Name: murine prostate micro-dissection and surgical castration
Uploaded: 2016-05-11T15:00:00
Description: Watch this Scientific Journal Video about Murine Prostate Micro-dissection and Surgical Castration at JoVE.com

Nogle af tallene i dette manuskript var generøst leveres af laboratoriet Bart O. Williams. Forfatterne er understøttet af National Cancer Institute giver U54CA143803, CA163124, CA093900 OG CA143055.

Denne protokol opfylder og følger de retningslinjer fastsat af Johns Hopkins University Institutional Animal Care og brug Udvalg. 1. Micro-dissekere en mus Prostata <ol><li> Aflive musen ved kuldioxid kvælning eller et andet godkendt metode i overensstemmelse med de institutionelle dyr pleje og brug retningslinjer. </li><li> Pin musen i liggende position til en dissektion bord ved at sætte en stift gennem hver af de fire poter. </li><li> Spray maven med 70% ethanol. </li><li> Hold huden på underlivet med p...

<ol>
	<li>Siegel, R. L., Miller, K. D., Jemal, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cancer+statistics.">Cancer statistics.</a> CA Cancer J Clin. 65 (1), 5-29 (2015).</li><li>Roehrborn, C. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pathology+of+benign+prostatic+hyperplasia.">Pathology of benign prostatic hyperplasia.</a> Int J Impot Res. 20 (suppl 3), 11-18 (2008).</li><li>Valkenburg, K. C., Williams, B. O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mouse+models+of+prostate+cancer.">Mouse models of prostate cancer.</a> Prostate Cancer. , 895238 (2011).</li><li>Shappell, S. 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S., Labrie, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Unique+features+of+the+basal+cells+of+human+prostate+epithelium.">Unique features of the basal cells of human prostate epithelium.</a> Microsc Res Tech. 51 (5), 436-446 (2000).</li><li>Prins, G. S., Huang, L., Birch, L., Pu, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+role+of+estrogens+in+normal+and+abnormal+development+of+the+prostate+gland.">The role of estrogens in normal and abnormal development of the prostate gland.</a> Ann N Y Acad Sci. 1089, 1-13 (2006).</li><li>Valkenburg, K. 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=Activation+of+Wnt/beta-catenin+signaling+in+a+subpopulation+of+murine+prostate+luminal+epithelial+cells+induces+high+grade+prostate+intraepithelial+neoplasia.">Activation of Wnt/beta-catenin signaling in a subpopulation of murine prostate luminal epithelial cells induces high grade prostate intraepithelial neoplasia.</a> Prostate. 74 (15), 1506-1520 (2014).</li><li>Lukacs, R. U., Goldstein, A. S., Lawson, D. A., Cheng, D., Witte, O. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Isolation,+cultivation+and+characterization+of+adult+murine+prostate+stem+cells.">Isolation, cultivation and characterization of adult murine prostate stem cells.</a> Nat Protoc. 5 (4), 702-713 (2010).</li><li>Michiel Sedelaar, J. P., Dalrymple, S. S., Isaacs, J. 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Prostata mikro-dissektion tillader lobe-specifikke eksperimenter og analyse af muse prostata (figur 1). I gensplejsede musemodeller, kan fænotyper ses i en lap, som ikke ses i andre. Også for histologisk analyse, denne protokol sikrer, at den maksimale mængde af ren prostata væv kan sektioneret og farves, uden andre urogenital tarmkanalen væv til stede i afsnittet. Endelig for encellede eksperimenter, denne protokol giver mulighed for isolation af næsten rene prostata celler. Epitelcelle berigelse...

Prostata er den mest almindelige sted for cancer i mænd i USA. Næsten 220.000 mænd hvert år vil blive diagnosticeret med prostatakræft, og ca. 27.000 mænd vil bukke under for deres sygdom en. Mænd har en 1 i 7 levetid risiko for prostatakræft diagnose i USA en. Godartet prostatahyperplasi (BPH), en aldersrelateret noncancerous forstørrelse af prostata, er også en udbredt tilstand, som påvirker mere end 80% af mænd over 80 2. Som sådan prostata er fokus for betydelig forskning. Musemodeller har vidt blevet anvendt til at undersøge sygdomme i prostata 3. Samlet set musen prostata er en fremragende repræsentant for den humane kirtel 4, men der er ligheder og forskelle mellem mus og menneske prostata anatomi og fysiologi 5. Hos begge arter er prostata placeret i bunden af ​​blæren, der omgiver urinrøret. Den humane prostatakirtel er en enkelt lovære, opdelt i fire zoner: central, overgang, perifer, og forreste fibromuskulær stroma. I modsætning hertil muse prostata består af tre parrede lapper på forskellige steder omkring urinrøret: anterior, ventrale, og dorsolaterale. På et cellulært niveau, den vigtigste forskel er, at den basale celle til luminal celle-forhold i human er ca. 1: 1, men det er 1: 4 i mus 6. Det murine stroma er også anderledes i mus i forhold til mennesker - mennesker har mere omfattende glat muskel, mens den muskel lag er meget tyndere i murine prostata. Den korrekte identifikation, dissektion, og håndtering af musen prostata er afgørende for mus prostata forskning. Hormonniveau drastisk påvirke udviklingen og homeostase af prostata hos både mennesker og mus. Mens østrogen synes at spille en rolle i prostata udvikling 7, det vigtigste hormon i prostata er testosteron. Testosteron er afgørende for glandular udvikling end vedligeholdelse. Efter enten fysisk eller kemisk kastration, ca. 90% af de luminale celler i den voksne prostata apoptose, og kirtlen skrumper. Hvis testosteron genindføres til en person under kastration betingelser, prostata er i stand til at regenerere sig selv til fuld kapacitet. Testosteron synes også at drive prostatacancer, hvilket er grunden til androgen deprivation terapi er en almindeligt anvendt terapeutisk strategi. Men mange prostatakræft bliver resistente over for androgen deprivation. Derudover mænd undergår androgen deprivation terapi til behandling af prostatacancer underkastes cyklusser af kastrerede og regenererede betingelser. Hos mus, kirurgisk kastration, sammen med hormonal regenerering af prostata ved at genindføre testosteron, er et vigtigt redskab til at studere kastration modstand og virkningerne af testosteron cykle på prostata. I denne artikel, vil vi diskutere og demonstrere de rigtige teknikker ved at søge efter disse datae og mikro-dissekere en mus prostata, såvel som til kirurgisk kastrere en mus.

<table border="0" cellpadding="0" cellspacing="0" width="741">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:244px;">Surgical tools</td>
			<td style="width:145px;">Roboz</td>
			<td style="width:113px;">Various</td>
			<td style="width:239px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Formaldehyde</td>
			<td>Sigma</td>
			<td>F8775</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;">OCT medium</td>
			<td>VWR</td>
			<td>25608-930</td>
			<td style="width:239px;">Must be placed on dry ice to freeze, and stored at -80C</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">PBS</td>
			<td>Life Technologies</td>
			<td align="right">10010</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Isoflurane</td>
			<td>Johns Hopkins</td>
			<td></td>
			<td>Also available from vendors online</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Cautery Pen</td>
			<td>Medline</td>
			<td>ESCT002</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Silk suture</td>
			<td>AD Surgical</td>
			<td>M-S330R19</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Surgical Wound Clips</td>
			<td>Roboz</td>
			<td>RS-9265</td>
			<td></td>
		</tr>
	</tbody>
</table>

murine prostate micro-dissection and surgical castration

Mouse models are used extensively to study prostate cancer and other diseases. The mouse is an excellent model with which to study the prostate and has been used as a surrogate for discoveries in human prostate development and disease. Prostate micro-dissection allows consistent study of lobe-specific prostate anatomy, histology, and cellular characteristics in the absence of contamination of other tissues. Testosterone affects prostate development and disease. Androgen deprivation therapy is a common treatment for prostate cancer patients, but many prostate tumors become castration-resistant. Surgical castration of mouse models allows for the study of castration resistance and other facets of hormonal biology on the prostate. This procedure can be coupled with testosterone reintroduction, or hormonal regeneration of the prostate, a powerful method to study stem cell lineages in the prostate. Together, prostate micro-dissection and surgical castration opens up a multitude of opportunities for robust and consistent research of prostate development and disease. This manuscript describes the protocols for prostate micro-dissection and surgical castration in the laboratory mouse.

Alle seks prostata lapper blev fjernet fra en mus via prostata mikro-dissektion (figur 1). Den komplette urogenitalkanalen (UGT) er sammensat af alle prostata lapper, blære, seminalis vesikler og urinrøret (figur 1A). Sædlederen tillægger urinrøret, men er unødvendigt for prostata mikro-dissektion, og kan således frigøres før fjernelse af UGT ved at skære urinrøret (figur 1b-1c). Urogenitalkanalen vil forblive sammen når fjer...

Watch this Scientific Journal Video about Murine Prostate Micro-dissection and Surgical Castration at JoVE.com

Murine Prostate Micro-dissection and Surgical Castration

Dette håndskrift beskriver protokoller for prostata mikro-dissektion og kirurgisk kastration i laboratoriet mus. Vi skildrer også repræsentative resultater produceret af disse protokoller. Endelig diskuterer vi fordele og udnyttelsen af ​​disse protokoller.

Murint Prostata Micro-dissektion og kirurgisk kastration

Mouse models are used extensively to study prostate cancer and other diseases. The mouse is an excellent model with which to study the prostate and has ...

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