Oxytocin Signaling Modulation in Rodent Neurobiology

All drugs used in this study are listed in the reagents and resource table.

REAGENT OR RESOURCE	SOURCE	IDENTIFIER
Antibodies
Chicken anti-GFP primary antibody	Abcam	ab13970
Mouse NeuN primary antibody	Millipore	MAB377
Rabbit vGluT2 primary antibody	Synaptic systems	135 103
Rabbit anti-dsRed primary antibody	Living Colors	632496
GAD67	Millipore	MAB5406
Guinea-pig anti-Fluorogold primary antibody	Protos Biotech Corp	NM-101
Mouse c-Fos polyclonal primary antibody	Santa-Cruz	sc-8047
Mouse monoclonal anti-OT primary antibody	Provided by Dr. Harold Gainer	PS 38
DAPI	Vector Laboratories	H-1200-10
Synaptophysin	Abcam	Ab32127
Rabbit polyclonal anti-vGluT2 primary antibody	SYSY	135403
Bacterial and virus strains
rAAV1/2-OTp-Venus	Lab made	N/A
rAAV1/2-OTp-mCherry	Lab made	N/A
rAAV1/2-OTp-FLEX-GFP	Lab made	N/A
CAV2-CMVp-Cre	Montpellier Vectorology Platform	N/A
rAAV1/2-OTp-ChR2-mCherry	Lab made	N/A
rAAV1/2-OTp-C1V1-mCherry	Lab made	N/A
rAAV1/2-EF1αp-FLEX-GFP	Lab made	N/A
rAAV1/2-EF1αp-FLEX-mCherry	Lab made	N/A
rAAV1/2-EF1αp-FLEX-hM3D(Gq)-mCherry	Lab made	N/A
rAAV2/9-hSyn-OT1.0-sensor	Lab made	N/A
Chemicals, peptides, and recombinant proteins
Fluorogold^TM	Santa-Cruz	sc-358883
Oxytocin Receptor Antagonist	Merck	L-368,899
Deschloroclozapine	MedChemExpress	1977-07-7
TGOT	Bachem®	50-260-164
CFA	Sigma	32160405
dOVT	Bachem	d(CH2)5-Tyr(Me)-[Orn8]- vasotocine
Retrobeads^TM	Lumafluor	N/A
RNAscope reagents and materials
RNAscope HiPlex12 detection Reagents (488, 550, 647)	ACD	324106
RNAscope HiPlex Probe Diluent	ACD	324301
RNAscope Wash Buffer Reagents	ACD	310091
RNAscope Protease III & IV Reagents	ACD	322340
RNAscope HiPlex Cleaving Stock Solution	ACD	324136
RNAscope HiPlex CRE-T2 Probe	ACD	312281-T2
RNAscope HiPlex Rn-Oxtr-T6 Probe	ACD	483671-T6
RNAscope HiPlex12 Positive Control Probe – Rn	ACD	324331
RNAscope HiPlex12 Negative Control Probe	ACD	324341
UltraPure 20X SSC Buffer	invitrogen	15557-044
ProLong Gold antifade reagent	invitrogen	P36930
ImmEdge Hydrophobic Barrier Pen	VECTOR LABORATORIES	H-40000
Tween 20	Sigma-Aldrich	P1379
Experimental Models: Organisms/Strains
Rattus Norvegicus (Wistar HAN)	Janvier	N/A
Rattus Norvegicus (Sprague Dawleys)	Charles River	N/A
OTR-IRES-Cre (Wistar HAN)	Lab made	N/A
Software and Algorithms
Graphpad prism 7.05	www.graphpad.com	N/A
Fiji	www.imagej.net/Fiji	N/A
Adobe Photoshop CS5	www.adobe.com	N/A
Adobe Illustrator 16.05	www.adobe.com	N/A
CorelDraw 2020	www.coreldraw.com	N/A
HiPlex Image Registration Software v1.0	ACD	300065
Matlab	Mathworks	N/A
Python	Open-source	N/A
Zen (3.1 blue edition and 3.0 SR black edition)	Zeiss	N/A
Other
Optic fiber implants	www.thorlabs.com	CFMC12L10
473 nm Blue Laser Generator	www.dreamlasers.com	SDL-473-XXXT
Programmable Pulse Stimulator (A.M.P.I.)	www.ampi.co.il	Master-9

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Iwasaki M., Lefevre A., Althammer F., Clauss Creusot E., Łąpieś O., Petitjean H., Hilfiger L., Kerspern D., Melchior M., Küppers S., Krabichler Q., Patwell R., Kania A., Gruber T., Kirchner M.K., Wimmer M., Fröhlich H., Dötsch L., Schimmer J., Herpertz S.C., Ditzen B., Schaaf C.P., Schönig K., Bartsch D., Gugula A., Trenk A., Blasiak A., Stern J.E., Darbon P., Grinevich V, & Charlet A. (2023). An analgesic pathway from parvocellular oxytocin neurons to the periaqueductal gray in rats. Nature Communications, 14, 1066.

Publication 2023

Ampi Buffer Drugs Fiber Fluorogold Gold Ires Peptides Prism Protease iii Pulse Virus

Corresponding Organization :

Other organizations : Institut des Neurosciences Cellulaires et Intégratives, Université de Strasbourg, Central Institute of Mental Health, Van Andel Institute, Georgia State University, University Hospital Heidelberg, Jagiellonian University

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Variable analysis

independent variables

Optogenetic manipulation using rAAV1/2-OTp-ChR2-mCherry and rAAV1/2-OTp-C1V1-mCherry
Chemogenetic manipulation using rAAV1/2-EF1αp-FLEX-hM3D(Gq)-mCherry
Pharmacological intervention using Oxytocin Receptor Antagonist L-368,899 and TGOT

dependent variables

Neuronal activity (c-Fos expression)
Oxytocin receptor expression (RNAscope)

control variables

Viral vectors: rAAV1/2-OTp-Venus, rAAV1/2-OTp-mCherry, rAAV1/2-OTp-FLEX-GFP, rAAV1/2-EF1αp-FLEX-GFP, rAAV1/2-EF1αp-FLEX-mCherry
Retrograde tracer: Fluorogold
Antibodies for immunohistochemistry (NeuN, vGluT2, dsRed, GAD67, Fluorogold, c-Fos, OT)
Experimental animal models: Wistar HAN and Sprague Dawley rats, OTR-IRES-Cre rats

positive controls

RNAscope HiPlex12 Positive Control Probe – Rn

negative controls

RNAscope HiPlex12 Negative Control Probe

Annotations

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