Article Figures & Data
Figures
- Figure 1.
Low (A) and high (B) magnification of the microphotographs illustrating single PRV152 (green), single H129 (red), and double PRV152+H129 (arrows) immunolabeling in the RPa and ROb following viral injections into IBAT. Gi, Gigantocellular reticular nucleus; py, pyramidal tract. n = 5. Low (C) and high (D) magnification of the microphotographs illustrating single PRV152 (green), single H129 (red), and double PRV152+H129 (arrows) immunolabeling in the PAG following viral injections into IBAT. 3N, Oculomotor nucleus; Aq, aqueduct; DpMe, deep mesencephalic nucleus; mlf, medial longitudinal fasciculus. Low (E) and high (F) magnification of the microphotographs illustrating single PRV152 (green), single H129 (red), and double PRV152+H129 (arrows) immunolabeling in the DM following viral injections into IBAT. 3V, Third ventricle. Low (G) and high (H) magnification of the microphotographs illustrating single PRV152 (green), single H129 (red), and double PRV152+H129 (arrows) immunolabeling in the LH following viral injections into IBAT. 3V, Third ventricle; TC, tuber cinereum area. Low (I) and high (J) magnification of the microphotographs illustrating single PRV152 (green), single H129 (red), and double PRV152+H129 (arrows) immunolabeling in the PVH following viral injections into IBAT. AHP, Anterior hypothalamic area, posterior part; opt; optic tract. Low (K) and high (L) magnification of the microphotographs illustrating single PRV152 (green), single H129 (red), and double PRV152+H129 (arrows) immunolabeling in the MPA following viral injections into IBAT. aca, Anterior commissure. Scale bars: A, C, E, G, I, K, 200 μm; B, D, F, H, J, L, 50 μm.
- Figure 2.
A, Brain areas with highest absolute numbers of PRV152- and H129-IR neurons. Note that the MPA contained predominantly SNS (PRV152) efferent output to IBAT as compared with a SS (H129) afferent input from the same fat pad; n = 5; *p < 0.05 versus PRV152. B, Percentile quantification of PRV152 and H129 double-labeled (-infected) in the hindbrain, midbrain, posterior hypothalamus and thalamus, and anterior hypothalamus; n = 5; *p < 0.05 versus hindbrain, midbrain, and posterior hypothalamus and thalamus.
- Figure 3.
A, The activation of IBAT thermogenesis produced by selective β3-adrenoceptor agonist CL316,243 microinjections intra-left IBAT in comparison with core body temperature and TIBAT after saline microinjections intra-right IBAT. B, Changes in TIBAT after CL316,243 microinjections intra-left IBAT and saline intra-right IBAT over a 60 min period; n = 12; *p < 0.05 versus saline, #p < 0.05 versus core body temperature.
- Figure 4.
Representative microphotographs of DRG at C4 vertebral level showing c-Fos immunostaining (A, D), FB labeling (B, E), and c-Fos+FB colocalization (C, F, arrows) after CL316,243 microinjections intra-left and saline intra-right IBAT. Distribution of c-Fos-IR and FB-labeled neurons in IBAT innervating DRG at C1–T4 vertebral levels contralateral (G) and ipsilateral (H) to CL316,243 microinjections. The total number of positively stained neurons per ganglion can be estimated by multiplying the number per section by 24; n = 12; *p < 0.05 versus saline counter-mate. Scale bar, 50 μm. I, Mean nerve activity from IBAT afferent fibers before (baseline), at 10 and at 20 min after CL316,243 or saline microinjections; n = 5 per group. CL316,243-evoked increase in IBAT sensory nerve activity. Bottom, Ten minute sample traces of IBAT afferent fibers before and after infusion of either CL316,243 (top) or saline (bottom); *p < 0.05 versus saline.
- Figure 5.
A diaphragmatic representation of the efferent sympathetic (green arrows) and afferent sensory (red arrows) brain-IBAT neural feedback circuit for control of IBAT functions. BAT, brown adipose tissue; C4–T2, Cervical and thoracic vertebral level; DH, dorsal horn; SG, sympathetic ganglia.
Tables
- Table 1.
Distribution of sympathetic (PRV152) and sensory (H129) neurons across the neuroaxis
PRV152 H129 PRV152 + H129 Percentage of PRV152 + H129 Hindbrain 7N 67.4 ± 12.1 57.6 ± 19.5 11.8 ± 3.0 10.4 ± 1.3 10N 16.0 ± 8.2 14.5 ± 1.2 2.0 ± 1.6 5.7 ± 4.7 12N 24.3 ± 5.2 17.0 ± 8.0 0.7 ± 0.3 1.9 ± 1.0 PnO 38.0 ± 16.0 27.4 ± 9.6 6.4 ± 2.5 11.3 ± 1.1 PnC 51.4 ± 15.1 42.6 ± 8.1 9.2 ± 2.5 10.6 ± 1.9 SuVe 74.2 ± 20.8 59.6 ± 16.1 13.2 ± 2.4 12.1 ± 1.8 MPB 36.6 ± 10.3 40.5 ± 8.4 7.6 ± 1.5 12.7 ± 1.1 LPBI 6.3 ± 2.9 5.3 ± 1.6 1.7 ± 0.8 7.3 ± 4.7 LPBC 23.6 ± 7.1 13.4 ± 5.3 3.0 ± 1.2 9.5 ± 4.3 MPBE 11.3 ± 5.6 11.3 ± 4.7 2.0 ± 0.7 8.0 ± 2.6 LC 29.8 ± 9.9 28.3 ± 5.7 5.0 ± 1.5 8.5 ± 2.4 Bar 16.3 ± 6.6 9.7 ± 4.6 3.8 ± 1.2 11.6 ± 4.8 Pr5 29.0 ± 13.3 26.8 ± 9.6 5.5 ± 2.3 11.3 ± 0.8 LDTg 13.3 ± 4.6 4.3 ± 0.7 1.7 ± 0.7 12.3 ± 7.0 Mo5 18.7 ± 3.3 20.7 ± 1.1 3.7 ± 0.5 10.2 ± 1.0 RMg 83.8 ± 25.9 87.2 ± 17.7 17.2 ± 4.1 11.4 ± 1.2 RPa 159.2 ± 34.0 148.8 ± 41.6 36.2 ± 9.2 12.6 ± 1.1 ROb 154.5 ± 55.9 116.0 ± 14.1 30.5 ± 5.0 12.9 ± 1.4 Gi 52.8 ± 28.8 39.0 ± 13.5 6.2 ± 3.0 6.4 ± 2.2 LPGi 31.0 ± 6.6 29.0 ± 6.8 7.0 ± 1.3 13.0 ± 1.2 DPGi 42.0 ± 17.3 42.3 ± 16.5 7.3 ± 2.8 8.5 ± 1.0 GiV 108.0 ± 19.7 127.2 ± 32.4 19.0 ± 3.0 9.2 ± 1.0 PCRt 14.0 ± 4.1 10.7 ± 3.9 1.3 ± 0.5 4.6 ± 2.0 IO 152.7 ± 41.6 129.3 ± 15.0 25.7 ± 6.1 9.8 ± 1.8 LRt 120.7 ± 36.1 96.0 ± 24.0 18.7 ± 2.9 9.4 ± 1.0 MdV 59.3 ± 39.9 43.0 ± 18.0 9.0 ± 5.9 6.8 ± 3.4 NTS 275.7 ± 60.4 201.2 ± 58.9 33.4 ± 4.2 7.3 ± 1.4 SolC 28.3 ± 5.2 21.0 ± 4.7 5.2 ± 1.1 10.5 ± 2.1 SolCe 7.2 ± 2.8 5.3 ± 2.0 1.3 ± 0.4 9.3 ± 2.6 SolDL 31.8 ± 15.2 27.9 ± 14.8 4.7 ± 1.9 7.7 ± 1.7 SolG 21.7 ± 10.9 15.3 ± 4.5 2.8 ± 0.4 7.1 ± 2.3 SolI 7.4 ± 3.0 4.8 ± 1.5 1.2 ± 0.2 9.3 ± 3.1 SolIM 25.3 ± 8.8 19.5 ± 4.1 3.7 ± 1.3 7.9 ± 2.2 SolM 97.3 ± 12.0 66.0 ± 14.5 12.7 ± 4.3 7.2 ± 2.0 SolV 11.7 ± 3.4 10.8 ± 3.5 2.0 ± 0.2 8.6 ± 1.3 SolVL 45.8 ± 18.3 32.1 ± 10.2 6.0 ± 2.8 7.5 ± 2.4 Pr 25.5 ± 11.2 15.0 ± 6.6 3.8 ± 2.8 6.2 ± 3.1 Midbrain PAG 260.6 ± 51.3 109.2 ± 28.8 32.0 ± 5.8 9.6 ± 1.5 DMPAG 32.4 ± 9.3 12.2 ± 4.7 2.8 ± 1.11 6.3 ± 1.2 DLPAG 26.0 ± 6.1 8.4 ± 2.6* 1.8 ± 0.7 5.1 ± 1.6 LPAG 60.0 ± 11.5 30.0 ± 6.9* 8.8 ± 1.3 11.5 ± 2.1 VLPAG 54.8 ± 11.6 18.2 ± 6.0 6.4 ± 1.4 10.3 ± 1.8 DR 23.6 ± 6.7 7.0 ± 2.2* 2.4 ± 0.6 7.7 ± 2.4 DpMe 54.3 ± 20.3 19.0 ± 6.4 6.0 ± 1.9 8.5 ± 3.1 R 18.0 ± 5.5 9.6 ± 2.9 2.6 ± 0.2 9.4 ± 3.3 VTA 32.5 ± 12.7 13.5 ± 4.6 3.5 ± 1.2 7.6 ± 2.8 Forebrain MPA 79.8 ± 8.9 6.4 ± 3.0* 2.2 ± 1.0 3.0 ± 1.7 MPO 36.4 ± 13.8 6.8 ± 2.6* 1.4 ± 0.9 5.0 ± 3.0 MnPO 17.4 ± 5.2 0.8 ± 0.8* 0.2 ± 0.2 2.0 ± 2.0 LPO 23.8 ± 10.5 1.5 ± 0.8 0.8 ± 0.4 1.7 ± 1.0 AVPO 32.6 ± 9.5 7.8 ± 5.9 1.4 ± 1.0 2.8 ± 1.7 AVPe 23.5 ± 7.9 3.3 ± 1.8 1.3 ± 0.8 3.3 ± 2.3 SCh 13.5 ± 6.9 4.5 ± 1.7 0.5 ± 0.4 1.2 ± 1.0 AHA 7.4 ± 2.2 3.8 ± 2.2 1 ± 0.4 9.2 ± 4.3 AHP 22.8 ± 9.1 9.8 ± 5.5 2.4 ± 1.3 5.8 ± 2.4 PVH 277.4 ± 55.5 237.8 ± 58.7 48.8 ± 8.7 11.2 ± 1.8 PaAP 15.0 ± 4.9 14.8 ± 7.2 3.5 ± 1.7 10.7 ± 3.2 PaDC 30.6 ± 7.3 36.4 ± 11.3 6.2 ± 1.7 8.9 ± 3.1 PaMP 137.8 ± 22.1 129.8 ± 32.8 23.6 ± 4.2 10.4 ± 1.7 PaMM 12.2 ± 4.9 7.2 ± 2.3 1.8 ± 1.1 6.8 ± 4.5 PaLM 36.0 ± 6.8 35.3 ± 10.7 5.8 ± 1.7 8.5 ± 1.4 PaV 16.7 ± 2.5 16.3 ± 4.2 3.3 ± 1.4 12.6 ± 5.8 PaPo 22.7 ± 12.0 25.4 ± 12.5 7.6 ± 5.2 8.4 ± 2.3 Arc 19.6 ± 7.1 8.2 ± 2.1 2.0 ± 0.9 5.9 ± 2.6 LH 101.6 ± 20.8 40.0 ± 3.9 11.8 ± 1.7 9.3 ± 0.6 VMH 6.4 ± 4.1 0 0 0 DM 83.3 ± 31.8 30.3 ± 5.8 8.7 ± 1.7 9.8 ± 2.3 PH 39.8 ± 12.9 15.8 ± 1.5 5.0 ± 1.1 11.1 ± 1.5 ZI 18 ± 11.0 8.6 ± 4.5 1.2 ± 0.8 2.6 ± 1.7 SubZI 31.6 ± 4.6 22.4 ± 8.9 6.8 ± 2.0 13.5 ± 3.9 7N, Facial nucleus; 10N, dorsal motor nucleus of the vagus nerve; 12N, hypoglossal nucleus; Arc, arcuate nucleus; AVPe, anteroventral periventricular nucleus; AVPO, anteroventral preoptic nucleus; Bar, Barrington's nucleus; GiV, gigantocellular reticular nucleus, ventral part; IO, inferior olive; LC, locus coeruleus; LDTg, laterodorsal tegmental nucleus; LPBC, lateral parabrachial nucleus, central part; LPBI, lateral parabrachial nucleus, internal part; LRt, lateral reticular nucleus; MdV, medullary reticular nucleus, ventral part; MnPO, median preoptic nucleus; Mo5, motor trigeminal nucleus; MPBE, medial parabrachial nucleus external part; PaAP, paraventricular hypothalamic nucleus, anterior parvicellular part; PaDC, paraventricular hypothalamic nucleus, dorsal cap; PaLM, paraventricular hypothalamic nucleus, lateral magnocellular part; PaMM, paraventricular hypothalamic nucleus, medial magnocellular part; PaPo, paraventricular hypothalamic nucleus, posterior part; PaV, paraventricular hypothalamic nucleus, ventral part; PCRt, parvicellular reticular nucleus; PH, posterior hypothalamic area; PnC, pontine reticular nucleus, caudal part; PnO, pontine reticular nucleus, oral part; Pr, prepositus nucleus; Pr5, principal sensory trigeminal nucleus; R, red nucleus; RMg, raphe magnus nucleus; SCh, suprachiasmatic nucleus; SolC, nucleus of the solitary tract, commissural part; SolM, nucleus of the solitary tract, medial part; SolVL, nucleus of the solitary tract, ventrolateral part.
Data are expressed as mean ± SEM (n = 5);
↵*p < 0.05 versus PRV152.
