lepr ires cre  (Jackson Laboratory)

Journal: bioRxiv

Article Title: A Preoptic Neurocircuit That Modulates Metabolic Flexibility

doi: 10.64898/2026.01.15.699760

Figure Lengend Snippet: (A) Immunostaining of axonal fibers in Vglut2-IRES-Cre mice transduced with Gq-DREADD (AAV-hSyn-DIO-hM3D(Gq)-mCherry). Axonal projections were identified in a variety of downstream regions, including the paraventricular thalamus (PVT), paraventricular hypothalamus (PVH), dorsomedial hypothalamus (DMH), periacqueductal grey (PAG), locus coeruleus (LC), and raphe pallidus (RPa). ( B) Optogenetic branch stimulation of hindbrain-projecting avPOA Vglut2 neurons. Blue bars correspond to regions targeted for optogenetic stimulation via cannula implantation and subsequent exposure to 455 nm blue light. (C) ( Left) Glucose tolerance testing of Vglut2-IRES-Cre animals transduced with a Cre-dependent ChR2 (AAV-DIO-ChR2(H134R)-EYFP) or EYFP (AAV-DIO-EYFP) with optic fibers placed unilaterally above either the POA, POA→DMH-projecting neurons, or POA→RPA-projecting neurons. (Right) AUC of GTT for each cohort described (ns = p > 0.05, ** = p < 0.005, student’s two-tailed t-test, mean ±SEM, n = 4 EYFP, n = 4 POA, n = 3 POA→DMH, n = 3 POA→RPa Vglut2-IRES-Cre animals). (D) UMAP plot and Leiden clustering of all avPOA Vglut2 neurons identified in Hrvatin et al., 2020. (E) UMAP plot of avPOA Vglut2 neurons colored by Adcyap1 and LepR expression. (F) Schematic showing four groups of animals: mCherry (mCherry-transduced Vglut2-IRES-Cre, n = 11), Vglut2 (Gq-DREADD transduced Vglut2-IRES-Cre ) (n=10), PACAP (Gq-DREADD-transduced PACAP-2A-Cre ) (n=11), and LepR (Gq-DREADD-transduced LepR-IRES-Cre (n=3) animals injected with 1 mg/kg CNO. (G) Glucose tolerance test of mCherry (avPOA Vglut2-mCherry ), Vglut2 (avPOA Vglut2-Gq ), PACAP (avPOA PACAP-Gq ), and LepR (avPOA LepR-Gq ), animals from (F). (H) AUC analysis for (G) (** = p < 0.05, student’s two-tailed t-test, mean ±SEM, n=7 avPOA Vglut2-mCherry , n = 10 avPOA Vglut2-Gq , n = 11 avPOA PACAP-Gq , n = 3 avPOA LepR-Gq animals). ( I) Volcano Plot of 120-minute polar metabolomics of PACAP-2A-Cre animals transduced with a Cre-dependent Gq-DREADD and injected with either PBS or CNO (1 mg/kg). Animals were treated as in . Selected overrepresented and underrepresented metabolites labeled in red and blue, respectively. p-values calculated using two-tailed t-test; red lines corresponding to p-value and Log 2 -Fold changes set at p < 0.05 and |Log 2 -Fold change| > 0.6.

Article Snippet: For natural, chemogenetic, and optogenetic torpor experiments, we used 8-12-week-old Vglut2-IRES-Cre (The Jackson Laboratory, Stock 028863), Adcyap1-2A-Cre (The Jackson Laboratory, Stock 030155), and Lepr-IRES-Cre (The Jackson Laboratory, Strain 008320) mice.

Techniques: Immunostaining, Transduction, Two Tailed Test, Expressing, Injection, Labeling

Journal: eLife

Article Title: tTARGIT AAVs mediate the sensitive and flexible manipulation of intersectional neuronal populations in mice

doi: 10.7554/eLife.66835

Figure Lengend Snippet: Representative image of GFP-IR (black) in the ( a ) hypothalamus of adult Lepr Cre ;Rosa26 LSL-eGFP-L10a ( Lepr Cre ; left), Slc17a6 FlpO ;Rosa26 FSF-eGFP- L10a ( Slc17a6 FlpO ; middle), and Slc17a6 FlpO ;Lepr Cre ;R26 RCFL-eGFP-L10a ( Slc17a6 FlpO ;Lepr Cre ; right) mice. ( b ) GFP-IR (black) in the ventral premamillary nucleus (PMv), periaqueductal gray (PAG), and nucleus of the solitary tract (NTS) in Slc17a6 FlpO ;Lepr Cre ;Rosa26 RCFL-eGFP-L10a animals. Scale bar = 100 μm.

Article Snippet: Genetic reagent ( M. musculus ) , Lepr IRES-Cre , Jackson Laboratories , Strain #: 032457 , Mouse: Lepr-IRES-Cre.

Techniques:

Journal: eLife

Article Title: tTARGIT AAVs mediate the sensitive and flexible manipulation of intersectional neuronal populations in mice

doi: 10.7554/eLife.66835

Figure Lengend Snippet: ( a ) Schematic of unilateral injection strategy for INTERSECT virus and injection site marker into the VMH of Slc17a6 FlpO ;Lepr Cre mice. Representative images showing mCherry-IR (red, left) and YFP-IR (green, right) in the VMH of three separate Slc17a6 FlpO ;Lepr Cre mice co-injected with a Cre-dependent hM3Dq-mCherry AAV (an injection site marker) and INTERSECT Flp-ON/Cre-ON ChR2-eYFP AAV.

Article Snippet: Genetic reagent ( M. musculus ) , Lepr IRES-Cre , Jackson Laboratories , Strain #: 032457 , Mouse: Lepr-IRES-Cre.

Techniques: Injection, Virus, Marker

Journal: eLife

Article Title: tTARGIT AAVs mediate the sensitive and flexible manipulation of intersectional neuronal populations in mice

doi: 10.7554/eLife.66835

Figure Lengend Snippet: ( a ) The tTARGIT system employs the combination of ‘Driver’ (AAV-hSYN-fDIO-tTA) and ‘Payload’ (AAV-TRE-DIO-Payload) AAVs. The Driver virus encodes a Flp-dependent tetracycline transactivator (tTA) under control of the human synapsin I (hSYN1) promoter and two tetracycline operators (TetO). The Payload virus encodes a Cre-dependent Payload transgene under control of the tetracycline response element (TRE). ( b,c ) Experimental scheme ( b ) and representative images ( c ) showing the detection of TdTomato (DSRed-IR, red) following the co-injection of AAV-hSYN-fDIO-tTA and AAV-TRE-DIO-ChR2-TdT into the VMH of (from left to right, as labeled) wild-type (WT), Slc17a6 FlpO , Lepr Cre , or Slc17a6 FlpO; Lepr Cre mice. ( d,e ) Experimental schematic ( d ) and representative images ( e ) showing the detection of TdTomato (DSRed-IR, red) following the injection of (from left to right, as labeled): (1) AAV-hSYN-fDIO-tTA, (2) AAV-TRE-DIO-ChR2-TdT, or the two viruses combined (1+2) into the VMH of Slc17a6 FlpO; Lepr Cre mice. Scale bars = 100 μm.

Article Snippet: Genetic reagent ( M. musculus ) , Lepr IRES-Cre , Jackson Laboratories , Strain #: 032457 , Mouse: Lepr-IRES-Cre.

Techniques: Virus, Control, Injection, Labeling

Journal: eLife

Article Title: tTARGIT AAVs mediate the sensitive and flexible manipulation of intersectional neuronal populations in mice

doi: 10.7554/eLife.66835

Figure Lengend Snippet: Representative images ( a ) and quantification ( b ) of mCherry-IR in Slc17a 6FlpO ;Lepr Cre and Lepr Cre -only mice co-injected with AAV-hSYN1-fDIO-tTA and AAV-TRE-DIO-hM3Dq-mCherry. Data presented as mean ± SEM. Scale bar = 100 μm. 3V = third ventricle.

Article Snippet: Genetic reagent ( M. musculus ) , Lepr IRES-Cre , Jackson Laboratories , Strain #: 032457 , Mouse: Lepr-IRES-Cre.

Techniques: Injection

Journal: eLife

Article Title: tTARGIT AAVs mediate the sensitive and flexible manipulation of intersectional neuronal populations in mice

doi: 10.7554/eLife.66835

Figure Lengend Snippet: ( a ) Experimental schematic showing unilateral co-injection of the Flp-dependent tTARGIT Driver AAV and ChR2-TdTomato-expressing tTARGIT Payload AAV targeting the VMH of Slc17a6 FlpO ;Lepr Cre mice, with optic fiber implantation. ( b,c ) Representative images and quantification ( d ) of ChR2-Tdtomato (red) and FOS-IR (cyan) with either no photostimulation (left) or 1 hr photostimulation (right); ( c ) a digital zoom on the boxed region in ( b ). Data presented as mean ± SEM. Scale bar = 100 μm.

Article Snippet: Genetic reagent ( M. musculus ) , Lepr IRES-Cre , Jackson Laboratories , Strain #: 032457 , Mouse: Lepr-IRES-Cre.

Techniques: Injection, Expressing

Journal: eLife

Article Title: tTARGIT AAVs mediate the sensitive and flexible manipulation of intersectional neuronal populations in mice

doi: 10.7554/eLife.66835

Figure Lengend Snippet: ( a ) The tTARGIT Flp-ON/Cre-OFF system is composed of a Flp-dependent tTA Driver virus (1) with a Payload virus (2) in which the transgene lies in the forward orientation unless Cre mediates its inversion. ( b–d ) Schematic of Flp-ON/Cre-OFF viral injection strategy, representative images ( c ) and quantification ( d ) of mCherry-IR (magenta) and GFP-IR (green) from three independent injections of the Flp-ON/Cre-OFF tTARGIT system using an hM3Dq-mCherry transgene into the VMH of Slc17a6 FlpO ;Lepr Cre ; Rosa26 LSL-eGFP-L10a mice (which express GFP in all Lepr Cre cells). Bottom images show zooms of the boxed regions in the top panels. White arrowheads indicate localization between GFP and mCherry. Data presented as mean± SEM. Scale bars = 100 μm.

Article Snippet: Genetic reagent ( M. musculus ) , Lepr IRES-Cre , Jackson Laboratories , Strain #: 032457 , Mouse: Lepr-IRES-Cre.

Techniques: Virus, Injection

Journal: eLife

Article Title: tTARGIT AAVs mediate the sensitive and flexible manipulation of intersectional neuronal populations in mice

doi: 10.7554/eLife.66835

Figure Lengend Snippet: ( a ) Experimental schematic showing the injection of the Driver AAV with the Payload AAV encoding a GFP-2A-SynmRuby transgene into the VMH of Slc17a6 FlpO; Lepr Cre animals. ( b,c ) Representative image of GFP-IR (green) showing viral transduction in the VMH ( b ) and projections ( c ) in the periaqueductal gray (PAG), arcuate nucleus (ARC), paraventricular hypothalamic nucleus (PVN), bed nucleus of the stria terminalis (BNST), and preoptic area (POA). ( d ) Cartoon showing the projection targets of LepRb VMH neurons.

Article Snippet: Genetic reagent ( M. musculus ) , Lepr IRES-Cre , Jackson Laboratories , Strain #: 032457 , Mouse: Lepr-IRES-Cre.

Techniques: Injection, Transduction

Journal: eLife

Article Title: tTARGIT AAVs mediate the sensitive and flexible manipulation of intersectional neuronal populations in mice

doi: 10.7554/eLife.66835

Figure Lengend Snippet: ( a ) Approach for activating LepRb VMH neurons by co-injecting the AAV-hSYN-fDIO-tTA Driver and the AAV-TRE-DIO-hM3Dq-mCherry Payload viruses into the VMH of Slc17a6 FlpO; Lepr Cre animals. ( b–d ) Representative images showing mCherry detection (DSRed-IR, red) and FOS-IR (cyan) in LepRb VMH-Dq mice administered Vehicle ( b ) or CNO (1 mg/kg) ( c,d ). The right-hand panel of ( c ) shows a digital zoom of the boxed region in the left panel. Scale bars = 100 μm. ( e–j ) Results from metabolic cage analysis of LepRb VMH-Dq mice (n = 6) treated with either vehicle (gray) or 1 mg/kg CNO (red) at 9:30 and 16:30 (dotted lines). Lines in left panels denotes mean value; shading denotes SEM. Each animal was treated with vehicle for 2 days followed by CNO for 2 days to allow pairing. Bar graphs to the right show the average for each mouse at each time point across 24 hr and separated by time in light cycle (day = light, night = dark). ( k, l ) Changes to intrascapular temperatures over 120 min in LepRb VMH-Dq mice (n = 8) or GFP-injected (n = 7) controls following vehicle (gray) or CNO (red; 1 mg/kg) administration at 30°C. ( l ) Shows area under the curve (AUC) for each treatment condition in ( k ). For metabolic cage studies, statistical significance was determined using either a paired t-test (full-day data) or a linear mixed model for effects by time of day. For interscapular temperature measurements, significance was determined by paired t-test. *p<0.05, **p<0.01, ***p<0.001. Figure 3—source data 1. Raw Data from metabolic cages.

Article Snippet: Genetic reagent ( M. musculus ) , Lepr IRES-Cre , Jackson Laboratories , Strain #: 032457 , Mouse: Lepr-IRES-Cre.

Techniques: Injection

Journal: eLife

Article Title: tTARGIT AAVs mediate the sensitive and flexible manipulation of intersectional neuronal populations in mice

doi: 10.7554/eLife.66835

Figure Lengend Snippet: ( a–c ) Body weight, food intake, and water intake of chow-fed Lepr Cre -only controls injected with AAV-hSYN1-fDIO-tTA and AAV-TRE-DIO-hM3Dq-mCherry (black lines; VEH n = 5; CNO n = 8) and LepRb VMH-Dq mice (red lines n = 8 both conditions) receiving 2 days normal drinking water (days 0–2), followed by 3 days exposure to CNO-laced drinking water (days 2–5), followed by an additional 2 days of normal drinking water. Onset and termination of CNO treatment are denoted by vertical lines in each panel. Vehicle controls (dashed red and black lines) did not receive CNO on days 2–5. ( d ) Comparisons between body weight (closed black circles) and food intake (gray squares) over time in LepRb VMH-Dq animals. Data is presented as mean ± SEM. Significance was determined by linear mixed model; ***p<0.001 between CNO and VEH LepRb VMH-Dq animals. # p<0.05, ### p<0.001 between CNO and VEH Lepr Cre -only Control animals. Figure 4—source data 1. Raw data from chronic CNO dosing.

Article Snippet: Genetic reagent ( M. musculus ) , Lepr IRES-Cre , Jackson Laboratories , Strain #: 032457 , Mouse: Lepr-IRES-Cre.

Techniques: Injection, Control

Journal: eLife

Article Title: tTARGIT AAVs mediate the sensitive and flexible manipulation of intersectional neuronal populations in mice

doi: 10.7554/eLife.66835

Figure Lengend Snippet:

Article Snippet: Genetic reagent ( M. musculus ) , Lepr IRES-Cre , Jackson Laboratories , Strain #: 032457 , Mouse: Lepr-IRES-Cre.

Techniques: Recombinant, Plasmid Preparation, Expressing, Knock-In, Software

Journal: eLife

Article Title: Leptin receptor neurons in the dorsomedial hypothalamus regulate diurnal patterns of feeding, locomotion, and metabolism

doi: 10.7554/eLife.63671

Figure Lengend Snippet: ( A ) Experimental schematic for chronic inhibition of DMH LepR by microinjection on day 0 of an AAV1 containing a Cre-dependent GFP-fused TeTx delivered bilaterally to the DMH of LepR-Cre+ male mice (TeTx; n=7) and Cre-negative littermate controls (control; n=5). ( B ) Stereological fluorescent images from a representative animal showing the rostral-caudal extent of GFP:TeTx expression. ( C ) Left: colorized, higher magnification view of the boxed orange region from ( B ). Middle: higher magnification view of the boxed orange region showing neuronal cell bodies targeted within the DMH. Right: higher magnification view of the boxed red region showing GFP:TeTx+ terminals of targeted DMH LepR neurons within the arcuate nucleus (ARC). ( D ) Mean daily food intake following viral microinjection. Two-way ANOVA: F (1,10) =4.658; p=0.0563 (main effect of TeTx); F (14,140) =4.886; p<0.0001 (time x TeTx interaction). ( E ) Mean daily food intake from week 1 relative to week 4. Two-way ANOVA: F (1,10) =5.575; p=0.0399 (main effect of TeTx); F (1,10) =39; p<0.001 (time x TeTx interaction). ( F ) Body weight expressed as %day 0 value. Two-way ANOVA: F (1,10) =20.18; p=0.0012 (main effect of TeTx). F (19,190) =14.67; p<0.0001 (time x TeTx interaction). ( G ) Fat, lean, and total mass 26 days after viral microinjection. Multiple t-tests; t fat =4.847; p=0.0014; t total =2.884; p=0.016. ( H ) Plasma leptin 21 days after viral microinjection. Unpaired t-test, t=5.17, p=0.0017. Data are mean ± SEM. For repeated measures, post hoc Sidak’s test for each time point is indicated on the graph. *p<0.05,**p<0.01, ***p<0.001, ****p<0.0001. Figure 1—source data 1. Longitudinal measures following DMH-LepR TeTx.

Article Snippet: Genetic reagent ( Mus musculus ) , B6; Lepr IRES-Cre/+ , Jackson Labs , RRID: IMSR_JAX:008320 , .

Techniques: Inhibition, Microinjection, Control, Expressing, Clinical Proteomics

Journal: eLife

Article Title: Leptin receptor neurons in the dorsomedial hypothalamus regulate diurnal patterns of feeding, locomotion, and metabolism

doi: 10.7554/eLife.63671

Figure Lengend Snippet: ( A ) Left: Representative image showing extensive overlap of pSTAT3 expression in GFP:TeTx-expressing DMH LepR in mice sacrificed 90 min after leptin administration (i.p. 5 mg/kg). Right: Higher magnification view of the boxed region from the left. ( B ) Change in body weight (unpaired t-test, t=8.483, p=0.0001) following a 24 hr (ZT2–ZT2’) fast 5 weeks following viral microinjection and before food was returned in ( C ). ( C ) Post-fast (24 hr) refeeding following i.p. injection of saline or leptin (3 mg/kg). Two-way ANOVA: F (1,4) =47.33; p=0.0023 (controls, main effect of leptin). F (1,6) =0.1203; p=0.7405 (TeTx, main effect of leptin). v-, c-, and dDMH = ventral, central, and dorsal compartments of the DMH, respectively. Data are mean ± SEM. For repeated measures, post hoc, Sidak’s test at each time point is indicated on the graph. *p<0.05, ***p<0.001, ****p<0.0001. Figure 2—source data 1. Fast-refeeding +/- Leptin.

Article Snippet: Genetic reagent ( Mus musculus ) , B6; Lepr IRES-Cre/+ , Jackson Labs , RRID: IMSR_JAX:008320 , .

Techniques: Expressing, Microinjection, Injection, Saline

Journal: eLife

Article Title: Leptin receptor neurons in the dorsomedial hypothalamus regulate diurnal patterns of feeding, locomotion, and metabolism

doi: 10.7554/eLife.63671

Figure Lengend Snippet: ( A ) Representative viral expression in DMH LepR from LepR-Cre+ male mice injected bilaterally with an AAV encoding Cre-dependent GFP (left) as a control (control; n=7) or Cre-dependent GFP:TeTx (right; TeTx; n=7). ( B ) Daily body weight expressed as %day 0 value. Two-way ANOVA: F (1,12) =37.82; p<0.0001 (main effect of TeTx); F (7,84) =25.17; p<0.0001 (time x TeTx interaction). Dashed boxes indicate periods wherein mice were housed in calorimetry 2 days after microinjection (this figure) and 30 days after microinjection . Shaded areas indicate dark cycle (ZT14–ZT24). ( C ) Food intake (2 hr binned). ( D ) Photoperiod-averaged food intake from (C). Two-way ANOVA: F (1,12) =3.151; p=0.1012 (main effect of TeTx); F (1,12) =5.725; p=0.034 (time x TeTx interaction). ( E ) Respiratory exchange ratio (RER; 2 hr binned). (F) Photoperiod-averaged RER from (E). Two-way ANOVA: F (1,12) =25.16; p=0.0003 (main effect of TeTx); F ( 1,12) =3.747; p=0.0768 (time x TeTx interaction). ( G ) Locomotor activity (LMA; 2 hr binned). ( H ) Photoperiod-averaged LMA from (G). Two-way ANOVA: F (1,12) =4.673; p=0.0516 (main effect of TeTx); F (1,12) =17.64; p=0.0012 (time x TeTx interaction).

Article Snippet: Genetic reagent ( Mus musculus ) , B6; Lepr IRES-Cre/+ , Jackson Labs , RRID: IMSR_JAX:008320 , .

Techniques: Expressing, Injection, Control, Microinjection, Activity Assay

Journal: eLife

Article Title: Leptin receptor neurons in the dorsomedial hypothalamus regulate diurnal patterns of feeding, locomotion, and metabolism

doi: 10.7554/eLife.63671

Figure Lengend Snippet: Two-hour binned continuous measures (left panels) and mean values across the light (L) and dark (D) periods (right panels) 30 days following microinjection of GFP:TeTx (TeTx; n=7) or GFP control (control; n=7) to the dorsomedial hypothalamic nucleus (DMH) of LepR-Cre+ male mice. Shaded areas indicate dark cycle (ZT14 – ZT24). ( A ) Food intake. Two-way ANOVA: F (1,12) =12; p=0.0047 (main effect of TeTx). F (87,1044) =2.354; p<0.0001 (time x TeTx interaction). ( B ) Mean food intake from (A) during L, D, and 24-hr periods. Two-way ANOVA: F (1,12) =9.567; p=0.0093 (main effect of TeTx). ( C ) Locomotor activity (LMA). Two-way ANOVA: F (1,12) =93.22; p<0.0001 (main effect of TeTx). ( D ) Mean LMA from (C) during L, D, and 24-hr periods. Two-way ANOVA: F (1,12) =110.4; p<0.0001 (main effect of TeTx). ( E ) Heat production. Two-way ANOVA: F (1,12) =1.006; p=0.3357 (main effect of TeTx). ( F ) Mean heat production from (E) during L and D periods. Two-way ANOVA: F (1,12) =1.209; p=0.2930 (main effect of TeTx). ( G ) Respiratory exchange ratio (RER). Two-way ANOVA: F (1,12) =2.789; p=0.1208 (main effect of TeTx). ( H ) Mean RER from (G) during L and D periods. Two-way ANOVA: F (1,12) =2.04; p=0.1788 (main effect of TeTx). Data are mean ± SEM. For repeated measures, post hoc, Sidak’s test at each time point is indicated on the graph. *p<0.05,**p<0.01, ***p<0.001, ****p<0.0001. Figure 3—source data 1. Female calorimetry.

Article Snippet: Genetic reagent ( Mus musculus ) , B6; Lepr IRES-Cre/+ , Jackson Labs , RRID: IMSR_JAX:008320 , .

Techniques: Microinjection, Control, Activity Assay

Journal: eLife

Article Title: Leptin receptor neurons in the dorsomedial hypothalamus regulate diurnal patterns of feeding, locomotion, and metabolism

doi: 10.7554/eLife.63671

Figure Lengend Snippet: ( A ) Average daily food intake following chronic inhibition of DMH LepR by bilateral microinjection on day 0 of a Cre-dependent GFP:TeTx delivered to LepR-Cre+ female mice (TeTx; n=9) and Cre-negative littermate controls (control; n=5). Two-way ANOVA: F (1,12) =0.3474; p=0.5665 (main effect of TeTx); F (13,156) =1.563, p=0.1014 (time x TeTx interaction). ( B ) Cumulative food intake for the first 11 days in A (inset). Two-way ANOVA: F (1,12) =1.503; p=0.2437 (main effect of TeTx); F (10,120) =1.427, p=0.1766 (time x TeTx interaction). ( C ) Daily body weight expressed as %day 0 value. Two-way ANOVA: F (1,12) =7.11; p=0.0205 (main effect of TeTx); F (14,168) =15.83, p<0.0001 (time x TeTx interaction). ( D ) Fat, lean, and total mass 14 days after viral microinjection. Multiple t-tests; t fat =3.268; p=0.0024; t total =4.705; p<0.0001. Data are mean ± SEM. *p<0.05,**p<0.01, ***p<0.001, ****p<0.0001.

Article Snippet: Genetic reagent ( Mus musculus ) , B6; Lepr IRES-Cre/+ , Jackson Labs , RRID: IMSR_JAX:008320 , .

Techniques: Inhibition, Microinjection, Control

Journal: eLife

Article Title: Leptin receptor neurons in the dorsomedial hypothalamus regulate diurnal patterns of feeding, locomotion, and metabolism

doi: 10.7554/eLife.63671

Figure Lengend Snippet: Two-hour binned continuous measures (left panels) and photoperiod-averaged values (right panels) 30 days following bilateral microinjection of Cre-dependent GFP:TeTx (TeTx; n=9) or GFP control (control; n=5) to the dorsomedial hypothalamic nucleus (DMH) of LepR-Cre+ female mice. Shaded areas indicate dark cycle (ZT14–ZT24). ( A ) Food intake. Two-way ANOVA: F (1,12) =12.32; p=0.0043 (main effect of TeTx). F (89,1068) =2.766, p<0.0001; F (89,1068) =2.766; p<0.0001 (time x TeTx interaction). ( B ) Mean food intake from (A) during L, D, and 24-hr periods. Two-way ANOVA: F (1,12) =16.26; p=0.0017 (main effect of TeTx). ( C ) Locomotor activity (LMA). Two-way ANOVA: F (1,12) =36.22; p<0.0001 (main effect of TeTx); F (89,1068) = 5.197; p<0.0001 (time x TeTx interaction). ( D ) Mean LMA from (C) during L, D, and 24-hr periods. Two-way ANOVA: F (1,12) =27.98; p=0.0002 (main effect of TeTx). ( E ) Heat production. Two-way ANOVA: F (1,12) =0.5405; p=0.4764 (main effect of TeTx); F (89,1068) =5.903; p<0.0001 (time x TeTx interaction).

Article Snippet: Genetic reagent ( Mus musculus ) , B6; Lepr IRES-Cre/+ , Jackson Labs , RRID: IMSR_JAX:008320 , .

Techniques: Microinjection, Control, Activity Assay

Journal: eLife

Article Title: Leptin receptor neurons in the dorsomedial hypothalamus regulate diurnal patterns of feeding, locomotion, and metabolism

doi: 10.7554/eLife.63671

Figure Lengend Snippet: ( A ) Experimental timeline. Six weeks following bilateral microinjection of Cre-dependent GFP:TeTx (TeTx; n=7) or GFP control (control; n=7) to the dorsomedial hypothalamic nucleus (DMH) of LepR-Cre+ male mice, mice were acclimated to time-restricted feeding (TRF) in their home cages for a 5-day lead-in before transfer into direct calorimetry. TRF was maintained in calorimetry for an additional 4 days, followed by ad lib feeding. ( B ) Two-hour binned continuous measures of food intake during TRF and transition back to ad lib feeding. Shaded areas indicate dark cycle (ZT14–ZT24). ( C ) Mean L:D food intake from (B) under TRF and ad lib feeding. Two-way ANOVA: F (1,12) =5.084; p=0.0436 (main effect of TeTx); F (3,36) =27.91; p<0.0001 (time x TeTx interaction). ( D ) Mean 24-hr food intake from (C) during TRF and ad lib feeding. Two-way ANOVA: F (1,12) =5.097; p=0.0434 (main effect of TeTx); F (1,12) =47.8; p<0.0001 (main effect of TRF); F (1,12) =19.58; p=0.0008 (TRF x TeTx interaction). Within treatment comparison (TRF vs. ad lib): control t (12) =1.759; p=0.1971; TeTx t (12) =8.018; p<0.0001. Data are mean ± SEM.  For repeated measures, post hoc, Sidak’s test at each time point is indicated on the graph. *p<0.05,**p<0.01, ***p<0.001, ****p<0.0001. Figure 4—source data 1. Body weight, RER, EE, and LMA during TRF.

Article Snippet: Genetic reagent ( Mus musculus ) , B6; Lepr IRES-Cre/+ , Jackson Labs , RRID: IMSR_JAX:008320 , .

Techniques: Microinjection, Control, Comparison

Journal: eLife

Article Title: Leptin receptor neurons in the dorsomedial hypothalamus regulate diurnal patterns of feeding, locomotion, and metabolism

doi: 10.7554/eLife.63671

Figure Lengend Snippet:

Article Snippet: Genetic reagent ( Mus musculus ) , B6; Lepr IRES-Cre/+ , Jackson Labs , RRID: IMSR_JAX:008320 , .

Techniques: Recombinant, Sequencing, Enzyme-linked Immunosorbent Assay, Software

Journal: eLife

Article Title: Leptin receptor neurons in the dorsomedial hypothalamus regulate diurnal patterns of feeding, locomotion, and metabolism

doi: 10.7554/eLife.63671

Figure Lengend Snippet: ( A ) Experimental schematic for chronic inhibition of DMH LepR by microinjection on day 0 of an AAV1 containing a Cre-dependent GFP-fused TeTx delivered bilaterally to the DMH of LepR-Cre+ male mice (TeTx; n=7) and Cre-negative littermate controls (control; n=5). ( B ) Stereological fluorescent images from a representative animal showing the rostral-caudal extent of GFP:TeTx expression. ( C ) Left: colorized, higher magnification view of the boxed orange region from ( B ). Middle: higher magnification view of the boxed orange region showing neuronal cell bodies targeted within the DMH. Right: higher magnification view of the boxed red region showing GFP:TeTx+ terminals of targeted DMH LepR neurons within the arcuate nucleus (ARC). ( D ) Mean daily food intake following viral microinjection. Two-way ANOVA: F (1,10) =4.658; p=0.0563 (main effect of TeTx); F (14,140) =4.886; p<0.0001 (time x TeTx interaction). ( E ) Mean daily food intake from week 1 relative to week 4. Two-way ANOVA: F (1,10) =5.575; p=0.0399 (main effect of TeTx); F (1,10) =39; p<0.001 (time x TeTx interaction). ( F ) Body weight expressed as %day 0 value. Two-way ANOVA: F (1,10) =20.18; p=0.0012 (main effect of TeTx). F (19,190) =14.67; p<0.0001 (time x TeTx interaction). ( G ) Fat, lean, and total mass 26 days after viral microinjection. Multiple t-tests; t fat =4.847; p=0.0014; t total =2.884; p=0.016. ( H ) Plasma leptin 21 days after viral microinjection. Unpaired t-test, t=5.17, p=0.0017. Data are mean ± SEM. For repeated measures, post hoc Sidak’s test for each time point is indicated on the graph. *p<0.05,**p<0.01, ***p<0.001, ****p<0.0001. Figure 1—source data 1. Longitudinal measures following DMH-LepR TeTx.

Article Snippet: Adult Lepr IRES-Cre/+ (LepR-Cre) mice (Jackson Laboratory no. 008320) or Cre-littermate controls were used for all experiments, as described below.

Techniques: Inhibition, Microinjection, Control, Expressing, Clinical Proteomics

Journal: eLife

Article Title: Leptin receptor neurons in the dorsomedial hypothalamus regulate diurnal patterns of feeding, locomotion, and metabolism

doi: 10.7554/eLife.63671

Figure Lengend Snippet: ( A ) Left: Representative image showing extensive overlap of pSTAT3 expression in GFP:TeTx-expressing DMH LepR in mice sacrificed 90 min after leptin administration (i.p. 5 mg/kg). Right: Higher magnification view of the boxed region from the left. ( B ) Change in body weight (unpaired t-test, t=8.483, p=0.0001) following a 24 hr (ZT2–ZT2’) fast 5 weeks following viral microinjection and before food was returned in ( C ). ( C ) Post-fast (24 hr) refeeding following i.p. injection of saline or leptin (3 mg/kg). Two-way ANOVA: F (1,4) =47.33; p=0.0023 (controls, main effect of leptin). F (1,6) =0.1203; p=0.7405 (TeTx, main effect of leptin). v-, c-, and dDMH = ventral, central, and dorsal compartments of the DMH, respectively. Data are mean ± SEM. For repeated measures, post hoc, Sidak’s test at each time point is indicated on the graph. *p<0.05, ***p<0.001, ****p<0.0001. Figure 2—source data 1. Fast-refeeding +/- Leptin.

Article Snippet: Adult Lepr IRES-Cre/+ (LepR-Cre) mice (Jackson Laboratory no. 008320) or Cre-littermate controls were used for all experiments, as described below.

Techniques: Expressing, Microinjection, Injection, Saline

Journal: eLife

Article Title: Leptin receptor neurons in the dorsomedial hypothalamus regulate diurnal patterns of feeding, locomotion, and metabolism

doi: 10.7554/eLife.63671

Figure Lengend Snippet: ( A ) Representative viral expression in DMH LepR from LepR-Cre+ male mice injected bilaterally with an AAV encoding Cre-dependent GFP (left) as a control (control; n=7) or Cre-dependent GFP:TeTx (right; TeTx; n=7). ( B ) Daily body weight expressed as %day 0 value. Two-way ANOVA: F (1,12) =37.82; p<0.0001 (main effect of TeTx); F (7,84) =25.17; p<0.0001 (time x TeTx interaction). Dashed boxes indicate periods wherein mice were housed in calorimetry 2 days after microinjection (this figure) and 30 days after microinjection . Shaded areas indicate dark cycle (ZT14–ZT24). ( C ) Food intake (2 hr binned). ( D ) Photoperiod-averaged food intake from (C). Two-way ANOVA: F (1,12) =3.151; p=0.1012 (main effect of TeTx); F (1,12) =5.725; p=0.034 (time x TeTx interaction). ( E ) Respiratory exchange ratio (RER; 2 hr binned). (F) Photoperiod-averaged RER from (E). Two-way ANOVA: F (1,12) =25.16; p=0.0003 (main effect of TeTx); F ( 1,12) =3.747; p=0.0768 (time x TeTx interaction). ( G ) Locomotor activity (LMA; 2 hr binned). ( H ) Photoperiod-averaged LMA from (G). Two-way ANOVA: F (1,12) =4.673; p=0.0516 (main effect of TeTx); F (1,12) =17.64; p=0.0012 (time x TeTx interaction).

Article Snippet: Adult Lepr IRES-Cre/+ (LepR-Cre) mice (Jackson Laboratory no. 008320) or Cre-littermate controls were used for all experiments, as described below.

Techniques: Expressing, Injection, Control, Microinjection, Activity Assay

Journal: eLife

Article Title: Leptin receptor neurons in the dorsomedial hypothalamus regulate diurnal patterns of feeding, locomotion, and metabolism

doi: 10.7554/eLife.63671

Figure Lengend Snippet: Two-hour binned continuous measures (left panels) and mean values across the light (L) and dark (D) periods (right panels) 30 days following microinjection of GFP:TeTx (TeTx; n=7) or GFP control (control; n=7) to the dorsomedial hypothalamic nucleus (DMH) of LepR-Cre+ male mice. Shaded areas indicate dark cycle (ZT14 – ZT24). ( A ) Food intake. Two-way ANOVA: F (1,12) =12; p=0.0047 (main effect of TeTx). F (87,1044) =2.354; p<0.0001 (time x TeTx interaction). ( B ) Mean food intake from (A) during L, D, and 24-hr periods. Two-way ANOVA: F (1,12) =9.567; p=0.0093 (main effect of TeTx). ( C ) Locomotor activity (LMA). Two-way ANOVA: F (1,12) =93.22; p<0.0001 (main effect of TeTx). ( D ) Mean LMA from (C) during L, D, and 24-hr periods. Two-way ANOVA: F (1,12) =110.4; p<0.0001 (main effect of TeTx). ( E ) Heat production. Two-way ANOVA: F (1,12) =1.006; p=0.3357 (main effect of TeTx). ( F ) Mean heat production from (E) during L and D periods. Two-way ANOVA: F (1,12) =1.209; p=0.2930 (main effect of TeTx). ( G ) Respiratory exchange ratio (RER). Two-way ANOVA: F (1,12) =2.789; p=0.1208 (main effect of TeTx). ( H ) Mean RER from (G) during L and D periods. Two-way ANOVA: F (1,12) =2.04; p=0.1788 (main effect of TeTx). Data are mean ± SEM. For repeated measures, post hoc, Sidak’s test at each time point is indicated on the graph. *p<0.05,**p<0.01, ***p<0.001, ****p<0.0001. Figure 3—source data 1. Female calorimetry.

Article Snippet: Adult Lepr IRES-Cre/+ (LepR-Cre) mice (Jackson Laboratory no. 008320) or Cre-littermate controls were used for all experiments, as described below.

Techniques: Microinjection, Control, Activity Assay

Journal: eLife

Article Title: Leptin receptor neurons in the dorsomedial hypothalamus regulate diurnal patterns of feeding, locomotion, and metabolism

doi: 10.7554/eLife.63671

Figure Lengend Snippet: ( A ) Average daily food intake following chronic inhibition of DMH LepR by bilateral microinjection on day 0 of a Cre-dependent GFP:TeTx delivered to LepR-Cre+ female mice (TeTx; n=9) and Cre-negative littermate controls (control; n=5). Two-way ANOVA: F (1,12) =0.3474; p=0.5665 (main effect of TeTx); F (13,156) =1.563, p=0.1014 (time x TeTx interaction). ( B ) Cumulative food intake for the first 11 days in A (inset). Two-way ANOVA: F (1,12) =1.503; p=0.2437 (main effect of TeTx); F (10,120) =1.427, p=0.1766 (time x TeTx interaction). ( C ) Daily body weight expressed as %day 0 value. Two-way ANOVA: F (1,12) =7.11; p=0.0205 (main effect of TeTx); F (14,168) =15.83, p<0.0001 (time x TeTx interaction). ( D ) Fat, lean, and total mass 14 days after viral microinjection. Multiple t-tests; t fat =3.268; p=0.0024; t total =4.705; p<0.0001. Data are mean ± SEM. *p<0.05,**p<0.01, ***p<0.001, ****p<0.0001.

Article Snippet: Adult Lepr IRES-Cre/+ (LepR-Cre) mice (Jackson Laboratory no. 008320) or Cre-littermate controls were used for all experiments, as described below.

Techniques: Inhibition, Microinjection, Control

Journal: eLife

Article Title: Leptin receptor neurons in the dorsomedial hypothalamus regulate diurnal patterns of feeding, locomotion, and metabolism

doi: 10.7554/eLife.63671

Figure Lengend Snippet: Two-hour binned continuous measures (left panels) and photoperiod-averaged values (right panels) 30 days following bilateral microinjection of Cre-dependent GFP:TeTx (TeTx; n=9) or GFP control (control; n=5) to the dorsomedial hypothalamic nucleus (DMH) of LepR-Cre+ female mice. Shaded areas indicate dark cycle (ZT14–ZT24). ( A ) Food intake. Two-way ANOVA: F (1,12) =12.32; p=0.0043 (main effect of TeTx). F (89,1068) =2.766, p<0.0001; F (89,1068) =2.766; p<0.0001 (time x TeTx interaction). ( B ) Mean food intake from (A) during L, D, and 24-hr periods. Two-way ANOVA: F (1,12) =16.26; p=0.0017 (main effect of TeTx). ( C ) Locomotor activity (LMA). Two-way ANOVA: F (1,12) =36.22; p<0.0001 (main effect of TeTx); F (89,1068) = 5.197; p<0.0001 (time x TeTx interaction). ( D ) Mean LMA from (C) during L, D, and 24-hr periods. Two-way ANOVA: F (1,12) =27.98; p=0.0002 (main effect of TeTx). ( E ) Heat production. Two-way ANOVA: F (1,12) =0.5405; p=0.4764 (main effect of TeTx); F (89,1068) =5.903; p<0.0001 (time x TeTx interaction).

Article Snippet: Adult Lepr IRES-Cre/+ (LepR-Cre) mice (Jackson Laboratory no. 008320) or Cre-littermate controls were used for all experiments, as described below.

Techniques: Microinjection, Control, Activity Assay

Journal: eLife

Article Title: Leptin receptor neurons in the dorsomedial hypothalamus regulate diurnal patterns of feeding, locomotion, and metabolism

doi: 10.7554/eLife.63671

Figure Lengend Snippet: ( A ) Experimental timeline. Six weeks following bilateral microinjection of Cre-dependent GFP:TeTx (TeTx; n=7) or GFP control (control; n=7) to the dorsomedial hypothalamic nucleus (DMH) of LepR-Cre+ male mice, mice were acclimated to time-restricted feeding (TRF) in their home cages for a 5-day lead-in before transfer into direct calorimetry. TRF was maintained in calorimetry for an additional 4 days, followed by ad lib feeding. ( B ) Two-hour binned continuous measures of food intake during TRF and transition back to ad lib feeding. Shaded areas indicate dark cycle (ZT14–ZT24). ( C ) Mean L:D food intake from (B) under TRF and ad lib feeding. Two-way ANOVA: F (1,12) =5.084; p=0.0436 (main effect of TeTx); F (3,36) =27.91; p<0.0001 (time x TeTx interaction). ( D ) Mean 24-hr food intake from (C) during TRF and ad lib feeding. Two-way ANOVA: F (1,12) =5.097; p=0.0434 (main effect of TeTx); F (1,12) =47.8; p<0.0001 (main effect of TRF); F (1,12) =19.58; p=0.0008 (TRF x TeTx interaction). Within treatment comparison (TRF vs. ad lib): control t (12) =1.759; p=0.1971; TeTx t (12) =8.018; p<0.0001. Data are mean ± SEM.  For repeated measures, post hoc, Sidak’s test at each time point is indicated on the graph. *p<0.05,**p<0.01, ***p<0.001, ****p<0.0001. Figure 4—source data 1. Body weight, RER, EE, and LMA during TRF.

Article Snippet: Adult Lepr IRES-Cre/+ (LepR-Cre) mice (Jackson Laboratory no. 008320) or Cre-littermate controls were used for all experiments, as described below.

Techniques: Microinjection, Control, Comparison

Journal: eLife

Article Title: Leptin receptor neurons in the dorsomedial hypothalamus regulate diurnal patterns of feeding, locomotion, and metabolism

doi: 10.7554/eLife.63671

Figure Lengend Snippet:

Article Snippet: Adult Lepr IRES-Cre/+ (LepR-Cre) mice (Jackson Laboratory no. 008320) or Cre-littermate controls were used for all experiments, as described below.

Techniques: Recombinant, Sequencing, Enzyme-linked Immunosorbent Assay, Software