【摘要】：精子進入雌性生殖道后,必須經(jīng)過獲能、頂體反應(yīng)等一系列復(fù)雜的結(jié)構(gòu)和功能的改變,即被激活后才具備受精能力。精子的激活受到許多生物因素的調(diào)節(jié),目前有關(guān)精子激活的分子機制及信號途徑遠未得到闡明。 雌性生殖道環(huán)境中存在一定量的雌激素,尤以卵泡液及宮頸粘液中雌激素的含量最高。有研究結(jié)果表明一些動物和人精子膜上可能存在雌激素受體或雌激素結(jié)合位點,因此,精子在穿越雌性生殖道時,雌性生殖道中的雌激素可能與精子相互作用,通過特定的信號途徑誘導(dǎo)精子激活。但是,雌激素對哺乳動物精子激活作用的普遍性還有待證明,雌激素對人精子的激活作用及其相關(guān)的信號機制也值得進一步探索。 為此,我們擬采用三色法頂體染色、流式細胞術(shù)、免疫印跡技術(shù)進行以下研究:第一,用不同濃度的17β-雌二醇(E_2)分別作用生育力正常男性和不明原因不育癥男性獲能或非獲能精子,檢測精子頂體反應(yīng)率(AR率)、精子內(nèi)游離鈣離子濃度([Ca~(2+)]_i)的變化,從而研究雌激素對人精子的激活作用及該作用對精子功能的意義。第二,比較游離雌二醇與非透膜性的雌激素-牛血清蛋白復(fù)合物(E_2-BSA)對人精子AR率、[Ca~(2+)]_i、蛋白質(zhì)酪氨酸磷酸化的影響的差異;并以無鈣液代替有鈣液再檢測E_2、E_2-BSA所致精子[Ca~(2+)]_i的變化,從而初步探討雌激素對人精子激活的可能機制。 實驗結(jié)果如下:
[Abstract]:After sperm enters the female reproductive tract, it must undergo a series of complex structural and functional changes, such as capacitation, acrosome reaction and so on, that is to say, the sperm is activated before it can be fertilized. Sperm activation is regulated by many biological factors. The molecular mechanism and signal pathway of sperm activation are far from elucidated. There is a certain amount of estrogen in female reproductive tract environment, especially in follicular fluid and cervical mucus. Studies have shown that estrogen receptor or estrogen binding sites may exist in some animal and human sperm membranes. Therefore, estrogen in the female reproductive tract may interact with spermatozoa when sperm passes through the female reproductive tract. Sperm activation is induced by specific signaling pathways. However, the universality of estrogen on mammalian sperm activation has yet to be proved. The activation of estrogen on human spermatozoa and its related signal mechanisms are also worth further exploration. To this end, we plan to use trichromatic acrosome staining, flow cytometry, immunoblotting techniques to carry out the following studies: first, The sperm acrosome reaction rate (AR rate) was detected by using different concentrations of 17 尾-estradiol (E 尾-estradiol) to affect capacitated or non-capacitated spermatozoa of normal fertility men and unexplained infertile men, respectively. The changes of intracellular free calcium concentration ([Ca~ (2)] I) in spermatozoa were used to study the activation of estrogen on human spermatozoa and the significance of this effect on sperm function. Secondly, the effects of free estradiol and non-membranous estrogen-bovine serum protein complex (E_2-BSA) on human sperm AR rate, [Ca~ (2)] _ I, protein tyrosine phosphorylation were compared. The changes of [Ca~ (2)] _ I induced by E _ (2) O _ (2) and E _ (2) O-BSA in human spermatozoa were detected with calcium-free solution instead of calcium-free solution, so as to explore the possible mechanism of estrogen-induced activation of human spermatozoa. The experimental results are as follows:
【學(xué)位授予單位】：四川大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2005
【分類號】：R33

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相關(guān)期刊論文 前2條

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本文編號：2465923