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Advan. Physiol. Edu. 27: 78-85, 2003; doi:0.1152/advan.00053.2002
1043-4046/03 $5.00
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ADV PHYSIOL EDUC 27:78-85, 2003
© 2003 American Physiological Society

TEACHING IN THE LABORATORY

UNDERSTANDING THE PHYSIOLOGY OF GLUTAMATE RECEPTORS BY USE OF A PROTOCOL FOR NEURONAL STAINING

João O. Malva, Ana P. Vieira and Catarina R. Oliveira

Center for Neuroscience and Cell Biology of Coimbra, Institute of Biochemistry, Faculty of Medicine, University of Coimbra, 3004–504 Coimbra, Portugal

Teaching students about the physiology of neurotransmitter receptors usually requires practical lessons with the use of sophisticated equipment and complex analysis of data. Here, we report our experience in teaching medical students with a simple, practical protocol that transforms the physiology of glutamate receptors into neuronal staining, observable under bright-field microscopy. Essentially, the students were challenged to selectively stain a subpopulation of cultured neurons expressing Ca2+-permeable {alpha}-amino-3-hydroxy-5-methylisoxazole-4-propionate (AMPA) receptors (a subgroup of ionotropic glutamate receptors). Neurons expressing this type of receptors were loaded with Co2+ (in substitution for Ca2+) after nondesensitizing activation of AMPA receptors. After precipitation, the Co2+ was revealed after treatment with silver. At the end of the procedure, the neurons expressing Ca2+-permeable AMPA receptors were visually identified under bright-field microscopy. The procedure allowed the visualization of the complete dendritic network of the stained neurons and allowed the students to learn very efficiently about the physiology of glutamate receptors.

Key words: neurons; cobalt staining; {alpha}-amino-3-hydroxy-5-methylisoxazole-4-propionate receptors; neurotransmitters






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