Name: in vitro measurements of tracheal constriction using mice
Uploaded: 2012-06-25T12:00:00
Description: Watch this Scientific Journal Video about In vitro Measurements of Tracheal Constriction Using Mice at JoVE.com

وقد تم تمويل هذا العمل من خلال منحة من مركز للإبداع في منع وعلاج أمراض مجرى الهواء، NINDS منحة (NS052574)، و من برنامج ساندلر للأبحاث الربو.

1. معدات ويبين المكونات الرئيسية للانكماش جهاز قياس تخطيطي في الشكل 2A). <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> وهناك حمام الأنسجة، وحمام الأنسجة يحافظ على حل الاوكسيجين الفسيولوجية في درجة حرارة دافئ...

<ol>
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بروتوكول المقدمة هنا يوفر إعداد فسيولوجية لتقييم مجرى الهواء وظيفة العضلات. نعمل عموما 3-4 محضرات الاستحمام الجهاز في وقت واحد، ومع ذلك، نظم الجاهزة المتوفرة من عدد من الموردين والتي تسمح للقياسات في وقت واحد يصل الى 8 التحضيرات (ADInstruments، أدوات دقيقة العالم، وجهاز هار...

<table border="1" style=";text-align:right;direction:rtl"><tbody><tr><td> اسم كاشف </td><td> شركة </td><td> فهرس العدد </td><td> تعليقات </td></tr><tr><td> التناظرية الرقمية المحول </td><td> ADInstruments </td><td> PowerLab 4/35 </td><td></td></tr><tr><td> كرباكول (كلوريد Carbamoylcholine) </td><td> سيغما الدريخ </td><td> C4832 </td><td> 10 -2 م في المياه (التي يمكن تخزينها في aliquots -20 درجة مئوية) </td></tr><tr><td> رسم برامج </td><td> ADInstrtuments </td><td> LabChart </td><td></td></tr><tr><td> التدفئة المروج </td><td> Haake </td><td> خلاط طاحونة MM400 </td><td></td></tr><tr><td> متساوي القياس قوة محول </td><td> كينت العلمية </td><td> TRN001 </td><td></td></tr><tr><td> منشط </td><td> عشب تقنيات </td><td> S88 جامعة السلطان قابوس الناتج المزدوجهي نبض محفز </td><td></td></tr><tr><td> أنسجة الحمام </td><td> WPI </td><td> 47264 </td><td></td></tr></tbody></table>

in vitro measurements of tracheal constriction using mice

Transgenic and knockout mice have been powerful tools for the investigation of the physiology and pathophysiology of airways1,2. In vitro tensometry of isolated tracheal preparations has proven to be a useful assay of airway smooth muscle (ASM) contractile response in genetically modified mice. These in vitro tracheal preparations are relatively simple, provide a robust response, and retain both functional cholinergic nerve endings and muscle responses, even after long incubations.
Tracheal tensometry also provides a functional assay to study a variety of second messenger signaling pathways that affect contraction of smooth muscle. Contraction in trachea is primarily mediated by parasympathetic, cholinergic nerves that release acetylcholine onto ASM (Figure 1). The major ASM acetylcholine receptors are muscarinic M2 and M3 which are Gi/o and Gq coupled receptors, respectively3,4,5. M3 receptors evoke contraction by coupling to Gq to activate phospholipase C, increase IP3 production and IP3-mediated calcium release from the sarcoplasmic reticulum3,6,7. M2/Gi/o signaling is believed to enhance contractions by inhibition of adenylate cyclase leading to a decrease in cAMP levels5,8,9,10. These pathways constitute the so called "pharmaco-contraction coupling" of airway smooth muscle11. In addition, cholinergic signaling through M2 receptors (and modulated by M3 signaling) involves pathways that depolarize the ASM which in turn activate L-type, voltage-dependent calcium channels (Figure 1) and calcium influx (so called "excitation-contraction coupling")4,7. More detailed reviews on signaling pathways controlling airway constriction can be found4,12. The above pathways appear to be conserved between mice and other species. However, mouse tracheas differ from other species in some signaling pathways. Most prominent is their lack of contractile response to histamine and adenosine13,14, both well-known ASM modulators in humans and other species5,15.
Here we present protocols for the isolation of murine tracheal rings and the in vitro measurement of their contractile output. Included are descriptions of the equipment configuration, trachea ring isolation and contractile measurements. Examples are given for evoking contractions indirectly using high potassium stimulation of nerves and directly by depolarization of ASM muscle to activate voltage-dependent calcium influx (1. high K+, Figure 1). In addition, methods are presented for stimulations of nerves alone using electric field stimulation (2. EFS, Figure 1), or for direct stimulation of ASM muscle using exogenous neurotransmitter applied to the bath (3. exogenous ACH, Figure 1). This flexibility and ease of preparation renders the isolated trachea ring model a robust and functional assay for a number of signaling cascades involved in airway smooth muscle contraction.

Watch this Scientific Journal Video about In vitro Measurements of Tracheal Constriction Using Mice at JoVE.com

In vitro Measurements of Tracheal Constriction Using Mice

وكانت الفئران المعدلة وراثيا مفيدة للغاية في وظيفة فسيولوجية يرجع إلى الجينات. كما بحث، هذه بصورة عامة، والدراسات الفنية للمجرى الهواء، على وجه الخصوص، شهدت تحولا ملحوظا نحو نماذج الفئران. هنا نقدم بروتوكولات لل<em&gt; في المختبر</em&gt; دراسات انقباض القصبة الهوائية لتقييم وظيفة العضلات الملساء في مجرى الهواء الفئران.

في قياسات المختبر من انقباض القصبة الهوائية باستخدام الفئران

Transgenic and knockout mice have been powerful tools for the investigation of the physiology and pathophysiology of airways1,2. In vitro tensometry of ...

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In vitro Measurements of Tracheal Constriction Using Mice

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