Abstract

Isoflurane (ISO) is a commonly used anesthetic that offers rapid recovery for laboratory animal research. Initial studies indicated no difference in arterial Pco2 () or pH between conscious (NO ISO) and 1% ISO-exposed CD-1 mice. Our laboratory investigated whether arterial blood sampling with 1% ISO is a suitable alternative to NO ISO sampling for monitoring ventilation in a commonly studied mouse strain. We hypothesized similar blood chemistry, breathing patterns, and cardiovascular responses with NO ISO and 1% ISO. C57BL/6J mice underwent unrestrained barometric plethysmography to quantify the pattern of breathing. Mice exposed to hypoxic and hypercapnic gas under 1% ISO displayed blunted responses; with air, there were no breathing differences. Blood pressure and heart rate were not different between NO ISO and 1% ISO-exposed mice breathing air. Oxygen saturation was not different between groups receiving 2% ISO, 1% ISO, or air. Breathing frequency stabilized at ~11 min of 1% ISO following 2% ISO exposure, suggesting that 11 min is the optimal time for a sample in C57BL/6J mice. Blood samples at 1% ISO and NO ISO revealed no differences in blood pH and in C57BL/6J mice. Overall, this method reveals similar arterial blood sampling values in awake and 1% ISO CD-1 and C57BL/6J mice exposed to air. Although this protocol may be appropriate in other mouse strains when a conscious sample is not feasible, caution is warranted first to identify breathing frequency responses at 1% ISO to tailor the protocol.NEW & NOTEWORTHY Conscious arterial blood sampling is influenced by extraneous factors and is a challenging method due to the small size of mice. Through a series of experiments, we show that arterial blood sampling with 1% isoflurane (ISO) is an alternative to awake sampling in C57BL/6J and CD-1 male mice breathing air. Monitoring breathing frequency during 1% ISO is important to the protocol and should be closely followed to confirm adequate recovery after the catheter implantation.

Highlights

  • Conscious arterial blood sampling is the gold standard to confirm ventilation changes in humans and animals

  • The same CD-1 mice had samples collected with not different between conscious (NO ISO) and ISO

  • Blood sampling revealed no difference in pH and PaCO2 between NO ISO and 1% isoflurane (1% ISO) CD-1 mice using the 5-min time point

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Summary

Introduction

Conscious arterial blood sampling is the gold standard to confirm ventilation changes in humans and animals. Other methods of arterial blood collection include cardiac puncture in mice [7, 37, 39], and following decapitation [6] These techniques are suitable for terminal experiments, we were interested in a protocol for chronic studies of ventilation that would be representative of quiet breathing in mice. Several measures are available for venous sampling (2a), but to monitor ventilation it is important to sample from the arterial system (reviewed in Ref. 8) Results from this investigation are applicable to respiratory physiology experiments, and to other studies that use a nonsurgical plane of anesthesia, such as noninvasive imaging and ultrasound [29, 32]

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