Napirei Markus, Ricken Albert, Eulitz Dirk, Knoop Heiko, Mannherz Hans Georg
Abteilung für Anatomie und Embryologie, Medizinische Fakultät, Ruhr-Universität Bochum, Universitätsstrasse 150, D-44780 Bochum, Germany.
Biochem J. 2004 Jun 15;380(Pt 3):929-37. doi: 10.1042/BJ20040046.
The tissue distribution of deoxyribonuclease 1 (DNASE1, DNase I), a Ca2+ and Mg2+/Mn2+-dependent secretory endonuclease, has previously been investigated. However, most of these studies did not account for the existence of different members of the DNASE1 gene family, did not differentiate between endogenous DNASE1 protein synthesis and its extracellular occurrence or were not performed with methods allowing both a sensitive and a specific detection. Now we re-examined the DNASE1 gene expression pattern by taking advantage of the Dnase1 knockout mouse model. Direct comparison of samples derived from wild-type (Dnase1+/+) and knockout (Dnase1-/-) mice allowed an unambiguous detection of Dnase1 gene expression at the mRNA and protein level. For the detection of Dnase1 activity, we developed a highly sensitive nuclease zymogram method. We observed high Dnase1 gene expression in the parotid and submandibular gland as well as in the kidney and duodenum, intermediate expression in the ileum, mesenterial lymph nodes, liver, ventral prostate, epididymis, ovary and stomach, and low expression in the sublingual, preputial, coagulation and pituitary gland. We report for the first time the lachrymal and thyroid glands, the urinary bladder and the eye to be Dnase1-expressing organs as well. Since Dnase1 knockout mice with the 129xC57Bl/6 mixed genetic background have indicated the protection against an anti-DNA autoimmune response as a new physiological function of Dnase1, knowledge of the physiological sites of its synthesis might prove helpful to find new therapeutic strategies.
脱氧核糖核酸酶1(DNASE1,DNase I)是一种依赖Ca2+和Mg2+/Mn2+的分泌性核酸内切酶,其组织分布此前已被研究过。然而,这些研究大多没有考虑DNASE1基因家族不同成员的存在,没有区分内源性DNASE1蛋白的合成与其细胞外的存在情况,或者没有采用能够同时进行灵敏且特异检测的方法。现在,我们利用Dnase1基因敲除小鼠模型重新研究了DNASE1基因的表达模式。对野生型(Dnase1+/+)和基因敲除型(Dnase1-/-)小鼠的样本进行直接比较,能够在mRNA和蛋白水平明确检测到Dnase1基因的表达。为了检测Dnase1的活性,我们开发了一种高度灵敏的核酸酶酶谱法。我们观察到Dnase1基因在腮腺、颌下腺以及肾脏和十二指肠中高表达,在回肠、肠系膜淋巴结、肝脏、腹侧前列腺、附睾、卵巢和胃中中度表达,而在舌下腺、包皮腺、凝血腺和垂体中低表达。我们首次报道泪腺、甲状腺、膀胱和眼睛也是表达Dnase1的器官。由于具有129xC57Bl/6混合遗传背景的Dnase1基因敲除小鼠已表明,抵御抗DNA自身免疫反应是Dnase1的一种新生理功能,了解其合成的生理部位可能有助于找到新的治疗策略。