Toll-like receptor (TLR)-mediated inflammatory processes play a critical role in the innate immune response during the initial interaction between the infecting microorganism and immune cells. This study aimed to investigate the possible microanatomical and histological differences in mandibular and bronchial lymph nodes in Akkaraman and Romanov lambs induced by lipopolysaccharide (LPS) and lipoteichoic acid (LTA) and study the gene, protein, and immunoexpression levels of TLR4, myeloid differentiation factor 88 (MyD88), and tumor necrosis factor-α (TNF-α) that are involved in the immune system. Microanatomical examinations demonstrated more intense lymphocyte infiltration in the bronchial lymph nodes of Akkaraman lambs in the LPS and LTA groups compared to Romanov lambs. TLR4, MyD88, and TNF-α immunoreactivities were more intense in the experimental groups of both breeds. Expression levels of MyD88 and TNF-α genes in the bronchial lymph node of Akkaraman lambs were found to increase statistically significantly in the LTA group. TLR4 gene expression level in the mandibular lymph node was found to be statistically significantly higher in the LTA + LPS group. In conclusion, dynamic changes in the immune cell populations involved in response to antigens such as LTA and LPS in the lymph nodes of both breeds can be associated with the difference in the expression level of the TLR4/MyD88/TNF-α genes.
Toll-like receptors are involved in the recognition of bacterial toxins, which cause infection in the respiratory system. This study aimed to evaluate microanatomical and histological alterations in the lungs of 24 healthy Akkaraman and Romanov lambs after the administration of lipoteichoic acid (LTA), lipopolysaccharide (LPS), and LTA + LPS and investigate the gene, protein, and immune expression levels of TLR4, MyD88, and TNF-α molecules, known to have immune functions. Microanatomical examinations showed thickened peribronchial and alveolar walls in the lungs of groups LTA, LPS, and LTA + LPS of both breeds due to immune cell infiltration. TLR4, MyD88, and TNF-α immunoexpressions were positive to varying degrees in the cytoplasm and nucleus of the bronchial and bronchiolar luminal epithelial cells, alveolar epithelial cells, and alveolar macrophages. TLR4 and TNF-α protein expressions were statistically different in the LPS-treated Romanov lambs, compared to the other groups. Among the Akkaraman lambs, TLR4 gene expression was significantly higher in group LPS, and among the Romanov lambs, TLR4, MyD88, and TNF-α gene expressions were significantly higher in group LTA + LPS. Therefore, TLR4, MyD88, and TNF-α molecules, involved in the immune response, were found to be expressed at different levels against LTA and LPS in the lungs of two different sheep breeds.
The aim of this study was to determine the genetic variation of MYF-5 and STAT5A genes in Anatolian water buffalo which was the only buffalo breed reared in Turkey by using the PCR-RFLP method. In this study, 120 Anatolian water buffalo were examined. After PCR amplification for MYF-5 gene, 512 bp PCR products were digested with TaqI enzyme. Although no AA genotype was found, the frequency of GG and AG genotypes were 0.77 and 0.33. PCR products of 215 bp for STAT5A gene were digested with AvaI enzyme and showed that all of the Anatolian water buffalo examined had monomorphic in terms of CC genotype. Anatolian water buffalo were found in Hardy-Weinberg equilibrium with respect to MYF-5-TaqI polymorphism.
This study, it was aimed to investigate the expression levels of transcription factor (TF) genes (MYF6, MYOD1, MYF5, MYOG) and proteins associated with muscle growth in the longissimus dorsi (LD) and gluteal (GL) muscles in sheep. For this purpose, two fat-tailed sheep breeds (Akkaraman (n = 10), İvesi (n = 10)) and two thin-tailed sheep breeds (Kivircik (n = 10) and Karayaka (n = 10)) from Turkey's native sheep breeds were examined. The expression level of RNAs and proteins isolated from fresh tissues and MYF6, MYOD1, MYF5, MYOG proteins were analyzed. As a result of the statistical analysis, in the LD tissue, respectively, MYOG and MYF5 genes in the Karayaka sheep breed; MYOD1 gene in Akkaraman sheep breed; MYF5 gene in Awassi sheep breed were found to be statistically significant (P < 0.05). In GL tissue, respectively, MYOG and MYF6 genes in Akkaraman sheep breed; MYOD1 gene in Karayaka sheep breed; MYF6 gene in Akkaraman and Awassi sheep breed were found to be statistically significant (P < 0.05). In the present study, it was found that the MYOG (fold change 6.87) and MYOD1 (fold change 15.41) genes were upregulated in the GL muscle of the fat-tailed Akkaraman sheep breed. In addition, in the thin-tailed Karayaka sheep breed, down-regulation of MYOD1 (fold change − 0.22) gene in LD muscle and up-regulation of MYOD1 (fold change 6.67) gene in GL muscle was found. As a result, it can be considered that MYOG and MYOD1 genes as candidate genes in molecular selection studies for our fat-tailed and thin-tailed indigenous breeds in terms of muscle development.
Öz: Bu çalışmada, Simental ve İsviçre Esmeri sığırlarda karaciğer ve kas dokusu MyH8 gen ekspresyonu ile canlı ağırlık ilişkisinin araştırılması amaçlanmıştır. Çalışmada Simental (n=40) ve İsviçre Esmeri (n=40) erkek sığırlar kullanılmıştır. Kesimden önce canlı ağırlık ölçümleri yapılmış, kas ve karaciğer dokusu alınarak sıvı azotta dondurulmuştur. Sırası ile dokulardan Trizol ile RNA izolasyonu, RNA'lardan c-DNA sentezi ve SYBR Green yöntemi ile ekspresyon analizi yapılmıştır. Canlı ağırlıklar ile kas ve karaciğerde MyH8 gen ekspresyonu arasında önemli bir korrelasyon bulunmamıştır (P>0.05). Simental ve İsviçre Esmeri arasında kas MyH8 gen ekspresyonu bakımından farklılık bulunmamıştır (P>0.05). Ancak karaciğer MyH8 gen ekspresyonu iki ırktada önemli bulunmuştur (P<0.001). İsviçre Esmeri karaciğer MyH8 ekspresyonu Simentale göre yüksek bulunmuştur. Sonuç olarak, çalışmada elde edilen veriler Simental ve İsviçre Esmeri MyH8 geni ekspresyonu ile canlı ağırlığı arasında anlamlı bir ilişkinin olmadığını ancak MyH8 geninin bu ırklarda farklı ekspresyon profili taşıma potansiyeli olduğunu ortaya koymaktadır.Abstract: The aim of this study was to investigate, the relationship between of MyH8 gene expression profile of the liver and muscle tissue with the live weight, in the Simmental and Brown Swiss cattle. Simental (n = 40) and Brown Swiss (n = 40) male cattle were used in this study. Before slaughter, live weight measurements, muscle and liver tissue were taken and tissues were frozen in liquid nitrogen. It was made respectively RNA isolation by Trizol from tissue, c-DNA synthesis from RNAs and expression analysis by SYBR Green method. There was no significant correlation between live weights and MyH8 gene expression in muscle and liver (P>0.05). There was no significant difference in muscle MyH8 gene expression between Simmental and Brown Swiss (P>0.05). However, the difference in MyH8 gene expression in the liver was significant for two breeds (P<0.001). MyH8 gene expression level in Brown Swiss liver was significantly higher than the Simmental. As a result, the data obtained from study indicate that there is no significant relationship between the expression level of the MyH8 gene and live weight in the Simmental and Brown Swiss, but the MyH8 gene has the potential to carry a different expression profile in these breeds.
In this study, it was aimed to investigate the relationship between different three SNP (C367400T, A496561G and G519663A) on the CACNA2D1 gene, and subclinical mastitis in Holstein breed cattle reared Kayseri and around Kayseri. The study consisted of 151 Holstein cattle breed all in their third lactation. Blood and milk samples were taken from the animals in the study group. Subclinical mastitis status of animals was determined by California Mastitis Test (CMT) from milk samples and total DNA was isolated from blood samples by phenol-chloroform extraction method. SNPs were genotyped from DNA samples by PCR-RFLP method. In the study, CMT data, and distributions of genotypes of the three SNPs on the CACNA2D1 gene were calculated. In the study, genotype distributions were determined in terms of C367400T, A496561G and G519663A SNPs found on the CACNA2D1 gene according to CMT status. The difference between the C367400T, A496561G and G519663A SNPs was not significant (P>0.05). In the study group examined the Chi-square (χ2) analysis conducted, it was observed that the Holstein cattle were in the Hardy-Weinberg equilibrium (HWE) in terms of C367400T and A496561G SNPs, deviation from HWE for the G519663A SNP. As a result, it was thought that the CACNA2D1 gene and these SNPs should be evaluated with more samples and different mastitis indicator data in studies on mastitis resistance.
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