BACKGROUND & AIMS: :Peptidylarginine deiminase 6 (PAD6) is an enzyme that is uniquely expressed in male and female germ cells. To study the function of this enzyme in vivo we generated mice deficient for PAD6. Here we show that inactivation of the PAD6 gene in mice leads to female infertility whereas male fertility is not affected. The absence of the PAD6 protein and consequently absence of citrullination activity in oocytes results in dispersal of the cytoskeletal sheets in oocytes, indicating an essential role of these germ cell-specific structures in zygote/embryo development. PAD6 deficient mice do not show any other overt phenotype. Thus, we identify citrullination as a new regulator of fertility.

背景与目标： ：肽基精氨酸脱亚氨酶6（PAD6）是一种在雄性和雌性生殖细胞中独特表达的酶。为了研究该酶在体内的功能，我们产生了PAD6缺陷的小鼠。在这里，我们显示，小鼠中PAD6基因的失活导致女性不育，而男性不育不受影响。卵母细胞中缺乏PAD6蛋白，因此缺乏瓜氨酸化活性，导致卵母细胞中细胞骨架片的分散，表明这些生殖细胞特异性结构在合子/胚胎发育中起着至关重要的作用。 PAD6缺陷的小鼠没有显示任何其他明显的表型。因此，我们确定瓜氨酸化是生育的新调节剂。

BACKGROUND & AIMS: :The identification of germ-line mutations in 2 genes (BRCA1 and BRCA2) responsible for the majority of hereditary ovarian cancers has led an increasing number of women carriers of these mutations to undergo prophylactic oophorectomy (PO) to reduce their risk of subsequent ovarian carcinoma. A large number of unexpected, clinically occult neoplasms are thus being discovered. Up to December 2004, the Medical Genetics Service of the National Cancer Institute in Milan, Italy, has tested 756 probands from breast and/or ovarian cancer families for BRCA1 and BRCA2 germ-line mutations. Molecular screening of family members led to the identification of 344 female carriers of BRCA1 (239) or BRCA2 (105) germ-line mutations. Of the 186 potentially eligible women (37 of whom had tested positive for BRCA1 and 13 for BRCA2 mutation), 50 (26.8%) chose to undergo PO. Six clinically occult primary gynecologic malignancies (2 stage IIIC serous carcinomas of the ovary, 3 in situ serous carcinomas of the fallopian tube, and 1 stage IIB invasive serous carcinoma of the fallopian tube) and 1 occult ovarian metastasis from breast carcinoma were identified in the PO specimens of 7 women (all BRCA1 mutated). Four of the patients with occult primary gynecologic cancers are alive without disease 129, 87, 38, and 7 months after PO, respectively. One of the 2 patients with primary ovarian cancer and the single patient with tubal invasive carcinoma are alive with recurrent disease 83 and 20 months after PO, respectively. In addition, one of the patients whose PO specimen did not show any malignancy presented with stage IIIC tubal carcinoma 77 months after PO. The relatively high number of tubal neoplasms found at PO in this group of patients underlines the linkage between mutation and the risk of developing tubal cancer, and stresses the need to include removal of the entire tubes at the time of PO and of thoroughly evaluating the specimens at the microscopic level. The upstaging of all 3 invasive carcinomas after staging surgery, and the late recurrence and persistence of 2 of them despite treatment indicate that small size of the tumors should not preclude therapy.

背景与目标： ：对负责大多数遗传性卵巢癌的2个基因（BRCA1和BRCA2）的种系突变的鉴定，导致越来越多的女性携带这些突变的人进行了预防性卵巢切除术（PO），以降低其患上卵巢癌的风险。因此，发现了大量意想不到的临床隐匿性肿瘤。截至2004年12月，意大利米兰国家癌症研究所的医学遗传学服务已对756个来自乳腺癌和/或卵巢癌家族的先证者进行了BRCA1和BRCA2种系突变测试。家庭成员的分子筛查导致识别出BRCA1（239）或BRCA2（105）种系突变的344个雌性携带者。在186名可能符合条件的妇女中（其中37人的BRCA1测试呈阳性，13人的BRCA2突变检测为阳性），其中50人（26.8％）选择接受PO。在临床中确定了6例临床隐匿的原发性妇科恶性肿瘤（2例卵巢IIIC浆液性癌，3例输卵管原位浆液性癌和1例IIB输卵管浸润性浆液性癌）和1例隐匿性卵巢癌卵巢转移。 7名妇女的PO标本（所有BRCA1突变）。分别在PO后129、87、38和7个月，有四名患有隐匿性原发性妇科癌症的患者还活着而没有疾病。 2例原发性卵巢癌患者中的1例和输卵管浸润性癌的1例患者在PO后分别存活83个月和20个月。此外，其中一名PO标本未显示任何恶性肿瘤的患者在PO后77个月出现IIIC期输卵管癌。该组患者在PO中发现的相对较多的输卵管肿瘤强调了突变与发生输卵管癌的风险之间的联系，并强调需要在PO时切除整个管并彻底评估标本在微观层面上。分期手术后所有3种浸润性癌的分期升级，并且尽管有治疗，但其中2种仍较晚复发和持续存在，这表明较小的肿瘤不应排除治疗的可能性。

BACKGROUND & AIMS: :A transmembrane extracellular matrix receptor of the integrin family, alpha 6 beta 4, is a component of the hemidesmosome, an adhesion complex of importance in epithelial cell-connective tissue attachment (Stepp, M. A., S. Spurr-Michaud, A. Tisdale, J. Elwell, and I. K. Gipson. 1990. Proc. Natl. Acad. Sci. USA. 87:8970-8974; Jones, J. C. R., M. A. Kurpakus, H. M. Cooper, and V. Quaranta. 1991. Cell Regulation. 2:427-438). Cytosolic components of hemidesmosomes include bullous pemphigoid (BP) antigens while extracellular components include a 125-kD component of anchoring filaments (CAF) and collagen type VII-containing anchoring fibrils. We have monitored the incorporation of the alpha 6 beta 4 integrins into forming hemidesmosomes in an in vitro wound-healing explant model. In epithelial cells recently migrated from the edges of unwounded sites over bare connective tissue, alpha 6 beta 4 first appears along the entire cell surface. At this stage, these cells contain little or no cytosolic hemidesmosomal components, at least as detectable by immunofluorescence using BP autoantibodies, whereas they are already positive for laminin and CAF. At a later stage, as cells become positive for cytosolic hemidesmosome components such as BP antigens as well as collagen type VII, alpha 6 beta 4 becomes concentrated along the basal pole of the epithelial cell where it abuts the connective tissue of the explant. Polyclonal antibodies to beta 4 do not interfere with the migration of epithelial cells in the explant. However, they prevent assembly of hemidesmosomal complexes and inhibit expression of collagen type VII in cells that have migrated over wound areas. In addition, they induce disruption of established hemidesmosomes in nonmigrating cells of the unwounded area of the explant. Monoclonal antibodies to alpha 6 have a more dramatic effect, since they completely detach epithelial cells in the unwounded area of the explant. Antibodies to CAF also detach epithelial cells in unwounded areas, apparently by inducing separation between epithelium and connective tissue at the lamina lucida of the basement membrane zone. These results suggest a model whereby polarization of alpha 6 beta 4 to the basal surface of the cells, perhaps induced by a putative anchoring filament-associated ligand, triggers assembly of hemidesmosome plaques.

背景与目标： ：整联蛋白家族的跨膜细胞外基质受体，α6 beta 4，是半桥粒的成分，半桥粒是在上皮细胞结缔组织附着中重要的粘附复合物（Stepp，MA，S。Spurr-Michaud，A。Tisdale， J. Elwell和IK Gipson。1990.美国国家科学院院刊87：8970-8974； Jones，JCR，MA Kurpakus，HM Cooper和V.Quaranta。1991.《细胞调节》 2：427- 438）。血小体的胞质成分包括大疱性类天疱疮（BP）抗原，而细胞外成分包括锚定丝（CAF）和含VII型胶原的锚定纤丝的125 kD成分。我们已经监测了在体外伤口愈合的外植体模型中将α6β4整合素整合到形成hemidemosomes中。在最近从裸结缔组织上方未受伤部位边缘迁移的上皮细胞中，α6 beta 4首先沿整个细胞表面出现。在这一阶段，这些细胞几乎不含有或不含胞质半桥粒成分，至少可以通过使用BP自身抗体的免疫荧光检测到，而对于层粘连蛋白和CAF则已经呈阳性。在稍后的阶段，随着细胞对胞质半桥体成分（例如BP抗原）和VII型胶原呈阳性，α6β4沿着上皮细胞的基极集中，并与外植体的结缔组织邻接。针对β4的多克隆抗体不会干扰外植体中上皮细胞的迁移。然而，它们阻止了半桥粒复合物的组装并抑制了在伤口区域上迁移的细胞中VII型胶原的表达。另外，它们在外植体未受伤区域的非迁移细胞中诱导已建立的半染色体的破坏。针对α6的单克隆抗体具有更引人注目的效果，因为它们可以完全脱离外植体未受伤区域中的上皮细胞。 CAF抗体还可以在未受伤的区域分离上皮细胞，这显然是通过诱导基底膜区域的透明层上皮与结缔组织之间的分离来实现的。这些结果表明了一个模型，其中α6β4极化到细胞的基底表面（可能是由假定的锚定细丝相关配体诱导的）触发了半血球斑块的组装。