机能实验学 离体蛙心灌流
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L-type calcium channel Ryanodine receptor
Nacl
Wash out
The underlying mechanisms…
Plasma membrane
myofibrils
Membrane depolarization
muscle contraction
Longitudinal tubule (纵管/肌浆网)
Transverse tubule (横管)
• To investigate regulatห้องสมุดไป่ตู้on of the frog heart contraction by different factors.
1. Na+,K+, and Ca2+ ;
2. Acetylcholine (Ach) and Norepinephrine (NE)
Procedures
A classic and the most popular experiment in our school
Ringer's solution Transducer
Canula Frog heart
BL-420 System
Objectives
• To learn how to intubate and perfuse the frog heart;
Dorsal view
注意：在主动脉下面穿2根细线备用
Procedures
4. Ventricle intubation
ventricle
aorta sinistra
arterial cone
canula
关键步骤：将蛙心套管的尖端从主动脉插入动脉圆锥 插入心室内的标准：套管内液面随心脏跳动而上下移动
Procedures
5. Connect the frog heart to the transducer;
vena cava clip
sinus venosus atrium
ventricle
Dorsal view
注意：1. 切勿损伤静脉窦 2. 张力换能器稍向下倾斜
Procedures
6. Record the contraction of frog heart using BL420 system;
Procedures
3. Exposure the frog heart and observe its structure;
ventricle
arterial cone
atrium
Aorta sinistra
vena cava
sinus venosus atrium
ventricle
Ventral view
Observation items
• Replace Ringer's solution with 0.65% NaCl; • Add 1-2 drops of CaCl2 (3%) into the canula； • Add 1-2 drops of KCl (2%) into the canula ； • Add 1-2 drops of NE (norepinephrine,1:10000)
1. Sacrifice the frog by destroying its brain and spine;
foramen magnum(枕骨大孔)
Posterior auricular glands （ 耳后腺）
注意：“麻醉”成功的标准：蛙的四肢松软无力
Procedures
2. Hold the frog to an operating table in a dorsal position;
into the canula ； • Add 1-2 drops of Ach (acetylcholine,1:10000)
into the canula；
注意：每个项目后都需用新鲜任氏液冲洗套管， 待蛙心活动稳定后再进行下一项
The underlying mechanisms…
• How and why do different chemicals change the contraction of frog heart?