Taar1 gene variants have a causal role in methamphetamine intake and response and interact with Oprm1
- Oregon Health & Science University, United States
- Oregon National Primate Research Center, United States
- University of Tennessee Health Sciences Center, United States
- Veterans Affairs Portland Health Care System, United States
Figures
Figure 1
Knock-in of Taar1+ converts high MA intake to low MA intake, but does not impact total volume of fluid consumed.
(a) MA consumption and (b) total volume consumed, when MA was offered vs. water for 18h/day at a concentration of 20 or 40 mg/l for control and knock-in mice. Presented are means (represented by …
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Figure 1—source data 1
MA consumption (mg/kg/18h) and total volume consumed (ml/18h) data for male and female, control MAHDR-Taar1m1J/m1J and knock-in MAHDR-Taar1+/+ mice.
- https://doi.org/10.7554/eLife.46472.004
Figure 2
Knock-in of Taar1+ restores a hypothermic response to MA.
Body temperatures obtained immediately before (T0) or 30–120 min (T30-120) after (a) saline or (b) 2 mg/kg MA treatment for control and knock-in mice. Presented are means ± SEM for data collapsed on …
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Figure 2—source data 1
Core body temperature (°C) data across time for saline and 2 mg/kg MA-treated male and female, control MAHDR-Taar1m1J/m1J and knock-in MAHDR-Taar1+/+ mice.
- https://doi.org/10.7554/eLife.46472.006
Figure 3
MA consumption and total volume consumed among BXD strains are strain-dependent.
MA consumption when MA was offered vs. water for 18h/day at a concentration of (a) 20 mg/l or (b) 40 mg/l. (c) Total volume consumed from the water and MA tubes during the 18h period when MA was …
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Figure 3—source data 1
MA consumption (mg/kg/18h) and total volume consumed (ml/18h) data for male and female, BXD mice, according to Taar1 and Oprm1 genotype.
- https://doi.org/10.7554/eLife.46472.008
Figure 4
MA consumption is impacted in BXD mice by a Taar1 x Oprm1 interaction.
(a) MA consumption and (b) total volume consumed, when MA was offered vs. water for 18h/day at a concentration of 20 or 40 mg/l for BXD strain mice with different combined Taar1/Oprm1 genotypes. …
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Figure 4—source data 1
MA consumption (mg/kg/18h) and total volume consumed (ml/18h) data for male and female BXD mice, according to Taar1 and Oprm1 genotype.
- https://doi.org/10.7554/eLife.46472.010
Figure 5
Body temperature among BXD strains treated with saline or MA is strain-dependent.
Body temperatures obtained just before (T0) or 30–120 min (T30-120) after injection for (a) saline-treated Taar1+/+ mice of either Oprm1 genotype, (b) saline-treated Taar1m1J/m1J mice of either Oprm1…
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Figure 5—source data 1
Core body temperature (°C) data across time for saline and 2 mg/kg MA-treated male and female, BXD mice, according to Taar1 and Oprm1 genotype.
- https://doi.org/10.7554/eLife.46472.013
Figure 6
MA-induced hypothermia is impacted in BXD mice by a Taar1 x Oprm1 interaction.
Body temperatures obtained just before (T0) or 30–120 min (T30-120) after (a) saline or (b) 2 mg/kg MA treatment for BXD strain mice with different combined Taar1/Oprm1 genotypes. Presented are …
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Figure 6—source data 1
Core body temperature (°C) data across time for saline and 2 mg/kg MA-treated male and female, BXD mice, according to Taar1 and Oprm1 genotype.
- https://doi.org/10.7554/eLife.46472.015
Figure 7
Genome-wide QTL scans identify a significant QTL on mouse chromosome 10 for both MA consumption and MA effect on body temperature.
Means for 18 BXD strains were used for mapping of (a) consumption of MA from a 40 mg/l solution, and 15 strains for mapping of (b) body temperature response to 2 mg/kg MA at 30 min post-treatment. …
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Figure 7—source data 1
BXD means for QTL mapping of MA consumption (mg/kg/18h) from the 40 mg/l concentration and for QTL mapping of 2 mg/kg MA-induced body temperature change.
- https://doi.org/10.7554/eLife.46472.017
Figure 8
Generation of MAHDR-Taar1+/+ knock-in and MAHDR-Taar1m1J/m1J control mice, and genotyping of Taar1 and Oprm1.
(a) Strategy for insertion of a point mutation into mouse Taar1 exon 1. The guide sequence is indicated in green text and the protospacer adjacent motif (PAM) is indicated by bold black text. The …
Tables
Table 1
Taar1/Oprm1 genotype combinations for the BXD strains.
https://doi.org/10.7554/eLife.46472.011| Taar1/Oprm1 genotype combination | MA intake | MA-induced change in body temperature |
|---|---|---|
| Taar1+/+/Oprm1B6/B6 | BXD 154, 184, 196, 212, 218 | BXD 154, 184, 196, 218 |
| Taar1m1J/m1J/Oprm1B6/B6 | BXD 113, 160, 171, 194 | BXD 113, 160, 171, 194 |
| Taar1+/+/Oprm1D2/D2 | BXD 161, 199, 205, 215 | BXD 161, 199, 205 |
| Taar1m1J/m1J/Oprm1D2/D2 | BXD 178, 186, 204, 210, 216 | BXD 178, 186, 210, 216 |
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Abbreviations: BXD, C57BL/6J (B6) x DBA2/J (D2); MA, methamphetamine; Oprm1, mu-opioid receptor gene; Oprm1B6/B6, homozygous for B6 Oprm1 allele; Oprm1D2/D2, homozygous for D2 Oprm1 allele; Taar1, trace-amine associated receptor 1 gene; Taar1+/+, homozygous for reference Taar1+ allele; Taar1m1J/m1J, homozygous for mutant Taar1m1J allele.
Key resources table
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Strain, strain background (Mus musculus, females) | CD1/NCrl | Charles River | Strain code: 022 | foster dams for generation of knock-in mice |
| Strain, strain background (Mus musculus, females and males) | BXD recombinant inbred strains | University of Tennessee Health Sciences Center | original breeders from Dr. Robert Williams; offspring for research produced by Dr. Tamara Phillips at VA Portland Health Care System (VAPORHCS) | |
| Genetic reagent (Mus musculus, females) | MAHDR | VAPORHCS | methamphetamine high drinking mice; created by Dr. Tamara Phillips; all are homozygous for SNP rs33645709 | |
| Genetic reagent (Mus musculus, females and males) | MAHDR-Taar1m1J/m1J control | this paper | created by Oregon Health & Science University Transgenic Mouse Models Shared Resource; offspring for research produced by Dr. Tamara Phillips at VAPORHCS | |
| Genetic reagent (Mus musculus, females and males) | MAHDR-Taar1+/+ knock-in | this paper | created by Oregon Health & Science University Transgenic Mouse Models Shared Resource; offspring for research produced by Dr. Tamara Phillips at VAPORHCS | |
| Biological sample (Mus musculus) | tail snip | other | obtained from breeders and research animals to determine genotype | |
| Sequence-based reagent | Taar1 gRNA | this paper | designed by ThermoFisher Scientific; reagent for generation of knock-in mice (Mus musculus) | |
| Sequence-based reagent | 100b oligo single strand DNA for Taar1+ (ossDNA) | this paper | MGI:MGI2148258 | synthesized by ThermoFisher Scientific; reagent for generation of knock-in mice (Mus musculus) |
| Sequence-based reagent | Cas9 mRNA | TriLink | Catalog number: L-7606 | reagent for generation of knock-in mice (Mus musculus) |
| Sequence-based reagent | primers for genotyping Oprm1 | this paper | original reference for these primers: Ferraro et al. (2005); based on Oprm1 sequence NM_001304955 (Mus musculus) | |
| Sequence-based reagent | DdeI restriction enzyme | ThermoFisher Scientific | Catalog number: ER1882 | reagent for Oprm1 genotyping (Helicobacter pylori, RFL3) |
| Commercial assay or kit | QuickExtract DNA Extraction Solution | Lucigen | Catalog number: QE09050 | |
| Commercial assay or kit | Hotstart DNA Polymerase Kit | Qiagen | Catalog number: 203205 | |
| Commercial assay or kit | Taqman kit for determining Taar1 alleles | ThermoFisher Scientific | Custom order | probes based on Taar1 sequence: NM_053205.1; SNP rs33645709 |
| Chemical compound, drug | ethidium bromide | Sigma Aldrich | Catalog number: E1510 | reagent for Oprm1 genotyping |
| Chemical compound, drug | (+)-methamphetamine hydrochloride | Sigma Aldrich | Catalog number: M8750 | |
| Chemical compound, drug | sterile 0.9% saline | Baxter Healthcare Corporation | Catalog number: 2F7124 | vehicle for methamphetamine |
| Software, algorithm | Statistica | TIBCO Software Inc | ||
| Software, algorithm | GeneNetwork | University of Tennessee | RRID:SCR_002388 | www.genenetwork.org |
| Software, algorithm | R | The R Foundation for Statistical Computing | RRID:SCR_001905 | www.r-project.org |
Additional files
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Transparent reporting form
- https://doi.org/10.7554/eLife.46472.019
