Name: 视频: 小鼠模型中的尿液消耗和膀胱冲洗:一种基于导管的小鼠膀胱排空和清洗程序 - 概念
Uploaded: 2023-04-30T13:52:27.000+00:00
Duration: 1 min 27 s
Description: 1.8K 次观看. 来源:Yu， C. et al. 通过电穿孔将质粒 DNA 递送到小鼠膀胱尿路的新方法。J. Vis. Exp. （2018 年） 在本视频中，我们描述了一种基于导管的手术，用于清空和冲洗小鼠模型的膀胱。然后，制备好的小鼠可用于进一步的实验程序。

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All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
&#160;1. Anesthetizing Animal
<ol>
	<li>Use a tabletop anesthesia system to anesthetize the animal with isoflurane. Turn on the O2&#160;tank and adjust the oxygen flowmeter to 1 L/min.</li>
	<li>Place an adult female C57BL/6J mouse in the induction chamber. Turn on and adjust the isoflurane vaporizer to 3% for induction. The mouse should be in anesthesia within 2 min.&#160;Administer buprenorphine analgesic (0.1 mg/kg) subcutaneously upon removal of the mouse from the induction chamber for preemptive analgesia. 
	NOTE: In this protocol, female mice are used, as they are easier to work with during urethra catheterization. The protocol also works for male mice, but extra caution is needed.</li>
	<li>Apply ophthalmic ointment to the eyes of the animal to prevent dryness. Place the anesthetized mouse facing up on a heating pad with its nose in the nose cone. Turn the air circuit switch to let isoflurane go through the nose cone.</li>
	<li>Restrain the head and the nose cone with adhesive tape. Adjust the isoflurane vaporizer to 2% for maintenance.</li>
	<li>Restrain the limbs with adhesive tape. Perform toe-pinch tests to make sure the animal is in deep anesthesia and then shave the abdominal area.</li>
</ol>
2. Urine Depletion and Bladder Rinsing
<ol>
	<li>Apply lubricant to the catheter (24G, length of 2.11 cm, outer diameter of 0.045 cm) and ensure that its outer surface is fully covered.</li>
	<li>Insert the catheter into the urethra opening and slowly push it until it reaches the bladder, at which time the urine should flow out through the catheter. Gently press the abdomen to help urine depletion.
	<ol>
		<li>Check if the catheter is inserted correctly into the bladder by observing automatic urine out-flow. Avoid piercing the urethra and bladder wall and apply lubricant multiple times if needed.</li>
	</ol>
	</li>
	<li>Discard the urine with a pipette into a waste beaker.</li>
	<li>Pipette 80 &#181;L of phosphate-buffered saline (PBS) into the outer end of the catheter, with the other end still in the bladder. Carefully attach a 1 mL syringe to the catheter. Gently push the syringe to inject the PBS into the bladder. Leave some PBS in the catheter to avoid creating air bubbles in the bladder.</li>
	<li>Remove the syringe. Wait for PBS to drain out of the bladder. Gently press the abdomen to evacuate PBS.</li>
	<li>Discard the PBS in the catheter with a pipette into the waste beaker.</li>
	<li>Repeat PBS washing (steps 2.4 to 2.6) two more times. 
	NOTE: It is essential to work gently during these washing steps. Blood in the catheter or failure of automatic out-flow of liquid usually indicates the urethra is pierced through</li>
</ol>

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>BD 1mL TB Syringe</td>
			<td>Fisher</td>
			<td>148232E</td>
			<td></td>
		</tr>
		<tr>
			<td>Exel International Disposable Safelet I.V. Catheters</td>
			<td>Fisher Scientific</td>
			<td>14-841-21</td>
			<td>24G, 2.11 cm length, 0.045 cm outer diameter</td>
		</tr>
		<tr>
			<td>K&#38;H Heated Resting Mat for Small Animals, 9 By 12 Inches</td>
			<td>Amazon</td>
			<td></td>
			<td>Cover the heat mat with paper towels</td>
		</tr>
		<tr>
			<td>Ophthalmic ointment</td>
			<td>Fisher</td>
			<td>NC0849514</td>
			<td></td>
		</tr>
		<tr>
			<td>PBS, pH7.4</td>
			<td>Life Technologies</td>
			<td>10010049</td>
			<td></td>
		</tr>
		<tr>
			<td>Pipet tips 200 &#181;L</td>
			<td>USA Scientific</td>
			<td>1111-0206</td>
			<td></td>
		</tr>
		<tr>
			<td>Ophthalmic ointment&#160;</td>
			<td>Fisher&#160;</td>
			<td>NC0849514</td>
			<td></td>
		</tr>
		<tr>
			<td>Pipet Tips, Universal Fit; 0.1 to 10 &#181;L</td>
			<td>Fisher</td>
			<td>02707438(CS)</td>
			<td></td>
		</tr>
		<tr>
			<td>Buprenorphine&#160;</td>
			<td>Sigma&#160;</td>
			<td>B9275-50MG</td>
			<td></td>
		</tr>
		<tr>
			<td>PIPETMAN NEO P2</td>
			<td>Gilson</td>
			<td>F144561</td>
			<td></td>
		</tr>
		<tr>
			<td>PIPETMAN NEO P200N</td>
			<td>Gilson</td>
			<td>F144565</td>
			<td></td>
		</tr>
		<tr>
			<td>Therio-gel Veterinary Lubricant</td>
			<td>PBS animal health</td>
			<td>353-736</td>
			<td></td>
		</tr>
		<tr>
			<td>V-1 Table top system anesthesia</td>
			<td>VetEquip</td>
			<td>901806</td>
			<td></td>
		</tr>
		<tr>
			<td>Isoflurane&#160;</td>
			<td>Vet one&#160;</td>
			<td>501017</td>
			<td></td>
		</tr>
		<tr>
			<td>Tape, Scotch</td>
			<td>Fisher</td>
			<td>19047257</td>
			<td></td>
		</tr>
	</tbody>
</table>

urine depletion and bladder rinsing in mouse model: a catheter-based procedure to empty and wash murine urinary bladder

Source: Yu, C. et al. <a href="https://www.jove.com/video/57649" target="_blank">Novel Method of Plasmid DNA Delivery to Mouse Bladder Urothelium by Electroporation.</a> J. Vis. Exp. (2018)
In this video, we describe a catheter-based procedure to empty and rinse the urinary bladder of a mouse model. The prepped mouse can then be used for further experimental procedures.

Urine Depletion and Bladder Rinsing in Mouse Model: A Catheter-based Procedure to Empty and Wash Murine Urinary Bladder

Source: Yu, C. et al. Novel Method of Plasmid DNA Delivery to Mouse Bladder Urothelium by Electroporation. J. Vis. Exp. (2018)
In this video, we describe ...

To empty and wash a mouse&#39;s urinary bladder, begin by taking an anesthetized female mouse in the supine position. Next, obtain an appropriately-sized catheter. Lubricate the tip of the catheter to reduce friction and ensure a smooth urethral insertion. Then, slowly and gently insert the lubricated catheter, passing it through the urethral opening, into the bladder.
Upon successful catheterization, the urine begins to drain out of the bladder into the catheter. Now, empty the urine-filled catheter. Using a pipette, add a suitable buffered saline into the outer end of the catheter while its opposite end is within the bladder. To the outer end of the catheter, then attach a syringe. Gently push the syringe to deliver the saline to the bladder.
Now, remove the syringe. Allow the saline, along with the residual urine, to drain out of the bladder through the catheter. Subsequently, vacate the residual urine mixed with saline from the catheter. Repeat the bladder washing with saline twice to ensure complete removal of the urine from the bladder. The prepped mouse is finally ready for further experimental procedures.

urine-depletion-bladder-rinsing-mouse-model-catheter-based-procedure

Research

JoVE Encyclopedia of Experiments

Cancer Research

Urinary Tract Cancer

In vivo studies

1.8K 次观看. 来源:Yu， C. et al. 通过电穿孔将质粒 DNA 递送到小鼠膀胱尿路的新方法。J. Vis. Exp. （2018 年） 在本视频中，我们描述了一种基于导管的手术，用于清空和冲洗小鼠模型的膀胱。然后，制备好的小鼠可用于进一步的实验程序。 

视频: 小鼠模型中的尿液消耗和膀胱冲洗:一种基于导管的小鼠膀胱排空和清洗程序 - 概念

Urinary Tract Infection in a Small Animal Model: Transurethral Catheterization of Male and Female Mice

Urinary tract infections (UTI) are extremely common worldwide, incurring significant morbidity and healthcare-associated expenses. Small animal models, which accurately reflect disease establishment and progression, permit dissection of host-pathogen interactions and generation of immunity to infection. In mice, intravesical instillation of uropathogenic E. coli, the causative agent in more than 85% of community acquired UTI, recapitulates many of the stages of infection observed in humans. Until recently, however, UTI could only be modeled in female animals. This limitation has hindered the study of sex-related differences in UTI, as well as other bladder pathologies, such as cancer. Here, we describe a method to instill male mice that allows direct comparison between female and male animals and provide a detailed protocol to assess bladder tissue by flow cytometry as a means to better understand host responses to infection. Together, these approaches will aid in the identification of host factors that contribute to sex biases observed in UTI and other bladder-associated diseases.

The established model of transurethral catheterization of mice allows the study of bladder pathologies, including urinary tract infection, but can only be performed in females. A new model of male transurethral instillation, presented here, will enable research in an area marked by strong clinical and epidemiological differences between the sexes.

小动物模型尿路感染的实验研究: 男性和女性的经尿道插管

Urinary tract infections (UTI) are extremely common worldwide, incurring significant morbidity and healthcare-associated expenses. Small animal models, ...

科研

JoVE Journal

免疫与感染

Establishment and Characterization of UTI and CAUTI in a Mouse Model

Urinary tract infections (UTI) are highly prevalent, a significant cause of morbidity and are increasingly resistant to treatment with antibiotics. Females are disproportionately afflicted by UTI: 50% of all women will have a UTI in their lifetime. Additionally, 20-40% of these women who have an initial UTI will suffer a recurrence with some suffering frequent recurrences with serious deterioration in the quality of life, pain and discomfort, disruption of daily activities, increased healthcare costs, and few treatment options other than long-term antibiotic prophylaxis. Uropathogenic Escherichia coli (UPEC) is the primary causative agent of community acquired UTI. Catheter-associated UTI (CAUTI) is the most common hospital acquired infection accounting for a million occurrences in the US annually and dramatic healthcare costs. While UPEC is also the primary cause of CAUTI, other causative agents are of increased significance including Enterococcus faecalis. Here we utilize two well-established mouse models that recapitulate many of the clinical characteristics of these human diseases. For UTI, a C3H/HeN model recapitulates many of the features of UPEC virulence observed in humans including host responses, IBC formation and filamentation. For CAUTI, a model using C57BL/6 mice, which retain catheter bladder implants, has been shown to be susceptible to E. faecalis bladder infection. These representative models are being used to gain striking new insights into the pathogenesis of UTI disease, which is leading to the development of novel therapeutics and management or prevention strategies.

The ability to model urinary tract infections (UTI) is crucial in order to be able to understand bacterial pathogenesis and spawn the development of novel therapeutics. This work&#8217;s goal is to demonstrate mouse models of experimental UTI and catheter associated UTI that recapitulate and predict findings seen in humans.

建立和UTI和CAUTI表征在小鼠模型

Urinary tract infections (UTI) are highly prevalent, a significant cause of morbidity and are increasingly resistant to treatment with antibiotics. ...

医学

A Decentralized (Ex Vivo) Murine Bladder Model with the Detrusor Muscle Removed for Direct Access to the Suburothelium during Bladder Filling

Previous studies have established the release of chemical substances from flat bladder mucosa sheets affixed in Ussing chambers and exposed to changes in hydrostatic pressure or mechanical stretch and from cultured urothelial cells upon hydrostatic pressure changes, stretch, cell swelling, or drag forces, and in bladder lumen at end of filling. Such findings led to the assumption that these mediators are also released in suburothelium (SubU)/lamina propria (LP) during bladder filling, where they affect cells deep in the bladder wall to ultimately regulate bladder excitability. There are at least two obvious limitations in such studies: 1) none of these approaches provide direct information about the presence of mediators in SubU/LP, and 2) the stimuli used are not physiological and do not recapitulate authentic filling of the bladder. Here, we discuss a procedure that enables direct access to the suburothelial surface of the bladder mucosa in the course of bladder filling. The murine detrusor-free preparation we created closely resembles filling of the intact bladder and allows pressure-volume studies to be performed on the bladder in the absence of confounding signaling from spinal reflexes and detrusor smooth muscle. Using the novel detrusor-free bladder model, we recently demonstrated that intravesical measurements of mediators cannot be used as a proxy to what has been released or present in the SubU/LP during bladder filling. The model enables examination of urothelium-derived signaling molecules that are released, generated by metabolism and/or transported into the SubU/LP during the course of bladder filling to transmit information to neurons and smooth muscle of the bladder and regulate its excitability during continence and micturition.

A Decentralized Ex Vivo Murine Bladder Model with the Detrusor Muscle Removed for Direct Access to the Suburothelium during Bladder Filling

The detrusor-free bladder model enables direct access to the suburothelium to study local mechanisms for regulation of biologically active mediator availability in suburothelium/lamina propria during storage and voiding of urine. The preparation closely resembles filling of an intact bladder and allows pressure-volume studies to be performed without systemic influences.

分散（Ex Vivo）的毛骨球模型，去除了除位肌肉，用于在膀胱填充期间直接进入苏布罗氦

Previous studies have established the release of chemical substances from flat bladder mucosa sheets affixed in Ussing chambers and exposed to changes in ...

Urine Depletion and Bladder Rinsing in Mouse Model: A Catheter-based Procedure to Empty and Wash Murine Urinary Bladder

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