In vivo labeling of parvalbumin-positive interneurons and analysis of electrical coupling in identified neurons.
GABAergic interneurons can pace the activity of principal cells and are thus critically involved in the generation of oscillatory and synchronous network activity. The specific role of various GABAergic subpopulations, however, has remained elusive. This is in part attributable to the scarcity of certain GABAergic neurons and the difficulty of identifying them in slices obtained from brain regions in which anatomical structures are not readily recognizable in the live preparation. To facilitate the functional analysis of GABAergic interneurons, we generated transgenic mice in which the enhanced green fluorescent protein (EGFP) was specifically expressed in parvalbumin-positive neurons. The high fidelity of expression obtained using bacterial artificial chromosome transgenes resulted in EGFP-labeled neurons in nearly all brain regions known to contain parvalbumin-expressing neurons. Immunocytochemical analysis showed that EGFP expression was primarily restricted to parvalbumin-positive cells. In addition to cell body labeling, EGFP expression was high enough in many neurons to enable the visualization of dendritic structures. With the help of these mice, we investigated the presence of electrical coupling between parvalbumin-positive cells in brain slices obtained from young and adult animals. In dentate gyrus basket cells, electrical coupling was found in slices from young [postnatal day 14 (P14)] and adult (P28 and P42) animals, but both strength and incidence of coupling decreased during development. However, electrical coupling between parvalbumin-positive multipolar cells in layer II/III of the neocortex remains unaltered during development. Yet another developmental profile of electrical coupling was found between layer II/III parvalbumin-positive cells and excitatory principal cells. Between these neurons, electrical coupling was found at P14 but not at P28. The results indicate that the presence and strength of electrical coupling is developmentally regulated with respect to brain area and cell type.
Meyer
Axel H
AH
Department of Clinical Neurobiology, Interdisciplinary Center for Neurosciences, University of Heidelberg, 69120 Heidelberg, Germany.
Katona
István
I
Blatow
Maria
M
Rozov
Andrei
A
Monyer
Hannah
H
D000200:Action Potentials / Q000502:physiology; D000367:Age Factors; D000818:Animals; D001921:Brain / Q000166:cytology / Q000378:metabolism; D022202:Chromosomes, Artificial, Bacterial / Q000378:metabolism; D017629:Gap Junctions / Q000378:metabolism; D015870:Gene Expression; D006624:Hippocampus / Q000166:cytology / Q000378:metabolism; D007150:Immunohistochemistry; D066298:In Vitro Techniques; D007395:Interneurons / Q000166:cytology / Q000378:metabolism; D008164:Luminescent Proteins / Q000235:genetics; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D009474:Neurons / Q000166:cytology / Q000502:physiology*; D010320:Parvalbumins / Q000096:biosynthesis* / Q000235:genetics; D018408:Patch-Clamp Techniques; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D009435:Synaptic Transmission / Q000502:physiology*; D005680:gamma-Aminobutyric Acid / Q000378:metabolism
D008164:Luminescent Proteins; D010320:Parvalbumins; D011993:Recombinant Fusion Proteins; D005680:gamma-Aminobutyric Acid