Transient expression of the Arf tumor suppressor during male germ cell and eye development in Arf-Cre reporter mice
Transient expression of the Arf tumor suppressor during male germ cell and eye development in Arf-Cre reporter mice
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DOI：
10.1073/pnas.0902310106
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发表时间：
2009-04-14
影响因子：
11.1
通讯作者：
Sherr, Charles J.
中科院分区：
文献类型：
--
作者：
Gromley, Adam;Churchman, Michelle L.;Sherr, Charles J.
Arf肿瘤抑制基因在小鼠雄性生殖细胞和眼睛发育过程中瞬时表达。随着小鼠年龄的增长，其失活损害精子发生，并导致眼睛玻璃体中细胞的出生后异常增殖，导致失明。在睾丸中，p19（Arf）的表达仅限于精原细胞，但在精母细胞中完全消失，这表明Arf在有丝分裂和减数分裂生殖细胞分裂之间的平衡中起着生理作用。一个敲入的等位基因编码Cre重组酶调节的小鼠细胞Arf启动子被用来跟踪Arf基因在体内的诱导。与表达Cre依赖性YFP的报告菌株杂交提供了Arf-Cre等位基因在雄性生殖细胞谱系中适当表达的原理证明。然而，Cre表达导致雄性不育，限制了基因敲入等位基因向雌性的生殖系传递。Arf缺失小鼠不能再吸收眼玻璃体内的玻璃体脉管系统，其中表达PDGF-β受体（Pdgfr β）的周细胞样细胞异常增殖并破坏视网膜和透镜。将Arf-Cre雌性与含有“缺失的”（FL）Arf等位基因的雄性杂交产生了Arf（Cre/FL）后代，其表现出视力的非穿透性缺陷，范围为全盲。将Arf（Cre/FL）等位基因与Pdgfr（β FL/FL）背景杂交使所有组织病理学正常化并完全恢复视力。
The Arf tumor suppressor is expressed transiently during mouse male germ cell and eye development. Its inactivation compromises spermatogenesis as mice age and leads to aberrant postnatal proliferation of cells in the vitreous of the eye, resulting in blindness. In the testis, expression of p19(Arf) is limited to spermatogonia but is extinguished completely in spermatocytes, suggesting that Arf plays a physiologic role in setting the balance between mitotic and meiotic germ cell division. A knock-in allele encoding Cre recombinase regulated by the mouse cellular Arf promoter was used to trace Arf gene induction in vivo. Interbreeding to a reporter strain that expresses Cre-dependent YFP provided proof-of-principle that the Arf-Cre allele was appropriately expressed in the male germ cell lineage. However, Cre expression resulted in male sterility, limiting germ line transmission of the knock-in allele to females. Arf-null mice fail to resorb the hyaloid vasculature within the ocular vitreous where pericyte-like cells that express the PDGF-beta receptor (Pdgfr beta) proliferate aberrantly and destroy the retina and lens. Interbreeding of Arf-Cre females to males containing "floxed'' (FL) Arf alleles yielded Arf(Cre/FL) progeny that exhibited variably penetrant defects in visual acuity ranging to total blindness. Crossing the Arf(Cre/FL) alleles onto a Pdgfr(beta FL/FL) background normalized all histopathology and restored vision fully.