Rousseau Jean-Philippe, Caravagna Céline
Department of Pediatrics, Laval University.
Department of Pediatrics, Laval University;
J Vis Exp. 2015 Nov 19(105):53071. doi: 10.3791/53071.
While it is well known that the central respiratory drive is located in the brainstem, several aspects of its basic function, development, and response to stimuli remain to be fully understood. To overcome the difficulty of accessing the brainstem in the whole animal, isolation of the brainstem and part of the spinal cord is performed. This preparation is maintained in artificial cerebro-spinal fluid where gases, concentrations, and temperature are controlled and monitored. The output signal from the respiratory network is recorded by a suction electrode placed on the fourth ventral root. In this manner, stimuli can be directly applied onto the brainstem, and the effect can be recorded directly. The signal recorded is linked to the inspiratory signal sent to the diaphragm via the phrenic nerve, and can be described as bursts (around 8 bursts per minute). Analysis of these bursts (frequency, amplitude, length, and area under the curve) allows precise characterization of the stimulus effect on the respiratory network. The main limitation of this method is the viability of the preparation beyond the early post-natal stages. Thus, this method greatly focuses on the study of the whole network without the peripheral inputs in the newborn rat.
虽然众所周知中枢呼吸驱动位于脑干,但其基本功能、发育以及对刺激的反应等几个方面仍有待充分了解。为了克服在完整动物中进入脑干的困难,需进行脑干和部分脊髓的分离。该标本置于人工脑脊液中,其中气体、浓度和温度均受到控制和监测。呼吸网络的输出信号通过置于第四腹根的吸引电极进行记录。通过这种方式,可以直接将刺激施加于脑干,并直接记录其效果。所记录的信号与经由膈神经发送至膈肌的吸气信号相关联,可描述为爆发(每分钟约8次爆发)。对这些爆发(频率、幅度、时长和曲线下面积)进行分析,能够精确表征刺激对呼吸网络的影响。该方法的主要局限性在于出生后早期阶段之后标本的存活能力。因此,此方法主要聚焦于对新生大鼠中无外周输入的整个网络的研究。