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. 2021 Jan 14;10:e55081. doi: 10.7554/eLife.55081

Figure 1. Experimental design and brain regions of interest.

(A) Sequence of experiments during a 30 day period: Female voles were bilaterally ovariectomized before MR and behavioral protocols. After being allowed to recover from surgery for 10 days, silastic capsules containing E2B (estradiol benzoate) were implanted via s.c. 4 days before cohabitation for sexual receptivity induction. Once couples went under cohabitation, they were housed together for the rest of the experiment and were only separated for PPT and MR scanning sessions. OVX: ovariectomy surgery. MR: magnetic resonance imaging scanning session. PPT: partner preference test. (B) Regions of interest (ROIs) for network functional connectivity analyses. Antero-posterior coronal slices of the prairie vole template overlayed with ROI masks with the resolution used in the analysis. Each color represents a different ROI. ACC: anterior cingulate cortex. AON: anterior olfactory nucleus. BLA: basolateral amygdala. BNST: bed nucleus of the stria terminalis. DG: dentate gyrus. dHIP: dorsal hippocampus. MeA: medial amygdala. MOB: main olfactory bulb. LS: lateral septum. mPFC: medial prefrontal cortex. NAcc: nucleus accumbens. PVN: paraventricular nucleus. RSC: retrosplenial cortex; VP: ventral pallidum. vHIP: ventral hippocampus. VTA: ventral tegmental area. (C) 3D views of ROI masks embedded within the prairie vole template.

Figure 1.

Figure 1—figure supplement 1. Representative rsfMRI time series.

Figure 1—figure supplement 1.

(a–p) Representative rsfMRI time series of regions of interest (ROIs). For each ROI, the time series are shown before (Raw) and after pre-processing (Pre-processed). ACC: anterior cingulate cortex. AON: anterior olfactory nucleus. BLA: basolateral amygdala. BNST: bed nucleus of the stria terminalis. LS: lateral septum. MeA: medial amygdala. MOB: main olfactory bulb. mPFC: medial prefrontal cortex. NAcc: nucleus accumbens. PVN: paraventricular nucleus. RSC: retrosplenial cortex. VP: ventral pallidum. VTA: ventral tegmental area. DG: dentate gyrus. dHIP: dorsal hippocampus. vHIP: ventral hippocampus.
Figure 1—figure supplement 2. Average functional connectivity correlation matrices between ROIs in all subjects shown by MR acquisition sessions: baseline, 24 hr, and 2 weeks.

Figure 1—figure supplement 2.

(A) Pearson’s correlation values of rsfMRI time-series connectivity matrices. (B) Partial-correlation values of rsfMRI time-series connectivity matrices. (C) Fisher z-transformed, rsfMRI time-series connectivity matrices used for functional connectivity analysis. ACC: anterior cingulate cortex. AON: anterior olfactory nucleus. BLA: basolateral amygdala. BNST: bed nucleus of the stria terminalis. LS: lateral septum. MeA: medial amygdala. MOB: main olfactory bulb. mPFC: medial prefrontal cortex. NAcc: nucleus accumbens. PVN: paraventricular nucleus. RSC: retrosplenial cortex. VP: ventral pallidum. VTA: ventral tegmental area. DG: dentate gyrus. dHIP: dorsal hippocampus. vHIP: ventral hippocampus.
Figure 1—figure supplement 3. Representative rsfMRI and anatomical raw data and examples of the registration steps.

Figure 1—figure supplement 3.

(a) Raw SE-EPI rsfMRI image; (b) raw anatomic image; (c) brain extraction and bias field correction of the anatomic image; (d) registration of the anatomic data (grayscale) to the prairie vole brain template (outlined in red); (e) final registration of a representative volume of the SE-EPI acquisition (grayscale) to the prairie vole brain template (outlined in red).
Figure 1—figure supplement 4. Average seed-based functional connectivity maps for each of the 16 ROIs here explored.

Figure 1—figure supplement 4.

For every subject, session, and ROI, the average time series from the ROI is extracted and the functional connectivity with every voxel of the brain is estimated as the Pearson’s correlation. Then connectivity maps are Fisher z-transformed, and a single-group t-test is performed, family-wise error rates are controlled with a false discovery rate lower than 0.05. Connectivity maps represent Pearson’s correlation values. ACC: anterior cingulate cortex. AON: anterior olfactory nucleus. BLA: basolateral amygdala. BNST: bed nucleus of the stria terminalis. LS: lateral septum. MeA: medial amygdala. MOB: main olfactory bulb. mPFC: medial prefrontal cortex. NAcc: nucleus accumbens. PVN: paraventricular nucleus. RSC: retrosplenial cortex. VP: ventral pallidum. VTA: ventral tegmental area. DG: dentate gyrus. dHIP: dorsal hippocampus. vHIP: ventral hippocampus.
Figure 1—figure supplement 5. Vole temperature during acquisition, discarded images and signal-to-noise ratio.

Figure 1—figure supplement 5.

(A) Prairie vole temperatures during MR acquisition sessions. A separate sub-sample was used to evaluate the design of the anesthesia protocol on physiological data in both male (n = 3) and female (n = 2) voles. Temperature mean was 34.93 ± 0.01°C. MRI data acquisition began when physiological measurements stabilized (approximately after ~15 min from bolus injection). (B) Two datasets from session 1 (baseline) showed signal loss in the posterior cortex and were discarded from the study. (C) Boxplots of the mean signal-to-noise ratio (SNR) values (n = 78) for each region of interest (16) used in the rsfMRI analyses. Top and bottom of the box are 25th and 75th percentiles, respectively, and the centerline shows data median. SNR estimates (n = 78) were calculated by dividing ROI mean signals by the standard deviation of the noise signal, as described elsewhere (Grandjean et al., 2020). ACC: anterior cingulate cortex. AON: anterior olfactory nucleus. BLA: basolateral amygdala. BNST: bed nucleus of the stria terminalis. LS: lateral septum; MeA: medial amygdala. MOB: main olfactory bulb. mPFC: medial prefrontal cortex. NAcc: nucleus accumbens. PVN: paraventricular nucleus. RSC: retrosplenial cortex. VP: ventral pallidum. VTA: ventral tegmental area. DG: dentate gyrus. dHIP: dorsal hippocampus. vHIP: ventral hippocampus.