Name: transurethral instillation procedure in adult male mouse
Uploaded: 2017-11-02T15:00:00
Description: Watch this Scientific Journal Video about Transurethral Instillation Procedure in Adult Male Mouse at JoVE.com

The authors do not have any acknowledgements.

All the procedures with animals, including the method and duration of anesthesia, as well as postoperative care, were discussed with a veterinarian and approved by the Institutional Animal Care and Use Committee (IACUC) at the affiliated institution. In this manuscript, all procedures were performed with 8 to 15-week-old C57BL/6 male mice and the protocol follows the animal care guidelines approved by the IACUC at the University of Colorado.
1. Preparation of Tubing
N...

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J., Bushman, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chronic+bacterial+infection+and+inflammation+incite+reactive+hyperplasia+in+a+mouse+model+of+chronic+prostatitis.">Chronic bacterial infection and inflammation incite reactive hyperplasia in a mouse model of chronic prostatitis.</a> Prostate. 67 (1), 14-21 (2007).</li><li>Boehm, B. J., Colopy, S. A., Jerde, T. J., Loftus, C. J., Bushman, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Acute+bacterial+inflammation+of+the+mouse+prostate.">Acute bacterial inflammation of the mouse prostate.</a> Prostate. 72 (3), 307-317 (2012).</li><li>Bjorling, D. E., Wang, Z. 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Y., Boldon, K., Bushman, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bacterial+cystitis+is+accompanied+by+increased+peripheral+thermal+sensitivity+in+mice.">Bacterial cystitis is accompanied by increased peripheral thermal sensitivity in mice.</a> J Urol. 179 (2), 759-763 (2008).</li><li>Hopkins, W. J., Gendron-Fitzpatrick, A., Balish, E., Uehling, D. 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This manuscript describes in detail a method of transurethral instillation in adult male mouse. Differential primary seeding area can be reached by using PE tubing of different diameters. PE-50 is recommended for successfully reaching the urinary bladder lumen, while a larger diameter PE-10 is used to reach the AP and SV (Figure 3A-B). In addition to the choice of size of PE tubing and target organ for inoculation, other factors, such as the volume of instilled inoculum, als...

The transurethral instillation approach has been used in previous studies as one of the methods to create rodent models of bladder pathologies1,2,3,4 and can be used to evaluate the effectiveness of locally delivered treatments in mice. Although animal models cannot fully recapitulate human pathologies, identification of the underlying mechanisms in animal studies provides a better understanding of the human LUT disorders such as interstitial cystitis/bladder pain syndrome, neurogenic cystitis, autoimmune cystitis, and prostatic inflammation5.
The transurethral instillation procedure performed on an adult male mouse is more technically challenging than intravesical instillation in an adult female mouse6. The naturally curved anatomy of the male urethra along with its small diameter make it technically difficult to accomplish the transurethral insertion of a catheter. Therefore, detailed instructions for the transurethral induction of mouse urinary tract infection6 and LUT inflammation7,8,9,10,11,12,13,14,15 via bladder-inserted catheter were previously outlined for female mice only. This manuscript aims to provide a step-by-step description of the technique for transurethral instillation of substances in male mice including the video clips, images, and illustrations. Transurethral instillation procedures in an adult male mouse can be performed with variable inoculates similar to that previously described in the female mouse7 and merged with additional techniques such as electromyogram recordings of visceromotor responses (VMR)16.

<table>
	<tbody>
		<tr>
			<td>BD INTRAMEDIC Polyethylene Tubing/ PE 10</td>
			<td>BD Medical</td>
			<td>63019-004</td>
			<td></td>
		</tr>
		<tr>
			<td>BD INTRAMEDIC Polyethylene Tubing/ PE 50</td>
			<td>BD Medical</td>
			<td>63019-048</td>
			<td></td>
		</tr>
		<tr>
			<td>BD Insyte-N Autoguard/ Angiocath</td>
			<td>BD Medical</td>
			<td>311411</td>
			<td></td>
		</tr>
		<tr>
			<td>Ethanol (EtOH)</td>
			<td>Fisher Scientific</td>
			<td>BP2818500</td>
			<td></td>
		</tr>
		<tr>
			<td>BD Needle Only 22 Gauge (G) 1 inch</td>
			<td>Becton Dickson</td>
			<td>205155</td>
			<td></td>
		</tr>
		<tr>
			<td>BD Needle Only 27 Gauge (G) 1/2 inch</td>
			<td>Becton Dickson</td>
			<td>305129</td>
			<td></td>
		</tr>
		<tr>
			<td>Luer stub, 22 G X 0.5 in / Blunt Needle</td>
			<td>Instech</td>
			<td>LS22</td>
			<td></td>
		</tr>
		<tr>
			<td>Luer stub, 27 G X 0.5 in / Blunt Needle</td>
			<td>Instech</td>
			<td>LS27</td>
			<td></td>
		</tr>
		<tr>
			<td>6&#34; Long x 2&#34; Wide x 1&#34; Thick, Aluminum Oxide Sharpening Stone / Oil Stone</td>
			<td>Norton</td>
			<td>40204034</td>
			<td></td>
		</tr>
		<tr>
			<td>Surgical Lubricant Sterile Bacteriostatic Folilpac</td>
			<td>Surgilube</td>
			<td>281020543</td>
			<td></td>
		</tr>
		<tr>
			<td>Isoflurane</td>
			<td>Fluriso</td>
			<td>13985-528-60</td>
			<td></td>
		</tr>
		<tr>
			<td>SomnoSuite Low-Flow Anesthesia System</td>
			<td>Kent Scientific</td>
			<td>SS-01</td>
			<td></td>
		</tr>
		<tr>
			<td>Dissecting Forceps, Curved</td>
			<td>vwr</td>
			<td>82027-392</td>
			<td></td>
		</tr>
		<tr>
			<td>BD 1 ml Syringe</td>
			<td>BD Medical</td>
			<td>309626</td>
			<td></td>
		</tr>
		<tr>
			<td>India Ink Solution., 0.2% in PBS buffer</td>
			<td>Alfa Aesar</td>
			<td>J61007</td>
			<td></td>
		</tr>
		<tr>
			<td>8~12 weeks old male C57B/6 Mice</td>
			<td>Jackson Lab</td>
			<td>#000664</td>
			<td></td>
		</tr>
	</tbody>
</table>

transurethral instillation procedure in adult male mouse

Transurethral instillation can be used to deliver different solutions with active ingredients (e.g., drugs, chemicals, bacteria, and viruses) locally into the urinary bladder to either induce animal models of bladder pathologies or evaluate the effectiveness of intravesical treatments. Most rodent models of lower urinary tract (LUT) pathologies are induced in female mice due to ease of intravesical instillation of the substances via the female urethra. However, due to anatomical differences between the female and male LUT, transurethral instillation in a male mouse has been deemed a very challenging procedure, and it has not been previously described. In this manuscript, we provide a detailed description of how to prepare polyethylene (PE) tubing for subsequent insertion into the urethra of a male mouse. In addition, we discuss the ideal types of PE tubing to be used depending on the desired site of inoculation. Furthermore, we describe point by point how to prepare an animal for a successful transurethral instillation to avoid injury to the urethra and ensure the delivery of the solution to the desired location. The procedure is started by retracting the prepuce and the glans to expose the opening of the urethral meatus. Next, the glans are grasped by blunt non-crushing forceps to stabilize the penis and the PE tubing. The PE tubing is first inserted into the urethral meatus parallel to the animal body, then its angle is adjusted by tilting the catheter to maneuver it to follow the natural curvature of the urethra. This technique can be used to induced murine models of bladder pathologies and/or evaluate the effectiveness of intravesical treatments in male mice.

Transurethral instillation of blue dye via PE 50 tubing resulted in instillation of the dye into the urinary bladder (Figure 4A), and via PE 10 tubing in colorization of the SV and AP (Figure 4B), respectively. To evaluate whether the performed transurethral instillation causes an instant reflux of the inoculum to the kidneys or not, the kidney and ureter were observed followed by instillation of 100 &#181;L of the blue dye and f...

Watch this Scientific Journal Video about Transurethral Instillation Procedure in Adult Male Mouse at JoVE.com

Transurethral Instillation Procedure in Adult Male Mouse

Transurethral instillation is a challenging procedure and has not been well described in the literature. The aim of this manuscript is to describe a technique for transurethral insertion of a catheter for intravesical delivery of liquids with active substances into the urinary bladder and/or prostate in adult male mice.

Transurethral instillation can be used to deliver different solutions with active ingredients (e.g., drugs, chemicals, bacteria, and viruses) locally into ...

The overall goal of this procedure is to use transurethral catheter insertion for the intravesical delivery of active substances into the urinary bladder and/or prostate of adult male mice. This method can help answer key questions in the field of bladder pathology about how to study the underlying mechanisms involved in lower urinary tract disease in males. The main advantages of this technique are that it can be used to induce animal models of bladder pathology and to evaluate the effectiveness of intravesicular treatment.Transurethra instillation in male is challenging due to the natural curvature of the urethra, the thinness of the urethral wall, and the need for repetitive active substance instillation. Before beginning the procedure, select the PE tubing size based on the target organ and the age of the animal, and cut an approximately 2.5-inch long piece from the tube. Dull the sharp edge of a compatibly sized needle on a commercially available oil stone, and carefully introduce the needle into the lumen of one end of the tubing.Then, immerse the needle and tubing in 70%ethanol, flushing both pieces with ethanol to confirm the absence of leakage. Next, using gentle pressure, massage the lower abdomen of the mouse to induce voiding of the bladder. Then, place the anesthetized animal on a warm heating blanket.After placing the nose cone, confirm a lack of response to toe pinch, and retract the prepuce. Using the thumb and index finger of one hand, apply pressure to the pubic region and pull out the glans penis with the thumb and index finger of the other hand. Remove the tubing and catheter from the ethanol.Rinse the tubing and needle in sterile saline and load the needle onto a 1-mL syringe containing the inoculum of interest. Lubricate the tip of the catheter and the glans with sterile surgical lubricant. Gently grasp the tip of the penis with dull forceps and apply gentle pressure to the glans to open the urethral meatus.Using a tripod finger grip, carefully insert the PE tubing into the meatus, adjusting the angle of the needle to follow the curvature of the urethra as necessary. When the tubing has reached the appropriate organ, depress the plunger to instill the full experimental volume of the inoculum. In these representative images, PE-52 tubing was used to facilitate the transurethral instillation of a blue dye into the mouse urinary bladder, and PE-10 tubing was used to colorize the seminal vesicles and the anterior prostate of a second animal.However, histological evaluation of the ureters or kidneys is highly recommended to confirm the absence of indirect contamination of the ureters or kidneys from transurethral instillations. Once mastered, this technique can be performed within 10 to 15 minutes if it is performed properly. After watching this video, you should have a good understanding of how to perform transurethra catheterization in an adult male mouse.

transurethral-instillation-procedure-in-adult-male-mouse

Research

JoVE Journal

Immunology and Infection

Transurethral Induction of Mouse Urinary Tract Infection

Uropathogenic bacterial strains of interest are grown on agar. Generally, uropathogenic E. coli (UPEC) and other strains can be grown overnight on Luria-Bertani (LB) agar at 37&deg;C in ambient air. UPEC strains grow as yellowish-white translucent colonies on LB agar. Following confirmation of appropriate colony morphology, single colonies are then picked to be cultured in broth. LB broth can be used for most uropathogenic bacterial strains. Two serial, overnight LB broth cultures can be employed to enhance expression of type I pili, a well-defined virulence factor for uropathogenic bacteria. Broth cultures are diluted to the desired concentration in phosphate buffered saline (PBS). Eight to 12 week old female mice are placed under isoflurane anesthesia and transurethrally inoculated with bacteria using polyethylene tubing-covered 30 gauge syringes. Typical inocula, which must be empirically determined for each bacterial/mouse strain combination, are 106 to 108 cfu per mouse in 10 to 50 microliters of PBS. After the desired infection period (one day to several weeks), urine samples and the bladder and both kidneys are harvested. Each organ is minced, placed in PBS, and homogenized in a Blue Bullet homogenizer. Urine and tissue homogenates are serially diluted in PBS and cultured on appropriate agar. The following day, colony forming units are counted.

This video will demonstrate methods to transurethrally induce mouse urinary tract infections and quantify the extent of resulting infections.

Uropathogenic bacterial strains of interest are grown on agar. Generally, uropathogenic E. coli (UPEC) and other strains can be grown overnight on ...

Retroviral Transduction of T-cell Receptors in Mouse T-cells

T-cell receptors (TCRs) play a central role in the immune system. TCRs on T-cell surfaces can specifically recognize peptide antigens presented by antigen presenting cells (APCs)1. This recognition leads to the activation of T-cells and a series of functional outcomes (e.g. cytokine production, killing of the target cells). Understanding the functional role of TCRs is critical to harness the power of the immune system to treat a variety of immunology related diseases (e.g. cancer or autoimmunity).
It is convenient to study TCRs in mouse models, which can be accomplished in several ways. Making TCR transgenic mouse models is costly and time-consuming and currently there are only a limited number of them available2-4. Alternatively, mice with antigen-specific T-cells can be generated by bone marrow chimera. This method also takes several weeks and requires expertise5. Retroviral transduction of TCRs into in vitro activated mouse T-cells is a quick and relatively easy method to obtain T-cells of desired peptide-MHC specificity. Antigen-specific T-cells can be generated in one week and used in any downstream applications. Studying transduced T-cells also has direct application to human immunotherapy, as adoptive transfer of human T-cells transduced with antigen-specific TCRs is an emerging strategy for cancer treatment6.
Here we present a protocol to retrovirally transduce TCRs into in vitro activated mouse T-cells. Both human and mouse TCR genes can be used. Retroviruses carrying specific TCR genes are generated and used to infect mouse T-cells activated with anti-CD3 and anti-CD28 antibodies. After in vitro expansion, transduced T-cells are analyzed by flow cytometry.

We present a protocol to produce antigen-specific mouse T-cells using retroviral transduction

T-cell receptors (TCRs) play a central role in the immune system. TCRs on T-cell surfaces can specifically recognize peptide antigens presented by antigen ...

Non-surgical Intratracheal Instillation of Mice with Analysis of Lungs and Lung Draining Lymph Nodes by Flow Cytometry

Phagocytic cells such as alveolar macrophages and lung dendritic cells (LDCs) continuously sample antigens from the alveolar spaces in the 
lungs. LDCs, in particular, are known to migrate to the lung draining lymph nodes (LDLNs) where they present inhaled antigens to T cells initiating an 
appropriate immune response to a variety of immunogens1,2. To model interactions between the lungs and airborne antigens in mice, antigens can be 
administered intranasally1,3,4, intratracheally5 or as aerosols6. Delivery by each route involves distinct technical skills and limitations that need to be 
considered before designing an experiment. For example, intranasal and aerosolized exposure delivers antigens to both the lungs and the upper respiratory 
tract. Hence antigens can access the nasal associated lymphoid tissue (NALT)7, potentially complicating interpretation of the results. In addition, swallowing, 
sneezing and the breathing rate of the mouse may also lead to inconsistencies in the doses delivered. Although the involvement of the upper respiratory tract may 
be preferred for some studies, it can complicate experiments focusing on events specifically initiated in the lungs. In this setting, the intratracheal (i.t) 
route is preferable as it delivers test materials directly into the lungs and bypasses the NALT. Many i.t injection protocols involve either blind intubation of the 
trachea through the oral cavity or surgical exposure of the trachea to access the lungs. Herein, we describe a simple, consistent, non-surgical method for i.t 
instillation. The opening of the trachea is visualized using a laryngoscope and a bent gavage needle is then inserted directly into the trachea to deliver the 
innoculum. We also describe procedures for harvesting and processing of LDLNs and lungs for analysis of antigen trafficking by flow cytometry.

We illustrate non-surgical delivery of test materials into the lungs of anesthetized mice via the trachea. This method permits lung exposure to bacterial and viral pathogens, cytokines, antibodies, beads, chemicals, or dyes. We further describe harvesting and processing of lungs and lung draining lymph nodes (LDLNs) for flow cytometry.

Phagocytic cells such as alveolar macrophages and lung dendritic cells (LDCs) continuously sample antigens from the alveolar spaces in the 
lungs. LDCs, ...

Culturing Microglia from the Neonatal and Adult Central Nervous System

Microglia are the resident macrophage-like cells of the central nervous system (CNS) and, as such, have critically important roles in physiological and pathological processes such as CNS maturation in development, multiple sclerosis, and spinal cord injury. Microglia can be activated and recruited to action by neuronal injury or stimulation, such as axonal damage seen in MS or ischemic brain trauma resulting from stroke. These immunocompetent members of the CNS are also thought to have roles in synaptic plasticity under non-pathological conditions. We employ protocols for culturing microglia from the neonatal and adult tissues that are aimed to maximize the viable cell numbers while minimizing confounding variables, such as the presence of other CNS cell types and cell culture debris. We utilize large and easily discernable CNS components (e.g. cortex, spinal cord segments), which makes the entire process feasible and reproducible. The use of adult cells is a suitable alternative to the use of neonatal brain microglia, as many pathologies studied mainly affect the postnatal spinal cord. These culture systems are also useful for directly testing the effect of compounds that may either inhibit or promote microglial activation. Since microglial activation can shape the outcomes of disease in the adult CNS, there is a need for in vitro systems in which neonatal and adult microglia can be cultured and studied.

We outline methods for the efficient and quick isolation/culture of viable microglia from the neonatal cerebral cortex and adult spinal cord. The dissection and plating of cortical microglia can be accomplished within 90 minutes, with the subsequent microglial harvest taking place ~ 10 days following the initial dissection.

Microglia are the resident macrophage-like cells of the central nervous system (CNS) and, as such, have critically important roles in physiological and ...

Complete Thymectomy in Adult Rats with Non-invasive Endotracheal Intubation

Thymectomy in neonatal rodents is an established and reliable procedure for immunological studies. However, in adult rats, complications of hemorrhage and pneumothorax from pleural disruption can result in a significant mortality rate. This protocol is a simple method of rat thymectomy that utilizes a mini-sternotomy and endotracheal intubation. Intubation is accomplished with a non-invasive and easily reproducible method and allows for positive pressure ventilation to prevent pneumothorax and a controlled airway that allows sufficient time for careful thymus dissection to minimize pleural disruption. A 1.5 cm sternal incision decreases contact with mediastinal vessels and pleura, while still providing full visualization of the thymus. Following exposure of the mediastinum, the thymus is removed by blunt dissection under magnification. The pleural space is then sealed by suture closure of the pre-tracheal muscles followed by the application of surgical glue. The thorax is then closed by suture closure of the sternum, followed by suture closure of the skin. All thymectomies were complete as evidenced by immunohistochemical (IHC) staining of mediastinal tissue, and absence of na&#239;ve T-cells by flow cytometry, and the procedure had a 96% survival rate. This method is suitable when complete thymectomy with minimal complications is desired for further immunological studies in athymic adult rats.

Rodent thymectomy is a valuable technique in immunological research. Here, a protocol for complete thymectomy in adult rats using a mini-sternotomy along with non-invasive intubation and positive pressure ventilation to minimize perioperative morbidity and mortality is described.

Thymectomy in neonatal rodents is an established and reliable procedure for immunological studies. However, in adult rats, complications of hemorrhage and ...

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.

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

Quantifying Vibrio cholerae Colonization and Diarrhea in the Adult Zebrafish Model

Vibrio cholerae is best known as the infectious agent that causes the human disease cholera. Outside the human host, V. cholerae primarily exists in the aquatic environment, where it interacts with a variety of higher aquatic species. Vertebrate fish are known to be an environmental host and are a potential V. cholerae reservoir in nature. Both V. cholerae and the teleost fish species Danio rerio, commonly known as zebrafish, originate from the Indian subcontinent, suggesting a long-standing interaction in aquatic environments. Zebrafish are an ideal model organism for studying many aspects of biology, including infectious diseases. Zebrafish can be easily and rapidly colonized by V. cholerae after exposure in water. Intestinal colonization by V. cholerae leads to the production of diarrhea and the excretion of replicated V. cholerae. These excreted bacteria can then go on to colonize new fish hosts. Here, we demonstrate how to assess V. cholerae-intestinal colonization in zebrafish and how to quantify V. cholerae-induced zebrafish diarrhea. The colonization model should be useful to researchers who are studying whether genes of interest may be important for host colonization and/or for environmental survival. The quantification of zebrafish diarrhea should be useful to researchers studying any intestinal pathogen who are interested in exploring zebrafish as a model system.

Quantifying Vibrio cholerae Colonization and Diarrhea in the Adult Zebrafish Model

Zebrafish are a natural Vibrio cholerae host and can be used to recapitulate and study the entire infectious cycle from colonization to transmission. Here, we demonstrate how to assess V. cholerae colonization levels and quantify diarrhea in zebrafish.

Vibrio cholerae is best known as the infectious agent that causes the human disease cholera. Outside the human host, V. cholerae primarily exists in the ...

Modeling Tuberculosis in Mycobacterium marinum Infected Adult Zebrafish

Mycobacterium tuberculosis is currently the deadliest human pathogen causing 1.7 million deaths and 10.4 million infections every year. Exposure to this bacterium causes a wide disease spectrum in humans ranging from a sterilized infection to an actively progressing deadly disease. The most common form is the latent tuberculosis, which is asymptomatic, but has the potential to reactivate into a fulminant disease. Adult zebrafish and its natural pathogen Mycobacterium marinum have recently proven to be an applicable model to study the wide disease spectrum of tuberculosis. Importantly, spontaneous latency and reactivation as well as adaptive immune responses in the context of mycobacterial infection can be studied in this model. In this article, we describe methods for the experimental infection of adult zebrafish, the collection of internal organs for the extraction of nucleic acids for the measurement of mycobacterial loads and host immune responses by quantitative PCR. The in-house-developed, M. marinum-specific qPCR assay is more sensitive than the traditional plating methods as it also detects DNA from non-dividing, dormant or recently dead mycobacteria. As both DNA and RNA are extracted from the same individual, it is possible to study the relationships between the diseased state, and the host and pathogen gene-expression. The adult zebrafish model for tuberculosis thus presents itself as a highly applicable, non-mammalian in vivo system to study host-pathogen interactions.

Modeling Tuberculosis in Mycobacterium marinum Infected Adult Zebrafish

Here, we present a protocol to model human tuberculosis in an adult zebrafish using its natural pathogen Mycobacterium marinum. Extracted DNA and RNA from the internal organs of infected zebrafish can be used to reveal the total mycobacterial loads in the fish and the host&#39;s immune responses with qPCR.

Mycobacterium tuberculosis is currently the deadliest human pathogen causing 1.7 million deaths and 10.4 million infections every year. Exposure to this ...

A Model of Self-limited Acute Lung Injury by Unilateral Intra-bronchial Acid Instillation

Selective intra-bronchial instillation of hydrochloric acid (HCl) to the murine left mainstem bronchus causes acute tissue injury with histopathologic findings similar to human acute respiratory distress syndrome (ARDS). The resulting alveolar edema, alveolar-capillary barrier damage, and leukocyte infiltration predominantly affect the left lung, preserving the right lung as an uninjured control and allowing animals to survive. This model of self-limited acute lung injury enables investigation of tissue resolution mechanisms, such as macrophage efferocytosis of apoptotic neutrophils and restitution of alveolar-capillary barrier integrity. This model has helped identify important roles for resolution agonists, including specialized pro-resolving mediators (SPMs), providing a foundation for the development of new therapeutic approaches for patients with ARDS.

Selective intra-bronchial acid instillation to the left lung in mice results in unilateral and self-limited acute lung injury that models human acute respiratory distress syndrome (ARDS) induced by gastric acid aspiration.

Selective intra-bronchial instillation of hydrochloric acid (HCl) to the murine left mainstem bronchus causes acute tissue injury with histopathologic ...

Experimental Viral Infection in Adult Mosquitoes by Oral Feeding and Microinjection

Mosquito-borne viruses (MBVs), which are infectious pathogens to vertebrates, are spread by many mosquito species, posing a severe threat to public health. Once ingested, the viruses must overcome the mosquito midgut barrier to reach the hemolymph, from where they might potentially spread to the salivary glands. When a mosquito bites, these viruses are spread to new vertebrate hosts. Similarly, the mosquito may pick up different viruses. In general, only a tiny portion of viruses may enter the salivary glands via the gut. The transmission efficiency of these viruses to the glands is be affected by the two physical barriers found in different mosquito species: midgut barriers and salivary glands barriers. This protocol presents a method for virus detection in salivary glands of Aedes aegypti&#39;s&#160;following oral feeding and intrathoracic injection infection. Furthermore, determining whether the guts and/or salivary glands hinder viral spread can aid in the risk assessments of MBVs transmitted by Aedes aegypti.

This methodology, which included oral feeding and intrathoracic injection infection, could effectively assess the influence of midgut and/or salivary gland barriers on arbovirus infection.

Mosquito-borne viruses (MBVs), which are infectious pathogens to vertebrates, are spread by many mosquito species, posing a severe threat to public ...