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Study Of The Relationship Between SEPT4, TEKT4and Idiopathic Asthenozoospermia

Posted on:2013-04-19Degree:MasterType:Thesis
Country:ChinaCandidate:X X FengFull Text:PDF
GTID:2234330371476043Subject:Clinical Laboratory Science
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Background and ObjectiveIn recent decades, the semen quality of men is downward, while male infertility has been an important problem in the the world. Sperm motility is an important indicator of semen quality.Sperm motility is essential for penetrating the cervical mucus to reach the fertilization site, penetrating the corona radiate and zona pellucida to reach oocytes and complete the whole process of fertilization. The sperm is divided into a, b, c and d sperm motility according to the ability of the sperm forward motility by World Health Organization. Asthenozoospermia (AZS),(a+b sperm motility<50%, or a sperm motility<25%in fresh ejaculate), according to World Health Organization(1999) Guidelines],is one of the main causes of human male subfertility and infertility.AZS is found in as many as24%of patients presenting for evaluation of male subfertility and may be a significant factor in another55%of patients with combined defects in sperm density, motility and morphology. The mechanism of causing asthenozoospermia is very complex,mainly related to the abnormal energy metabolism of sperm, the abnormal signal transduction pathways,and the abnormalities of sperm structure. Because sperm motility depends mainly on sperm flagellar motion, understanding the structural characteristics of the sperm flagellumand analyzing the cause and the mechanism of asthenozoospermia is particularly important to provide effective clinical treatment strategies for the subfertility and infertility men. The important component of sperm flagellum,which is a major component of sperm,is axoneme, outer dense fibers and mitochondrial sheath. Recent studies showed that there are250kinds of sperm axoneme and its appendages which are essential for flagellar motion and whose mutations or expression abnormalities can be caused the sperm flagellar structure and function, resulting in sperm motility lowly. To understand the causes and mechanism of the idiopathic asthenozoospermia,we explore the relationship between the sperm flagellum gene SEPT4, TEKT4and idiopathic asthenozoospermia,and provid reliable experimental and theoretical basis for the dignosis and the treatment of idiopathic asthenozoospermia.Materials and method1Objects30sperm samples,(normal control group), were collected from normozoospermic donors of Human Sperm Bank in Henan Province from September2010to February2011.During the same time, there were32sperm samples of idiopathic asthenozoospermia patients(idiopathic asthenozoospemia group) from the andrology outpatient clinics of the Third Affiliated Hospital, Zhengzhou University. The age of the two groups ranged from20to38, and there was no significant difference between the two groups.2Methods2.1reverse transcription polymerase chain reaction testsSperm samples from normal male and asthenozoospermia patients, were used for detecting the changes of SEPT4mRNA and TEKT4mRNA by reverse transcription polymerase chain reaction.2.2Western blottig testsSperm samples from normal male and asthenozoospermia patients, were used for detecting the changes of SEPT4protein and TEKT4protein by Western blotting.2.3Immunocytochemistry experimentssperm samples from normal male and asthenozoospermia patients, were used for analysising the difference distribution and expression of SEPT4protein by immunocytochemistry.3Statistics analysisThe data obtained from the tests was described by x±s and analyzed statistically by spss17.0version.The distribution and expressions of SEPT4and TEKT4by RT-PCR,western blotting and immunocytochemistry were analysised by two independent samples t-test.Results1Distribution and experssions of SEPT4between normal male and idiopathic asthenozoospermia patientsDistribution of SEPT4protein was localized in the annulus of the sperm flagella by imunocytochemistry.The expressions of SEPT4in idiopathic asthenozoospermia patients were significantly decreased compared with the control group (t=3.452, P=0.001).2Experssions of SEPT4mRNA between normal male and idiopathic asthenozoospermia patientsThe expressions of SEPT4mRNA in idiopathic asthenozoospermia patients were significantly decreased compared with the control group by reverse transcription polymerase chain reaction (t=3.521,P=0.001).3Experssions of SEPT4protein between normal male and idiopathic asthenozoospermia patientsBy Western blotting, the expressions of SEPT4protein in idiopathic astheno-zoospermia patients were significantly decreased compared with the control group (t= 5.872, P=0.001).4Experssions of TEKT4mRNA between normal male and idiopathic asthenozoospermia patientsBy reverse transcription polymerase chain reaction, the expressions of TEKT4mRNA in idiopathic asthenozoospermia patients were significantly decreased Compared with the control group.(t=4.325,p=0.001).5Experssions of TEKT4protein between normal male and idiopathic asthenozoospermia patientsCompared with the control group, the expressions of TEKT4protein in idiopathic asthenozoospermia patients were significantly decreased by Western blotting (t=5.939, P=0.001).ConclusionThe decreasing expression of SEPT4and TEKT4could be one of the causes of idiopathic asthenozoospermia.
Keywords/Search Tags:idiopathic, asthenozoospermia, SEPT4, TEKT4, RT-PCRWestern blotting, imunocytochemistry
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