title: Linking genetically defined neurons to behavior through a broadly applicable silencing allele.
abstract: Tools for suppressing synaptic transmission gain power when able to target highly selective neuron subtypes, thereby sharpening attainable links between neuron type, behavior, and disease; and when able to silence most any neuron subtype, thereby offering broad applicability. Here, we present such a tool, RC::PFtox, that harnesses breadth in scope along with high cell-type selection via combinatorial gene expression to deliver tetanus toxin light chain (tox), an inhibitor of vesicular neurotransmission. When applied in mice, we observed cell-type-specific disruption of vesicle exocytosis accompanied by loss of excitatory postsynaptic currents and commensurately perturbed behaviors. Among various test populations, we applied RC::PFtox to silence serotonergic neurons, en masse or a subset defined combinatorially. Of the behavioral phenotypes observed upon en masse serotonergic silencing, only one mapped to the combinatorially defined subset. These findings provide evidence for separability by genetic lineage of serotonin-modulated behaviors; collectively, these findings demonstrate broad utility of RC::PFtox for dissecting neuron functions.
authors.affiliation: Department of Genetics, Harvard Medical School, Boston, MA 02115, USA.
authors.forename: Jun Chul
authors.initials: JC
authors.lastname: Kim
authors.forename: Melloni N
authors.initials: MN
authors.lastname: Cook
authors.forename: Megan R
authors.initials: MR
authors.lastname: Carey
authors.forename: Chung
authors.initials: C
authors.lastname: Shen
authors.forename: Wade G
authors.initials: WG
authors.lastname: Regehr
authors.forename: Susan M
authors.initials: SM
authors.lastname: Dymecki
mesh_terms: D000161:Acoustic Stimulation / Q000379:methods; D000704:Analysis of Variance; D000818:Animals; D000831:Animals, Newborn; D051792:Basic Helix-Loop-Helix Proteins / Q000378:metabolism; D001522:Behavior, Animal / Q000502:physiology*; D001703:Biophysics; D002531:Cerebellum / Q000033:anatomy & histology / Q000166:cytology / Q000378:metabolism; D003213:Conditioning, Psychological / Q000502:physiology; D004558:Electric Stimulation / Q000379:methods; D005106:Exploratory Behavior / Q000502:physiology; D005239:Fear / Q000502:physiology; D018756:GABA Antagonists / Q000494:pharmacology; D008040:Genetic Linkage / Q000502:physiology*; D049452:Green Fluorescent Proteins / Q000235:genetics; D066298:In Vitro Techniques; D018782:Maze Learning / Q000502:physiology; D051379:Mice; D008822:Mice, Transgenic; D046529:Microscopy, Electron, Transmission / Q000379:methods; D008959:Models, Neurological; D009419:Nerve Tissue Proteins / Q000378:metabolism; D009474:Neurons / Q000502:physiology* / Q000648:ultrastructure; D018408:Patch-Clamp Techniques; D010641:Phenotype; D010721:Phosphinic Acids / Q000494:pharmacology; D011412:Propanolamines / Q000494:pharmacology; D011506:Proteins / Q000235:genetics; D022661:RNA, Untranslated; D045522:Recombinases / Q000235:genetics; D013216:Reflex, Startle / Q000235:genetics; D012701:Serotonin / Q000378:metabolism; D009435:Synaptic Transmission / Q000235:genetics / Q000502:physiology*; D013744:Tetanus Toxin / Q000737:chemistry / Q000378:metabolism; D050988:Vesicle-Associated Membrane Protein 2 / Q000378:metabolism
chemical_list: D051792:Basic Helix-Loop-Helix Proteins; D018756:GABA Antagonists; D049452:Green Fluorescent Proteins; D009419:Nerve Tissue Proteins; D010721:Phosphinic Acids; D011412:Propanolamines; D011506:Proteins; D022661:RNA, Untranslated; D045522:Recombinases; D012701:Serotonin; D013744:Tetanus Toxin; D050988:Vesicle-Associated Membrane Protein 2; C500943:Atoh1 protein, mouse; C110853:Gt(ROSA)26Sor non-coding RNA, mouse; C085075:CGP 55845A
references.citation: J Neurosci. 2003 Jul 30;23(17):6759-67
references.pmid: 12890769
references.citation: Gene. 1991 Dec 15;108(2):193-9
references.pmid: 1660837
references.citation: J Neurosci. 1999 Jul 15;19(14):6157-68
references.pmid: 10407051
references.citation: Neuron. 2006 Apr 20;50(2):205-18
references.pmid: 16630833
references.citation: Neuropsychopharmacology. 2000 Jun;22(6):650-9
references.pmid: 10788764
references.citation: Genes Brain Behav. 2003 Dec;2(6):365-80
references.pmid: 14653308
references.citation: Science. 1994 Sep 23;265(5180):1875-8
references.pmid: 8091214
references.citation: Science. 2008 Feb 29;319(5867):1260-4
references.pmid: 18218862
references.citation: Dev Dyn. 2005 Nov;234(3):633-50
references.pmid: 16145671
references.citation: Psychopharmacology (Berl). 1995 Jan;117(1):41-8
references.pmid: 7724701
references.citation: Proc Natl Acad Sci U S A. 1990 Apr;87(7):2662-5
references.pmid: 1969639
references.citation: Neuron. 2008 Mar 13;57(5):634-60
references.pmid: 18341986
references.citation: Behav Brain Res. 2000 Jul;112(1-2):145-50
references.pmid: 10862945
references.citation: Psychopharmacology (Berl). 2001 Jul;156(2-3):234-58
references.pmid: 11549226
references.citation: Nat Genet. 2000 Jun;25(2):139-40
references.pmid: 10835623
references.citation: Proc Natl Acad Sci U S A. 1992 Apr 1;89(7):3020-4
references.pmid: 1557407
references.citation: J Neurosci. 1982 Oct;2(10):1499-506
references.pmid: 6750049
references.citation: Nat Genet. 2003 Sep;35(1):70-5
references.pmid: 12923530
references.citation: Physiol Rev. 2000 Apr;80(2):717-66
references.pmid: 10747206
references.citation: J Neurosci. 2000 Feb 15;20(4):1374-85
references.pmid: 10662828
references.citation: Acta Physiol (Oxf). 2006 May-Jun;187(1-2):329-44
references.pmid: 16734770
references.citation: Neuron. 2005 Oct 6;48(1):31-43
references.pmid: 16202707
references.citation: Neuropsychopharmacology. 1999 Aug;21(2 Suppl):52S-60S
references.pmid: 10432489
references.citation: Eur J Pharmacol. 2000 Sep 15;404(1-2):1-12
references.pmid: 10980257
references.citation: Chin J Physiol. 1996;39(3):139-46
references.pmid: 8955560
references.citation: J Neurosci. 1999 Dec 1;19(23):10348-56
references.pmid: 10575032
references.citation: Genesis. 2000 Nov-Dec;28(3-4):93-8
references.pmid: 11105049
references.citation: Acta Physiol Scand Suppl. 1964;:SUPPL 232:1-55
references.pmid: 14229500
references.citation: Neuron. 2004 Aug 5;43(3):345-57
references.pmid: 15294143
references.citation: Behav Neurosci. 1995 Jun;109(3):404-12
references.pmid: 7662151
references.citation: Genes Dev. 2000 Jun 1;14(11):1377-89
references.pmid: 10837030
references.citation: Methods Enzymol. 1993;225:890-900
references.pmid: 8231893
references.citation: Genesis. 2007 Sep;45(9):593-605
references.pmid: 17868096
references.citation: Proc Natl Acad Sci U S A. 1997 Apr 15;94(8):3789-94
references.pmid: 9108056
references.citation: Neuron. 2004 May 27;42(4):553-66
references.pmid: 15157418
references.citation: BMC Dev Biol. 2001;1:4
references.pmid: 11299042
references.citation: Nat Neurosci. 2008 Apr;11(4):417-9
references.pmid: 18344997
references.citation: J Neurosci. 1996 Dec 15;16(24):7859-67
references.pmid: 8987814
references.citation: Mol Pharmacol. 1998 Apr;53(4):649-55
references.pmid: 9547354
references.citation: Cell. 2005 May 6;121(3):479-92
references.pmid: 15882628
references.citation: J Neurosci. 2007 Sep 12;27(37):9817-23
references.pmid: 17855595
references.citation: Mamm Genome. 2007 Aug;18(8):559-72
references.pmid: 17629744
references.citation: Brain Res. 1998 Apr 20;790(1-2):178-84
references.pmid: 9593883
references.citation: Neuron. 2007 Apr 5;54(1):17-34
references.pmid: 17408575
references.citation: Science. 2004 Oct 29;306(5697):879-81
references.pmid: 15514160
references.citation: Behav Neurosci. 1993 Apr;107(2):377-88
references.pmid: 8484901
references.citation: Psychopharmacology (Berl). 1997 Jul;132(2):125-34
references.pmid: 9266609
references.citation: Neuropsychopharmacology. 2003 Dec;28(12):2138-47
references.pmid: 12888784
references.citation: Mt Sinai J Med. 2006 Nov;73(7):941-9
references.pmid: 17195879
references.citation: Neuron. 2002 Dec 19;36(6):1115-26
references.pmid: 12495626
references.citation: Nat Neurosci. 2008 Sep;11(9):1001-3
references.pmid: 19160496