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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 吳瑞菁 | zh_TW |
| dc.contributor.advisor | Jui-Ching Wu | en |
| dc.contributor.author | 孫里葶 | zh_TW |
| dc.contributor.author | Li-Ting Sun | en |
| dc.date.accessioned | 2024-08-16T17:58:58Z | - |
| dc.date.available | 2024-08-17 | - |
| dc.date.copyright | 2024-08-16 | - |
| dc.date.issued | 2024 | - |
| dc.date.submitted | 2024-07-14 | - |
| dc.identifier.citation | Eisenberg, M.L., et al., Male infertility. Nature Reviews Disease Primers, 2023. 9(1).
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Hu, J., et al., Distinct roles of two myosins in C. elegans spermatid differentiation. PLOS Biology, 2019. 17(4): p. e3000211. Duerr, J.S., Antibody Staining in C. Elegans Using "Freeze-Cracking". Journal of Visualized Experiments, 2013(80). 林怡秀, 利用線蟲為模型探討PP1去磷酸酶GSP-1與GSP-2在雄性減數分裂中染色體分離的角色, in 醫學檢驗暨生物技術學研究所. 2016, 國立臺灣大學. p. 1-56. Hsu, J.-Y., et al., Mitotic Phosphorylation of Histone H3 Is Governed by Ipl1/aurora Kinase and Glc7/PP1 Phosphatase in Budding Yeast and Nematodes. Cell, 2000. 102(3): p. 279-291. Kondo, T. and A. Kimura, Impaired chromosome segregation results in sperms with excess centrosomes in emb-27 APC6 mutant C. elegans. 2018, Cold Spring Harbor Laboratory. Gorjánácz, M., et al., Caenorhabditis elegans BAF-1 and its kinase VRK-1 participate directly in post-mitotic nuclear envelope assembly. The EMBO Journal, 2007. 26(1): p. 132-143. Flora, P., et al., Role of Chromatin Modifications in Drosophila Germline Stem Cell Differentiation. 2017, Springer International Publishing. p. 1-30. Park, J.-A., et al., Deacetylation and Methylation at Histone H3 Lysine 9 (H3K9) Coordinate Chromosome Condensation during Cell Cycle Progression. Molecules and Cells, 2011. 31(4): p. 343-350. Cornes, E., et al., piRNAs initiate transcriptional silencing of spermatogenic genes during C. elegans germline development. Developmental Cell, 2022. 57(2): p. 180-196.e7. Mazzetto, M., et al., Characterization of the distribution and dynamics of chromatin states in the C. elegans germline reveals substantial H3K4me3 remodeling during oogenesis. Genome Research, 2024. 34(1): p. 57-69. Schedl, T. and J. Kimble, fog-2, a germ-line-specific sex determination gene required for hermaphrodite spermatogenesis in Caenorhabditis elegans. Genetics, 1988. 119(1): p. 43-61. Depina, A.S. and G.M. Langford, Vesicle transport: The role of actin filaments and myosin motors. Microscopy Research and Technique, 1999. 47(2): p. 93-106. Varkey, J.P., et al., The Caenorhabditis elegans spe-26 gene is necessary to form spermatids and encodes a protein similar to the actin-associated proteins kelch and scruin. Genes & Development, 1995. 9(9): p. 1074-1086. | - |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/94750 | - |
| dc.description.abstract | 晚期成精作用代表了精細胞從減數分裂前期轉入中期，並得以進行分裂的階段。在這個轉變過程中，染色體和細胞質組件經歷廣泛的重新排列。雄性特有的轉譯調節因子CPB-1在粗線期的細胞質中高度表達。為了研究CPB-1在雄性生育中的功能，我們產生了一個可誘導降解CPB-1的線蟲株，以達成特定條件下的CPB-1降解。
在缺乏CPB-1的情況下，形成了初級精母細胞，但未能繼續進行分裂。對性腺的形態進行檢查顯示，CPB-1的缺乏不影響一般粗線期的染色體排列。然而，精元細胞核未能形成核小體，這是在初級精母細胞形成之前實現轉錄沉默的階段。此外，CPB-1缺失的初級精母細胞表現出轉錄活性。 我們的研究結果表明，CPB-1通過調節核小體階段的形成，在轉錄沉默中發揮作用。此外，CPB-1缺失的精原細胞缺乏細胞質顆粒，並表現出空泡狀結構，表明細胞膜胞器的異常。支持這一點的是，在缺乏CPB-1的情況下，精子特異性胞器纖維體-膜細胞器複合體(FB-MO)的結構出現高度混亂。FB-MO中的主要物質MSP蛋白有顯著的減少，並且混亂的MO結構與類似空泡的結構共定位。 綜上所述，CPB-1調控了成精作用的兩個方面的關鍵因子的轉譯：首先，調控生成核小體階段，以在形成初級精母細胞之前實現轉錄沉默；其次，在細胞質中組裝精子特異性胞器FB-MO。這些發現表明CPB-1在男性生育中參與了關鍵作用。 | zh_TW |
| dc.description.abstract | Late spermatogenesis represents a stage at which the spermatogenic cells transition into the M phase and are committed for division. During this transition, chromosomes and cytoplasmic components undergo extensive rearrangement. The male-specific translation regulator CPB-1 is highly expressed in the cytoplasm during the pachytene spermatogenic germline stage. To investigate the function of CPB-1 in male fertility, we generated an inducible CPB-1 degradation strain to enable controlled CPB-1 depletion.
In the absence of CPB-1, primary spermatocytes were formed but failed to progress to division. Examination of germline morphology revealed that CPB-1 depletion did not affect the general early pachytene chromosome arrangement. However, spermatogenic nuclei failed to form the karyosome, a stage that allows transcription silencing before primary spermatocyte formation. Furthermore, CPB-1-depleted primary spermatocytes exhibited transcriptional activity. Our findings suggest that CPB-1 plays a role in transcriptional silencing by regulating the formation of the karyosome stage. Additionally, CPB-1-depleted spermatocytes lacked cytoplasmic granules and exhibited vacuole-like structures, indicating disorganization of cellular membranous organelles. Supporting this, the sperm-specific organelle FB-MO was highly disorganized in the absence of CPB-1. The levels of MSP, a major cargo in FB-MO, were severely reduced, and the disorganized MO structures colocalized with vacuole-like structures. In summary, CPB-1 controls the translation of key regulators in two critical aspects of spermatogenesis: first, the formation of the karyosome stage to achieve transcriptional silence before primary spermatocyte formation; second, the assembly of the sperm-specific organelle FB-MO in the cytoplasm. These findings highlight the crucial role of CPB-1 in male fertility. | en |
| dc.description.provenance | Submitted by admin ntu (admin@lib.ntu.edu.tw) on 2024-08-16T17:58:58Z No. of bitstreams: 0 | en |
| dc.description.provenance | Made available in DSpace on 2024-08-16T17:58:58Z (GMT). No. of bitstreams: 0 | en |
| dc.description.tableofcontents | 誌謝 i
中文摘要 ii Abstract iii Table of contents v Chapter 1-Introduction 1 1.1 The role of spermatogenesis in infertility. 1 1.2 Caenorhabditis elegans serves as an ideal model for investigating spermatogenesis. 2 1.3 Prophase chromosome dynamics in C. elegans spermatocytes. 3 1.4 Spermatocytes undergo transcriptional silencing during karyosome formation. 5 1.5 The crucial role of sperm-specific organelle FB-MO in C. elegans spermatogenesis. 6 1.6 Nascent protein synthesis is required for spermatogenesis. 8 1.7 Male-specific translation factors are crucial for meiotic division in spermatogenesis. 9 1.8 CPB-1 plays a role in spermatogenesis through translation control. 11 1.9 The conditional CPB-1 depletion in male germline via the AID system. 12 1.10 CPB-1 depletion leads to meiotic arrest and abnormal primary spermatocyte morphology. 13 Chapter 2-Material and Method 15 2.1 Worm Strain 15 2.2 Worm Culture 15 2.3 KNAA Treatment 16 2.4 Immunoblot Assay 17 2.5 Time-lapse Live Image Microscopy 18 2.7 Quantification Method for Vacuole-Like Structures 19 2.8 Immunofluorescence Staining Assay 20 2.9 Post-pachytene cell in syncytial gonad (Rachis) count 22 2.10 Nascent RNA detection 22 Chapter 3-Result 24 3.1 CPB-1 depletion leads to chromosomal and cytoplasmic defects during primary spermatocytes development. 24 3.2 CPB-1-depleted primary spermatocytes exhibit M phase markers. 26 3.3 Transcription remains active in CPB-1 depleted primary spermatocytes. 28 3.4 Primary spermatocytes bud out at diplotene in CPB-1 depletion. 30 3.5 Transcriptional inhibition could not rescue the meiotic arrest caused by CPB-1 depletion. 32 3.6 The chromosome program during pachytene remains normal in CPB-1 depletion. 33 3.7 Vacuole-like structure formation correlates with duration of CPB-1 depletion. 35 3.8 Vacuole-like structure formation is not caused by meiotic arrest. 36 3.9 CPB-1 depletion causes disorganization of FB-MO structures. 37 3.10 The disorganized FB-MO structures cause vacuole-like structures formation in CPB-1 depletion. 38 Chapter 4-Discussion 41 4.1 The potential role of CPB-1 in karyosome formation in male germline. 42 4.2 The potential role of karyosome formation in C. elegans spermatogenesis. 43 4.3 Potential mechanisms of MSP downregulation in CPB-1 depleted spermatocytes. 45 4.4 Cytoplasmic implications of CPB-1 depletion in pachytene stage spermatocytes. 46 Figures 48 Figure 1: Model of hypothesis in this study. 49 Figure 2: 3xFLAG-CPB-1 in germline. 51 Figure 3: The phenotypes caused by CPB-1 Depletion. 53 Figure 4: Primary spermatocyte can enter the M phase but unable to divide. 56 Figure 5: CPB-1 depleted primary spermatocytes remain transcription activity 58 Figure 6: CPB-1 depleted primary spermatocytes bud out during Diplotene. 60 Figure 7: Transcriptional inhibition could not rescue the meiotic arrest caused by CPB-1 Depletion. 63 Figure 8: Chromosome program remains normal in CPB-1 depleted pachytene stage. 65 Figure 9: Vacuole-like structure formation correlates with duration of CPB-1 depletion. 67 Figure 10: Vacuole-like structure formation is not directly caused by meiotic arrest. 69 Figure 11: FB-MO structures disorganized in CPB-1 depletion. 71 Figure 12: The disorganized FB-MO structures cause vacuole-like structures formation in CPB-1 depletion. 74 Figure 13: Model of CPB-1 in spermatogenesis. 76 Tables 77 Table 1. Strains used in this study 77 Reference 78 | - |
| dc.language.iso | en | - |
| dc.subject | 精子生成 | zh_TW |
| dc.subject | CPEB | zh_TW |
| dc.subject | CPB-1 | zh_TW |
| dc.subject | 核小體生成 | zh_TW |
| dc.subject | 轉錄沉默 | zh_TW |
| dc.subject | 精子特異性胞器FB-MO | zh_TW |
| dc.subject | CPEB | en |
| dc.subject | Karyosome formation | en |
| dc.subject | Spermatogenesis | en |
| dc.subject | Transcriptional silencing | en |
| dc.subject | CPB-1 | en |
| dc.subject | FB-MO | en |
| dc.title | 雄性特異性轉譯調節因子CPB-1在線蟲精子生成中參與調控轉錄關閉和精子特異性胞器組成 | zh_TW |
| dc.title | Male-specific translation regulator CPB-1 regulates transcriptional silencing and sperm-specific organelle assembly in C. elegans spermatogenesis. | en |
| dc.type | Thesis | - |
| dc.date.schoolyear | 112-2 | - |
| dc.description.degree | 碩士 | - |
| dc.contributor.oralexamcommittee | 吳師誠;賴韻如;蔡欣祐 | zh_TW |
| dc.contributor.oralexamcommittee | Shih-Chen Wu;Yun-Ju Lai;Hsin-Yue Tsai | en |
| dc.subject.keyword | 精子生成,CPEB,CPB-1,核小體生成,轉錄沉默,精子特異性胞器FB-MO, | zh_TW |
| dc.subject.keyword | Spermatogenesis,CPEB,CPB-1,Karyosome formation,Transcriptional silencing,FB-MO, | en |
| dc.relation.page | 87 | - |
| dc.identifier.doi | 10.6342/NTU202401754 | - |
| dc.rights.note | 未授權 | - |
| dc.date.accepted | 2024-07-15 | - |
| dc.contributor.author-college | 醫學院 | - |
| dc.contributor.author-dept | 醫學檢驗暨生物技術學系 | - |
| 顯示於系所單位： | 醫學檢驗暨生物技術學系 | |
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