Membrane potential dye imaging of ventromedial hypothalamus neurons from adult mice to study glucose sensing

• Published in: Journal of visualized experiments Vol. 81; no. 81; pp. 1 - 9
• Main Authors: Vazirani, Reema P, Fioramonti, Xavier, Routh, Vanessa H
• Format: Journal Article
• Language: English
• Published: United States JoVE 2013; MyJove Corporation
• Subjects: Animal biology; Animals; Development Biology; Fluorescent Dyes - chemistry; Glucose - metabolism; Life Sciences; Male; Membrane Potentials - physiology; Mice; Mice, Inbred C57BL; Neurons - cytology; Neurons - metabolism; Neurons - physiology; Neurons and Cognition; Neuroscience; Spectrometry, Fluorescence - methods; Staining and Labeling - methods; Ventromedial Hypothalamic Nucleus - cytology; Ventromedial Hypothalamic Nucleus - metabolism; Ventromedial Hypothalamic Nucleus - physiology; inbred c57bl; nucleus/cytology/metabolism/ physiology; glucose/ metabolism; fluorescence/ methods; neurons/cytology/metabolism/ physiology; ventromedial hypothalamic; fluorescent dyes/ chemistry; mice; staining and labeling/ methods; membrane potentials/physiology; spectrometry; animals; male
• ISSN: 1940-087X; 1940-087X
• DOI: 10.3791/50861

Studies of neuronal activity are often performed using neurons from rodents less than 2 months of age due to the technical difficulties associated with increasing connective tissue and decreased neuronal viability that occur with age. Here, we describe a methodology for the dissociation of healthy hypothalamic neurons from adult-aged mice. The ability to study neurons from adult-aged mice allows the use of disease models that manifest at a later age and might be more developmentally accurate for certain studies. Fluorescence imaging of dissociated neurons can be used to study the activity of a population of neurons, as opposed to using electrophysiology to study a single neuron. This is particularly useful when studying a heterogeneous neuronal population in which the desired neuronal type is rare such as for hypothalamic glucose sensing neurons. We utilized membrane potential dye imaging of adult ventromedial hypothalamic neurons to study their responses to changes in extracellular glucose. Glucose sensing neurons are believed to play a role in central regulation of energy balance. The ability to study glucose sensing in adult rodents is particularly useful since the predominance of diseases related to dysfunctional energy balance (e.g. obesity) increase with age.