We are grateful for funding from the NCI Ovarian Cancer SPORE Program P50CA228991, post-doctoral training program 5T32OD011089, and the Richard W. TeLinde Endowment, Johns Hopkins University. The project was also partly funded by the subsidies for current expenditures to Private Institutions of Higher Education from the Promotion and Mutual Aid Corporation for Private Schools of Japan.

All procedures and experiments involving mice were performed according to protocols approved by Johns Hopkins Animal Care and Use Committee. For all procedures, appropriate PPE was worn, including gloves and disposable isolation gowns. Precautions were taken when handling sharps, which were properly disposed of in red box sharps containers immediately after use. See the Table of Materials for details about all the materials and equipment used in this protocol.
1. Inducti...

<ol>
	<li>Ajayi, A. F., Akhigbe, R. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Staging+of+the+estrous+cycle+and+induction+of+estrus+in+experimental+rodents:+an+update.">Staging of the estrous cycle and induction of estrus in experimental rodents: an update.</a> Fertility Research and Practice. 6, 5 (2020).</li><li>Kim, S. W., Kim, Y. Y., Kim, H., Ku, S. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Animal+models+closer+to+intrauterine+adhesive+pathology.">Animal models closer to intrauterine adhesive pathology.</a> Annals of Translational Medicine. 8 (18), 1125 (2020).</li><li>Shi, D., Vine, D. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Animal+models+of+polycystic+ovary+syndrome:+a+focused+review+of+rodent+models+in+relationship+to+clinical+phenotypes+and+cardiometabolic+risk.">Animal models of polycystic ovary syndrome: a focused review of rodent models in relationship to clinical phenotypes and cardiometabolic risk.</a> Fertility and Sterility. 98 (1), 185-193 (2012).</li><li>Greco, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ultrasound+biomicroscopy+in+small+animal+research:+applications+in+molecular+and+preclinical+imaging.">Ultrasound biomicroscopy in small animal research: applications in molecular and preclinical imaging.</a> Journal of Biomedicine and Biotechnology. 2012, 519238 (2012).</li><li>Palsdottir, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Interobserver+agreement+of+transvaginal+ultrasound+and+magnetic+resonance+imaging+in+local+staging+of+cervical+cancer.">Interobserver agreement of transvaginal ultrasound and magnetic resonance imaging in local staging of cervical cancer.</a> Ultrasound in Obstetrics and Gynecology. 58 (5), 773-779 (2021).</li><li>Gabrielson, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vivo+imaging+with+confirmation+by+histopathology+for+increased+rigor+and+reproducibility+in+translational+research:+A+review+of+examples,+options,+and+resources.">In vivo imaging with confirmation by histopathology for increased rigor and reproducibility in translational research: A review of examples, options, and resources.</a> ILAR Journal. 59 (1), 80-98 (2018).</li><li>Peterson, R. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Continuing+education+course+#1:+Non-invasive+imaging+as+a+problem-solving+tool+and+translational+biomarker+strategy+in+toxicologic+pathology.">Continuing education course #1: Non-invasive imaging as a problem-solving tool and translational biomarker strategy in toxicologic pathology.</a> Toxicologic Pathology. 39 (1), 267-272 (2011).</li><li>Pfeifer, L. F., Adams, G. P., Pierson, R. A., Singh, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ultrasound+biomicroscopy:+A+non-invasive+approach+for+in+vivo+evaluation+of+oocytes+and+small+antral+follicles+in+mammals.">Ultrasound biomicroscopy: A non-invasive approach for in vivo evaluation of oocytes and small antral follicles in mammals.</a> Reproduction, Fertility and Development. 26 (1), 48-54 (2013).</li><li>Cheung, A. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Three-dimensional+ultrasound+biomicroscopy+for+xenograft+growth+analysis.">Three-dimensional ultrasound biomicroscopy for xenograft growth analysis.</a> Ultrasound in Medicine and Biology. 31 (6), 865-870 (2005).</li><li>Snyder, C. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Complementarity+of+ultrasound+and+fluorescence+imaging+in+an+orthotopic+mouse+model+of+pancreatic+cancer.">Complementarity of ultrasound and fluorescence imaging in an orthotopic mouse model of pancreatic cancer.</a> BMC Cancer. 9, 106 (2009).</li><li>Wu, G., Wang, L., Yu, L., Wang, H., Xuan, J. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+use+of+three-dimensional+ultrasound+micro-imaging+to+monitor+prostate+tumor+development+in+a+transgenic+prostate+cancer+mouse+model.">The use of three-dimensional ultrasound micro-imaging to monitor prostate tumor development in a transgenic prostate cancer mouse model.</a> The Tohoku Journal of Experimental Medicine. 207 (3), 181-189 (2005).</li><li>Rinaldi, S. F., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ultrasound-guided+intrauterine+injection+of+lipopolysaccharide+as+a+novel+model+of+preterm+birth+in+the+mouse.">Ultrasound-guided intrauterine injection of lipopolysaccharide as a novel model of preterm birth in the mouse.</a> The American Journal of Pathology. 185 (5), 1201-1206 (2015).</li><li>Wang, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ultrasonography+in+experimental+reproductive+investigations+on+rats.">Ultrasonography in experimental reproductive investigations on rats.</a> Journal of Visualized Experiments. 130, e56038 (2017).</li><li>Suryo Rahmanto, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Inactivation+of+Arid1a+in+the+endometrium+is+associated+with+endometrioid+tumorigenesis+through+transcriptional+reprogramming.">Inactivation of Arid1a in the endometrium is associated with endometrioid tumorigenesis through transcriptional reprogramming.</a> Nature Communications. 11, 2717 (2020).</li><li>Pani, F., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pre-existing+thyroiditis+ameliorates+papillary+thyroid+cancer:+Insights+from+a+new+mouse+model.">Pre-existing thyroiditis ameliorates papillary thyroid cancer: Insights from a new mouse model.</a> Endocrinology. 162 (10), bqab144 (2021).</li></ol>

The authors have no conflicts of interest to disclose.

This protocol examines the utility of ultrasound for assessing uterine morphological changes in the progression of adenocarcinoma in the uterus in mice. In this study, by following the induction of endometrial cancer in mice longitudinally, the anatomical details detected by ultrasound were found to be indicators of gross and histological pathology. This opens the door for the use of longitudinal studies with smaller numbers of mice monitored by ultrasound at multiple time points to follow the progression of uterine canc...

Mice remain one of the most important animal models for reproductive disorders1,2,3. There are several genetically modified or induced rodent models of ovarian and uterine cancers. These studies typically rely on multiple cohorts euthanized at different time points to capture longitudinal trends in morphologic and pathologic changes. This prevents the ability to acquire continuous data on cancer development in an individual mouse. Additionally, without knowing the individual mouse disease progression state, intervention studies are based on predetermined time points and averaged findings of previous cohorts rather than individual thresholds for the detection of progression in a specific animal4,5. Therefore, imaging approaches that allow for longitudinal assessment in live animals are needed to facilitate preclinical models for testing new drugs or compounds and accelerate the understanding of pathobiology while also increasing the rigor and reproducibility6.
Ultrasound imaging (US) is an appealing method for the longitudinal monitoring of mouse uterine cancer progression because it is relatively facile and inexpensive compared to other imaging methods, is easy to perform, and can have remarkable resolution6,7. This non-invasive modality can capture features to the micron scale in awake mice or with mice under brief sedation using a 5-10 min exam. Ultrasound microscopy has been validated as a method to measure mouse ovarian follicle development 8 and the growth of implanted or induced neoplasia9,10,11. High-frequency US has also been used for percutaneous intrauterine injections12 and observing rat uterine change over the estrus cycle13. High-frequency US can be used with mice held on specialized stationary platforms using a rail system to hold the transducer/probe to capture high-resolution images with standardized position and pressure; however, this equipment is not available at all institutions. Hand-held transducer scanning methods can be adopted with less dedicated equipment and used for both clinical diagnostics and research applications in mice.
The question remains as to whether US imaging with hand-held, high-frequency probes could be used to monitor uterine cancer development over multiple weeks. Similar to the intestines, the rodent uterus is a thin-walled, slender structure that is very mobile within the abdomen and is contiguous through multiple tissue depths, making imaging more challenging than with relatively immobile organs such as the kidneys. This study sought to establish the correlation between tissues observed by ultrasound and histopathology, define landmarks for locating the mouse uterus, and determine the feasibility of the longitudinal assessment of endometrial cancer. This study presents data showing a qualitative correspondence between the appearance of uteri imaged by US and histopathology, as well as serial imaging of mice over several weeks. These results indicate that hand-held US can be used to monitor endometrial cancer development in mice, thus creating an opportunity for collecting individual mouse longitudinal data to study uterine cancer without the need for dedicated equipment.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Reagents and Equipment Used for Animal Care</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Rodent Diet (2018, 625 Doxycycline)</td>
			<td>Envigio</td>
			<td>TD.01306</td>
			<td>Mouse Feed</td>
		</tr>
		<tr>
			<td>Reagents and Equipment Used for Ultrasound Imaging</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>10 mL injectable 0.9% NaCl&#160;</td>
			<td>Hospira, Inc</td>
			<td>RL-7302</td>
			<td>Isotonic Fluid</td>
		</tr>
		<tr>
			<td>Absorbent Pad with Plastic Backing</td>
			<td>Daigger</td>
			<td>EF8313</td>
			<td>Absorbant Pads</td>
		</tr>
		<tr>
			<td>Anesthesia Induction Chambers</td>
			<td>Harvard Apparatus</td>
			<td>75-2029</td>
			<td>Induction Chamber</td>
		</tr>
		<tr>
			<td>Anesthetic absorber kit with absorber canister, holder, tubing, &#38; adapters</td>
			<td>CWE, Inc</td>
			<td>13-20000</td>
			<td>Nose Cone and Tubing</td>
		</tr>
		<tr>
			<td>Aquasonic Clear Ultrasound Gel (0.25 Liter)</td>
			<td>Parker Laboratoies</td>
			<td>08-03</td>
			<td>Ultrasound Gel</td>
		</tr>
		<tr>
			<td>BD Plastipak 3 mL Syringe</td>
			<td>BD Biosciences</td>
			<td>309657</td>
			<td>Syringe</td>
		</tr>
		<tr>
			<td>F/Air Scavenger Charcoal Canister</td>
			<td>OMNICON</td>
			<td>80120</td>
			<td>Scavenging System for Anesthesia</td>
		</tr>
		<tr>
			<td>Isoflurane, USP</td>
			<td>Vet One</td>
			<td>502017</td>
			<td>Anesthesia Agent</td>
		</tr>
		<tr>
			<td>M1050 Non-Rebreathing Mobile Anesthesia Machine</td>
			<td>Scivena Scientific</td>
			<td>M1050</td>
			<td>Anestheic Vaporizer</td>
		</tr>
		<tr>
			<td>MX550S, 25-55 MHz Transducer, 15mm, Linear</td>
			<td>VisualSonics</td>
			<td>MX550S</td>
			<td>Ultrasound Transducer (Probe)</td>
		</tr>
		<tr>
			<td>Nair Hair Aloe &#38; Lanolin Hair Removal Lotion - 9.0 oz</td>
			<td>Nair</td>
			<td></td>
			<td>Depilliating Cream</td>
		</tr>
		<tr>
			<td>Philips Norelco Multigroomer All-in-One Trimmer Series 7000</td>
			<td>Philips North America</td>
			<td>MG7750</td>
			<td>Clippers</td>
		</tr>
		<tr>
			<td>PrecisionGlide 25 G 1&#34; Needle</td>
			<td>BD Biosciences</td>
			<td>305125</td>
			<td>Needle</td>
		</tr>
		<tr>
			<td>Puralube Ophthalmic Ointment</td>
			<td>Dechra</td>
			<td>17033-211-38</td>
			<td>Lubricating Eye Drops</td>
		</tr>
		<tr>
			<td>Vevo 3100 Imaging System</td>
			<td>VisualSonics</td>
			<td>Vevo 3100</td>
			<td>Ultrasound Machine</td>
		</tr>
		<tr>
			<td>Vevo LAB 5.6.1</td>
			<td>VisualSonics</td>
			<td>Vevo LAB 5.6.1</td>
			<td>Ultrasound Analysis Software</td>
		</tr>
		<tr>
			<td>Vinyl Heating Pad with cover, 12 x 15&#34;</td>
			<td>Sunbeam</td>
			<td>731-500-000R</td>
			<td>Heating Pad</td>
		</tr>
		<tr>
			<td>Wd Elements 2TB Basic Storage</td>
			<td>Western Digital Elements</td>
			<td>WDBU6Y0020BBK-WESN</td>
			<td>Data Storage</td>
		</tr>
		<tr>
			<td>Reagents and Equipment Used for Immunohistochemistry</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>10% w/v Formalin</td>
			<td>Fischer Scientific</td>
			<td>SF98-4</td>
			<td>Tissue Fixation Buffer</td>
		</tr>
		<tr>
			<td>Animal-Free Blocker and Diluent, R.T.U.</td>
			<td>Vector Laboratories Inc.</td>
			<td>&#160;SP5035</td>
			<td>Antibody Blocker</td>
		</tr>
		<tr>
			<td>Charged Super Frost Plus Glass Slides</td>
			<td>VWR</td>
			<td>4831-703</td>
			<td>Tissue Mounting Slides</td>
		</tr>
		<tr>
			<td>Citrate Buffer</td>
			<td>MilliporeSigma</td>
			<td>&#160;C9999-1000ML</td>
			<td>Epitope Retrival Buffer (pTEN)</td>
		</tr>
		<tr>
			<td>Cytoseal &#8211; 60</td>
			<td>Thermo Scientific</td>
			<td>8310-4</td>
			<td>Resin for Slide Sealing</td>
		</tr>
		<tr>
			<td>Gold Seal Cover Glass</td>
			<td>Thermo Scientific</td>
			<td>3322</td>
			<td>Coverslide</td>
		</tr>
		<tr>
			<td>Harris Modified Hematoxylin</td>
			<td>MilliporeSigma</td>
			<td>HHS32-1L</td>
			<td>Counterstain Buffer</td>
		</tr>
		<tr>
			<td>Hybridization Incubator (Dual Chamber)</td>
			<td>Fischer Scientific</td>
			<td>13-247-30Q</td>
			<td>Oven to Melt Parraffin</td>
		</tr>
		<tr>
			<td>ImmPACT DAB Substrate, Peroxidase (HRP)</td>
			<td>Vector Laboratories Inc.</td>
			<td>SK-4105</td>
			<td>Signal Development Substrate</td>
		</tr>
		<tr>
			<td>ImmPRESS HRP Goat Anti-Rabbit IgG Polymer Detection Kit, Peroxidase</td>
			<td>Vector Laboratories Inc.</td>
			<td>MP-7451</td>
			<td>Secondary IHC Antibody</td>
		</tr>
		<tr>
			<td>Oster 5712 Digital Food Steamer</td>
			<td>Oster</td>
			<td>5712</td>
			<td>Vegetable Steamer for Epitope Retrival</td>
		</tr>
		<tr>
			<td>rabbit mAB anti-ARID1a</td>
			<td>abcam</td>
			<td>ab182560</td>
			<td>Primary IHC Antibody (1:1,000)</td>
		</tr>
		<tr>
			<td>rabbit mAB anti-PTEN</td>
			<td>Cell Signaling</td>
			<td>9559</td>
			<td>Primary IHC Antibody (1:100)</td>
		</tr>
		<tr>
			<td>Scotts Tap Water Substitute</td>
			<td>MilliporeSigma</td>
			<td>S5134-100ML</td>
			<td>&#34;Blueing&#34; Buffer</td>
		</tr>
		<tr>
			<td>Tissue Path IV Cassette</td>
			<td>Fischer Scientific</td>
			<td>22272416</td>
			<td>Tissue Fixation Cassette</td>
		</tr>
		<tr>
			<td>Trilogy Buffer</td>
			<td>Cell Marque</td>
			<td>&#160;920P-10</td>
			<td>Epitope Retrival Buffer (ARID1a)</td>
		</tr>
	</tbody>
</table>

non-invasive ultrasound assessment of endometrial cancer progression in pax8-directed deletion of the tumor suppressors arid1a and pten in mice

Uterine cancers can be studied in mice due to the ease of handling and genetic manipulation in these models. However, these studies are often limited to assessing pathology post-mortem in animals euthanized at multiple time points in different cohorts, which increases the number of mice needed for a study. Imaging mice in longitudinal studies can track the progression of disease in individual animals, reducing the number of mice needed. Advances in ultrasound technology have allowed for the detection of micrometer-level changes in tissues. Ultrasound has been used to study follicle maturation in ovaries and xenograft growth but has not been applied to morphological changes in the mouse uterus. This protocol examines the juxtaposition of pathology with in vivo imaging comparisons in an induced endometrial cancer mouse model. The features observed by ultrasound were consistent with the degree of change seen by gross pathology and histology. Ultrasound was found to be highly predictive of the observed pathology, supporting the incorporation of ultrasonography into longitudinal studies of uterine diseases such as cancer in mice.

Pax8-Cre-Arid1a-Pten double deletion (iPAD) transgenic mice were maintained on a mixed genetic background (129S, BALB/C, C57BL/6), as previously described14. The mice were all fed a doxycycline feed for 2 weeks to induce Cre recombinase. In previous work by our group, doxycycline was dosed by gavage14; however, in this current study, the doxycycline feed induction method worked efficiently and reduced the stress of gavage for the mice. It is important to c...

Watch this Scientific Journal Video about Non-Invasive Ultrasound Assessment of Endometrial Cancer Progression in Pax8-Directed Deletion of the Tumor Suppressors Arid1a and Pten in Mice at JoVE.com

Non-Invasive Ultrasound Assessment of Endometrial Cancer Progression in Pax8-Directed Deletion of the Tumor Suppressors Arid1a and Pten in Mice

This protocol describes a method for monitoring the progression of morphological changes over time in the uterus in an inducible mouse model of endometrial cancer using ultrasound imaging with correlation to gross and histological changes.

Non-Invasive Ultrasound Assessment of Endometrial Cancer Progression in Pax8-Directed Deletion of the Tumor Suppressors Arid1a and Pten in Mice

Uterine cancers can be studied in mice due to the ease of handling and genetic manipulation in these models. However, these studies are often limited to ...

This study shows that manual guided ultrasound is well-suited to monitor the development of uterine pathology in an induced mouse model of endometrial cancer. Ultrasound imaging is noninvasive and thus allows for longitudinal studies of individual mice across days and weeks. This allows for the capture of individual variation and ultimately for fewer animals to be used for study.This method can be readily applied to monitor pathologic changes and other mirroring models, including those for neoplasia and chronic disease of the urinary and digestive tract. Begin setting up the ultrasound machine for imaging the uterus or ovary by selecting a transducer or probe with a range of 32 to 56 megahertz. Go to the Applications tab from the home screen and select Mouse Small Abdomen Mode.Then click on Scan to return to the home screen and wait for a live image to be displayed. Next, select B Mode from the options on the left toolbar before clicking on More Controls option to view additional tools for image refinement, such as image gain and depth or to adjust the clip acquisition settings, such as the number of frames per second. Once the image settings are selected, return to the home screen by clicking on B Mode.Place a three to 10-milliliter syringe filled with a sterile isotonic fluid solution between a heating pad and an absorbent pad for several minutes to preheat it to 35 to 40 degrees Celsius. If the machine does not have a warmer, place a bottle of ultrasound gel on the heating pad. To inject one to two milliliters of solution into the peritoneal cavity of a mouse, secure the mouse by the scruff in one hand, exposing the ventrum.Then hold the mouse at approximately a 20-degree angle with the nose pointed to the floor. Insert a 25-gauge tuberculin syringe needle through the skin and abdominal wall of the caudal right quadrant of the abdomen. To avoid injection into the vasculature or the GI tract, pull back on the syringe plunger with minimal pressure to look for blood.Position the mouse in ventral recumbency on the absorbent pad over a heating pad before securely placing a rodent nose cone over the mouse's nose and muzzle. Place a small amount of pre-warmed ultrasound gel abaxial to the spine on either side of the anesthetized mouse between the last rib and pelvis. Put a small amount of gel on the ultrasound probe.Next, place the probe parallel to the vertebrae with the front of the probe on the cranial side. With a mouse in ventral recumbency, slowly scan the area for the kidney landmark. With the kidney in view, pull the probe caudal to find the ovary, which is a slightly hyperechoic oval to round structure within a very hyperechoic ovarian fat pad, bordered cranial-ventrally by the kidney and dorsal-laterally by the dorsal abdominal wall.Improve the image contrast by adjusting the signal gain using the slider at the bottom of the control screen. To improve the imaging of the kidney, apply pressure to the contra-lateral abdomen using a finger. Vary the pressure and angle from parallel to the spine to approximately 20 degrees ventral.Once the organ of interest is in view, capture a video by clicking on Save Clip or Start Recording. Once done, click Stop Recording to retain images at a preset number of frames. Hold the probe caudally until the ovary is in the most cranial aspect of the field of view.Vary the probe pressure and angle to view and image the uterus. Repeat video and frame collection for each organ of interest. Find the uterus running longitudinally along the dorsal abdominal wall with the lateral leg musculature in view.Monitor the tissue for a peristaltic motion to differentiate the intestinal loops from the uterine stationary horns. Place the mouse in dorsal recumbency to collect the images from a ventral approach. Ensure the eye lubrication is sufficient and the muzzle is secured in the nose cone.Apply a small amount of pre-warmed ultrasound gel to the ventral abdomen before applying the probe at the midline and cranial of the pubis to locate the bladder as a hypoechoic landmark. To find the uterine horns, pull the probe lateral to the bladder. Apply light digital pressure from either or both sides of the mouse.Once the uterine horns are in the field of view, hold the probe perpendicular to the mouse and scan both sides of the abdomen to capture transverse views of both horns. Rotate the probe to capture sagittal views. After the ultrasound, wipe the mouse with a paper towel before returning it to its cage to recover.Once the mouse is fully awake after two to five minutes, return the mouse to the animal room. The ultrasound image comparison of the mouse abdominal cavity before and after the saline injection showed the improved visualization of the abdominal organs, including the uterus and ovaries after the saline injection. A ventral approach was used to study the cross-sections of the uterine horns adjacent to the hypoechoic bladder.Immunohistochemistry with antibodies directed against Arid1a and Pten was used to study the adequate tumor suppressor protein loss accomplished in the uterine epithelial cells compared to the neighboring stromal tissue. Ultrasound images of a mouse uterus with early pathology changes were detected after three weeks of doxycycline treatment, showing the hypoechoic and dilated lumen of the uterus. In the gross pathology and histopathology study of the same uterus, a smaller but dilated thin-walled uterus and early endometrial hyperplasia with the preservation of the distinct lumen were observed respectively.The mice euthanized at later time points showed larger uterus with a smaller lumen and a thickened wall. When learning to use manual placement of probes, excessive pressure or rapid movement can make organ identification difficult. Use gentle, deliberate motions and focus on identifying the organs which are landmarks first.At study endpoint, mice are sacrificed and tissue is collected to confirm gene knockdown and characterize tissue changes. This technique allows investigators to monitor intraabdominal lesions over time. And in our case, we used it to monitor the uterine cancer progression in the iPad model, which we then confirmed by histopathology.

non-invasive-ultrasound-assessment-endometrial-cancer-progression

Research

JoVE Journal

Cancer Research

1.3K Görüntüleme.  Johns Hopkins University School of Medicine. Bu çalışma, manuel kılavuzlu ultrasonun, endometriyal kanserin indüklenmiş bir fare modelinde uterus patolojisinin gelişimini izlemek için çok uygun olduğunu göstermektedir. Ultrason görüntüleme noninvazivdir ve bu nedenle günler ve haftalar boyunca bireysel farelerin uzunlamasına çalışmalarına izin verir. Bu, bireysel varyasyonun yakalanmasına ve sonuçta çalışma için daha az hayvanın kullanılmasına izin verir.Bu yöntem, patolojik değişiklikleri ve neopl...