Bu eser Belçika Federal Hükümeti (IUAP P6/28), Araştırma Vakfı-Flanders (FWO) (G.0565.07 ve G.0686.09), Astellas Avrupa Vakfı Ödülü 2009 ve KU Leuven Araştırma Konseyi bağışları ile desteklenmiştir (GOA 2009/07, EF/95/010 ve PFV/10/006). PU ve BİZ Araştırma Vakfı-Flanders (FWO) ve doktora öğrencisinin vardır. MB Marie Curie akademisyendir. FWO DDR temel-klinik fellow.

1. Laboratuvar Hayvanları <ol><li> Hayvanlar (fare, sıçan) 12 saat ışık-karanlık döngüsü, su ve standart gıda granül ad libitum erişimi ile özel bir hayvan tesisi içinde muhafaza edilir. Hayvanların yaş ve cinsiyet Hem ihtiyaçlarına göre standardize edilmelidir önemli parametrelerdir. 12 haftalık dişi hayvanlarda 5,6 - Biz genellikle 10 sistometri gerçekleştirin. </li><li> Tüm hayvan deneyleri Avrupa Birliği Topluluğu Konseyi kurallarına uygun olarak yürütülen ve yerel etik kurul tarafından onaylanmıştır. </li></ol> 2. Anestezi <ol><li> Bu yarı-ömrü kısa olması nedeniyle doza kolay olduğundan İzofluran anestezi, küçük bir cerrahi işlemlerin gerçekleştirilmesi için kullanılır. Izofluran kullanırken süpürücü Uygun atık gaz gereklidir. <ol class="lalpha"><li> Hayvanlar saf oksijen izofluran% 5 (1 l / dk) ile gazlandırılan kapalı bir kutu, yerleştirilir. </li><li> Uygun l indüksiyon, anestezi sonrası1 l / 1 dak oksijen - - izofluran% 1.5 evel 0,8 oranında küçük bir maske (bir kesim 10 ml şırınga) tarafından yapılmaktadır. </li></ol></li><li> Fareler için, fonksiyonel üretan anestezi kayıtları (cfr. Kızılötesi) sırasında kullanılır. Diğer birçok anestetik aksine, işeme refleks bu anestezik madde 4 ile bastırılmış değildir, çünkü anestezi fonksiyonel üretan kayıt sırasında kullanılır. Üretan 1.2 g / kg vücut ağırlığı (sc) 60 dakikalık kayıtların başlamasından önce deri altından tatbik edilmektedir. Eğer gerekirse, uygun anestezi elde etmek için gerekli olan, üretan ve subkütan olarak ek doz (0.1 g / kg) verilerek titre edilebilir. Bu belirtmek gerekir, ancak, bu ek anestezi işeme verimlilik ve dolayısıyla işeme frekans 7,8 değiştirebilir. Dolayısıyla, mesane kayıtları yakından takip ve kontrol davranışı önemli sapmaların tespit edildiği anda atılmalıdır. Ederken Anestezikler uygulanmamalıdır testibileşiklerin veri yorumlanmasına önlemek için uygulanır. </li></ol> 3. Cerrahi Prosedür - Mesane Kateteri İmplantasyonu <ol><li> Cerrahi aletler kullanmadan önce otoklavda sterilize edilir. , Hayvanın karın Tıraş etanol% 70 ile dezenfekte ve mesane maruz düşük bir orta hat laparotomi uygulandı. </li><li> Cerrahi mikroskop altında, bir emilmeyen, monofilaman sütür (boyut 6-0) kullanarak mesane kubbesi bir çanta-string sütür koyun. </li><li> Çanta dize içinde küçük bir sistostomi (biz tercihen 18 G iğne kullanın) gerçekleştirin ve bu delikten sonunda küçük bir manşet (dikkatle tüp ısıtılmasıyla elde edilen) ile, PE 50 Polietilen kateter yerleştirin. Bizim ellerde, ince borular (PE 10) daha yüksek ve değişken direnç var. Tüp, etanol% 70 solüsyon koyarak veya bir otoklavda sterilize ile dezenfekte edilebilir. Takılmadan önce boru tuzlu su ile yıkanması gerekir. </li><li> Çanta str sabitleyinBir cerrahın düğüm ile tüp etrafında ing. Çan şeklinde ucu kadar yavaşça dışarıya doğru dikiş altında konumlandırılmış kateter çekin. Sızıntı önlemek için dikiş doğru 18 G kateter bir parça itin. </li><li> Yavaşça sızıntı olup olmadığını kontrol etmek - salin küçük hacimli (200 ul 100) Infuse. Sızıntı varsa, bir ek sütür konulmalıdır. </li><li> Kesi üst kısmında kateter için bir geçit bırakarak monofilaman sütürler kullanılarak karın kaslarının kapatın. </li><li> Tünel içi boş bir metal çubuk kullanılarak interskapular için tüp, böylece deney sırasında boru ısırmak için hayvanlar önler. </li><li> Cilt emilmeyen, monofilaman sütürler kullanılarak kapatılır. Implante kateter geçirgenliğini kontrol etmek için tuzlu su ile temizlenir. </li><li> Hayvanlar uyandırmak önce, analjezikler (buprenorfin, 0.05 mg / kg) deri altından verilmiştir. </li></ol> 4. Kurulum ve Sistometriler <ol><li> Deneysel Questi bağlı olarak,(ve tür kullanılan) üzerinde sistometri uyanık (kısıtlama ya da serbestlik) veya üretan-anestezi uygulanmış hayvanlarda gerçekleştirilir. Sistometri bir ölçülü ortamda sakin olmaya eğilimi bilinçli sıçanlarda, mükemmel mümkündür Oysa fareler 3,5,6,9 kullanırken, biz genellikle üretan anestezisi (subkütan 1.2 mg / kg) altında sistometri gerçekleştirin. </li><li> Anestezi farelerde vücut sıcaklığı sürekli olarak izlenir ve 36.5 &#39;da tutulmaktadır ° C ± 0.2 ° C arasında bir sıcaklık probu ve bir ısıtma lambası aracılığı ile. </li><li> Set-up üreticinin talimatlarına göre her deneyden önce kalibre edilir. </li><li> Implante kateter bir tarafında ve (Harvard Apparatus) başka bir enfüzyon pompası üzerinde basınç transdüktör (Edwards Lifesciences) bağlı olan 3-yollu bir musluk için bir luer saplama ile bağlıdır. Basınç dönüştürücü veri toplama sistemi (DI-730-USB, Dataq araçlar) için bir amplifikatör (78534c monitör, Hewlett Packard) ve bir ortak ile bağlandığındamputer. Windaq / Lite yazılım kayıt için de kullanılabilir. </li><li> Enfüzyon pompası, örneğin salin ya da PBS içinde infüzyon sağlayan başlatıldığında, 100 ul / dakika (sıçanlar) ya da 20 ul / dk (farelerde). Intravesikal basınç kaydedilirken gibi mesane, sabit bir oranda doldurulur. </li><li> Şekil 5 tipik bir basınç izleme gösteriyor: belirli bir hacim / basıncına ulaşılana kadar sıvı infüzyonu sırasında, mesane yavaş bir baskı birikimini miksiyon eşiği vardır. Bir kez bu eşiği, mesane sözleşme ulaşılması ve daha sonra üriner sfinkter açılır, üretra yoluyla idrarın geçişine izin. Bu nedenle, mesane boşaltılır, daralma duracak ve basınç &quot;bazal&quot; seviyesine tekrar düşecek. </li><li> Bir sıçan metabolik kafesinde ve bir dijital denge üstüne yerleştirilir gibi, idrar hacmi idrar hacmi ile ilgili bilgi vererek, eşzamanlı olarak ölçülebilir. Mil içinde küçük işeme hacimleri nedeniyle,ce, bu sistemi kullanarak çok zor olabilir. Bu nedenle, idrar hacmi işeme önce ve hemen sonra tartılmış küçük bir filtre kağıdı kullanılarak tespit edilir. </li><li> Genellikle, sistem 30 dakika arasında bir süre kayıt ardından, 30 dakika için denge sağlar. Sonra, ilaç sistemik veya intravesikal tatbik edilebilir ve bir 30 dakika daha kaydedilir. </li><li> Kaydedilmiş veriler nicel analiz için özel bir yazılım alınabilir. Csv dosyaları olarak ihraç edilebilir. Biz genellikle (originlab Corporation, MA, ABD) Menşe kullanın. </li><li> Deneylerden sonra, hayvanlar servikal dislokasyon (fare) veya CO 2 zehirlenmesi (sıçan) tarafından kurban edilir. </li></ol> 5. Temsilcisi Sonuçlar <img alt="Şekil 1" src="/files/ftp_upload/3869/3869fig1.jpg" /> Şekil 1. Laparotomi bakış. A) yatar pozisyonda sıçan yerleştirin. B) Tıraş ve antiseptically cerrahi sitesi hazırlamak. Cilt C) İnsizyonunun. D) Kesi of karın kasları, mesane ve pozlama. <img alt="Şekil 2," src="/files/ftp_upload/3869/3869fig2.jpg" /> Şekil 2. A purse-string sütür. <img alt="Şekil 3" src="/files/ftp_upload/3869/3869fig3.jpg" /> 3. Kateter implantasyonu Şekil. <img alt="Şekil 4," src="/files/ftp_upload/3869/3869fig4.jpg" /> Şekil 4,. Kateterin tip tünel. Bilinçli bir sıçan ve anestezi uygulanmış bir farenin salin intravesikal basınç perfüzyon sırasında elde edilen ölçüm örnekleri Şekil 5 &#39;de gösterilmiştir. Birden çok parametreler basınç sinyali (intercontractile aralık, temel eşik basınç ve basınç gibi) dan elde edilebilir. Bu parametrelerin tanımları için kapsamlı, Andersson, et al. (Ref 4) ve Yoshiyama ve diğerleri bakınız. (<html> ref 10). Biz son zamanlarda hardal yağı moleküler hedeflerin (MO), uzun inflamasyon ve bu mesane 11,12 olarak visseral organların hiperaljezi deneysel modellerinde kullanılan bir yüksek reaktif bileşiği tanımlamak için sistometri kullandık. 10 mM MO İntravezikal infüzyon vahşi tip farelerde işeme sıklığı güçlü bir artış (intercontractile aralığında düşüş) (Şekil 6A, B) ve işeme hacmi 6 azalma indüklenen. İlginçtir, MO MO reseptör TRPA1 arasında eksik farelerde benzer değişiklikler indüklenen. Buna karşılık, MO WT farelere kıyasla daha TRPV1 KO fareleri içinde mesane parametreleri çok daha zayıftır değişimlere neden olan ve Trpa1/Trpv1 KO fareleri içinde herhangi bir etkisi yoktu. Birlikte Peptid (CGRP) 6 İlişkin Kalsitonin Gen verildiği ölçümleri ile, bu veriler TRPV1 MO tarafından indüklenen visseral tahriş önemli bir rol oynayabileceğini göstermektedir göstermektedir. ve_content &quot;&gt; <img alt="Şekil 5," src="/files/ftp_upload/3869/3869fig5.jpg" /> Şekil 5. Bilinçli bir dişi sıçan (A) ve anestezi dişi fare (B) kaydedilmiş intravezikal basınç Temsilcisi izleri. En düşük basınç &quot;taban baskısı&quot; (kırmızı oklar) olarak tanımlanır. Doldurma aşamasının sonunda, basınç mavi oklarla işaretlenmiştir. Basınç farkı bölünmesiyle bu noktaların arasındaki infüzyon sıvı hacmi sağlayan mesane duvarının uyum (uyum = infüzyon hacmi / (eşik basıncı hesabı -. Bazal çizgi basıncı) &quot;intercontractile aralığı&quot; (ICI) arasındaki zaman iki işeme kasılmalar. <img alt="Şekil 6" src="/files/ftp_upload/3869/3869fig6.jpg" /> Vahşi tip ve Trpa1, TRPV1 ve Trpa1/Trpv1 knockout farelerde sistometri desen üzerine hardal yağı intravezikal uygulama Şekil 6. Etkileri.(A) ve 10 mM salin infüzyonu, MO yanıt olarak WT, Trpa1 KO, TRPV1 KO ve Trpa1/Trpv1 KO fareleri içinde kaydedilmiş intravesikal basınç değişiklikleri temsil eden örnekler. Öncesi ve MO intravezikal infüzyon sırasında ortalama anlık işeme sıklığı (B) Zaman kursu. Tüm fareler için, veri salin infüzyonu sırasında elde edilen ortalama frekans için normalize edildi. Bu veriler, Elsevier izni ile Everaerts ve ark. (Ref 6) uyarlanmıştır.

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Çıkar çatışması ilan etti.

Burada sunulan sistometri tekniği hayvan modellerinde mesane fonksiyonu in vivo ölçümleri performans sağlar. Sıçanlar belkide en çok kullanılan hayvan modeli vardır. Fare işlemek için daha zor, ama genetik olarak manipüle hayvanları kullanmanın avantajı sunuyoruz. Çünkü implante kateter ve intravezikal basıncı etkileyebilir karın içi basınç değişiklikleri gevşemesi sonucunda çok aktif olma eğilimi bilinçli fareler kullanarak teknik zorluk, biz tam sistometrisi sırasında üretan ...

<table border="1"><tbody><tr><td> Reaktif Adı </td><td> Ticari adı, şirket, </td><td> Katalog numarası </td><td> Yorumlar </td></tr><tr><td> üretan </td><td> Üretan, Sigma-Aldrich </td><td> 315419 </td><td> Grup 2B kanserojen </td></tr><tr><td> izofluran </td><td> Isoba, Schering-Plough Hayvan Sağlığı </td><td></td><td></td></tr><tr><td> polietilen kateter </td><td> Intramedic polietilen boru PE50, Becton Dickinson </td><td> 427411 </td><td></td></tr><tr><td> Cerrahi mikroskop </td><td> Op-Mi 6, Carl Zeiss </td><td> Op-Mi 6 </td><td></td></tr><tr><td> purse-string sütür </td><td> Prolen 6/0, Ethicon </td><td> 8610H </td><td></td></tr><tr><td> fasya ve cilt sütür </td><td> Ethilon 4/0 veya 5/0, Ethicon </td><td> 662G veya 661G </td><td></td></tr><tr><td> postoperatif analjezik </td><td> Temgesic, Schering-Plough Hayvan Sağlığı </td><td></td><td> fareler için doz: 0.05 mg / kg &#39; </td></tr><tr><td> amplifikatör </td><td> 78534c monitör, Hewlett Packard </td><td></td><td></td></tr><tr><td> analitik dengeleri ve denge veri toplama yazılımı </td><td> FZ 300i, A &amp; D </td><td> FZ-300i </td><td></td></tr><tr><td> infüzyon pompaları </td><td> Pompa 33, Harvard Apparatus </td><td> HA33 </td><td></td></tr><tr><td> sistometri kayıt sistemi </td><td> Dataq aletleri, DI-730 serisi ve Windaq / Lite </td><td> DI-730-USB Windaq / Lite </td><td></td></tr><tr><td> sıcaklık kaydı </td><td> Fluke 52 KJ termometre </td><td> 52 KJ </td><td></td></tr><tr><td> basınç dönüştürücüler </td><td> Edwards Lifesciences, basınç izleme seti </td><td> T322247A </td><td></td></tr></tbody></table>

the use of cystometry in small rodents: a study of bladder chemosensation

<p class="jove_content"&gt; Idrarı depolamak ve verimli uygun bir anda çıkarmak için güçlü bir dinamik rezervuar olarak alt üriner sistem (AÜS) fonksiyonları. Idrar depolanması sırasında, ancak, mesane atık ürünler için uzun süre maruz kalır. Sıkı bir bariyer gibi hareket ederek, LUT, Ürotelyumun, epitel zarının zararlı maddelerin emilimini önler. Ayrıca, zararlı kimyasallar mesane nosiseptif innervasyon uyarmak ve mesane içeriğini sınırdışı kasılmaları işeme başlatabilir. İlginçtir, zararlı kimyasallar mesane hassasiyeti intravezikal nörojenik mesane aşırı aktivitesi tedavi TRPV1 agonisti kapsaisin beslerken tarafından, klinik pratikte başarıyla kullanılmıştır<sup&gt; 1</sup&gt;. Bu kimyasal duyusal organ olarak mesane görüntüleme avantajı altını çiziyor ve daha ileri klinik araştırmalar için ister. Ancak, etik konular ciddi insan denekler, LUT klinik farmakoloji moleküler temelleri çözülmeye gerekli olan invaziv ölçümler, gerçekleştirmek için olanakları sınırlı. Bu sınırlama üstesinden gelmek için bir yol birkaç hayvan modellerinin kullanımı<sup&gt; 2</sup&gt;. Burada fare ve sıçanlarda sistometri, kontrollü mesane perfüzyon koşullarında intravezikal basıncı ölçmek sağlayan bir tekniğin uygulanmasını açıklar.</p&gt;<p class="jove_content"&gt; Laparotomi sonra bir kateter mesane kubbesi implante ve interskapular bölgeye subkutan tünel. Üretra işeme için serbest bırakılırken sonra mesane, kontrollü bir oranda doldurulabilir. Dolum ve işeme tekrarlayan döngüleri sırasında intravezikal basınç implante kateter yoluyla ölçülebilir. Bunun gibi, basınç değişiklikleri belirleyen ve analiz edilebilir. Ayrıca, işeme hacmi aynı anda ölçülmesini sağlar dışı işeme kasılmalar ayırt işeme kasılmalar<sup&gt; 3</sup&gt;.</p&gt;<p class="jove_content"&gt; Önemlisi, kemirgenler ve insanlar arasında miksiyon kontrolü farklılıklar nedeniyle, bu hayvanlarda sistometrik ölçümler sadece sınırlı ötelenme değeri<sup&gt; 4</sup&gt;. Yine de, deneysel klinik öncesi ortamlarda mesane patofizyolojisi ve farmakoloji çalışmasında oldukça aracı bulunmaktadır. Bu tekniği kullanarak son araştırmalar mesane Mechano-ve kemo-duyusal özelliklerinde yeni moleküler oyuncuların rol ortaya çıkarmıştır.</p

Watch this Scientific Journal Video about The Use of Cystometry in Small Rodents: A Study of Bladder Chemosensation at JoVE.com

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

Sistometriler küçük hayvanların mesane fonksiyonu ölçmek için etkili bir tekniktir<em&gt; In vivo</em&gt;. Mesane sürekli idrar işeme için serbest bırakılmalıdır ise, intravezikal kateter yoluyla kontrol fiyatla demlenir. Bu intravezikal basıncı ve işeme hacmi kaydedilir iken, tekrarlayan için doldurma ve mesane boşaltılmasını sağlar.

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 ...

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Research

JoVE Journal

Medicine

22.4K Görüntüleme.  KU Leuven, Belgium. Sistometriler küçük hayvanların mesane fonksiyonu ölçmek için etkili bir tekniktir In vivo. Mesane sürekli idrar işeme için serbest bırakılmalıdır ise, intravezikal kateter yoluyla kontrol fiyatla demlenir. Bu intravezikal basıncı ve işeme hacmi kaydedilir iken, tekrarlayan için doldurma ve mesane boşaltılmasını sağlar.

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

Making Sense of Listening: The IMAP Test Battery

The ability to hear is only the first step towards making sense of the range of information contained in an auditory signal. Of equal importance are the abilities to extract and use the information encoded in the auditory signal. We refer to these as listening skills (or auditory processing AP). Deficits in these skills are associated with delayed language and literacy development, though the nature of the relevant deficits and their causal connection with these delays is hotly debated.
When a child is referred to a health professional with normal hearing and unexplained difficulties in listening, or associated delays in language or literacy development, they should ideally be assessed with a combination of psychoacoustic (AP) tests, suitable for children and for use in a clinic, together with cognitive tests to measure attention, working memory, IQ, and language skills. Such a detailed examination needs to be relatively short and within the technical capability of any suitably qualified professional. Current tests for the presence of AP deficits tend to be poorly constructed and inadequately validated within the normal population. They have little or no reference to the presenting symptoms of the child, and typically include a linguistic component. Poor performance may thus reflect problems with language rather than with AP. To assist in the assessment of children with listening difficulties, pediatric audiologists need a single, standardized child-appropriate test battery based on the use of language-free stimuli.
We present the IMAP test battery which was developed at the MRC Institute of Hearing Research to supplement tests currently used to investigate cases of suspected AP deficits. IMAP assesses a range of relevant auditory and cognitive skills and takes about one hour to complete. It has been standardized in 1500 normally-hearing children from across the UK, aged 6-11 years. Since its development, it has been successfully used in a number of large scale studies both in the UK and the USA. IMAP provides measures for separating out sensory from cognitive contributions to hearing. It further limits confounds due to procedural effects by presenting tests in a child-friendly game-format. Stimulus-generation, management of test protocols and control of test presentation is mediated by the IHR-STAR software platform. This provides a standardized methodology for a range of applications and ensures replicable procedures across testers. IHR-STAR provides a flexible, user-programmable environment that currently has additional applications for hearing screening, mapping cochlear implant electrodes, and academic research or teaching.

A test battery (IMAP) for performing an in-depth assessment of auditory and cognitive abilities contributing to listening skills is described. It is quick to administer, child-friendly and free from linguistic confounds. Stimulus generation and protocol management are controlled via a software platform (IHR-STAR) to ensure replicable procedures.

Dinlemede Making Sense IMAP Test Batarya:

The ability to hear is only the first step towards making sense of the range of information contained in an auditory signal. Of equal importance are the ...

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Mouse Bladder Wall Injection

Mouse bladder wall injection is a useful technique to orthotopically study bladder phenomena, including stem cell, smooth muscle, and cancer biology. Before starting injections, the surgical area must be cleaned with soap and water and antiseptic solution. Surgical equipment must be sterilized before use and between each animal. Each mouse is placed under inhaled isoflurane anesthesia (2-5% for induction, 1-3% for maintenance) and its bladder exposed by making a midline abdominal incision with scissors. If the bladder is full, it is partially decompressed by gentle squeezing between two fingers. The cell suspension of interest is intramurally injected into the wall of the bladder dome using a 29 or 30 gauge needle and 1 cc or smaller syringe. The wound is then closed using wound clips and the mouse allowed to recover on a warming pad. Bladder wall injection is a delicate microsurgical technique that can be mastered with practice.

Mouse bladder wall injection is a useful approach to orthotopically study bladder stem cell and cancer biology. This delicate microsurgical method can be mastered with careful technique and practice.

Fare mesane Duvar Enjeksiyon

Mouse bladder wall injection is a useful technique to orthotopically study bladder phenomena, including stem cell, smooth muscle, and cancer biology. ...

Biomarkers in an Animal Model for Revealing Neural, Hematologic, and Behavioral Correlates of PTSD

Identification of biomarkers representing the evolution of the pathophysiology of Post Traumatic Stress Disorder (PTSD) is vitally important, not only for objective diagnosis but also for the evaluation of therapeutic efficacy and resilience to trauma. Ongoing research is directed at identifying molecular biomarkers for PTSD, including traumatic stress induced proteins, transcriptomes, genomic variances and genetic modulators, using biologic samples from subjects' blood, saliva, urine, and postmortem brain tissues. However, the correlation of these biomarker molecules in peripheral or postmortem samples to altered brain functions associated with psychiatric symptoms in PTSD remains unresolved. Here, we present an animal model of PTSD in which both peripheral blood and central brain biomarkers, as well as behavioral phenotype, can be collected and measured, thus providing the needed correlation of the central biomarkers of PTSD, which are mechanistic and pathognomonic but cannot be collected from people, with the peripheral biomarkers and behavioral phenotypes, which can.
Our animal model of PTSD employs restraint and tail shocks repeated for three continuous days - the inescapable tail-shock model (ITS) in rats. This ITS model mimics the pathophysiology of PTSD 17, 7, 4, 10. We and others have verified that the ITS model induces behavioral and neurobiological alterations similar to those found in PTSD subjects 17, 7, 10, 9. Specifically, these stressed rats exhibit (1) a delayed and exaggerated startle response appearing several days after stressor cessation, which given the compressed time scale of the rat's life compared to a humans, corresponds to the one to three months delay of symptoms in PTSD patients (DSM-IV-TR PTSD Criterian D/E 13), (2) enhanced plasma corticosterone (CORT) for several days, indicating compromise of the hypothalamopituitary axis (HPA), and (3) retarded body weight gain after stressor cessation, indicating dysfunction of metabolic regulation.
The experimental paradigms employed for this model are: (1) a learned helplessness paradigm in the rat assayed by measurement of acoustic startle response (ASR) and a charting of body mass; (2) microdissection of the rat brain into regions and nuclei; (3) enzyme-linked immunosorbent assay (ELISA) for blood levels of CORT; (4) a gene expression microarray plus related bioinformatics tools 18. This microarray, dubbed rMNChip, focuses on mitochondrial and mitochondria-related nuclear genes in the rat so as to specifically address the neuronal bioenergetics hypothesized to be involved in PTSD.

We describe a rat model of post traumatic stress disorder (PTSD) that reveals the persistent alterations in neuroendocrine function and the delayed long-term, exaggerated fear response, characteristic of PTSD patients. The animal model and methods described here are useful for correlating biomarkers in brain nuclei, which are mechanistic but cannot be measured in patients, with biomarkers in peripheral white blood cells, which can.

TSSB, Nöral Hematolojik ve Davranış Bağlantılı Revealing için Hayvan Modelinde Biyobelirteçler

Identification of biomarkers representing the evolution of the pathophysiology of Post Traumatic Stress Disorder (PTSD) is vitally important, not only for ...

Human Neuroendocrine Tumor Cell Lines as a Three-Dimensional Model for the Study of Human Neuroendocrine Tumor Therapy

Neuroendocrine tumors (NETs) are rare tumors, with an incidence of two per 100,&#x200A;000 individuals per year, and they account for 0.5% of all human malignancies.1 Other than surgery for the minority of patients who present with localized disease, there is little or no survival benefit of systemic therapy. Therefore, there is a great need to better understand the biology of NETs, and in particular define new therapeutic targets for patients with nonresectable or metastatic neuroendocrine tumors. 3D cell culture is becoming a popular method for drug screening due to its relevance in modeling the in vivo tumor tissue organization and microenvironment.2,3 The 3D multicellular spheroids could provide valuable information in a more timely and less expensive manner than directly proceeding from 2D cell culture experiments to animal (murine) models.
To facilitate the discovery of new therapeutics for NET patients, we have developed an in vitro 3D multicellular spheroids model using the human NET cell lines. The NET cells are plated in a non-adhesive agarose-coated 24-well plate and incubated under physiological conditions (5% CO2, 37 &deg;C) with a very slow agitation for 16-24 hr after plating. The cells form multicellular spheroids starting on the 3rd or 4th day. The spheroids become more spherical by the 6th day, at which point the drug treatments are initiated. The efficacy of the drug treatments on the NET spheroids is monitored based on the morphology, shape and size of the spheroids with a phase-contrast light microscope. The size of the spheroids is estimated automatically using a custom-developed MATLAB program based on an active contour algorithm. Further, we demonstrate a simple method to process the HistoGel embedding on these 3D spheroids, allowing the use of standard histological and immunohistochemical techniques. 
This is the first report on generating 3D spheroids using NET cell lines to examine the effect of therapeutic drugs. We have also performed histology on these 3D spheroids, and displayed an example of a single drug's effect on growth and proliferation of the NET spheroids. Our results support that the NET spheroids are valuable for further studies of NET biology and drug development.

We present a simple agarose overlay platform to grow 3D multicellular spheroids using neuroendocrine cancer cell lines. This method provides a very convenient way to examine the effect of therapeutic drugs on the neuroendocrine tumor cells. It could also help us establish human neuroendocrine tumor spheroids for cancer therapy.

İnsan Nöroendokrin Tümör Terapi Çalışmaları Üç Boyutlu Model İnsan nöroendokrin tümör hücre hatlarında

Neuroendocrine tumors (NETs) are rare tumors, with an incidence of two per 100,&#x200A;000 individuals per year, and they account for 0.5% of all human ...

Urinary Bladder Distention Evoked Visceromotor Responses as a Model for Bladder Pain in Mice

Approximately 3-8 million people in the United States suffer from interstitial cystitis/bladder pain syndrome (IC/BPS), a debilitating condition characterized by increased urgency and frequency of urination, as well as nocturia and general pelvic pain, especially upon bladder filling or voiding. Despite years of research, the cause of IC/BPS remains elusive and treatment strategies are unable to provide complete relief to patients. In order to study nervous system contributions to the condition, many animal models have been developed to mimic the pain and symptoms associated with IC/BPS. One such murine model is urinary bladder distension (UBD). In this model, compressed air of a specific pressure is delivered to the bladder of a lightly anesthetized animal over a set period of time. Throughout the procedure, wires in the superior oblique abdominal muscles record electrical activity from the muscle. This activity is known as the visceromotor response (VMR) and is a reliable and reproducible measure of nociception. Here, we describe the steps necessary to perform this technique in mice including surgical manipulations, physiological recording, and data analysis. With the use of this model, the coordination between primary sensory neurons, spinal cord secondary afferents, and higher central nervous system areas involved in bladder pain can be unraveled. This basic science knowledge can then be clinically translated to treat patients suffering from IC/BPS.

Approximately 3-8 million people in the United States suffer from interstitial cystitis/bladder pain syndrome (IC/BPS), a debilitating condition characterized in part by pelvic pain. In order to study nervous system contributions to the condition, a physiological model of urinary bladder pain is used in mice and rats.

Farelerde Mesane Ağrı için bir model olarak Mesane Distansiyonunun Uyarılmış Visceromotor Cevapları

Approximately 3-8 million people in the United States suffer from interstitial cystitis/bladder pain syndrome (IC/BPS), a debilitating condition ...

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 ...

Evaluation of the Efficacy of the H. pylori Protein HP-NAP as a Therapeutic Tool for Treatment of Bladder Cancer in an Orthotopic Murine Model

Bladder cancer is one of the most common malignancies of the urogenital tract. Intravesical injection of Bacillus Calmette-Gu&#233;rin (BCG) is the gold standard treatment for the high-grade non-muscle invasive bladder cancer (NMIBC). However, since the treatment-related side effects are relevant, newer biological response modifiers with a better benefit/side effects ratio are needed.
The tumour microenvironment can influence both tumour development and therapy efficacy. In order to obtain a good model, it is desirable to implant tumour cells in the organ from which the cancer originates.
In this protocol, we describe a method for establishing a tumour in the bladder cavity of female mice and subsequent delivery of therapeutic agents; the latter are exemplified by our use of Helicobacter pylori neutrophil activating protein (HP-NAP). A preliminary chemical burn of the mucosa, followed by the injection of mouse urothelial carcinoma cell line MB49 via urethral catheterization, enables the cells to attach to the bladder mucosa. After a period, required to allow an initial proliferation of the cells, mice are treated with HP-NAP, administrated again via catheterization. The anti-tumour activity of HP-NAP is evaluated comparing the tumour volume, the extent of necrosis and the degree of vascularization between vehicle- and HP-NAP-treated animals.

Evaluation of the Efficacy of the H. pylori Protein HP-NAP as a Therapeutic Tool for Treatment of Bladder Cancer in an Orthotopic Murine Model

Here the method to establish a syngeneic mouse model of orthotopic bladder tumour to evaluate the anti-tumour efficacy of the bacterial protein HP-NAP is described.

Etkinliğinin Değerlendirilmesi<em&gt; H. pylori</emOrtotopik Fare Modelinde Mesane Kanseri Tedavisinde Terapötik Aracı Olarak&gt; Protein HP-NAP

Bladder cancer is one of the most common malignancies of the urogenital tract. Intravesical injection of Bacillus Calmette-Gu&#233;rin (BCG) is the gold ...

Use of a Piglet Model for the Study of Anesthetic-induced Developmental Neurotoxicity (AIDN): A Translational Neuroscience Approach

Anesthesia cannot be avoided in many cases when surgery is required, particularly in children. Recent investigations in animals have raised concerns that anesthesia exposure may lead to neuronal apoptosis, known as anesthesia-induced developmental neurotoxicity (AIDN). Furthermore, some clinical studies in children have suggested that anesthesia exposure may lead to neurodevelopmental deficits later in life. Nonetheless, an ideal animal model for preclinical study has yet to be developed. The neonatal piglet represents a valuable model for preclinical study, as they share a striking number of developmental similarities with humans.
The anatomy and physiology of piglets allow for implementation of rigorous human perioperative conditions in both survival and non-survival procedures. Femoral artery catheterization allows for close monitoring, thus enabling prompt correction of any deviation of the piglet's vital signs and chemistries. In addition, there are multiple developmental similarities between piglets and human neonates. The techniques required to use piglets for experimentation will require experience to master. A pediatric anesthesiologist is a critical member of the investigative team. We describe, in a general sense, the appropriate use of a piglet model for neurodevelopmental study.

Use of a Piglet Model for the Study of Anesthetic-induced Developmental Neurotoxicity AIDN: A Translational Neuroscience Approach

Anesthesia-induced developmental neurotoxicity (AIDN) research has focused on rodents, which are not broadly applicable to humans. Non-human primate models are more relevant, but are cost-prohibitive and difficult to use for experimentation. The piglet, in contrast, is a clinically relevant, practical animal model ideal for the study of anesthetic neurotoxicity.

Anesteziye Bağlı Gelişimsel Nörotoksisite (AIDN) Çalışması İçin Bir Piglet Modelinin Kullanımı: Bir Translasyonel Sinirbilimi Yaklaşımı

Anesthesia cannot be avoided in many cases when surgery is required, particularly in children.  Recent investigations in animals have raised concerns that ...

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

Partial bladder outlet obstruction (pBOO) has a high prevalence, causes significant patient burden, and immense health care costs. The most common animal model to investigate bladder remodeling in pBOO are female rodents undergoing partial obstruction at the proximal urethra. Variability in the degree of obstruction and animal mortality are major concerns with proximal obstruction. Furthermore, dissecting around the proximal urethra and bladder neck jeopardizes bladder innervation.
We developed a nerve-sparing mid-urethral obstruction (NeMO) model for pBOO avoiding the disadvantages of the traditional model. We approached the urethra just inferior to the pubic symphysis, which obviated the need for laparotomy as well as for dissection in this area; also, the striated urethral sphincter remained untouched. We performed NeMO in female Sprague-Dawley rats (12 obstructions, 6 sham animals) as well as in female C57/bl6 mice (20 obstructions, 18 sham animals). After two weeks, we evaluated bladder function, bladder mass, and body mass.
We had no mortalities among obstructed- or sham-operated female rats; as described for the traditional proximal pBOO-method, we tied the suture around the proximal urethra and a temporarily placed 0.9 mm metal rod. NeMO induced an 85% increase in bladder mass after two weeks, average residual urine volume was 0.4 mL in partially obstructed rats while only 0.03 mL in sham animals. In mice, we tested 3 sizes of cannulas that we placed along the urethra when tying the suture. We found that using a 27-gauge cannula resulted in over 50% animal mortality; placing the 25-gauge cannula did not yield the desired response in increasing bladder mass; utilizing a 26-gauge cannula yielded favorable results with minimal animal mortality (1/8) yet a significant 2-fold increase in bladder mass.

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

Traditional modeling of partial bladder outlet obstruction in rodents is fraught with animal mortality. A denervation injury from dissection around the proximal urethra and bladder neck is also of major concern. We developed and evaluated a safe and reliable mid-urethral obstruction model, avoiding the shortcomings of the traditional model.

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 ...