 1.6 to 8.4, P= 0.001; OR= 3.2, 95% CI= 1.2 to 8.6, P= 0.03; OR= 3.5, 95%CI= 1.6 to 7.7, P= 0.001) respectively. A allele of the LEP gene is more prevalent among subjects with hyperglycemia (OR= 1.9, 95%CI= 1.2 to 3.0, P=0.006). G-2548A variant of the LEP gene may not be considered as a genetic risk factor for hypertension in Saudi obese patients. However, the genotypes (GA and AA) and -2548AA allele of this gene may represent important risk factors predisposing healthy subjects to develop T2DM irrespective of the status of blood pressure.

Leptin G-2548A Gene Polymorphism is Positively Associated with Increased Plasma Leptin and Glucose Levels in Obese Saudi Patients Irrespective Status of Blood Pressure

The association between LEP G-2548A gene polymorphism with increased plasma leptin and glucose levels and blood pressure in a sample of obese Saudi patients has been evaluated. This is a cross-sectional study involved 206 Saudi adult subjects (94 males and 112 females), randomly selected from the primary health care centers, Riyadh, Saudi Arabia. The study sample was categorized into three groups: 50 normotensive ND controls (age: 47.9±5.4 yr.; BMI 22.9±2.1 Kg/m2), 80 obese normotensive ND (age: 47.7±6.0 yr.; BMI 34.1±4.2 Kg/m2) and 76 obese hypertensives with T2D patients (age: 49.4±5.9 yr.; BMI: 35.1±4.7 Kg/m2). A blood sample was collected from the participants for PCR-RFLP, chemical autoanalyzer Konelab, and Luminex instruments. Analyses of LEP G-2548A gene polymorphism were determined using polymerase chain reaction (PCR), followed by restriction fragment length polymorphism (RFLP) with 2U of HhaI restriction enzyme. Plasma leptin and insulin levels were measured using the Luminex instrument. Fasting plasma glucose, total cholesterol, HDL-cholesterol, and triglycerides were measured using a chemical autoanalyzer Konelab machine. Also, blood pressure and anthropometric data were measured. The association analysis with metabolic parameters showed that homozygous AA of the LEP gene had significantly higher plasma glucose levels and HOMA-IR compared with homozygous GG (6.8±0.55 vs. 5.8±0.30; p< 0.04; 4.1±0.84 vs. 2.6±0.67; p=0.03) respectively. Besides, heterozygous GA had significantly higher plasma leptin levels compared with homozygous GG (40.0±2.6 vs. 29.6±2.6; P= 0.04). GA, AA, GA+AA genotypes of the LEP G-2548A gene polymorphism are more prevalent among individuals with hyperglycemia (OR= 3.7, 95% CI= 1.6 to 8.4, P= 0.001; OR= 3.2, 95% CI= 1.2 to 8.6, P= 0.03; OR= 3.5, 95%CI= 1.6 to 7.7, P= 0.001) respectively. A allele of the LEP gene is more prevalent among subjects with hyperglycemia (OR= 1.9, 95%CI= 1.2 to 3.0, P=0.006). G-2548A variant of the LEP gene may not be considered as a genetic risk factor for hypertension in Saudi obese patients. However, the genotypes (GA and AA) and -2548AA allele of this gene may represent important risk factors predisposing healthy subjects to develop T2DM irrespective of the status of blood pressure.