Fig. 9
Functions of LSv-projecting PVH neurons. a–d Behaviors induced by photostimulation of PVH→LSv fibers from selective LSv-projecting neurons. a Diagram showing delivery of retrograde AAVrg-Cre-Venus vectors to the LSv (b, left panel), and Cre-dependent AAV-Flex-ChR2-eGFP vectors to the PVH (b, right panel), and implantation of optic fibers targeting the LSv of adult wild-type mice. Short pulse (10 ms, 5 Hz, 5 mW mm2) photostimulation of PVH→LSv fibers from selective LSv-projecting neurons induced self-grooming (c, n = 3; light on vs pre-light p = 0.0003, light on vs post-light p = 0.0005) and long pulse (100 ms, 5 Hz, 5 mW/mm2) induced jumping (d, n = 3; light on vs pre-light p = 0.0005, light on vs post-light p = 0.0005); and while the short pulse stimulation caused marginal reduction, the long pulse caused dramatic reduction, on refeeding after 12 hr fasting (e, n = 3; Off vs 5 HZ, 100 ms p = 0.0018). **P < 0.01, one-way ANOVA tests. f–m Responses in the activity of LSv-projecting PVH neurons to water spray and loud sound. e Diagram showing delivery of retrograde AAVrg-Cre-Venus vectors to the LSv, and Cre-dependent AAV-Flex-GCaMP6m or AAV-Flex-GFP g vectors to the PVH, and implantation of optic fibers targeting the PVH of adult wild-type mice. Response to water spray in control mice with GFP expression in traced LSv-projecting neurons h and in mice with GCaMP6m expression in those neurons i; responses to loud sound in mice with GCaMP6m expression in traced LSv-projecting neurons j. k–m Representative traces showing changes in Ca2+ signal during a period with feeding bouts in mice with GFP expression k or GCaMP6m expression l in LSv-projecting neurons. m Summary of changes in Ca2+ signal in GFP and GCaMP6m mice. Mice used in feeding were fasted to increase the number of eating bouts (n = 20 each, ***p < 0.001, unpaired Student’s t tests). The red trace in h–j indicating averages of individual traces shown in gray. The red traces in k and l showing Ca2+-dependent signals, whereas the gray traces showing Ca2+-independent signals. Scale bar = 100 µm. The white line enclosed areas in b (right panel) outlining the PVH. LV: lateral ventricle; LSv: ventral part of lateral septum; fx: fornix; 3 V: the third ventricle; aca: anterior commissure area; AcbSh: accumbens shell
