Hyperglycemic Clamp and Hypoglycemic Clamp in Conscious Mice
1Department of Neuroscience for Metabolic Control, Graduate School of Medical Science, Kumamoto University.
Journal of Visualized Experiments : Jove
|February 12, 2024
Summary
This study introduces a new clamp method for mice to precisely measure insulin release and counter-regulatory responses (CRR) in diabetes research. This technique stabilizes blood glucose levels, improving the assessment of glucose metabolism and hormone dynamics.
Area of Science:
- Metabolic Research
- Endocrinology
- Animal Models
Background:
- Diabetes Mellitus (DM) involves impaired insulin release (Type 1) or sensitivity (Type 2).
- Current methods for studying insulin release and counter-regulatory responses (CRR) in rodent models have limitations in accurately assessing these dynamics due to persistent blood glucose fluctuations.
- Insulin therapy for DM can lead to hypoglycemia, necessitating understanding of CRR.
Purpose of the Study:
- To describe a novel clamp method for maintaining stable blood glucose levels in conscious mice.
- To enable precise measurement of insulin release during hyperglycemia and CRR hormone release during hypoglycemia.
- To provide a standardized experimental approach for researchers studying systemic glucose metabolism.
Main Methods:
- Surgical implantation of carotid artery and jugular vein catheters in mice for blood collection and infusion.
- Development of hyperglycemic clamp technique to maintain blood glucose at 250 mg/dL via glucose infusion, measuring insulin.
- Development of hypoglycemic clamp technique to maintain blood glucose at 50 mg/dL via variable glucose and insulin infusion, measuring CRR hormones.
Main Results:
- The described clamp methods allow for stable control of blood glucose levels at both hyperglycemic (250 mg/dL) and hypoglycemic (50 mg/dL) setpoints in conscious mice.
- This stable glucose control facilitates accurate measurement of insulin secretion and counter-regulatory hormone release.
- The consistent surgical and experimental setup for both clamps enhances comparability.
Conclusions:
- The developed hyperglycemic and hypoglycemic clamp techniques offer a robust and reproducible method for investigating glucose metabolism in rodent models.
- This approach overcomes limitations of previous methods by stabilizing blood glucose, allowing for more accurate assessment of insulin dynamics and CRR.
- This technique is valuable for advancing research in systemic glucose metabolism and diabetes pathophysiology.
