中华鳖输卵管Telocyte的特性研究
发布时间：2020-10-27 22:18
输卵管是脊椎动物重要的生殖器官,参与和调控卵子的运输、成熟、储存、受精与早期胚胎发育,但各种动物输卵管的组成与结构存在差异。中华鳖是我国养殖面积最大的爬行动物,不仅具有重要的经济价值,而且具有特殊的精子储存现象。中华鳖输卵管由漏斗部、蛋白分泌部、峡部、子宫部和阴道部5段组成,其中峡部、子宫部和阴道部前段是发生精子储存的部位。新近在哺乳动物器官中发现了一种特异的细胞—Telocyte(暂时翻译为远细胞),参与体内许多器官(尤其是管状器官)的功能活动与调节。但Telocyte在非哺乳动物的分布与作用未见资料报道。本文利用形态学方法和细胞生物学技术,观测鉴定Telocyte在中华鳖输卵管精子储存部位(包括峡部、子宫部和阴道部)的分布,分析其超微结构特点,阐明Telocyte及其突起与周围环境和细胞的形态联系。进一步在分子水平上探究Telocyte标记物-CD34 mRNA在输卵管的表达。首次在哺乳类之外的动物体内鉴定发现telocyte的分布及其特性,研究结果为阐明中华鳖精子储存机理提供细胞分子基础。实验Ⅰ中华鳖输卵管阴道部telocytes的超微结构与免疫组化特征Telocytes是2005年新发现的一种间质细胞,由Popescu和他的同事命名,目前已经在哺乳动物不同类型的组织和器官中被确认存在,但阴道部中未见资料报道(www.telocytes.com)。本研究利用透射电镜和免疫组化探查中华鳖输卵管阴道部telocytes的分布、超微结构以及它们与周围细胞的形态关系。结果显示,telocytes主要位于单层柱状上皮下方结缔组织中,靠近分泌腺细胞处可见Tcs及其突起,有些Tcs及其突起与肥大细胞和胶原纤维密切相关。Tcs及其突起呈CD34免疫反应阳性。在细胞水平上显示Tcs在中华鳖输卵管阴道部的定位,它们的细胞标记与哺乳类具有同源性。研究结果为理解阴道功能及防治阴道疾病提供了新视角。实验Ⅱ中华鳖输卵管子宫部telocytes的超微特征Telocytes(Tcs)与体内许多机能调节和某些疾病发生有关。Tcs具有极其细长的突起(telopodes,Tps),可伸达远端,有人将其翻译为远(端)细胞。Tps由细段(podomers)和膨大部(podoms)交替排列,呈串珠样结构。Tcs的研究报道全部为人和哺乳动物的资料,已经在心、肺、肾、消化道等器官组织中被鉴定发现。应用透射电镜对中华鳖子宫部Tcs的超微结构特性进行了观察鉴定,结果显示中华鳖输卵管子宫部具有Tcs分布,而且主要位于子宫固有膜结缔组织中,胞体结构完整,突起细长,与神经末梢、毛细血管、胶原纤维束以及子宫腺密切相关。这种结构关系表明,Tcs及其突起具有参与中华鳖子宫收缩及其腺体分泌的形态学特征。实验Ⅲ中华鳖输卵管峡部telocytes的特征Telocytes拥有一个很小的细胞体,很细长的telopodes(Tps),且podomers与podoms交替排布。Tps释放3种胞外小泡:外来体,核外颗粒体及多泡小体,这些都参与旁分泌信号传导。本研究通过透射电镜及免疫组化研究中华鳖输卵管峡部内telocytes的超微形态特征及其结构功能。峡部具有不同形状的telocytes,这些形态取决于突起的数量。分布位置与输卵管子宫部相似,一般位于黏膜固有层。重要的发现是,telocytes参与细胞外囊泡的分泌和转运。该研究第一次揭示了中华鳖输卵管峡部内TCs的存在及其分布,并在超微水平上显示Tcs具有分泌和转运细胞外囊泡的功能。实验ⅣCD34 mRNA在中华鳖输卵管的表达造血祖细胞CD34 Telocytes的一种特异标记物。本文应用qRT-PCR技术分析了CD34-mRNA在中华鳖输卵管的表达动态。CD34基因表达于输卵管蛋白分泌部、峡部、子宫部和阴道部4段,与之对应,透射电镜下大量的Telocytes细胞分布于中华鳖输卵管壁内。CD34 mRNA的表达水平具有区域性差异,子宫部最高,阴道部次之,峡部中等,而蛋白分泌部表达最低。这种表达区域性差异可能与输卵管节律性蠕动的调控有关。
【学位单位】：南京农业大学
【学位级别】：博士
【学位年份】：2015
【中图分类】：S917.4
【文章目录】：
ACKNOWLEDGEMENTS
ABSTRACT
摘要
List of Abbreviations
CHAPTER 1 Introduction
1 The need for studying the Telocytes in oviduct
2 Research goals
CHAPTER 2 Literature Review
PartⅠ Chinese soft shelled turtle, Pelodiscus sinensis
1 Introduction
2 Description
3 Scientific classification
4 Habitat
5 Distribution
6 Movement
7 Diet
8 Relationship with humans
9 Life cycle
PartⅡ The female reproductive tract of the Chinese soft shelled turtle
PartⅢ Telocytes
1 Brief Introduction of Telocytes
1.1 Identification of Telocytes
1.2 Ultrasture of Telocytes
1.3 Table1-1 Telocyte distribution in various organs
1.4 Telocytes and fibroblasts
1.5 Table1-2 Comparison of the characteristics of fibroblasts and telocytes
1.6 The Functions of telocytes
1.7 Role of telocytes in Pathology
1.8 Telocytes and regenerative medicine
References
CHAPTER 3 （ExperimentⅠ） Ultrastructural and immunohistochemical characteristics of telocytes in the vagina of the Chinese soft-shelled turtle, Pelodiscus sinensis
1 Introduction
2 Materials and methods
2.1 Ethical Approval
2.2 Experimental Animals
2.3 Transmission electron microscopy （TEM）
2.4 Immunohistochemistry
3 Results
4 Discussion
5 Conclusion
References
CHAPTER 4 （ExperimentⅡ） Identification and characterization of telocytes in the uterus of the oviduct in Chinese soft shelled turtle, Pelodiscus Sinensis
1 Introduction
2 Materials and methods
2.1 Ethical Approval
2.2 Experimental Animals
2.3 Transmission electron microscopy （TEM）
2.4 Immunohistochemistry
3 Results
4 Discussions
5 Conclusion
References
CHAPTER 5 （ExperimentⅢ） Identification and Ultrastructural characterization of telocytes in the Isthmus of the oviduct of Chinese soft shelled turtle, Pelodiscus sinensis
1 Introduction
2 Materials and methods
2.1 Ethical Approval
2.2 Experimental Ajahnals
2.3 Transmission electron microscopy （TEM）
2.4 Immunohistochemistry
3 Results
4 Discussion
5 Conclusion
References
CHAPTER 6 （ExperimentⅣ） The Expression of CD34 mRNA in the Oviduct of Chinese soft shelled turtle,Pelodiscus sinensis
1 Introduction
2 Materials and Methods
2.1 Ethical approval and Experimental Animals
2.2 Sterilization of glass-ware and plastic-ware
2.3 Total RNA extraction
2.4 Complimentary DNA （cDNA） construction
2.5 Real-Time Quantitative Polymerase Chain Reaction （RT-qPCR）
2.6 Statistical analysis
3 Results
4 Discussion
5 Conclusion
References
Summary
Conclusion
Innovation
Future Research Directions
PUBLICATIONS
【参考文献】
本文编号：2859139
【学位单位】：南京农业大学
【学位级别】：博士
【学位年份】：2015
【中图分类】：S917.4
【文章目录】：
ACKNOWLEDGEMENTS
ABSTRACT
摘要
List of Abbreviations
CHAPTER 1 Introduction
1 The need for studying the Telocytes in oviduct
2 Research goals
CHAPTER 2 Literature Review
PartⅠ Chinese soft shelled turtle, Pelodiscus sinensis
1 Introduction
2 Description
3 Scientific classification
4 Habitat
5 Distribution
6 Movement
7 Diet
8 Relationship with humans
9 Life cycle
PartⅡ The female reproductive tract of the Chinese soft shelled turtle
PartⅢ Telocytes
1 Brief Introduction of Telocytes
1.1 Identification of Telocytes
1.2 Ultrasture of Telocytes
1.3 Table1-1 Telocyte distribution in various organs
1.4 Telocytes and fibroblasts
1.5 Table1-2 Comparison of the characteristics of fibroblasts and telocytes
1.6 The Functions of telocytes
1.7 Role of telocytes in Pathology
1.8 Telocytes and regenerative medicine
References
CHAPTER 3 （ExperimentⅠ） Ultrastructural and immunohistochemical characteristics of telocytes in the vagina of the Chinese soft-shelled turtle, Pelodiscus sinensis
1 Introduction
2 Materials and methods
2.1 Ethical Approval
2.2 Experimental Animals
2.3 Transmission electron microscopy （TEM）
2.4 Immunohistochemistry
3 Results
4 Discussion
5 Conclusion
References
CHAPTER 4 （ExperimentⅡ） Identification and characterization of telocytes in the uterus of the oviduct in Chinese soft shelled turtle, Pelodiscus Sinensis
1 Introduction
2 Materials and methods
2.1 Ethical Approval
2.2 Experimental Animals
2.3 Transmission electron microscopy （TEM）
2.4 Immunohistochemistry
3 Results
4 Discussions
5 Conclusion
References
CHAPTER 5 （ExperimentⅢ） Identification and Ultrastructural characterization of telocytes in the Isthmus of the oviduct of Chinese soft shelled turtle, Pelodiscus sinensis
1 Introduction
2 Materials and methods
2.1 Ethical Approval
2.2 Experimental Ajahnals
2.3 Transmission electron microscopy （TEM）
2.4 Immunohistochemistry
3 Results
4 Discussion
5 Conclusion
References
CHAPTER 6 （ExperimentⅣ） The Expression of CD34 mRNA in the Oviduct of Chinese soft shelled turtle,Pelodiscus sinensis
1 Introduction
2 Materials and Methods
2.1 Ethical approval and Experimental Animals
2.2 Sterilization of glass-ware and plastic-ware
2.3 Total RNA extraction
2.4 Complimentary DNA （cDNA） construction
2.5 Real-Time Quantitative Polymerase Chain Reaction （RT-qPCR）
2.6 Statistical analysis
3 Results
4 Discussion
5 Conclusion
References
Summary
Conclusion
Innovation
Future Research Directions
PUBLICATIONS
【参考文献】
相关期刊论文 前1条
1 ZHOU Jin;WANG Yan;ZHU Ping;SUN HongYu;MOU YongChao;DUAN CuiMi;YAO AnNing;LV ShuangHong;WANG ChangYong;;Distribution and characteristics of telocytes as nurse cells in the architectural organization of engineered heart tissues[J];Science China(Life Sciences);2014年02期
本文编号：2859139
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