Name: partial bile duct ligation in the mouse: a controlled model of localized obstructive cholestasis
Uploaded: 2018-03-28T13:00:00
Description: Watch this Scientific Journal Video about Partial Bile Duct Ligation in the Mouse: A Controlled Model of Localized Obstructive Cholestasis at JoVE.com

Dr. Khan acknowledges grant support from NIH/NICHD K12HD052892, the Alpha-1 Foundation, the Hillman Foundation, and the Pittsburgh Liver Research Center. Dr. Michalopoulos acknowledges grant support from NIH/NIDDK P01DK096990. We appreciate the excellent and generous technical assistance of Anne Orr. We are also grateful to Dr. David A. Geller for providing access to the Transplant/Rodent Microsurgery lab.

All procedures were conducted in accordance with ethical guidelines of the Institutional Animal Care and Use Committee of the University of Pittsburgh.
1. Basic Techniques and Common Procedures
<ol>
	<li>Autoclave all microsurgical equipment before use.</li>
	<li>To avoid killing the mouse by anesthesia, check its breathing pattern carefully.</li>
	<li>Touch organs and intestines as little as possible to avoid damaging organs and causing bleeding. Press bleeding points with wet cotton swabs or gauze sponges.</li...

<ol>
	<li>Higgins, G. M., Anderson, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+pathology+of+the+liver+-+Restoration+of+the+liver+of+the+white+rat+following+partial+surgical+removal.">Experimental pathology of the liver - Restoration of the liver of the white rat following partial surgical removal.</a> Arch Pathol. 12, 186-202 (1931).</li><li>Cameron, G. R., Oakley, C. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ligation+of+the+common+bile+duct.">Ligation of the common bile duct.</a> The Journal of Pathology and Bacteriology. 35 (5), 769-798 (1932).</li><li>Kountouras, J., Billing, B. H., Scheuer, P. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Prolonged+bile+duct+obstruction:+a+new+experimental+model+for+cirrhosis+in+the+rat.">Prolonged bile duct obstruction: a new experimental model for cirrhosis in the rat.</a> Br J Exp Pathol. 65 (3), 305-311 (1984).</li><li>Michalopoulos, G. K., Barua, L., Bowen, W. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transdifferentiation+of+rat+hepatocytes+into+biliary+cells+after+bile+duct+ligation+and+toxic+biliary+injury.">Transdifferentiation of rat hepatocytes into biliary cells after bile duct ligation and toxic biliary injury.</a> Hepatology. 41 (3), 535-544 (2005).</li><li>Dipaola, 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=Identification+of+intramural+epithelial+networks+linked+to+peribiliary+glands+that+express+progenitor+cell+markers+and+proliferate+after+injury+in+mice.">Identification of intramural epithelial networks linked to peribiliary glands that express progenitor cell markers and proliferate after injury in mice.</a> Hepatology. 58 (4), 1486-1496 (2013).</li><li>Guyot, C., Combe, C., Desmouliere, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+common+bile+duct+ligation+in+rat:+A+relevant+in+vivo+model+to+study+the+role+of+mechanical+stress+on+cell+and+matrix+behaviour.">The common bile duct ligation in rat: A relevant in vivo model to study the role of mechanical stress on cell and matrix behaviour.</a> Histochem Cell Biol. 126 (4), 517-523 (2006).</li><li>Zhang, 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=Effect+of+bile+duct+ligation+on+bile+acid+composition+in+mouse+serum+and+liver.">Effect of bile duct ligation on bile acid composition in mouse serum and liver.</a> Liver Int. 32 (1), 58-69 (2012).</li><li>Tag, C. G., 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=Bile+duct+ligation+in+mice:+induction+of+inflammatory+liver+injury+and+fibrosis+by+obstructive+cholestasis.">Bile duct ligation in mice: induction of inflammatory liver injury and fibrosis by obstructive cholestasis.</a> J Vis Exp. (96), e52438 (2015).</li><li>Osawa, 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=Systemic+mediators+induce+fibrogenic+effects+in+normal+liver+after+partial+bile+duct+ligation.">Systemic mediators induce fibrogenic effects in normal liver after partial bile duct ligation.</a> Liver Int. 26 (9), 1138-1147 (2006).</li><li>Khan, Z., 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=Bile+Duct+Ligation+Induces+ATZ+Globule+Clearance+in+a+Mouse+Model+of+alpha-1+Antitrypsin+Deficiency.">Bile Duct Ligation Induces ATZ Globule Clearance in a Mouse Model of alpha-1 Antitrypsin Deficiency.</a> Gene Expr. 17 (2), 115-127 (2017).</li><li>Birkhead, H. A., Briggs, G. B., Saunders, L. Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Toxicology+of+cefazolin+in+animals.">Toxicology of cefazolin in animals.</a> J Infect Dis. 128, 378 (1973).</li><li>Kunst, M. W., Mattie, H., van Furth, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Antibacterial+efficacy+of+cefazolin+and+cephradine+in+neutropenic+mice.">Antibacterial efficacy of cefazolin and cephradine in neutropenic mice.</a> Infection. 7 (1), 30-34 (1979).</li><li>Traul, K. 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=Safety+studies+of+post-surgical+buprenorphine+therapy+for+mice.">Safety studies of post-surgical buprenorphine therapy for mice.</a> Lab Anim. 49 (2), 100-110 (2015).</li><li>Carlson, J. 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=Accumulation+of+PiZ+alpha+1-antitrypsin+causes+liver+damage+in+transgenic+mice.">Accumulation of PiZ alpha 1-antitrypsin causes liver damage in transgenic mice.</a> J Clin Invest. 83 (4), 1183-1190 (1989).</li><li>Sifers, R. N., Finegold, M. J., Woo, S. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Alpha-1-antitrypsin+deficiency:+accumulation+or+degradation+of+mutant+variants+within+the+hepatic+endoplasmic+reticulum.">Alpha-1-antitrypsin deficiency: accumulation or degradation of mutant variants within the hepatic endoplasmic reticulum.</a> Am J Respir Cell Mol Biol. 1 (5), 341-345 (1989).</li><li>Rudnick, D. A., Shikapwashya, O., Blomenkamp, K., Teckman, J. 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S., Alpini, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Activation+of+the+cholehepatic+shunt+as+a+potential+therapy+for+primary+sclerosing+cholangitis.">Activation of the cholehepatic shunt as a potential therapy for primary sclerosing cholangitis.</a> Hepatology. 49 (6), 1795-1797 (2009).</li><li>Gurpinar, E., 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=A+novel+sulindac+derivative+inhibits+lung+adenocarcinoma+cell+growth+through+suppression+of+Akt/mTOR+signaling+and+induction+of+autophagy.">A novel sulindac derivative inhibits lung adenocarcinoma cell growth through suppression of Akt/mTOR signaling and induction of autophagy.</a> Mol Cancer Ther. 12 (5), 663-674 (2013).</li><li>Alpini, G. G., Francis, H., Marzioni, M., Venter, J., Phinizy, J., LeSage, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Madame Curie Bioscience Database. , (2013).</li><li>Kirkland, J. G., 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=Reversible+surgical+model+of+biliary+inflammation+and+obstructive+jaundice+in+mice.">Reversible surgical model of biliary inflammation and obstructive jaundice in mice.</a> J Surg Res. 164 (2), 221-227 (2010).</li><li>Delhove, J. 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=Longitudinal+in+vivo+bioimaging+of+hepatocyte+transcription+factor+activity+following+cholestatic+liver+injury+in+mice.">Longitudinal in vivo bioimaging of hepatocyte transcription factor activity following cholestatic liver injury in mice.</a> Sci Rep. 7, 41874 (2017).</li><li>Li, C., 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=Homing+of+bone+marrow+mesenchymal+stem+cells+mediated+by+sphingosine+1-phosphate+contributes+to+liver+fibrosis.">Homing of bone marrow mesenchymal stem cells mediated by sphingosine 1-phosphate contributes to liver fibrosis.</a> J Hepatol. 50 (6), 1174-1183 (2009).</li><li>Xu, J., 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=Factors+released+from+cholestatic+rat+livers+possibly+involved+in+inducing+bone+marrow+hepatic+stem+cell+priming.">Factors released from cholestatic rat livers possibly involved in inducing bone marrow hepatic stem cell priming.</a> Stem Cells Dev. 17 (1), 143-155 (2008).</li><li>Sharma, 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=Propitious+role+of+bone+marrow-derived+mononuclear+cells+in+an+experimental+bile+duct+ligation+model:+potential+clinical+implications+in+obstructive+cholangiopathy.">Propitious role of bone marrow-derived mononuclear cells in an experimental bile duct ligation model: potential clinical implications in obstructive cholangiopathy.</a> Pediatr Surg Int. 29 (6), 623-632 (2013).</li></ol>

Although cBDL is the most common experimental technique for obstructive cholestasis, it can present multiple challenges in rodents. Depending on the number of post-operative days required to achieve an endpoint, the animals can experience significant morbidity and mortality as time progresses. The degree of tissue injury and biliary necrosis is further exacerbated when working with some transgenic mouse models of liver disease, which demonstrate abnormal liver parenchyma and function at baseline. Compared to cBDL, report...

Surgical models of acute liver injury and repair, such as partial hepatectomy or common bile duct ligation (cBDL), have been used for decades in rodents to study liver regeneration and pathobiology1,2. In cBDL, the common bile duct (which drains all bile produced in the liver) is ligated, resulting in obstructive cholestasis, inflammation, and fibrosis in the first 2-3 weeks following surgery, with eventual progression to cirrhosis3. Bile duct proliferation, transdifferentiation, and induction of resident liver progenitor cells are also features of cBDL4,5. Although cBDL elucidates specific mechanisms of obstructive cholangiopathy, which are well-characterized with reproducible outcomes in wild-type mice and in those strains with normal livers6,7,8, the procedure can be technically demanding in some transgenic mouse models of liver diseases, requiring more animals than anticipated due to the higher morbidity and mortality of biliary obstruction over time. This technique is also beneficial for studying transgenic mice with underlying liver disease, which might not otherwise tolerate the stress of prolonged time course experiments following cBDL.
pBDL can provide a reasonable alternative to help overcome some of these challenges. Initial pBDL studies in wild-type mice aimed to distinguish locally acting and systemic mediators of inflammation9. In pBDL, the main biliary confluence of the right and left hepatic bile ducts above the common bile duct is carefully visualized at the hilum, and only the left hepatic bile duct is ligated to generate a localized obstructive cholestasis. pBDL offers several advantages to cBDL. In pBDL, the unligated right lobe serves as an identically matched internal control for the ligated left lobe, subjected to the same systemic effects, such as nutritional status or circulating factors. Although systemic pro-inflammatory mediators were present, Osawa et al. observed less fibrosis and necrosis in the unligated lobes, but increased inflammatory cells and transforming growth factor-&#946; expression in the ligated lobe9.
More recently, pBDL has been redefined to study liver parenchymal effects of retained bile10. By using the same animal for internal control (unligated &#34;sham&#34;) and experimental groups, pBDL effectively halves the number of animals needed per experiment, which in turn is more cost-effective. The mice tolerate the effects of pBDL much better, so that the ideal time course experiment (&#8805;2 weeks) can be achieved in the ligated lobe. Most importantly, pBDL can distinguish direct intrahepatic effects of obstructive cholestasis and retained bile components in the ligated lobe, compared to the unligated control lobe. Overall, pBDL is an attractive method to study lobe-specific effects of localized cholestasis in the liver.

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			<td height="21" style="height:21px;width:343px;">Microscope-Leica Wild M650, 6&#8211;40&#215; magnification</td>
			<td style="width:336px;">Leica Microsystems</td>
			<td style="width:195px;">n/a</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Tec 3 isoflurane funnel fill vaporizer</td>
			<td>General Anesthetic Services</td>
			<td>n/a</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Stevens Tenotomy Scissors</td>
			<td>Accurate Surgical &#38; Scientific Instruments Corporation</td>
			<td>ASSI.9193</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Adson Forceps</td>
			<td>Fine science tools</td>
			<td>11006-12</td>
			<td></td>
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			<td height="21" style="height:21px;">Mosquito classic delicate hemostatic forceps</td>
			<td>Codman</td>
			<td>30-4472</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;">Micro-retractor</td>
			<td>Murdock; Roboz Surgical Instrument</td>
			<td>RS-6550</td>
			<td></td>
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			<td height="21" style="height:21px;">Nonwoven gauze sponges</td>
			<td>Fisherband</td>
			<td>870-PC-DBL</td>
			<td></td>
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			<td height="21" style="height:21px;">Puritan 6&#8221; Small Cotton Swab w/Wooden Handle</td>
			<td>Puritan Medical Products Company</td>
			<td>868-WCS</td>
			<td></td>
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			<td height="21" style="height:21px;">Dumont SS Forceps - Standard Tips/Straight/Inox/13.5cm</td>
			<td>Fine science tools</td>
			<td>11203-23</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;">Dumont SS Forceps - Standard Tips/Angled 45&#176;/Inox/13.5cm</td>
			<td>Fine science tools</td>
			<td>11203-25</td>
			<td></td>
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			<td height="21" style="height:21px;">Vannas Spring Scissors - 3mm Cutting Edge</td>
			<td>Fine science tools</td>
			<td>15000-00</td>
			<td></td>
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			<td height="21" style="height:21px;">Microneedle holder</td>
			<td>Aesculap Surgical Instruments</td>
			<td>FD231R</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;">Micro AROSuture, sterile 10-0 nylon suture, 70 &#181;m, TAP points</td>
			<td>AROSurgical Instruments Corporation</td>
			<td>T4A10N07</td>
			<td></td>
		</tr>
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			<td height="21" style="height:21px;">4-0 Vicryl</td>
			<td>Ethicon</td>
			<td>J662H</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;">5-ml Syringe</td>
			<td>BD</td>
			<td>309646</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Falcon tissue culture dish, 60 &#215; 15 mm</td>
			<td>Corning</td>
			<td>353002</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:343px;">Isoflurane, United States Pharmacopeia (USP) liquid for inhalation, 250 ml</td>
			<td>Piramal Healthcare</td>
			<td>NDC 66794-017-25</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Buprenex injectable (buprenorphine hydrochloride)</td>
			<td>Reckitt Benckiser Pharmaceuticals</td>
			<td>NDC 12496-0757-1</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Cefazolin for injection , USP</td>
			<td>APP Pharmaceuticals</td>
			<td>NDC 63323-238-61</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">PVP scrub solution (povidone&#8211;iodine 7.5%)</td>
			<td>Medline</td>
			<td>NDC 12496-0757-1</td>
			<td></td>
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			<td height="21" style="height:21px;">70% Ethanol</td>
			<td>Decon Labs</td>
			<td>2716</td>
			<td></td>
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			<td height="42" style="height:42px;width:343px;">Phosphate Buffered Saline Without Calcium or Magnesium</td>
			<td style="width:336px;">LONZA</td>
			<td style="width:195px;">17-516F</td>
			<td></td>
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			<td height="21" style="height:21px;width:343px;">Sirius Red</td>
			<td style="width:336px;">Sigma</td>
			<td style="width:195px;">365548</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:343px;">Hematolxylin</td>
			<td style="width:336px;">Fisher (Richard-Allan Scientific)</td>
			<td style="width:195px;">22-050-111 (7211)</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:343px;">Eosin</td>
			<td style="width:336px;">Anatech (Fisher)</td>
			<td style="width:195px;">832 (NC9686037)</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:343px;">Formalin</td>
			<td style="width:336px;">Fisher</td>
			<td style="width:195px;">SF100-20</td>
			<td></td>
		</tr>
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partial bile duct ligation in the mouse: a controlled model of localized obstructive cholestasis

In rodents, complete bile duct ligation (cBDL) of the common bile duct is an established surgical technique for studying obstructive cholestasis and bile duct proliferation. However, long-term experiments can lead to increased morbidity and mortality. In select mouse strains with underlying liver disease, meaningful comparisons can be made even with ligation of a single lobe of the liver, which can reduce animal losses and expenses. Here, we describe partial bile duct ligation (pBDL) in the mouse, in which only the left hepatic bile duct is ligated, causing biliary obstruction in the left lobe but not the remaining lobes. With careful microsurgical technique, pBDL experiments can be cost-effective, since the unligated lobe serves as an internal control to the ligated lobes, when subjected to the same conditions in the same animal. Unlike cBDL, a separate sham-operated control group is not necessary. pBDL is highly useful to directly compare localized versus systemic effects of cholestasis and other retained bile components. pBDL can also be repurposed as a novel method to investigate mechanisms related to medications and cell migration.

For this typical experiment, pBDL was performed in triplicate PiZ transgenic mice, which recapitulate the human liver disease found in alpha-1 antitrypsin (A1AT) deficiency. The PiZ mouse overexpresses a transgene consisting of multiple genomic fragments of DNA that contain the coding regions of the mutant human A1AT gene and promoter, together with introns and ~2 kilobases of upstream and downstream flanking regions14,15. Normal ...

Watch this Scientific Journal Video about Partial Bile Duct Ligation in the Mouse: A Controlled Model of Localized Obstructive Cholestasis at JoVE.com

Partial Bile Duct Ligation in the Mouse: A Controlled Model of Localized Obstructive Cholestasis

Here, we present partial bile duct ligation as a surgical model of liver injury and regeneration in rodents.

In rodents, complete bile duct ligation (cBDL) of the common bile duct is an established surgical technique for studying obstructive cholestasis and bile ...

The overall goal of this surgical intervention is to observe the effects of localized obstructive cholestasis. This method can help answer key questions in the hepatology field about how to identify mechanisms of liver regeneration. The main advantage of this technique is that meaningful comparisons can be made between the ligated experimental lobe and the unligated control lobe within the same animal.Demonstrating the procedure today will be Dr.Toshimasa Nakao, a post-doc from the transplant surgery laboratory. After confirming a lack of response to toe pinch in an anesthetized eight to 12-week-old C57 black 6 mouse, shave the abdominal region with hair clippers and place the animal under a surgical microscope. Secure the limbs to the table and use sterile swabs to sequentially disinfect the surgical area with 0.3 to 0.5 milliliters of povidone-iodine and 70%ethanol.Next, make a midline incision from the xiphoid to the pubis and use a micro retractor to open the abdominal cavity. Use mosquito forceps to retract the xiphoid toward the head of the mouse and use soft clay to hold the forceps and xiphoid in this position. Hydrate the abdominal cavity with one to two milliliters of sterilized 37 degrees Celsius PBS and use wet cotton swabs to gently move the small intestine toward the head of the mouse.Cut the ligaments between the small intestine and the retroperitoneum to avoid damaging the tissue and wrap the small and large intestines with a wet non-woven gauze sponge. Place the bowel tissue on the caudal side of the abdominal cavity and use a small wet non-woven gauze sponge to gently retract the entire liver toward the xiphoid to expose the hepatic hilum and the hepatoduodenal ligament. Locate the hepatic hilum, the common bile duct on the ventral side of the portal vein, and the left and right hepatic bile ducts which drain into the main biliary confluence at the hepatic hilum.Using a 10-0 nylon suture and the blunt side of a suture needle, ligate the left hepatic bile duct lobe. When the suture has been placed, return the small and large intestines to their anatomical location and confirm a lack of intestinal torsion or intraabdominal bleeding. Then remove the mosquito forceps, separately close the muscle and skin layers with continuous 4-0 Vicryl sutures, and place the mouse under a heat lamp with monitoring until full recovery.Comparison of the serum liver biochemistries between partial bile duct ligated mice and complete bile duct ligated mice transgenic for insoluble liver protein aggregation reveals minimal liver injury or cholestasis in the partial bile duct ligated mice and severe injury and enzyme buildup in the complete bile duct ligated transgenic animals. As mice subjected to partial bile duct ligation developed lobe-specific obstructive cholestasis over time, the ligated lobe appears slightly paler in color 14 days after surgery compared to the unligated lobes which maintained their normal dark reddish brown color. The ligated lobe also demonstrates an increase in bile duct proliferation and fibrosis compared to the unligated lobes in response to ligation-induced localized cholestasis.Once mastered, this technique can be completed in 10 to 12 minutes if it is performed properly. Following this procedure, other methods like lineage tracing can be used to answer additional questions about stem cell homing to sites of liver injury and regeneration. After watching this video, you should have a good understanding of how to make meaningful comparisons using partial bile duct ligation to identify local versus systemic effects.

Research

JoVE Journal

Biology

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