Article Figures & Data
Figures
- Figure 1.
Identifying aromatase-expressing (arom+) neurons in the mouse brain. a, The aromatase enzyme mediates the conversion of testosterone into estradiol. b, Crossing aromatase-cre and Ai9 tdTomato reporter mice ensures that the tdTomato fluorescent protein is coexpressed with aromatase. c, A large population of aromatase-expressing cells is present in the posterodorsal MeA [arrow in (1) horizontal, (2) sagittal, and (3) coronal view].
- Figure 2.
Mapping the inputome of arom+ neurons in the MeA. a, Conditional AAV vectors were injected in aromatase-cre mice to express TVA, RG, and mCherry in arom+ neurons of the MeA. b, Fourteen days after AAV injection, G-deleted rabies (GFP) was injected at the same stereotaxic coordinates to infect arom+ starter neurons; followed by 10 d for retrograde movement of rabies from arom+ neurons in the MeA to upstream neurons. c, Infected starter neurons in the MeApd identified by simultaneous tdtomato and EGFP expression. d, The number of neurons labeled with GFP by rabies infection outside the MeA scales linearly with the number of starter neurons infected in the MeApd. e, View through the dorsal aspect of the brain showing location of starter neurons (red) and inputome (green), demonstrating that input cells are almost entirely ipsilateral to the injection site (see graph). f, Lateral view of the same cells shown in panel e showing that most inputs originate from subcortical regions.
- Figure 3.
Input cell numbers vary across subcortical brain regions. A sampling of input cells observed in one individual after alignment to the Allen mouse brain common coordinate framework demonstrates that inputs can be localized to subcortical regions brain-wide and that input cell density varies among brain regions. a, ventral tegmental area (VTA). b, bed nucleus of the stria teminalis (BNST). c, hippocampal formation (HPF). d, medial preoptic area (MPOA). e, ventromedial hypothalamus (VMH). f, caudoputamen (CP). g, central amygdala (CeA). h, accessory olfactory bulb (AOB). Scale bars, 500 μm.
- Figure 4.
Coarse population count of the MeA arom+ inputome. Each input cell was assigned to 1 of 10 nonoverlapping regions spanning the full volume of the reference brain. Defined regions are as follows: cerebral nuclei (CNU), hypothalamus (H), hippocampal formation (HPF), cortical subplate (CTXsp), olfactory areas (OF), thalamus (TH), midbrain (MB), isocortex, hindbrain (HB), and cerebellum (CB). a, Graph showing the percentage of the average total observed input neurons found in each brain region relative to the volume of that brain region shows that some regions have either much higher or much lower density of inputs than would be expected if the cells were evenly distributed across the brain (straight line). b, Percentages of the average total number of input cells found in each defined brain region. c, Pie chart illustrating the proportions of inputome cells found in eight brain regions. Extended Data Figures 4-1, 4-2, 4-3, and 4-4 support Figure 4.
- Figure 5.
MeA arom+ inputome originating in the cerebral nuclei (CNU). a, Left, A pie chart indicates the fraction of all labeled neurons in the CNU and the brain atlas depicts the position of CNU regions. Right, Bar graph depicts the percentages of input cells found in the striatum (STR) and pallidum (PAL) relative to the whole-brain inputome; pie chart shows the relative proportion of those cells within the cerebral nuclei alone. b, Locations of input neurons within the striatum: line graph shows relative density of cells in subregions of the striatum; bar graph shows relative distribution of inputs relative to the whole-brain inputome; pie chart shows the relative proportion of those cells within parts of the striatum. Regions shown: medial amygdala (MEA), central amygdala (CEA), caudoputamen (CP), intercalated amygdalar nucleus (IA), anterior amygdalar area (AAA), nucleus accumbens (ACB), lateral septal nucleus (LS), fundus of striatum (FS), olfactory tubercle (OT), septofimbrial nucleus (SF), bed nucleus of the accessory olfactory tract (BA), and septohippocampal nucleus (SH). c, Locations of input neurons within the pallidum: bar graph shows relative distribution of inputs relative to the whole-brain inputome; pie chart shows the relative proportion of those cells within parts of the pallidum. Regions shown: external globus pallidus (GPe), substantia innominata (SI), internal globus pallidus (GPi), BNST, diagonal band nucleus (NDB), magnocellular nucleus (MA), medial septal nucleus (MS), and triangular nucleus of septum (TRS). Extended Data Figure 5-1 supports Figure 5.
- Figure 6.
MeA arom+ inputome originating in the hypothalamus (HY). a, Left, A pie chart indicates the fraction of all labeled neurons in the hypothalamus, and the brain atlas depicts the position of hypothalamic regions. Right, Bar graph depicts the percentages of input cells found in the five subregions of the hypothalamus relative to the whole-brain inputome; pie chart shows the relative proportion of those cells within those hypothalamic regions. Regions shown: lateral zone (LZ), medial zone (MEZ), periventricular region (PVR), periventricular zone (PVZ), and median eminence (ME). b, Locations of input neurons within the lateral zone of the hypothalamus: bar graph shows relative distribution of inputs relative to the whole-brain inputome; and pie chart shows the relative proportion of those cells within parts of the medial zone. Regions shown: lateral hypothalamic area (LHA), Tuberal nucleus (TU), zona incerta (ZI), lateral preoptic area (LPO), subthalamic nucleus (STN), parasubthalamic nucleus (PSTN), retrochiasmatic area (RCH), perifornical nucleus (PeF), and preparasubthalamic nucleus (PST). c, Locations of input neurons within the medial zone of the hypothalamus: bar graph shows relative distribution of inputs relative to the whole-brain inputome; pie chart shows the relative proportion of those cells within parts of the medial zone. Regions shown: ventromedial hypothalamic nucleus (VMH), PMv, medial mammillary nucleus (MM), medial preoptic nucleus (MPN), dorsal premammillary nucleus (PMd), anterior hypothalamic nucleus (AHN), posterior hypothalamic nucleus (PH), tuberomammillary nucleus (TM), supramammillary nucleus (SUM), paraventricular hypothalamic nucleus (PVHd), and lateral mammillary nucleus (LM). Extended Data Figure 6-1 supports Figure 6.
- Figure 7.
MeA arom+ inputome originating in the hippocampal formation (HPF) and the cortical subplate (CTXsp). a, Left, A pie chart indicates the fraction of all labeled neurons in the hippocampus and the brain atlas depicts the position of hippocampal regions. Right, Bar graph depicts the percentages of input cells found in the regions of the hippocampal formation relative to the whole-brain inputome; and pie chart shows the relative proportion of those cells within those regions. The lower inset image illustrates that input cells are located primarily in ventral portions of the HPF. Regions shown: field CA1 (CA1), field CA2 (CA2), field CA3 (CA3), entorhinal area (ENT), hippocampal-amygdalar transition area (HATA), prosubiculum (ProS), dentate gyrus (DG), presubiculum (PRE), subiculum (SUB), area prostriata (APr), postsubiculum (POST), parasubiculum (PAR), induseum griseum (IG), and fasciola cinerea (FC). b, Left, A pie chart indicates the fraction of all labeled neurons in the cortical subplate, and the brain atlas depicts the position of cortical subplate regions. Right, Bar graph depicts the percentages of input cells found in the regions of the cortical subplate relative to the whole-brain inputome; and pie chart shows the relative proportion of those cells within those regions. Regions shown: posterior amygdalar nucleus (PA), anterior BMA (BMAa), posterior BMA (BMAp), anterior BMA (BLAa), ventral basolateral amygdalar nucleus (BLAv), posterior basolateral amygdalar nucleus (BLAp), lateral amygdalar nucleus (LA), ventral endopiriform nucleus (EPv), dorsal endopiriform nucleus (EPd), and claustrum (CLA). Extended Data Figure 7-1 supports Figure 7.
- Figure 8.
MeA arom+ inputome originating in the olfactory areas (OLF) and the thalamus (TH). a, Left, A pie chart indicates the fraction of all labeled neurons in olfactory areas; and the brain atlas depicts the position of olfactory areas. Right, Bar graph depicts the percentages of input cells found in subregions of the olfactory area relative to the whole-brain inputome; and pie chart shows the relative proportion of those cells within those regions. Regions shown: cortical amygdalar area (COA), piriform area (PIR), accessory olfactory bulb (AOB) , piriform-amygdalar area (PAA), postpiriform transition area (TR), nucleus of the lateral olfactory tract (NLOT), main olfactory bulb (MOB), taenia tecta (TT), anterior olfactory nucleus (AON), and dorsal peduncular area (DP). b, Left, A pie chart indicates the fraction of all labeled neurons in the thalamus; and brain atlas depicts the position of thalamic regions. Right, Bar graph depicts the percentages of input cells found in subregions of the thalamus relative to the whole-brain inputome; and pie chart shows the relative proportion of those cells within those regions. Regions shown: reticular nucleus (RT), ventral group of the dorsal thalamus (VENT), dorsal geniculate group (GENd), midline group of the dorsal thalamus (MTN), ventral geniculate group (GENv), intralaminar nuclei of the dorsal thalamus (ILM), peripeduncular nucleus (PP), lateral group of the dorsal thalamus (LAT), medial group of the dorsal thalamus (MED), epithalamus (EPI), anterior group of the dorsal thalamus (ATN), subparafascicular area (SPA), and subparafascicular nucleus (SPF). Extended Data Figure 8-1 supports Figure 8.
- Figure 9.
Sex differences in the inputome of aromatase-expressing cells in the MeA. The average percentage of synaptic inputs to MeA arom+ neurons compared for each region in males and females. Only regions that provide 0.5% of the overall input are shown, and regions identified as having a statistically significant sex difference are shown in color (p < 0.05, permutation post hoc test following ANOVA with repeated measures on “sex,” “region,” and “individual”). These regions include field CA1 (CA1), the accessory olfactory bulb (AOB), piriform area (PIR), piriform-amygdalar area (PAA), postpiriform transition area (TA), basolateral amygdalar nucleus-anterior (BLAa), basolateral amygdalar nucleus-ventral (BLAv), diagonal band nucleus (NDB), lateral preoptic area (LPO), and ventral premammillary nucleus (PMv). Regions lying above the line of unity represent a male bias, and neurons lying to the right of the unity line indicate a female bias. Error bars indicate the SEMs (male, vertical; female, horizontal) for each region. Extended Data Figure 9-1 supports Figure 9.
- Figure 10.
The inputome of MeA arom+ neurons. a, Illustration showing the relative locations of the major inputs to the aromatase-expressing MeA cells throughout the brain as defined by our data. Major regions are shaded to demonstrate that the included regions were analyzed in groups and are considered components of a larger analysis region. These inputs were selected based on their relative abundance in the data and behavioral significance as represented in previous literature. b, A Venn diagram showing the relative distribution of the regions displayed in a sorted by their reported roles in the production or modulation of memory, parenting and aggression, metabolism, sociosexual behaviors, and fear and anxiety behaviors.
Extended Data
Table 1-1
Ipsilateral bias across brain regions. Download Table 1-1, XLSX file.
Figure 4-1
Input cell density in regions brain wide. Percentage of input cells in each coarse region identified in the Allen Brain Atlas normalized to the volume of that region to determine cell densities in each region are presented as graph (a) and pie chart (b). The densest concentration of input cells is found in the cortical subplate (CTXsp). Download Figure 4-1, TIF file.
Figure 4-2
MeA arom+ inputome originating in the midbrain (MB). a, Bar graph depicts the percentages of input cells found in major divisions of the midbrain relative to the whole-brain inputome; and pie chart shows the relative proportion of those cells within those regions. Regions shown: motor-related midbrain regions (MBmot), sensory-related midbrain regions (MBsec), and behavior-related midbrain regions (MBsta). b, Inputs from motor-related midbrain regions: bar graph depicts the percentages of input cells found in major divisions of these regions relative to the whole-brain inputome; and pie chart shows the relative proportion of input cells within those regions. Regions shown: midbrain reticular nucleus (MRN), superior colliculus—motor (SCm), VTA, substantia nigra (SNr), periaqueductal gray (PAG), lateral terminal nucleus of accessory optic tract (LT), cuneiform nucleus (CUN), red nucleus (RN), retrorubal area of midbrain reticular nucleus (RR), paratrochlear nucleus (Pa4), pretectal region (PRT), medial terminal nucleus of accessory optic tract (MT), dorsal terminal nucleus of accessory optic tract (DT), anterior tegmental nucleus (AT), ventral tegmental nucleus (VTN), trochlear nucleus (IV), Edinger-Westphal nucleus (EW), medial accessory oculomotor nucleus (MA3), oculomotor nucleus (III), and paranigral nucleus (PN). c, Inputs from sensory-related midbrain regions: bar graph depicts the percentages of input cells found in major divisions of these regions relative to the whole-brain inputome; and pie chart shows the relative proportion of input cells within those regions. Regions shown: inferior colliculus (IC), brachium of inferior colliculus (NB), subcommissural organ (SCO), midbrain trigeminal nucleus (MEV), parabigeminal nucleus (PBG), nucleus sagulum (SAG), and superior colliculus (sensory; SCs). **Less than 0.01% of MeA arom+ inputs originate in behavior-related midbrain area; most of those cells are found in the dorsal raphe nucleus (DR), pedunculopontine nucleus (PPN), and interpeduncular nucleus (IPN). Download Figure 4-2, TIF file.
Figure 4-3
MeA arom+ inputome originating in the hindbrain (HB). a, Whole hindbrain: bar graph depicts the percentages of input cells found in major divisions of the hindbrain relative to the whole-brain inputome; and pie chart shows the relative proportion of input cells within those regions. Regions shown: motor-related medulla (MY-mot), sensory-related medulla (MY-sen), behavioral state medulla (MY-sat), motor-related pons (P-mot), sensory-related pons (P-sen), and behavioral state pons (P-sat). b, Only one of the behavior-state portions of the medulla, the nucleus raphe magnus (RM) contains inputs to arom+ cells in the MeA. c, Bar graph depicts the percentages of input cells found in major divisions of the motor-related medulla relative to the whole-brain inputome; and pie chart shows the relative proportion of input cells within those regions. Two regions of the motor-related medulla provide inputs: the gigantocellular reticular nucleus (GRN) and the facial motor nucleus (VII). d, The caudal part of the pontine reticular nucleus (PRNc) is the only part of the motor-related pons to provide inputs to arom+ MeA cells. e, In the sensory-related pons, inputs originate from the parabrachial nucleus (PB) and the nucleus of the lateral lemniscus (NLL). Download Figure 4-3, TIF file.
Figure 4-4
MeA arom+ inputome originating in the isocortex and the cerebellum (CB). a, Bar graph depicts the percentages of input cells found in major divisions of the isocortex relative to the whole-brain inputome; and pie chart shows the relative proportion of input cells within those regions. Regions shown: Secondary motor area (MOs), agranular insular area (AI), orbital area (ORB), temporal association area (Tea), ectorhinal area (ECT), visceral area (VISC), anterior cingulate area (ACA), infralimbic area (ILA), primary motor area (MOp), supplementary somatosensory area (SSs), gustatory areas (GU), auditory areas (AUD), visual areas (VIS), primary somatosensory area (SSp), prelimbic area (PL), perirhinal area (PERI), posterior parietal association areas (PTLps), and retrosplenial area (RSP). b, Inputs from the cerebellum are only found in the cerebellar cortex (CBX). Download Figure 4-4, TIF file.
Figure 5-1
Input cell density in the cerebral nuclei (CNU). a, Input cell densities of constituent regions of the cerebral nuclei. b, Input cell densities of constituent regions in the striatum. c, Input cell densities of constituent regions in the pallidum. Results are presented as both bar chart and pie charts. Download Figure 5-1, TIF file.
Figure 6-1
Input cell density in the hypothalamus (HY). a, Input cell densities of constituent regions in the hypothalamus. b, Input cell densities of constituent regions in the lateral zone of the hypothalamus. c, Input cell densities of constituent regions in the medial zone of the hypothalamus. Results are presented as both bar chart and pie charts. Download Figure 6-1, TIF file.
Figure 7-1
Input cell density in the hippocampal formation (HPF) and the cortical subplate (CTXsp). a, Input cell densities of constituent regions in the hippocampal formation. b, Input cell densities of constituent regions in the cortical subplate. Results are presented as both bar chart and pie charts. Download Figure 7-1, TIF file.
Figure 8-1
Input cell density in the olfactory areas (OLF) and the thalamus (TH). a, Input cell densities of constituent regions in the olfactory areas. b, Input cell densities of constituent regions in the thalamus. Results are presented as both bar chart and pie charts. Download Figure 8-1, TIF file.
Table 2-1
Hypothalamic inputs to arom+ cells in the MeA. Download Table 2-1, XLSX file.
Table 3-1
Statistical analysis of observed input cells across brain regions. Download Table 3-1, DOCX file.
Figure 9-1
Comparison of sex differences in observed inputs to MeAArom+ Cells. a, Graph comparing the fraction of labeled neurons in the MeA, local to the injection site, in females and males. b, graph showing the d′ values for sex differences in the regions presented in Figure 9. Download Figure 9-1, TIF file.
Table 4-1
Statistical analysis of sex differences in major input regions. Download Table 4-1, XLS file.
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