HOXA 13 REGULATION OF GENITOURINARY DEVELOPMENT

说明(改编自应用程序) 转录因子HOXA 13的无义突变导致严重 生殖器(GU)区畸形。在人类中，Hoxa 13的缺失 功能导致家族性手足生殖器综合征(HFGS)，导致 尿道下裂。前列腺，子宫，苗勒管，阴道膀胱，输尿管。和 直肠缺陷。在小鼠身上，这些发育途径是保守的，因为 靶向干扰‘Hoxa 13’导致胚胎畸形或发育不全 前列腺、子宫、尿路、输尿管、输尿管、阴道、直肠和 膀胱。这些结构的发展令人感兴趣的是 它们中的许多是由未分化的间充质凝聚而成。最近， 我证明了HOXA 13调节肢体间充质凝聚和 脐带血管(UV)通过控制直接细胞间黏附的基因， 细胞凋亡和同一性。因为在HFGS中受影响的许多结构 源自间充质冷凝。我假设遗传途径 受Hoxal13调控，调节肢体和紫外线的发育也具有 直接关系到顾区的正常发展。 为了验证这一假设，一只带有框内GFP报告的Hoxa 13突变小鼠 将会被使用。Hoxa 13 GFP等位基因产生一个强烈的Hoxa 13特有的信号 在受影响的古族地区。这一信号有助于富集 通过荧光激活细胞分选(FACS)获得均一细胞群 从表达HOXA基因的解剖组织中提取13.流式细胞仪纯化间充质细胞 来自受影响区域的突变胚胎和对照胚胎将被鉴定 在体外研究基因表达、细胞黏附、细胞凋亡和 可以解释特定畸形的增殖。这些FAC 丰富的细胞也将被用作基于RNA和cDNA的基因的来源。 识别受Hoxa 13调控的其他基因的发现方法 在GU发展中发挥作用。一旦确定，染色体定位 HOXA 13特定的靶基因将通过老鼠/仓鼠的辐射来确定 混合DNA面板。任何一种的地图位置和胚胎表达模式 新发现的基因将提供给研究界， 为鉴定良性疾病中涉及的未知遗传成分提供资源 前列腺增生症、恶性肿瘤和GU畸形。上位者 Hoxa 13和双羟睾酮之间的关系将通过 Hoxa-13绿色荧光蛋白报告细胞系的体外筛选 识别激活Hoxa 13的基因。这种独特的实验设计允许 从细胞和组织的角度研究胃溃疡的发育 受Hoxa 13功能丧失的影响。从这个角度来看，洞察力遗传 并将获得正常GU发育所需的细胞机制。

DESCRIPTION (adapted from the application) Nonsense mutations in the transcription factor Hoxa 13 result in severe malformations of the genitourinary (GU) region. In humans, loss of Hoxa 13 function causes familial hand-foot-genital syndrome (HFGS), resulting in hypospadia. prostate, uterine, Mullerian duct, vaginal bladder, ureter. and rectal defects. In mice, these developmental pathways are conserved, as targeted disruption of' Hoxa 13 results in malformation or agenesis of the prostate, uterus, urethra, ureter, MuIlerian ducts, vagina, rectum, and bladder. What is intriguing about the development of these structures is that many of them form from condensations of undifferentiated mesenchyme. Recently, I demonstrated that Hoxa 13 regulates mesenchymal condensation in the limb and umbilical vasculature (UV) by controlling genes that-direct cell-cell adhesion, apoptosis, and identity. Because many of the structures affected in HFGS are derived from mesenchymal condensations. I hypothesize that the genetic pathways regulated by Hoxal13, to mediate limb and UV development also function to direct normal development of the GU region. To test this hypothesis, a Hoxa 13 mutant mouse with an in-frame GFP reporter will be used. The Hoxa 13 GFP allele produces a strong, Hoxa 13-specific signal in the affected GU regions. This signal facilitates the enrichment of homogeneous populations of cells by fluorescence-activated cell sorting (FACS) from dissected tissues expressing Hoxa 13. Mesenchyme cells purified by FACS from the affected regions of mutant and control embryos will be characterized in vitro for changes in gene expression, cell adhesion, apoptosis, and proliferation that could account for specific malformations. These FACS enriched cells will also be used as a resource for RNA-and cDNA-based gene discovery methodologies to identify additional genes regulated by Hoxa 13 that play a role in GU development. Once identified, the chromosomal localization of Hoxa 13 specific target genes will be determined using mouse/hamster radiation hybrid DNA panels. The map positions and embryonic expression patterns of any newly identified genes will be made available to the research community, providing a resource to identify unknown genetic components involved in benign prostate hyperplasias, malignancies, and GU malformations. The epistatic relationship between Hoxa 13, and dhydroxytestosterone will be determined using a Hoxa 13 GFP reporter cell line in conjunction with in vitro screen, to identify genes that activate Hoxa 13. This unique experimental design allows study of GU development from the, perspective of the cells and tissues most affected by loss of Hoxa 13 function. From this perspective, insight genetic and cellular mechanisms required for normal GU development will be gained.