From synapse to behavior: rapid modulation of defined neuronal types with engineered GABAA receptors.
In mammals, identifying the contribution of specific neurons or networks to behavior is a key challenge. Here we describe an approach that facilitates this process by enabling the rapid modulation of synaptic inhibition in defined cell populations. Binding of zolpidem, a systemically active allosteric modulator that enhances the function of the GABAA receptor, requires a phenylalanine residue (Phe77) in the gamma2 subunit. Mice in which this residue is changed to isoleucine are insensitive to zolpidem. By Cre recombinase-induced swapping of the gamma2 subunit (that is, exchanging Ile77 for Phe77), zolpidem sensitivity can be restored to GABAA receptors in chosen cell types. We demonstrate the power of this method in the cerebellum, where zolpidem rapidly induces significant motor deficits when Purkinje cells are made uniquely sensitive to its action. This combined molecular and pharmacological technique has demonstrable advantages over targeted cell ablation and will be invaluable for investigating many neuronal circuits.
Wulff
Peer
P
Department of Clinical Neurobiology, University of Heidelberg, Im Neuenheimer Feld 364, 69120 Heidelberg, Germany. p.wulff@abdn.ac.uk
Goetz
Thomas
T
Leppä
Elli
E
Linden
Anni-Maija
AM
Renzi
Massimiliano
M
Swinny
Jerome D
JD
Vekovischeva
Olga Y
OY
Sieghart
Werner
W
Somogyi
Peter
P
Korpi
Esa R
ER
Farrant
Mark
M
Wisden
William
W
D000818:Animals; D001345:Autoradiography; D001522:Behavior, Animal / Q000187:drug effects*; D004594:Electrophysiology; D005260:Female; D018755:GABA Agonists / Q000494:pharmacology; D005838:Genotype; D049452:Green Fluorescent Proteins / Q000235:genetics; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D008297:Male; D051379:Mice; D018345:Mice, Knockout; D009474:Neurons / Q000187:drug effects / Q000502:physiology*; D018408:Patch-Clamp Techniques; D004856:Postural Balance / Q000187:drug effects / Q000502:physiology; D015202:Protein Engineering; D011689:Purkinje Cells / Q000187:drug effects / Q000502:physiology; D011725:Pyridines / Q000494:pharmacology; D011963:Receptors, GABA-A / Q000235:genetics / Q000502:physiology*; D018080:Receptors, GABA-B / Q000235:genetics / Q000502:physiology; D013569:Synapses / Q000187:drug effects / Q000502:physiology*; D000077334:Zolpidem
D018755:GABA Agonists; C485504:Gabbr2 protein, mouse; D011725:Pyridines; D011963:Receptors, GABA-A; D018080:Receptors, GABA-B; D049452:Green Fluorescent Proteins; D000077334:Zolpidem