Quick Marsha L, Done Joseph D, Thumbikat Praveen
Department of Urology, Northwestern University Feinberg School of Medicine, USA.
J Vis Exp. 2013 Jan 16(71):e50158. doi: 10.3791/50158.
Uropathogenic Escherichia coli (UPEC) are pathogens that play an important role in urinary tract infections and bacterial prostatitis. We have recently shown that UPEC have an important role in the initiation of chronic pelvic pain, a feature of Chronic prostatitis/Chronic pelvic pain syndrome (CP/CPPS). Infection of the prostate by clinically relevant UPEC can initiate and establish chronic pain through mechanisms that may involve tissue damage and the initiation of mechanisms of autoimmunity. A challenge to understanding the pathogenesis of UPEC in the prostate is the relative inaccessibility of the prostate gland to manipulation. We utilized a previously described intraurethral infection method to deliver a clinical strain of UPEC into male mice thereby establishing an ascending infection of the prostate. Here, we describe our protocols for standardizing the bacterial inoculum as well as the procedure for catheterizing anesthetized male mice for instillation of bacteria. CP/CPPS is primarily characterized by the presence of tactile allodynia. Behavior testing was based on the concept of cutaneous hyperalgesia resulting from referred visceral pain. An irritable focus in visceral tissues reduces cutaneous pain thresholds allowing for an exaggerated response to normally non-painful stimuli (allodynia). Application of normal force to the skin result in abnormal responses that tend to increase with the intensity of the underlying visceral pain. We describe methodology in NOD/ShiLtJ mice that utilize von Frey fibers to quantify tactile allodynia over time in response to a single infection with UPEC bacteria.
尿路致病性大肠杆菌(UPEC)是在尿路感染和细菌性前列腺炎中起重要作用的病原体。我们最近发现,UPEC在慢性盆腔疼痛的引发中起重要作用,慢性盆腔疼痛是慢性前列腺炎/慢性盆腔疼痛综合征(CP/CPPS)的一个特征。临床相关的UPEC感染前列腺可通过可能涉及组织损伤和自身免疫机制启动的机制引发并导致慢性疼痛。理解UPEC在前列腺发病机制中的一个挑战是前列腺相对难以进行操作。我们利用先前描述的尿道内感染方法将一株临床UPEC菌株导入雄性小鼠体内,从而建立前列腺的上行感染。在此,我们描述了标准化细菌接种物的方案以及对麻醉的雄性小鼠进行插管以注入细菌的程序。CP/CPPS的主要特征是存在触觉异常性疼痛。行为测试基于由牵涉性内脏痛引起的皮肤痛觉过敏的概念。内脏组织中的一个易激惹灶会降低皮肤疼痛阈值,从而导致对通常无疼痛的刺激产生夸大的反应(异常性疼痛)。对皮肤施加正常压力会导致异常反应,这种反应往往会随着潜在内脏疼痛的强度增加而增强。我们描述了在NOD/ShiLtJ小鼠中使用von Frey纤维来量化单次感染UPEC细菌后随时间变化的触觉异常性疼痛的方法。