An in vivo model to study blood-testis barrier dynamics

研究血睾屏障动力学的体内模型

基本信息

批准号：
7485598
负责人：
C. Yan Cheng
金额：
$ 7.09万
依托单位：
POPULATION COUNCIL
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-08-15 至 2009-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7485598
关键词：
Abbreviations Address Adherens Junction Adult Amino Acids Apical Bacitracin Biochemical Biological Biology Blood-Testis Barrier Bos taurus Bovine Serum Albumin Butadiene Cadmium Cattle Cells DNA Nucleotidylexotransferase Development Dimethyl Sulfoxide Disruption Eagles Electrical Resistance Epidermal Growth Factor Epithelial Event Extracellular Signal Regulated Kinases Fibroblast Growth Factor 2 Fluorescein Fluorescein-5-isothiocyanate Fluoresceins Follicle Stimulating Hormone Gap Junctions Gentamicins Germ Cells Glycerol Goals HEPES Human Imidazole Immunohistochemistry In Situ Nick-End Labeling In Vitro Infertility Inhibitory Concentration 50 Insulin Integral Membrane Protein Interferons Isothiocyanates Kinetics Label Laboratory Study Liquid substance Luteinizing Hormone MAP Kinase Gene MDCK cell Mediating Mitogen-Activated Protein Kinases Mitogens Modeling Molecular Movement Nutrient Oligodendroglia Pathway interactions Peptides Permeability Phase Physiological Polymerase Chain Reaction Proteins Rattus Recovery Regulation Research Personnel Reverse Transcriptase Polymerase Chain Reaction Reverse Transcription STF Spermatids Spermatocytes Spermatogenesis Spermatogonia Staging Structure of rete testis Study models Testis Tight Junctions TimeLine Title Transferrin Transforming Growth Factors Tubular formation Tumor Necrosis Factor-alpha Tumor Necrosis Factors U-0126 base cell motility cytokine cytotoxic extracellular human MAPK14 protein human TNF protein in vivo in vivo Model inhibitor/antagonist innovation junctional adhesion molecule kinase inhibitor membrane-associated guanylate kinase mitogen-activated protein kinase p38 myotubularin novel occludin outer surface lipoprotein response sertoli cell spermatogenic epithelium structure synthetic peptide toxicant upstream kinase

项目摘要

DESCRIPTION (provided by applicant): During spermatogenesis, developing preleptotene spermatocytes residing in the basal compartment of the seminiferous epithelium must traverse the blood-testis barrier (BTB) at stage VIII of the epithelial cycle in adult rat testes, entering the adluminal compartment for further development. Without this timely movement of developing germ cells across the BTB, spermatogenesis will be disrupted, leading to infertility. While this cellular phenomenon pertinent to spermatogenesis is known for decades, the mechanism(s) that regulates BTB dynamics to facilitate germ cells to traverse the BTB is entirely unknown. This by and large is due to the lack of a suitable in vivo model to study BTB dynamics. In this application, the P.I. proposes to develop and extensively characterize a novel model to meet this need. In brief, local administration of a 22-amino acid synthetic peptide based on the second extracellular loop of occludin, a tight junction (TJ)-integral membrane protein at the BTB, to adult rat testes was shown to induce reversible disruption of BTB in the seminiferous epithelium. Also, this event was associated with changes in the expression of several target proteins (e.g., transforming growth factor ¿-3, TGF-¿3, and tumor necrosis factor a, TNFa) that mimicked the Sertoli cell TJ-barrier restructuring events in vitro. Perhaps the most important of all, this local occludin peptide treatment also induced reversible germ cell loss (in particular spermatids and spermatocytes, but not spermatogonia) from the seminiferous epithelium. The P.I. seeks to extensively characterize this novel in vivo model by delineating the detailed timeline of cellular, molecular and biochemical changes in the seminiferous epithelium correlating with the status of spermatogenesis and the integrity of the BTB. Results of these studies will yield a reliable study model for investigators in the field to understand the mechanism and regulation of BTB restructuring during spermatogenesis. Our primary goal is to develop an innovative in vivo model to study BTB dynamics which is significantly different from currently available study models using toxicants (e.g., cadmium and glycerol). First, the peptide that can induce BTB restructuring is non-cytotoxic. Second and perhaps most importantly, the disrupted BTB can be "resealed", making this model uniquely suitable to study the biology and regulation of BTB re-assembly during spermatogenesis.

描述（由申请人提供）：在精子发生过程中，位于生精上皮基底室中的发育中的前细线精母细胞必须在成年大鼠睾丸上皮周期的第八阶段穿过血睾屏障（BTB），进入腔内室进一步如果发育中的生殖细胞不及时穿过 BTB，精子发生就会受到干扰，从而导致不育。与精子发生相关的现象已为人所知数十年，但调节 BTB 动力学以促进生殖细胞穿过 BTB 的机制完全未知，这大体上是由于缺乏合适的体内模型来研究 BTB 动力学。在本申请中，P.I. 提议开发并广泛描述一种新模型来满足这一需求，简而言之，基于 occludin 的第二个细胞外环（紧密连接）局部施用 22 个氨基酸的合成肽。 (TJ)-BTB 整合膜蛋白，对成年大鼠的测试表明，可诱导生精上皮中 BTB 的可逆性破坏。此外，该事件与多种靶蛋白（例如转化生长因子 ¿）的表达变化有关。 -3，TGF-¿ 3 和肿瘤坏死因子 a，TNFa）在体外模拟支持细胞 TJ 屏障重组事件，也许最重要的是，这种局部 occludin 肽治疗还诱导可逆性生殖细胞损失（特别是精子细胞和精母细胞）。 P.I. 试图通过描绘细胞、分子和生化变化的详细时间线来主要描述这种新颖的体内模型。这些研究结果将为该领域的研究人员提供可靠的研究模型，以了解精子发生过程中 BTB 重组的机制和调节。研究 BTB 动力学的创新体内模型与目前使用有毒物质（例如镉和甘油）的研究模型显着不同。其次，也许最重要的是，破坏的 BTB 可以“重新密封”，使得该模型特别适合研究精子发生过程中 BTB 重新组装的生物学和调节。