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（由适用提供）：在精子发生过程中，在半率上皮的基本隔室中发展了前体精子细胞，必须在成年大鼠测试中的上皮循环的VIII阶段横穿血液 - 遗传屏障（BTB），并进入成人隔室以进一步开发。如果没有这种生殖细胞在BTB中的及时运动，将会破坏精子发生，从而导致不育症。尽管这种与精子发生相关的细胞现象已知数十年，但调节BTB动力学以促进生殖细胞横穿BTB的机制完全未知。总体而言，这是由于缺乏适合研究BTB动力学的体内模型。在此应用程序中开发和广泛表征一个新型模型以满足这种需求的建议。简而言之，基于塞巴丁的第二个细胞外循环（BTB处的紧密连接连接（TJ） - 整合性膜蛋白）的局部给药，显示出成年大鼠测试，可诱导半生产上皮中BTB的可逆破坏。同样，此事件与几种靶蛋白的表达变化有关（例如，转化生长因子�-3，TGF-€3和肿瘤坏死因子A，TNFA）模仿了Sertoli细胞TJ-barrier，从而在体外恢复了事件。也许最重要的是，这种局部塞链蛋白肽处理还引起了半成分的上皮细胞的可逆生殖细胞损失（特别是精子和精子细胞，而不是精子的生殖细胞损失）。 P.I.试图通过描述半生层上皮的细胞，分子和生化变化的详细时间表，以广泛的表征这种新颖的体内模型。这些研究的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恢复的机理和调节。我们的主要目标是开发一种创新的体内模型来研究BTB动力学，该模型与当前使用毒物（例如镉和甘油）的研究模型有显着不同。首先，可以诱导BTB恢复的胡椒是无毒性的。其次也是最重要的是，被破坏的BTB可以被“重新安装”，这使得该模型独特地适合研究精子发生过程中BTB重新组装的生物学和调节。