(a) Schematic of Bdnf transcript production. Transcription is initiated from promoters upstream of individual 5’-untranslated regions (UTRs) and spliced to the common coding exon IX. Each transcript …
(a-d) Representative images depicting male genitalia injuries observed in Bdnf-e1 breeder cages. Breeder males frequently show inflamed, infected, and/or mutilated genitalia (b–d) compared to normal …
Bdnf-e1 -/- virgin females enter all stages of the estrous cycle, including proestrous (P), estrous (E), diestrous (D), and metestrous (M), similar to WT females. Bdnf-e1 -/- virgins enter estrous ~e…
(a–b) qPCR demonstrating relative expression levels of oxytocin (a) and oxytocin receptor (b) transcripts in the HYP of 4 week old Bdnf-e1, -e2, -e4, and -e6 female mice (n = 4–5 per genotype; 1-way …
(a) Confocal z-projections of Oxt and Ntrk2 transcripts in brain slices containing the PVN from an adult female visualized with RNAscope in situ hybridization. TrkB transcripts (green) are highly …
(a-d) Confocal z-projections of PVN from OxtCre; TrkBflox/flox (a, d), TrkBflox/flox (b), and OxtCre (c) adult mice. Immunolabeling of OXT (a, b, c, d), TrkB (a’, b’, c’), and merged images (a”, b”, …
(a-c) Confocal z-projections of Oxt (a, b, c) and Ntrk2 (a’, b’, c’) transcripts in brain sections containing the PVN from OxtCre; TrkBflox/flox (a), OxtCre (b), and TrkBflox/flox (c) visualized …
(a-c) Maternal (a), abnormal parenting (b), and retrieval (c) indices for WT and OxtCre virgins during foreign pup retrieval test. OxtCre virgin females show no differences on these indices compared …
(a) Locus of the ribosomal protein Rpl22 in the Ribotag mouse and breeding strategy used to obtain OxtCre; Rpl22HA mice. (b) qPCR analysis validating significant enrichment of Oxt transcripts in IP …
(a) qPCR analysis validating significant enrichment (Myo5a, Ank2, Peg3, and Kmt2a) and de-enrichment (Agrp, Cartpt) of select transcripts in IP compared to Input fractions. (b–d) Representative …
(a) Plot depicting log2 fold changes of DEGs in Oxt IP vs. Oxt Input using RNA-seq V2 system compared to the SoLo RNA-seq system. Fold changes are highly correlated between kits. (b) Plot depicting …
(a) Strategy for TRAP in OXT neurons from control and Bdnf-e1 -/- mice (n = 3 per genotype) using AAV-DIO-RiboTag and RNA-seq. (b) Confocal image showing HA (white) and tdTom (red) expression in …
(a) Plot depicting log2 fold changes of DEGs in Oxt IP vs. Oxt Input compared to the log2 fold changes of DEGs in control IP vs. Bdnf-e1 -/- IP. As expected, there is no correlation between the two …
Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional Information |
---|---|---|---|---|
Genetic reagent (Mus musculus) | Bdnf-e1; Bdnf-e2 mice | PMID: 26585288 | ||
Genetic reagent (Mus musculus) | Oxt-Cre mice | Jackson Labs | RRID:IMSR_JAX:024234 | |
Genetic reagent (Mus musculus) | tdTomato mice | Jackson Labs | RRID:IMSR_JAX:007914 | |
Genetic reagent (Mus musculus) | Ntrk2 flox/ flox mice | PMID: 18511296 | ||
Genetic reagent (Mus musculus) | Cort-Cre mice | Jackson Labs | RRID:IMSR_JAX:010910 | |
Genetic reagent (Mus musculus) | RPL22-HA mice | Jackson Labs | RRID:IMSR_JAX:011029 | |
Genetic reagent (Adeno-assoicated virus) | AAV1-DIO-RPL22HA-GFP | PMID: 25855171 | ||
Antibody | anti-mouse HA | Covance | RRID:AB_291262 | |
Antibody | goat anti-mouse Alexa 647 | Thermo-Fisher Scientific | RRID:AB_141693 | |
Antibody | anti-mouse OXT | PMID: 3880813 | ||
Antibody | anti-rabbit cFOS | Millipore | RRID:AB_2631318 | |
Antibody | donkey anti-rabbit Alexa 488 | Thermo-Fisher Scientific | RRID:AB_141708 | |
Antibody | anti-rabbit TrkB H-181 | Santa Cruz | RRID:AB_2155274 | |
Commercial assay or kit | Ovation RNA-Seq V2 kit | Nugen | Cat#: 7102 | |
Commercial assay or kit | Ovation SoLo RNA-seq System Mouse | Nugen | Cat#: 0502–32 | |
Commercial assay or kit | KAPA Library Quantification Kit | KAPA Biosystems | Cat#: KR0405 | |
Commercial assay or kit | Miseq Reagent Kit v3 | Illumina | Cat#: MS-102–3001 | |
Commercial assay or kit | Ribogreen RNA assay kit | Invitrogen | Cat#: R11490 | |
Commercial assay or kit | RNAscope Fluorescent Multiplex V1 | Advanced Cell Diagnostics | Cat#: 320850 | |
Chemical compound, drug | A/G magnetic beads | Pierce | Cat#: 88803 | |
Chemical compound, drug | 10% NBF | Sigma | Cat#: HT501128 | |
Chemical compound, drug | Fluoromount G | Southern Biotechnology | Cat#: 0100–01 | |
Sequence- based reagent | Taqman probe Oxt | Life Technologies | Cat#: Mm01329577_g1 | |
Sequence- based reagent | Taqman probe Oxtr | Life Technologies | Cat#: Mm01182684_m1 | |
Sequence- based reagent | Taqman probe Gapdh | Life Technologies | Cat#: 4352932E | |
Sequence- based reagent | Taqman probe Myo5a | Life Technologies | Cat#: Mm00487823_m1 | |
Sequence- based reagent | Taqman probe Ank2 | Life Technologies | Cat#: Mm00618325_m1 | |
Sequence- based reagent | Taqman probe Peg3 | Life Technologies | Cat#: Mm01337379_m1 | |
Sequence- based reagent | Taqman probe Kmt2a | Life Technologies | Cat#: Mm01179235_m1 | |
Sequence- based reagent | Taqman probe Agrp | Life Technologies | Cat#: Mm00475829_g1 | |
Sequence- based reagent | Taqman probe Cartpt | Life Technologies | Cat#: Mm04210469_m1 | |
Sequence- based reagent | RNAscope probe Oxt | Advanced Cell Diagnostics | Cat#: 493171 | |
Sequence- based reagent | RNAscope probeNtrk2 | Advanced Cell Diagnostics | Cat#: 423611 | |
Sequence- based reagent | RNAscope probeGabra2 | Advanced Cell Diagnostics | Cat#: 435011 | |
Sequence- based reagent | RNAscope probeCckar | Advanced Cell Diagnostics | Cat#: 313751 | |
Sequence- based reagent | RNAscope probeNtrk2 (Exon S) | Advanced Cell Diagnostics | Cat#: 539481 | |
Sequence- based reagent | RNAscope probe Kmt2a | Advanced Cell Diagnostics | Cat#: 408951 | |
Sequence- based reagent | RNAscope probePeg3 | Advanced Cell Diagnostics | Cat#: 492581 | |
Sequence- based reagent | RNAscope probe Ank2 | Advanced Cell Diagnostics | Cat#: 413221 |
Table Legends (Excel tables attached as separate tabs in a single file).
Table S1. Differential gene expression analysis of OXT Input vs. IP samples for all expressed genes. Table S2. Comparison of significant OXT neuron gene enrichment using Nugen Ovation RNA-seq System V2 kit vs. Nugen Ovation SoLo RNA-seq system. Table also includes comparison of differential gene expression between OXT neurons, Cort inhibitory interneurons, and Ntsr1 layer VI corticothalamic neurons versus their respective input RNA samples, as well as an overall OXT-IP versus pooled Cort-IP and Ntsr1-IP samples to assess specificity for these genes that were enriched in OXT neurons. Table S3. Gene Ontology analysis in OXT neurons. Enriched and depleted GO terms from differential expression analysis of OXT Input vs. IP. Table S4. Differential gene expression analysis of OXT-IP vs. Cort-IP and Ntsr1-IP samples for all expressed genes. Table S5. Gene ontology enrichment analysis for genes differentially expressed in OXT-IP vs. Cort-IP and Ntsr1-IP samples Table S6. OXT enriched mouse model genes in SFARI. List of genes enriched in OXT neurons implicated in genetic animal models of autism spectrum disorder (ASD) by the Simons Foundation Autism Research Initiative (SFARI). Table S7. OXT enriched human genes in SFARI. List of genes enriched in OXT neurons implicated in ASD by the SFARI Human Gene Module using the subset of mouse-expressed genes with human homologs. Table S8. Differential gene expression analysis of control IP vs. Bdnf-e1 -/- IP samples. Table S9. Gene Ontology analysis in OXT neurons with disruption of BDNF signaling. GO terms enriched from differential expression analysis of control IP vs. Bdnf-e1 -/- IP samples. Table S10. BDNF-dependent OXT genes in SFARI. List of genes perturbed in OXT neurons following disruption of BDNF signaling that are implicated in autism spectrum disorder (ASD) by the Simons Foundation Autism Research Initiative (SFARI).