Linking genetically defined neurons to behavior through a broadly applicable silencing allele.
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.
Kim
Jun Chul
JC
Department of Genetics, Harvard Medical School, Boston, MA 02115, USA.
Cook
Melloni N
MN
Carey
Megan R
MR
Shen
Chung
C
Regehr
Wade G
WG
Dymecki
Susan M
SM
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
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
J Neurosci. 2003 Jul 30;23(17):6759-67
12890769
Gene. 1991 Dec 15;108(2):193-9
1660837
J Neurosci. 1999 Jul 15;19(14):6157-68
10407051
Neuron. 2006 Apr 20;50(2):205-18
16630833
Neuropsychopharmacology. 2000 Jun;22(6):650-9
10788764
Genes Brain Behav. 2003 Dec;2(6):365-80
14653308
Science. 1994 Sep 23;265(5180):1875-8
8091214
Science. 2008 Feb 29;319(5867):1260-4
18218862
Dev Dyn. 2005 Nov;234(3):633-50
16145671
Psychopharmacology (Berl). 1995 Jan;117(1):41-8
7724701
Proc Natl Acad Sci U S A. 1990 Apr;87(7):2662-5
1969639
Neuron. 2008 Mar 13;57(5):634-60
18341986
Behav Brain Res. 2000 Jul;112(1-2):145-50
10862945
Psychopharmacology (Berl). 2001 Jul;156(2-3):234-58
11549226
Nat Genet. 2000 Jun;25(2):139-40
10835623
Proc Natl Acad Sci U S A. 1992 Apr 1;89(7):3020-4
1557407
J Neurosci. 1982 Oct;2(10):1499-506
6750049
Nat Genet. 2003 Sep;35(1):70-5
12923530
Physiol Rev. 2000 Apr;80(2):717-66
10747206
J Neurosci. 2000 Feb 15;20(4):1374-85
10662828
Acta Physiol (Oxf). 2006 May-Jun;187(1-2):329-44
16734770
Neuron. 2005 Oct 6;48(1):31-43
16202707
Neuropsychopharmacology. 1999 Aug;21(2 Suppl):52S-60S
10432489
Eur J Pharmacol. 2000 Sep 15;404(1-2):1-12
10980257
Chin J Physiol. 1996;39(3):139-46
8955560
J Neurosci. 1999 Dec 1;19(23):10348-56
10575032
Genesis. 2000 Nov-Dec;28(3-4):93-8
11105049
Acta Physiol Scand Suppl. 1964;:SUPPL 232:1-55
14229500
Neuron. 2004 Aug 5;43(3):345-57
15294143
Behav Neurosci. 1995 Jun;109(3):404-12
7662151
Genes Dev. 2000 Jun 1;14(11):1377-89
10837030
Methods Enzymol. 1993;225:890-900
8231893
Genesis. 2007 Sep;45(9):593-605
17868096
Proc Natl Acad Sci U S A. 1997 Apr 15;94(8):3789-94
9108056
Neuron. 2004 May 27;42(4):553-66
15157418
BMC Dev Biol. 2001;1:4
11299042
Nat Neurosci. 2008 Apr;11(4):417-9
18344997
J Neurosci. 1996 Dec 15;16(24):7859-67
8987814
Mol Pharmacol. 1998 Apr;53(4):649-55
9547354
Cell. 2005 May 6;121(3):479-92
15882628
J Neurosci. 2007 Sep 12;27(37):9817-23
17855595
Mamm Genome. 2007 Aug;18(8):559-72
17629744
Brain Res. 1998 Apr 20;790(1-2):178-84
9593883
Neuron. 2007 Apr 5;54(1):17-34
17408575
Science. 2004 Oct 29;306(5697):879-81
15514160
Behav Neurosci. 1993 Apr;107(2):377-88
8484901
Psychopharmacology (Berl). 1997 Jul;132(2):125-34
9266609
Neuropsychopharmacology. 2003 Dec;28(12):2138-47
12888784
Mt Sinai J Med. 2006 Nov;73(7):941-9
17195879
Neuron. 2002 Dec 19;36(6):1115-26
12495626
Nat Neurosci. 2008 Sep;11(9):1001-3
19160496