مجله بیوتکنولوژی کشاورزی (Sep 2022)
mRNA expression profile of DNAH1 gene in testis tissue of Raini Cashmere goat
Abstract
ObjectiveIt has been shown that mutation in DNAH1 (Dynein Axonemal Heavy Chain 1) gene is associated with MMAF (multiple morphological anomalies of the flagella) and PCD (primary ciliary dyskinesia) phenotypes. MMAF has been widely studied in male reproductive traits and is often associated with abnormal sperm, which seriously affects male reproductive traits and even leads to infertility. Indeed, as a MMAF-dependent gene, many studies have identified the DNAH1 gene as a key and essential player in male and female gonadal development and have investigated the clinical applications of the DNAH1 gene. PCD is often associated with ciliary motility disorder, which is also an autosomal recessive genetic disease. Animals with severe PCD show infertility. This study aimed to study the expression pattern of the DNAH1 gene in the testicular tissue of Raini Cashmere goat using Real-Time PCR.Materials and MethodsSampling was done from the testicular tissue of three heads (3 replicates from each head) of Raini Cashmere goats in the slaughterhouse. The isolated samples were placed in 1.5 mL microtubes. Total RNA was extracted from testicular tissue. Agarose gel electrophoresis and UV spectrophotometry were used to evaluate the quality and quantity of extracted RNA. Then cDNA was made and Real-Time PCR was performed. Also, the GAPDH housekeeping gene was used as a control. Melting curves were analyzed and data obtained from Real-Time PCR were performed.Results The results of Real-Time PCR curves and observing the results of electrophoresis of PCR products on 2% agarose gel showed that the DNAH1 gene is expressed in testicular tissue. For the DNAH1 gene, a band of 140 bp was observed and for the GAPDH gene, a band of 143 bp was observed, and the accuracy of the amplification test was confirmed.ConclusionsAccording to the results of the present study and the results of other researchers, it can be concluded that the DNAH1 gene plays an important role in fertility. In the present study, it was found that the DNAH1 gene is expressed in testicular tissue. Therefore, the DNAH1 gene is most likely essential for male fertility, and the results of this research have provided a way for future studies to describe the role of the DNAH1 gene as a candidate gene for better fertility and normal physiology in domestic animals, especially goats.
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