Mouse Models of Epididymitis Induced by Pathogen-Associated Molecular Patterns

摘要

Here, two distinct surgical injection models are described for inducing inflammatory stimuli in the initial segment (interstitial injection) and cauda epididymidis (intravasal injection) of mice. These methods enable studies of the region-specific responses in the different regions of the epididymis.

摘要

Epididymitis is a highly prevalent disease in the outpatient urologic clinic and a relevant factor in male infertility. Bacterial infections, including sexually transmitted bacteria and common uropathogens, represent the most common etiological factors of epididymitis. Nevertheless, the pathophysiological mechanisms underlying the onset, natural history, and outcomes of bacterial epididymitis remain poorly understood. In this regard, rodent models are valuable tools for investigating the mechanistic responses of the epididymis when challenged with various inflammatory or infectious stimuli. Studies in rats and mice showed that bacterial-derived pathogen-associated molecular patterns (PAMPs), such as lipopolysaccharide (LPS) and lipoteichoic acid (LTA) from Gram-negative and Gram-positive bacteria, respectively, trigger acute inflammation in the epididymis. The severity of these inflammatory responses differs according to the epididymal region affected and is associated with poor sperm parameters. Herein, two distinct surgical approaches are described in detail for inducing epididymitis in mice through the injection of PAMPs in the interstitial compartment of the initial segment or the luminal compartment of the vas deferens toward the cauda epididymidis, thus allowing investigation of the inflammatory responses of the epididymal proximal and distal regions. It is expected that these experimental models of epididymitis will enable future studies aimed at advancing the understanding of the mechanisms governing the epididymal responses to invading pathogens, opening novel ways for therapeutic interventions to mitigate the repercussions of epididymitis on male fertility.

引言

The epididymis is a mucosal male reproductive organ composed of a single, highly convoluted tubule that connects the efferent ducts to the vas deferens. The epididymis is essential for sperm maturation (acquisition of sperm motility and fertilizing ability), concentration, transport, storage, and protection until the ejaculation^1,2. Classically, the rodent epididymis has four anatomical regions: the initial segment, caput, corpus, and cauda¹ (Figure 1). Furthermore, each region of the epididymis can be subdivided into additional segments separated by inters....

方案

All animal experiments were performed in accordance with the Guide for the Care and Use of Laboratory Animals (National Institute of Health, USA) and National Council for the Control of Animal Experimentation (CONCEA, Brazil). Procedural approval was granted by the Ethics Committee for the Use of Experimental Animals (CEUA-IBB/UNESP, Brazil; protocol number: 6272050320; 2269040625). Adult C57BL/6 male mice (90-120 days) were used in these studies. The reagents and the equipment used in this study are listed in the Table of Materials.

1. Interstitial injection in the initial segment of the epididymis

1. Mouse preoperative preparatio....

结果

Here, two distinct surgical injection models for inflammatory stimuli in the initial segment (interstitial injection) and cauda epididymidis (intravasal injection) in mice are described. The former triggers inflammatory responses in the proximal epididymis, whereas the latter triggers them in the distal epididymis.

The interstitial injection is performed through an incision in the abdomen, making use of the possibility of moving.......

讨论

Here, two distinct surgical injection procedures for the induction of region-specific epididymitis in mice are presented. They are used to perform experimental models of epididymitis specifically targeting the initial segment via interstitial injection or the cauda epididymidis via retrograde intravasal injection. The detailed and visual guide increases the clarity and accessibility of these protocols, seeking to enhance the reproducibility of these methods.

The diameter of t.......

材料

本文使用的材料清单

姓名	公司	目录编号	评论
斜角钝头手术钳及nbsp;	邦瑟	xtycga1012
斜角手术钳	邦瑟	PWGHVV1012
蓝埃文斯解	西格玛-奥尔德里奇	E2129
斗牛犬镊子夹具;	邦瑟	JTKPQS1012
氯己定	里奥库尔特;云母	Riohex 0.25%
弯曲手术钳	邦瑟	506e6e1012
电动修剪笔形状	-	-
汉密尔顿注射器	汉密尔顿	80400
头带放大镜	OptiVISOR（视角视觉）	-
氯胺酮	文科动物健康	凯他莱克斯
LPS 来自大肠杆菌 055：B55	生前者	TLRL-pb5lps
金黄色葡萄球菌的LTA	生前者	Tlrl-pslta
针架	邦瑟	8pihbf1012
5/0号手术缝线	沙隆医疗	N550CTI20
无菌生理盐水	-	-
直式手术产钳	邦瑟	ejaiuz1012
木苯丙嗪	新泰克	西拉津