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研究生:修明
研究生(外文):Azhar Mahmud
論文名稱:ANAPC2表現與精蟲外型相關性研究
論文名稱(外文):The correlation of sperm morphology to the expression pattern of ANAPC2
指導教授:鄧燕妮
指導教授(外文):Yen-ni Teng
學位類別:碩士
校院名稱:國立臺南大學
系所名稱:生物科技學系碩士班
學門:生命科學學門
學類:生物科技學類
論文種類:學術論文
論文出版年:2013
畢業學年度:101
語文別:英文
論文頁數:38
外文關鍵詞:spermiogenesisinfertilityANAPC2
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ABSTRACT
For many years, tremendous efforts have been made to identify genetic defects or polymorphisms that are associated with human infertility, which is a major health problem worldwide. We have identified a sterility related male germ-cell specific gene, ANAPC2. Previous studies showed that anaphase-promoting complex or cyclosome play an important role to regulate meiosis, and has been implicated in post-mitotic function including dendrite formation in neurons, metabolic, and learning and memory processes. Because ANAPC2 was expressed at the sperm head and neck during spermiogenesis, we proposed that it will have an important role in sperm morphology.
Through more detailed observation, we found that trough immuno-fluorescence staining, ANAPC2 located in acrosome and neck region, during terminal differentiation. In human mature sperm, ANAPC2 was expressed specifically in the equatorial segment and neck area. Semen analysis showed ANAPC2 signals significantly decreased in the head and neck of sperm with abnormal morphology. Statistical analysis showed a significant ANAPC2 expression differences between normal and abnormal head and neck region. The real time polymerase chain reaction concludes that the most significant differences between normal and abnormal groups of sperm showed between control and asthenoteratozoospermia. Thus, our studies inferred that ANAPC2 is critical during mammalian sperm elongation.
Key words: infertility, ANAPC2, spermiogenesis
TABLE OF CONTENT
ABSTRACT ............................................................................................................................. i
ACKNOWLEDGEMENT ..................................................................................................... ii
TABLE OF CONTENT ........................................................................................................ iii
LIST OF FIGURES................................................................................................................ v
I. INTRODUCTION .......................................................................................................... 1
1.1 Male infertile .................................................................................................................. 1
1.2 The process of spermatogenesis ..................................................................................... 2
1.3 Anaphase promoting complex-2 (ANAPC2) ................................................................. 4
1.4 Objective of the study .................................................................................................... 6
II. MATERIALS AND METHODS ................................................................................... 7
2.1 Mouse sperm preparation ............................................................................................... 7
2.2 Immuno-fluorescence Assay (IFA) ................................................................................ 8
2.3 Subjects ........................................................................................................................ 10
2.4 Semen Analysis ............................................................................................................ 10
2.5 Total RNA isolation ...................................................................................................... 11
2.6 Real-time Polymerase Chain Reaction (RT PCR)........................................................ 12
2.7 Statistical analysis ........................................................................................................ 13
iv
III. RESULTS ....................................................................................................................... 14
3.1 Expression pattern of a novel gene in mouse and human sperm ................................. 14
3.2 Expression pattern of ANAPC2 in human spermatozoa with abnormal morphology . 15
3.3 Statistical Analysis ....................................................................................................... 16
IV. DISCUSSION ............................................................................................................... 17
4.1 The functional role of ANAPC2 in spermiogenesis ..................................................... 17
4.2 ANAPC2 in mammalian spermatogenesis ................................................................... 18
4.3 The role of ANAPC2 in male infertility ....................................................................... 19
V. CONCLUSION............................................................................................................. 21
References ............................................................................................................................. 22
Figures and Table ................................................................................................................. 26
CURRICULUM VITAE ...................................................................................................... 32
v
LIST OF FIGURES
Figure 1. Fluorescence analysis of mouse mature sperm and germ cells .............................. 26
Figure 2. ANAPC2 expression in human mature sperm and germ cells................................ 27
Figure 3. Sperm head analysis................................................................................................ 28
Figure 4. Sperm neck analysis................................................................................................ 28
Figure 5. Anova analysis by real-time polymerase chain reaction (RT-PCR)........................ 29
Figure 6. Human spermatozoa with various types of morphological defects. ....................... 30
vi
LIST OF TABLE
Table 1. Core and regulatory subunits of the ANAPC..........................................................31
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