The work was funded by a CHIR operating grant (DJB).

Aşağıdaki deneysel protokol kurumun hayvan bakımı komitesi tarafından onaylandı. Dişi sıçanlarda 1. NEMO <ol><li> Hazırlık <ol><li> Yaklaşık 200 g 3 altında% izofluran anestezisi ağırlığında dişi Sprague-Dawley sıçan yerleştirin hayvan tartmak ve tıraş ve bir iyot cerrahi yıkama kullanılarak prepubik parkı 3 kez dezenfekte. </li><li> aşağı bir ısıtma yastığı, şerit arka bacak ve kuyruk üzerinde ve ayrıca yumuşak, kaymasını önlemek üzere aşağı bantlanmış 20-gauge anjiyokateterinden ile mesaneye kateter. </li></ol></li><li> teşrih <ol><li> üretra deliğine doğru aşikar pubis eklemi bir 8 mm uzunlamasına bir deri yarığı gerçekleştirmek için bir neşter kullanarak. Daha sonra yara kapanmasını kolaylaştırmak için her iki tarafta derisini kaldırın. </li><li> uzunlamasına ince makasla yayarak stentli üretra tanımlamak için açık açık teşrih. Açıklığı artırmak için ince kendini tutan deri kancaları yerleştirinüretra. </li><li> Gerçekleştirmek bazı yumuşak uzunlamasına üretra ve vajina arasında düzlemi geliştirmek için kavisli bir makas üretraya dorsal kullanarak yayılma. Sonra, açık açık üretra arkasında 4-0 ipek sütür geçmek ve bir cerrahın düğüm çift atış hazırlayın. </li></ol></li><li> Kısmi üretral tıkanıklığı <ol><li> , Angiocatheter kaldırma üretra 0.9 mm&#39;lik bir metal çubuk paralel yerleştirin ve ikinci hala kolayca sonra kayarak ve böylece daha sonra, üretra ve metal çubuk çevresinde bir düğüm sıkın. </li><li> 3 daha çubuk çıkarmadan önce nazik gerginliği kullanarak atış ile düğüm sabitleyin. sütür uzun yaklaşık 3 mm veya tıkanıklık sonradan serbest bırakma deney parçası ise 4 mm biter kesin. </li></ol></li><li> Yara kapatma <ol><li> emilebilen örgülü sütür 4-0 gömülü tek dikişle paraüretral bezleri yaklaşmaktadır. </li><li> Aynı emilebilen SUT kullanarak gömülü yatay yatak dikiş ile cilt kapatınobilyaları. Not: Bir cilt-delici dikiş için emilebilir dikiş kullanmak alışılmamış görünse de, bu tür yara enfeksiyonları gibi komplikasyonlar karşılaşmak yoktu ve bu dikiş alınması önlenebilir. </li></ol></li><li> bakım sonrası <ol><li> Enjekte buprenorfin 0.1 mg / kg deri altından, bir kurtarma kafeste sıçan yerleştirmeden önce. </li><li> Tek Housed fareye ilk 24 saat bazı yumuşak kurtarma diyet sunun. Bu süre geçtikten sonra, prosedür önce olduğu gibi aynı ikili tekrar ev hayvanların daha iyi olması ve aynı bakım grubuna aittir sağladı. </li><li> mesane büyüklüğü ve genel iyi sağlık için günlük sıçanlar kontrol; en azından haftada kendi ağırlığı kaydedin. </li></ol></li></ol> Kadın Farelerde 2. NEMO <ol><li> Hazırlık <ol><li> yaklaşık 18 g ağırlığında dişi C57BL / 6 fare ve aşama 1.1.1 detaylı devam edin. </li><li> 24 ayar ang kullanmak dışında adım 1.1.2 deki gibi detaylı Devamiocatheter. </li></ol></li><li> teşrih <ol><li> üretra deliğine doğru aşikar pubis eklemi 6 mm uzunlamasına cilt insizyonu gerçekleştirmek için bir neşter kullanarak. </li><li> uzunlamasına ince makasla yayarak stentli üretra tanımlamak için açık açık teşrih. benzer künt diseksiyon ile üretra üzerinde ventral çalışan nöro-vasküler demet kaldırın. </li><li> Gerçekleştirmek bazı yumuşak uzunlamasına üretra ve vajina arasında düzlemi geliştirmek için kavisli bir makas üretraya dorsal kullanarak yayılma. Sonra, açık açık üretra arkasında 5-0 emilemeyen örgülü sütür geçmek ve bir cerrahın düğüm çift atış hazırlayın. </li></ol></li><li> Kısmi üretral tıkanıklığı <ol><li> 1 ml şırınga ile angiocatheter bağlayın ve mesane içine, normal tuzlu su, 0.1 mL enjekte edilir. </li><li> üretraya 26 numara bir kanül paralel yerleştirin. angioca geri çekerken yavaşça üretra ve kanül etrafında düğüm sıkıninci. </li><li> 3 daha kanül çıkarmadan önce nazik gerginliği kullanarak atış ile düğüm sabitleyin. </li><li> hayvanın mesane hafif basınç uygulanması üzerine meatusta görünen idrar için test; idrar ifade edemeyen eğer kravat gevşetin. Aksi takdirde, ameliyat dikiş ipliği, yaklaşık 3 mm long uçları kesilmiş. </li></ol></li><li> Yara kapatma <ol><li> 5-0 örgülü emilebilir sütür kullanarak gömülü yatay yatak dikiş ile cilt kapatın. </li></ol></li><li> bakım sonrası <ol><li> Kurtarma kafes içinde fare yerleştirmeden önce 0.1 mg / kg deri altından buprenorfın enjekte edilir. </li><li> Tek Housed fareye ilk 48 saat bazı yumuşak kurtarma diyet sunun. Bu dönemden sonra, işlem öncesi aynı grupta yine ev hayvanları onlar iyi yapıyor ve aynı tedavi grubuna ait koşuluyla. NOT: Aşırı engellendiği ele zaman geçersiz yoktur ve n içinde ölecektir işlemden sonra ele gelen şişmiş mesane 24 saat ile Farelerengelleyici ligature çıkarılması için anestezi altında geri alınmıştır 2 gün sürece 1 - dahili. </li><li> mesane büyüklüğü ve genel iyi sağlık için günlük fareler kontrol; en azından haftada kendi ağırlığı kaydedin. </li></ol></li></ol>

<ol>
	<li>Andersson, K. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Storage+and+voiding+symptoms:+pathophysiologic+aspects.">Storage and voiding symptoms: pathophysiologic aspects.</a> Urology. 62 (5), 3-10 (2003).</li><li>Boyle, P., 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=The+prevalence+of+lower+urinary+tract+symptoms+in+men+and+women+in+four+centres.+The+UrEpik+study.">The prevalence of lower urinary tract symptoms in men and women in four centres. The UrEpik study.</a> BJU Int. 92 (4), 409-414 (2003).</li><li>Coyne, K. S., Wein, A., Nicholson, S., Kvasz, M., Chen, C. I., Milsom, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Economic+burden+of+urgency+urinary+incontinence+in+the+United+States:+a+systematic+review.">Economic burden of urgency urinary incontinence in the United States: a systematic review.</a> J Manag Care Pharm. 20 (2), 130-140 (2014).</li><li>Buttyan, R., Chen, M. W., Levin, R. M. <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+bladder+outlet+obstruction+and+molecular+insights+into+the+basis+for+the+development+of+bladder+dysfunction.">Animal models of bladder outlet obstruction and molecular insights into the basis for the development of bladder dysfunction.</a> Eur. Urol. 32 (Suppl 1), 32-39 (1997).</li><li>Mattiasson, A., Uvelius, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Changes+in+contractile+properties+in+hypertrophic+rat+urinary+bladder.">Changes in contractile properties in hypertrophic rat urinary bladder.</a> J Urol. 128 (6), 1340-1342 (1982).</li><li>Chang, 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=Alteration+of+the+PKC-mediated+signaling+pathway+for+smooth+muscle+contraction+in+obstruction-induced+hypertrophy+of+the+urinary+bladder.">Alteration of the PKC-mediated signaling pathway for smooth muscle contraction in obstruction-induced hypertrophy of the urinary bladder.</a> Lab Invest. 89 (7), 823-832 (2009).</li><li>Moore, J. A., Brading, A. 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The authors have nothing to disclose.

Hayvan ölümleri Birçok raporda 10, 11, 13% 15 veya daha fazlasında proksimal üretra de pBOO geleneksel modelin önemli sorunlardan biridir. NEMO dişi sıçanlarda uygulandığında çok az ölüm oranı gibi görünmektedir. Farelerde proksimal üretral tıkanma teknik olarak daha zor sıçanlarda daha ve böylece daha meyilli komplikasyonlar etmektir. Bizim orta üretral yaklaşım için yer tutucu olara...

Kısmi mesane çıkışı tıkanması (pBOO) yüksek oranda bulunur ve şiddetli mesane fonksiyon bozukluğu 1 neden olabilir; tayfı, iyi huylu prostat hiperplazisi için alınan üretral striktür fazla posterior üretral valfleri veya hipospadyas gibi konjenital malformasyonlar, arasında değişmektedir. Erkekler altmış yıldan fazla% 30 ve 2 eski etkileyen ikincisi. Büyük hasta yükü ve pBOO taşıdığı büyük sağlık masrafları artan dış akış direnci 3 cevaben okuyan mesane yeniden koymak önemli araştırma çaba garanti. 2006 yılından itibaren 2015 yılına kadar 220 üzerinde PubMed makaleleri idrar kesesi üzerinde pBOO etkisiyle ilgili yayımlandı endekslendi. PBOO için hayvan modelleri, örneğin sıçan 5, tavşan 6, domuz 7 ve Mous gibi çeşitli türler 4 üzere tasarımlanmış olmakla berabere 8, 9, ancak proksimal üretra kısmi engelleme uygulanan dişi fareler burada en yaygın olarak kullanılan bir hayvan modelidir; Mesane boynuna hayvanların karın erişim, mesanenin exteriorizing ve diseksiyon bu geleneksel proksimal üretral tıkanıklığı tekniği ile kaçınılmazdır. Tıkanıklığı ve hayvan ölüm derecesinde değişkenlik sadece 11 Bu prosedür 10 ile ilişkili endişelerden bazılarını; Biz sahte ameliyat hayvanlarda, idrar yoluna diseksiyon mesane boynu 12, sinir liflerinin kaybı ile ilişkili fizyolojik değişikliklere neden olduğunu göstermiştir. Bu bulgu, bir kadın kemirgenlerde yakın üretra erişmek kapsar en yaygın olarak kullanılan pBOO hayvan modeli, etki, mesane bağlantılı bir yapısal ve işlevsel değişiklikler ile denervasyon hasara yol açtığını, göstermektedirsahte ve tıkalı hayvanlar. Bu nedenle, alternatif bir yaklaşım kaçınarak denervasyon yaralanması ihtiyaç vardı. Bizim laboratuvar geliştirilmiş ve sham ameliyat edilen hayvanlar ameliyat edilmemiş kontrol hayvanları ayırt iken, bu, organ kütlesini ve rezidüel idrar artış olarak mesanede beklenen tıkanıklığı ile uyumlu değişiklikler indüklenmesinde etkili olan bir sinir-koruyucu Orta üretral tıkanıklık (NEMO) yaklaşımı değerlendirildi . Bu diseksiyon seviyesine yakınlığında uzandığına olarak da, çizgili üretral sfinkter bakir kalmıştır. Bundan başka, mesane kütlesi içinde tıkanma ile uyarılmış değişkenlik geleneksel Üretra tıkanıklığında önemli ölçüde daha düşük olduğu ve hayvan ölüm olmamıştır. Bugüne kadar tarif edilmiş fareler için tüm pBOO modelleri 50 yaklaşık% mortalite ile ilişkili ise de başarılı bir tıkanmış hayvanlarda% 10&#39;dan az mortalite ile dişi farelerde NEMO uygulanır. Farelerde pBOO bağlamında mesane yenileme incelenmesi applicabil yararlanacaktırTransgenik modifikasyonların bütün spektrum üretebilir. Kadın kemirgenlerde orta idrar yoluna Kesme geleneksel yakın obstrüksiyon modelinde gözlenen idrar kesesi, istenmeyen ve karıştırıcı yapısal veya fonksiyonel değişiklikler indüklemez. Bununla birlikte, orta üretral düzeyinde bir kısmi engelleme indükleyici hala mesane hipertrofi indükler ve pBOO için bir hayvan modeli beklendiği gibi, artık idrar artmıştır. Önemli olarak, farelerde NEMO performans büyük kemirgenlerde neredeyse kullanılamaz transgenik yöntemlere pBOO mesane biçimlenme soruşturma açar.

<table border="0" cellpadding="0" cellspacing="0" width="1064">
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		<tr height="40">
			<td height="40" style="height:40px;width:216px;">Female rat&#160;</td>
			<td style="width:123px;">Charles-River Canada</td>
			<td style="width:559px;">Sprague Dawley Rat (Crl:SD); Strain Code: 400</td>
			<td style="width:167px;">about 200g</td>
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			<td style="width:559px;">Isoflorane; product code CP0406V2</td>
			<td style="width:167px;">&#160;250ml bottle</td>
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			<td height="40" style="height:40px;width:216px;">Disposable scalpell</td>
			<td style="width:123px;">Ted Pella, Inc.</td>
			<td style="width:559px;">#15 Sterile, Stainless Steel Scalpels; catalogue number 549-9-15S</td>
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			<td style="width:559px;">Stainless Steel - AISI 316L Wire Diameter:0.9mm; order code 911-684-65</td>
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			<td style="width:123px;">goodfellow.com</td>
			<td style="width:559px;">Stainless Steel - AISI 316L Wire Diameter:0.9mm; order code 911-684-65</td>
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			<td style="width:123px;">esutures.com</td>
			<td style="width:559px;">5-0 Vicryl undyed 27&#34; RB-1 taper; product number J213H</td>
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		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Normal Saline</td>
			<td style="width:123px;">Baxter</td>
			<td style="width:559px;">0.9% Sodium Chloride Injection, USP, 100 mL MINI-BAG; product code 2B0043</td>
			<td style="width:167px;"></td>
		</tr>
	</tbody>
</table>

nerve-sparing mid-urethral obstruction (nemo) in female small rodents

Kısmi mesane çıkışı tıkanması (pBOO), oldukça yaygın olarak görülen önemli hasta yükünü ve muazzam sağlık maliyetlerini de beraberinde getirmektedir. pBOO içinde mesane yenileme araştırmak için en yaygın hayvan modeli proksimal üretra kısmi tıkanıklık geçiren kadın kemirgenler. tıkanıklığı ve hayvan ölüm derecesindeki Değişkenlik proksimal tıkanıklığı ile önemli sorunlardır. Ayrıca, proksimal üretra ve mesane boynunda kesme mesane inervasyonu tehlikeye atar. Biz geleneksel modelin dezavantajları kaçınarak pBOO için sinir koruyucu orta üretral tıkanma (Nemo) model geliştirdi. Biz laparotomi yanı sıra bu alanda nasıl tedavi edilmesi gerektiği obviated pubis eklemi, sadece alt üretra yaklaştı; Ayrıca, çizgili üretral sfinkter bakir kalmıştır. Bu dişi Sprague-Dawley farelerinde (12 engel, 6 sahte hayvanlar) ve aynı zamanda dişi C57 / BL6 fareleri NEMO gerçekleştirildi (20 tıkanıklığıs, 18 sahte hayvanlar). İki hafta sonra, mesane fonksiyonu, mesane kitlesi ve vücut kitle değerlendirdi. Biz obstructed- veya yapay olarak ameliyat dişi sıçanların arasında hiçbir ölümlerinin vardı; Geleneksel yakın pBOO-yöntemi için tarif edildiği gibi, biz idrar yoluna dikiş bağlı ve bir geçici 0.9 mm delikli metal çubuk yerleştirilir. NEMO iki hafta sonra, mesane kütlesinde% 85&#39;lik bir artışa neden ortalama rezidüel idrar hacmi, sadece 0.03 mL iken plasebo hayvanlarda kısmen tıkanmış sıçanlarda 0.4 ml idi. farelerde, dikiş bağlama biz üretra boyunca yerleştirilmiş kanüller 3 boyutları test edilmiştir. Bir 27-gauge kanül kullanılarak% 50 hayvan üzerinde mortalite sonuçlandığını tespit; artan mesane kütlesinde istenen yanıtı sağlamak değil, 25-gauge bir kanül yerleştirilmesi; 26 numara bir kanül kullanılarak en az bir hayvan ölüm (1/8) henüz mesane kitlesinde önemli bir 2-kat bir artış ile olumlu sonuçlar vermiştir.

Dişi sıçanlarda sinir-koruyucu Orta üretral tıkanması (NEMO) ölümlülük Şu ana kadar 40&#39;ın üzerinde dişi fareler üzerinde bir orta üretral tıkanma gerçekleştirilen ve hiçbir ölümlerinin vardı. mesane kütlesi <p class="jove_content" fo:keep-toge...

Watch this Scientific Journal Video about Nerve-sparing Mid-urethral Obstruction NeMO in Female Small Rodents at JoVE.com

Nerve-sparing Mid-urethral Obstruction NeMO in Female Small Rodents

kemirgenlerde kısmi mesane çıkışı tıkanıklığı geleneksel modelleme hayvan ölüm ile doludur. Proksimal üretra ve mesane boynuna diseksiyon A denervasyon yaralanması büyük sorun oluşturmaktadır. Biz geleneksel modelin eksiklikleri kaçınarak, gelişmiş ve güvenli ve güvenilir bir orta üretral tıkanma modelini değerlendirdi.

Kadın Küçük Kemirgenler Sinir koruyucu Orta üretral Tıkanıklığı (Nemo)

Partial bladder outlet obstruction (pBOO) has a high prevalence, causes significant patient burden, and immense health care costs. The most common animal ...

nerve-sparing-mid-urethral-obstruction-nemo-in-female-small-rodents

Research

JoVE Journal

Medicine

Nerve-sparing Mid-urethral Obstruction (NeMO) in Female Small Rodents

9.5K Görüntüleme.  The Hospital for Sick Children.  kemirgenlerde kısmi mesane çıkışı tıkanıklığı geleneksel modelleme hayvan ölüm ile doludur. Proksimal üretra ve mesane boynuna diseksiyon A denervasyon yaralanması büyük sorun oluşturmaktadır. Biz geleneksel modelin eksiklikleri kaçınarak, gelişmiş ve güvenli ve güvenilir bir orta üretral tıkanma modelini değerlendirdi. 

Video: Nerve-sparing Mid-urethral Obstruction NeMO in Female Small Rodents

Protocol for Long Duration Whole Body Hyperthermia in Mice

Hyperthermia is a general term used to define the increase in core body temperature above normal. It is often used to describe the increased core body temperature that is observed during fever. The use of hyperthermia as an adjuvant has emerged as a promising procedure for tumor regression in the field of cancer biology. For this purpose, the most important requirement is to have reliable and uniform heating protocols. We have developed a protocol for hyperthermia (whole body) in mice. In this protocol, animals are exposed to cycles of hyperthermia for 90 min followed by a rest period of 15 min. During this period mice have easy access to food and water. High body temperature spikes in the mice during first few hyperthermia exposure cycles are prevented by immobilizing the animal. Additionally, normal saline is administered in first few cycles to minimize the effects of dehydration. This protocol can simulate fever like conditions in mice up to 12-24 hr. We have used 8-12 weeks old BALB/Cj female mice to demonstrate the protocol.

This paper describes a protocol for whole body hyperthermia in mice that can stimulate fever like conditions up to 12-24 hr.

Farelerde Uzun Süre Whole Body Hipertermi için protokol

Hyperthermia is a general term used to define the increase in core body temperature above normal. It is often used to describe the increased core body ...

Tıp

The Use of Cystometry in Small Rodents: A Study of Bladder Chemosensation

The lower urinary tract (LUT) functions as a dynamic reservoir that is able to store urine and to efficiently expel it at a convenient time. While storing urine, however, the bladder is exposed for prolonged periods to waste products. By acting as a tight barrier, the epithelial lining of the LUT, the urothelium, avoids re-absorption of harmful substances. Moreover, noxious chemicals stimulate the bladder's nociceptive innervation and initiate voiding contractions that expel the bladder's contents. Interestingly, the bladder's sensitivity to noxious chemicals has been used successfully in clinical practice, by intravesically infusing the TRPV1 agonist capsaicin to treat neurogenic bladder overactivity1. This underscores the advantage of viewing the bladder as a chemosensory organ and prompts for further clinical research. However, ethical issues severely limit the possibilities to perform, in human subjects, the invasive measurements that are necessary to unravel the molecular bases of LUT clinical pharmacology. A way to overcome this limitation is the use of several animal models2. Here we describe the implementation of cystometry in mice and rats, a technique that allows measuring the intravesical pressure in conditions of controlled bladder perfusion.
	After laparotomy, a catheter is implanted in the bladder dome and tunneled subcutaneously to the interscapular region. Then the bladder can be filled at a controlled rate, while the urethra is left free for micturition. During the repetitive cycles of filling and voiding, intravesical pressure can be measured via the implanted catheter. As such, the pressure changes can be quantified and analyzed. Moreover, simultaneous measurement of the voided volume allows distinguishing voiding contractions from non-voiding contractions3.
	Importantly, due to the differences in micturition control between rodents and humans, cystometric measurements in these animals have only limited translational value4. Nevertheless, they are quite instrumental in the study of bladder pathophysiology and pharmacology in experimental pre-clinical settings. Recent research using this technique has revealed the key role of novel molecular players in the mechano- and chemo-sensory properties of the bladder.

Cystometry is an efficient technique to measure bladder function of small animals in vivo. The bladder is continuously infused at rates controlled through an intravesical catheter, whereas the urethra is left free for micturition. This allows for repetitive filling and emptying of the bladder, while intravesical pressure and voided volume are recorded.

Küçük Kemirgenler sistometri ve Kullanımı: Mesane Chemosensation Çalışması

	The lower urinary tract (LUT) functions as a  dynamic reservoir that is able to store urine and to efficiently expel it at a  convenient time. While ...

Orthotopic Small Bowel Transplantation in Rats

Small bowel transplantation has become an accepted clinical option for patients with short gut syndrome and failure of parenteral nutrition (irreversible intestinal failure). In specialized centers improved operative and managing strategies have led to excellent short- and intermediate term patient and graft survival while providing high quality of life 1,3. Unlike in the more common transplantation of other solid organs (i.e. heart, liver) many underlying mechanisms of graft function and immunologic alterations induced by intestinal transplantation are not entirely known6,7. Episodes of acute rejection, sepsis and chronic graft failure are the main obstacles still contributing to less favorable long term outcome and hindering a more widespread employment of the procedure despite a growing number of patients on home parenteral nutrition who would potentially benefit from such a transplant. The small intestine contains a large number of passenger leucocytes commonly referred to as part of the gut associated lymphoid system (GALT) this being part of the reason for the high immunogenity of the intestinal graft. The presence and close proximity of many commensals and pathogens in the gut explains the severity of sepsis episodes once graft mucosal integrity is compromised (for example by rejection). To advance the field of intestinal- and multiorgan transplantation more data generated from reliable and feasible animal models is needed. The model provided herein combines both reliability and feasibility once established in a standardized manner and can provide valuable insight in the underlying complex molecular, cellular and functional mechanisms that are triggered by intestinal transplantation. We have successfully used and refined the described procedure over more than 5 years in our laboratory 8-11. The JoVE video-based format is especially useful to demonstrate the complex procedure and avoid initial pitfalls for groups planning to establish an orthotopic rodent model investigating intestinal transplantation.

Small bowel transplantation has become an accepted treatment option for patients with irreversible intestinal failure. Our experimental model of orthotopic small bowel transplantation in rats serves as a reliable tool to address underlying immunologic and inflammatory processes that complicate intestinal transplantation.

Sıçanlarda Ortotopik Küçük Bağırsak Nakli

Small  bowel transplantation has become an accepted clinical option for patients with  short gut syndrome and failure of parenteral nutrition ...

Retinal Detachment Model in Rodents by Subretinal Injection of Sodium Hyaluronate

Subretinal injection of sodium hyaluronate is a widely accepted method of inducing retinal detachment (RD). However, the height and duration of RD or the occurrence of subretinal hemorrhage can affect photoreceptor cell death in the detached retina. Hence, it is advantageous to create reproducible RDs without subretinal hemorrhage for evaluating photoreceptor cell death. We modified a previously reported method to create bullous and persistent RDs in a reproducible location with rare occurrence of subretinal hemorrhage. The critical step of this modified method is the creation of a self-sealing scleral incision, which can prevent leakage of sodium hyaluronate after injection into the subretinal space. To make the self-sealing scleral incision, a scleral tunnel is created, followed by scleral penetration into the choroid with a 30 G needle. Although choroidal hemorrhage may occur during this step, astriction with a surgical spear reduces the rate of choroidal hemorrhage. This method allows a more reproducible and reliable model of photoreceptor death in diseases that involve RD such as rhegmatogenous RD, retinopathy of prematurity, diabetic retinopathy, central serous chorioretinopathy, and age-related macular degeneration (AMD).

Creating experimental retinal detachments with a reproducible and sustained height of detachment, and without subretinal hemorrhage, is important for studying the pathophysiology of photoreceptor cell loss in retinal disease and evaluating potential therapeutic interventions. Here, we report such a method in detail.

Sodyum Hyaluronat subretinal Enjeksiyon Kemirgenler Retina Dekolmanı Modeli

Subretinal injection of sodium hyaluronate is a widely accepted method of inducing retinal detachment (RD). However, the height and duration of RD or the ...

Remote Limb Ischemic Preconditioning:  A Neuroprotective Technique in Rodents

Sublethal ischemia protects tissues against subsequent, more severe ischemia through the upregulation of endogenous mechanisms in the affected tissue. Sublethal ischemia has also been shown to upregulate protective mechanisms in remote tissues. A brief period of ischemia (5-10 min) in the hind limb of mammals induces self-protective responses in the brain, lung, heart and retina. The effect is known as remote ischemic preconditioning (RIP). It is a therapeutically promising way of protecting vital organs, and is already under clinical trials for heart and brain injuries. This publication demonstrates a controlled, minimally invasive method of making a limb &#8211; specifically the hind limb of a rat &#8211; ischemic. A blood pressure cuff developed for use in human neonates is connected to a manual sphygmomanometer and used to apply 160 mmHg pressure around the upper part of the hind limb. A probe designed to detect skin temperature is used to verify the ischemia, by recording the drop in skin temperature caused by pressure-induced occlusion of the leg arteries, and the rise in temperature which follows release of the cuff. This method of RIP affords protection to the rat retina against bright light-induced damage and degeneration.

Remote ischemic preconditioning (RIP) is a method of conditioning tissues against damaging stress. We have established a method of remote ischemia at the hind limb, by inflating a sphygmomanometer cuff for 5-10 min. The neuroprotective capabilities of RIP have been demonstrated in a model of retinal degeneration in rodents.

Uzaktan Ekstremite İskemik Önkoşullanma: Kemirgenler Bir Nöroprotektiftir Tekniği

Sublethal ischemia protects tissues against subsequent, more severe ischemia through the upregulation of endogenous mechanisms in the affected tissue. ...

Performing Subretinal Injections in Rodents to Deliver Retinal Pigment Epithelium Cells in Suspension

The conversion of light into electrical impulses occurs in the outer retina and is accomplished largely by rod and cone photoreceptors and retinal pigment epithelium (RPE) cells. RPE provide critical support for photoreceptors and death or dysfunction of RPE cells is characteristic of age-related macular degeneration (AMD), the leading cause of permanent vision loss in people age 55 and older. While no cure for AMD has been identified, implantation of healthy RPE in diseased eyes may prove to be an effective treatment, and large numbers of RPE cells can be readily generated from pluripotent stem cells. Several interesting questions regarding the safety and efficacy of RPE cell delivery can still be examined in animal models, and well-accepted protocols used to inject RPE have been developed. The technique described here has been used by multiple groups in various studies and involves first creating a hole in the eye with a sharp needle. Then a syringe with a blunt needle loaded with cells is inserted through the hole and passed through the vitreous until it gently touches the RPE. Using this injection method, which is relatively simple and requires minimal equipment, we achieve consistent and efficient integration of stem cell-derived RPE cells in between the host RPE that prevents significant amount of photoreceptor degeneration in animal models. While not part of the actual protocol, we also describe how to determine the extent of the trauma induced by the injection, and how to verify that the cells were injected into the subretinal space using in vivo imaging modalities. Finally, the use of this protocol is not limited to RPE cells; it may be used to inject any compound or cell into the subretinal space.

Here we present a community accepted protocol in multimedia format for subretinally injecting a bolus of RPE cells in rats and mice. This approach can be used for determining rescue potentials, safety profiles, and survival capacities of grafted RPE cells upon implantation in animal models of retinal degeneration.

Süspansiyon Retina pigment epiteli Hücreleri sunun Kemirgenler Subretinal Enjeksiyonları Sahne

The conversion of light into electrical impulses occurs in the outer retina and is accomplished largely by rod and cone photoreceptors and retinal pigment ...

A Murine Model of Irreversible and Reversible Unilateral Ureteric Obstruction

Obstruction of the kidney may affect native or transplanted kidneys and results in kidney injury and scarring. Presented here is a model of obstructive nephropathy induced by unilateral ureteric obstruction (UUO), which can either be irreversible (UUO) or reversible (R-UUO). In the irreversible UUO model, the ureter may be obstructed for variable periods of time in order to induce increasingly severe renal inflammation and interstitial fibrotic scarring. In the reversible R-UUO model the ureter is obstructed to induce hydronephrosis, tubular dilation and inflammation. After a suitable period of time the ureteric obstruction is then surgically reversed by anastomosis of the severed previously obstructed ureter to the bladder in order to allow complete decompression of the kidney and restoration of urinary flow to the bladder. The irreversible UUO model has been used to investigate various aspects of renal inflammation and scarring including the pathogenesis of disease and the testing of potential anti-inflammatory or anti-fibrotic therapies. The more challenging model of R-UUO has been used by some investigators and does offer significant research potential as it allows the study of inflammatory and immune processes and tissue remodeling in an injured and scarred kidney following the removal of the injurious stimulus. As a result, the R-UUO model offers investigators the opportunity to explore the resolution of kidney inflammation together with key aspects of tissue repair. These experimental models are of relevance to human disease as patients often present with obstruction of the renal tract that requires decompression and are commonly left with significant residual kidney impairment that has no current treatment options and may lead to eventual end stage kidney failure.

The murine model of irreversible unilateral ureteric obstruction (UUO) is presented together with the model of reversible UUO in which the ureteric obstruction is relieved by anastomosis of the severed ureter into the bladder. These models enable the study of renal inflammation and scarring as well as tissue remodeling.

Geri dönüşümsüz ve Tersinir Tek taraflı Üreterik Obstrüksiyon bir fare modeli

Obstruction of the kidney may affect native or transplanted kidneys and results in kidney injury and scarring. Presented here is a model of obstructive ...

Transcutaneous Assessment of Renal Function in Conscious Rodents

Glomerular filtration rate (GFR) is the gold standard to assess overall kidney function. However, traditional methods to evaluate GFR are cumbersome and time-consuming. In addition, serial blood or urine samples are required, with the associated stress for the experimental animals. A recent technique significantly reduces the investment in time and resources, minimizing the invasiveness and the animal stress, but being equally valid as the traditional approaches. The method measures transcutaneously renal function. Using an optical device and the exogenous renal marker fluorescein isothiocyanate (FITC)-sinistrin, this technique is capable of measuring the elimination kinetics of the marker through the skin. With neither blood nor urine samples nor the associated laboratory assays needed, the results of the transcutaneous measurement are almost instantaneously available. The method has been already validated in different species and successfully applied in several models of renal pathology. Moreover, due to its minimally invasive characteristics, it is suitable for sequential measurements within the same animal. Here is provided a detailed protocol to carry out the transcutaneous assessment of renal function in rodents.

Determination of glomerular filtration rate (GFR) is the gold standard to assess overall kidney function. However, traditional procedures to measure this parameter are cumbersome and require a large investment of time. Here we describe a faster and minimally invasive method to determinate GFR transcutaneously.

Bilinçli Kemirgenler Böbrek Fonksiyon Transkutanöz Değerlendirmesi

Glomerular filtration rate (GFR) is the gold standard to assess overall kidney function. However, traditional methods to evaluate GFR are cumbersome and ...

An Efficient Sieving Method to Isolate Intact Glomeruli from Adult Rat Kidney

Preservation of glomerular structure and function is pivotal in the prevention of glomerulonephritis, a category of kidney disease characterized by proteinuria which can eventually lead to chronic and end-stage renal disease. The glomerulus is a complex apparatus responsible for the filtration of plasma from the body. In disease, structural integrity is lost and allows for the abnormal leakage of plasma contents into the urine. A method to isolate and examine glomeruli in culture is critical for the study of these diseases. In this protocol, an efficient method of retrieving intact glomeruli from adult rat kidneys while conserving structural and morphological characteristics is described. This process is capable of generating high yields of glomeruli per kidney with minimal contamination from other nephron segments. With these glomeruli, injury conditions can be mimicked by incubating them with a variety of chemical toxins, including protamine sulfate, which causes foot process effacement and proteinuria in animal models. Degree of injury can be assessed using transmission electron microscopy, immunofluorescence staining, and western blotting. Nephrin and Wilms Tumor 1 (WT1) levels can also be assessed from these cultures. Due to the ease and flexibility of this protocol, the isolated glomeruli can be utilized as described or in a way that best suits the needs of the researcher to help better study glomerular health and structure in diseased states.

The main focus of this protocol is to efficiently isolate viable primary glomeruli cultures with minimal contaminants for use in a variety of downstream applications. The isolated glomeruli retain structural relationships between component cell types and can be cultured ex vivo for a short time.

Yetişkin sıçan böbrek üzerinden olduğu gibi Glomeruli yalıtmak için verimli bir eleme yöntemi

Preservation of glomerular structure and function is pivotal in the prevention of glomerulonephritis, a category of kidney disease characterized by ...

Studying Left Ventricular Reverse Remodeling by Aortic Debanding in Rodents

To better understand the left ventricular (LV) reverse remodeling (RR), we describe a rodent model wherein, after aortic banding-induced LV remodeling, mice undergo RR upon removal of the aortic constriction. In this paper, we describe a step-by-step procedure to perform a minimally invasive surgical aortic debanding in mice. Echocardiography was subsequently used to assess the degree of cardiac hypertrophy and dysfunction during LV remodeling and RR and to determine the best timing for aortic debanding. At the end of the protocol, terminal hemodynamic evaluation of the cardiac function was conducted, and samples were collected for histological studies. We showed that debanding is associated with surgical survival rates of 70-80%. Moreover, two weeks after debanding, the significant reduction of ventricular afterload triggers the regression of ventricular hypertrophy (~20%) and fibrosis (~26%), recovery of diastolic dysfunction as assessed by the normalization of left ventricular filling and end-diastolic pressures (E/e&#39;&#160;and LVEDP). Aortic debanding is a useful experimental model to study LV RR in rodents. The extent of myocardial recovery is variable between subjects, therefore, mimicking the diversity of RR that occurs in the clinical context, such as aortic valve replacement. We conclude that the aortic banding/debanding model represents a valuable tool to unravel novel insights into the mechanisms of RR, namely the regression of cardiac hypertrophy and the recovery of diastolic dysfunction.

Here we describe a step-by-step protocol of surgical aorta debanding in the well-established mice model of aortic-constriction. This procedure not only allows studying the mechanisms underlying the left ventricular reverse remodeling and regression of hypertrophy but also to test novel therapeutic options that might accelerate myocardial recovery.

Kemirgenlerde Aort Debanding tarafından Sol Ventrikül Ters Tadilatı Eğitimi

To better understand the left ventricular (LV) reverse remodeling (RR), we describe a rodent model wherein, after aortic banding-induced LV remodeling, ...