The p75 neurotrophin receptor in AgRP neurons is necessary for homeostatic feeding and food anticipation
- University of Virginia, United States
- The Ohio State University College of Medicine, United States
Figures
Figure 1 with 2 supplements
Fasting Ngfr-KO mice reveals activity and feeding deficits.
(A) Schematic of nighttime fasting (left). Mice are fasted just before lights out at ZT12, weighed at ZT0, and fed for 3 hr between ZT4 and ZT7 (right). n = 8 WT, 9 KO. *p=0.0195 by Student’s …
Figure 1—figure supplement 1
Germline loss of p75NTR does not alter body weight, activity, or daily food intake.
(A) Weekly body weight measurements; n = 7/group. Not significant by two-way repeated measures ANOVA with Bonferroni multiple comparisons. (B) 24 hr (total) and 12 hr (night, day), measurements of …
Figure 1—figure supplement 2
Refeeding food intake normalized to total body weight.
(A) Daytime refeeding normalized to overnight fasted body weight. *p=0.0149 by Student’s t-test. (B) Nighttime refeeding normalized to overday fasted body weight. Not significant by Student’s t-test.
Figure 2 with 2 supplements
Ngfr-KO mice have blunted food anticipatory activity in response to daytime, but not nighttime, restricted feeding.
(A) Schematic of daytime restricted feeding paradigm. Mice are fasted just before lights out at ZT12, weighed at ZT0, fed for 3 hr between ZT4 and ZT7, and then fasted again until the next day’s …
Figure 2—figure supplement 1
Glucose homeostasis and ketone production are intact in Ngfr-KO mice during daytime restricted feeding; Normalized food intake.
Blood glucose (A) and β-hydroxybutyrate (B) levels measured at ZT0 during daytime restricted feeding; n = 6 WT, 8 KO. Not significant by two-way repeated measures ANOVA with Bonferroni multiple …
Figure 2—figure supplement 2
Daytime restricted feeding selectively dampens FAA, while nighttime restricted feeding blunts several activity measures.
(A) 24 hr plot of day five restriction locomotor activity (left) quantified into several discrete time periods (right); n = 6/group. *p=0.046 by Student’s t-test. (B) 24 hr plot of night five …
Figure 3 with 1 supplement
Adult loss of p75NTR leads to reduced homeostatic feeding and absent FAA.
(A) Ad libitum food intake (left) and body weight (right) 4 weeks after tamoxifen injection; n = 7/group. Not significant by Student’s t-test. (B) Mice are fasted just before lights out at ZT12 and …
Figure 3—figure supplement 1
Adult food intake normalized to total body weight.
(A) Ad libitum food intake normalized to total body weight. Not significant by Student’s t-test. n = 7/group. (B) Daytime refeeding normalized to overnight fasted body weight. *p=0.0401 by Student’s …
Figure 4 with 1 supplement
p75NTR is expressed in AgRP neurons and is necessary for the arcuate hypothalamus response to fasting.
(A) Representative images of immunofluorescent staining for p75NTR in an Npy-GFP expressing reporter mouse. (B) Immunofluorescent staining for c-Fos (left) in 16 hr overnight fasted mice …
Figure 4—figure supplement 1
p75NTR is necessary for the arcuate hypothalamus response to fasting in germline and adult knockout mice.
(A) Immunofluorescent staining for c-Fos (left) in 16 hr overnight fasted Ngfr-KO and control mice. Quantification (right) as the percent increase of c-Fos+ cells/dapi+ cells in each fasted Ngfr-WT …
Figure 5 with 2 supplements
AgRP neuron specific loss of p75NTR leads to reduced daytime restricted feeding and FAA.
(A) Ad libitum food intake (left) and body weight (right); n = 10 WT, 13 KO. Not significant by Student’s t-test. (B) Mice are fasted just before lights out at ZT12 and fed for 3 hr between ZT4 and …
Figure 5—figure supplement 1
AgRP food intake normalized to total body weight.
(A) Ad libitum food intake normalized to total body weight. Not significant by Student’s t-test. (B) Daytime refeeding normalized to overnight fasted body weight. *p=0.021 by Student’s t-test. (C) …
Figure 5—figure supplement 2
Ghrelin-induced feeding is intact in AgRP-Ngfr-KO mice.
(A) Three hour food intake following central infusion of either vehicle or ghrelin (1 ug) in ad libitum fed AgRP-Ngfr-KO mice or controls (left). Mice are weighed prior to infusion (right). n = 10 …
Figure 6 with 1 supplement
AgRP-Ngfr-KO mice have intact fasting JNK signaling, but blunted CREB activation.
(A) phospho-c-Jun immunofluorescence in fasted AgRP-Ngfr-KO mice and littermate controls. Scale bar = 50 µM. (B) Quantification of the number of phospho-c-Jun+ cells/dapi+ cells in fed and fasting …
Figure 6—figure supplement 1
AgRP-Ngfr-KO mice have blunted activation of fasting CREB in AgRP neurons.
(A) Immunofluorescent staining for pCREB in 16 hr overnight fasted mice co-expressing tdTomato from the AgRP promoter. Red lines indicate the approximate boundaries of AgRP-tdTomato expression. …
Tables
Table 1
Serum chemistries exhibit similar daytime changes in fed and fasted Ngfr-KO mice.
Serum was collected at ZT4 (Zeitgeber Time) in 12–16 week old ad libitum fed mice or following a 16 hr overnight fast. Data are presented as mean ± SEM. *p<0.05 compared to WT by Student’s t-test. …
| Ngfr-WT | Ngfr-KO | ||
|---|---|---|---|
| ZT4 | |||
| Body Weight (g) | Fed | 29.8 ± 0.7 | 26.6 ± 1.2* |
| Fasted | 23.8 ± 1.3# | 22.0 ± 0.7# | |
| Glucose (mg/dl) | Fed | 380 ± 17.5 | 385 ± 16.4 |
| Fasted | 245 ± 19.1# | 307 ± 25.7# | |
| Insulin (ng/ml) | Fed | 0.23 ± 0.1 | 0.37 ± 0.1 |
| Fasted | 0.10 ± 0.1 | 0.10 ± 0.1# | |
| Ketones (mM) | Fed | 0.0 ± 0.02 | 0.0 ± 0.02 |
| Fasted | 2.1 ± 0.36# | 2.1 ± 0.10# | |
| Leptin (ng/ml) | Fed | 2.1 ± 0.5 | 0.9 ± 0.2 |
| Fasted | 0.8 ± 0.16# | 0.9 ± 0.14 | |
| Corticosterone (ng/ml) | Fed | 117 ± 12.4 | 126.7 ± 13.4 |
| Fasted | 170 ± 5.3# | 180 ± 4.1# | |
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Table 1—source data 1
p values by Student’s t-test for each pair-wise comparison in Table 1.
- https://cdn.elifesciences.org/articles/52623/elife-52623-table1-data1-v2.docx
Table 2
Serum chemistries exhibit similar nighttime changes in fed and fasted Ngfr-KO mice.
Serum was collected at ZT16 in 12–16 week old ad libitum fed mice or following a 16 hr overday fast. Data are presented as mean ± SEM. *p<0.05 compared to WT by Student’s t-test. #p<0.05 compared to …
| Ngfr-WT | Ngfr-KO | ||
|---|---|---|---|
| ZT16 | |||
| Body Weight (g) | Fed | 29.4 ± 0.4 | 23.9 ± 0.5* |
| Fasted | 26.2 ± 0.5# | 23.3 ± 0.7* | |
| Glucose (mg/dl) | Fed | 299 ± 13.0 | 312 ± 29.2 |
| Fasted | 145 ± 25.6# | 160 ± 13.6# | |
| Insulin (ng/ml) | Fed | 2.0 ± 0.5 | 0.88 ± 0.2 |
| Fasted | 0.16 ± 0.1# | 0.03 ± 0.1# | |
| Ketones (mM) | Fed | 0.1 ± 0.03 | 0.2 ± 0.11 |
| Fasted | 1.6 ± 0.10# | 1.4 ± 0.13# | |
| Leptin (ng/ml) | Fed | 3.3 ± 0.9 | 1.5 ± 0.2 |
| Fasted | 0.8 ± 0.15# | 0.8 ± 0.12# | |
| Corticosterone (ng/ml) | Fed | 123 ± 8.3 | 146 ± 8.5 |
| Fasted | 163 ± 8.2# | 176 ± 3.7# | |
-
Table 2—source data 1
1 p values by Student’s t-test for each pair-wise comparison in Table 2.
- https://cdn.elifesciences.org/articles/52623/elife-52623-table2-data1-v2.docx
Key resources table
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Gene Mus musculus | Ngfr | NCBI Gene ID: 18053 | ||
| Strain, strain background (Mus musculus, males) | B6.129S4-Ngfrtm1Jae/J Also known as p75NGFR | Jackson Labs | RRID:IMSR_JAX:002213 | |
| Strain, strain background (Mus musculus, males) | Ngfrtm1.1Vk/BnapJ Also known as p75NTR-Fxflox | Bogenmann et al., 2011 | RRID:IMSR_JAX:031162 | Gift from Brian Pierchala |
| Strain, strain background (Mus musculus, males) | B6.Cg-Ndor1Tg(UBC-cre/ERT2)1Ejb/1J Also known as UBC-Cre-ERT2 | Jackson Labs | RRID:IMSR_JAX:007001 | |
| Strain, strain background (Mus musculus, males) | Agrptm1(cre)Lowl/J Also known as Agrp-IRES-cre | Jackson Labs | RRID:IMSR_JAX:012899 | |
| Strain, strain background (Mus musculus, males) | B6.FVB-Tg(Npy-hrGFP)1Lowl/J Also known as NPY-GFP | Jackson Labs | RRID:IMSR_JAX:006417 | |
| strain, strain background (Mus musculus, males) | B6.Cg-Gt(ROSA)26Sortm9(CAG-tdTomato)Hze/J Also known as Ai9 | Jackson Labs | RRID:IMSR_JAX:007909 | |
| Antibody | p75NTR, goat monoclonal | Neuromics | Cat # GT15057 | IF 1:5000 |
| Antibody | c-Fos, rabbit polyclonal | Synaptic Systems | Cat # 226 003 | IF 1:1000 |
| Antibody | phospho-cJun, rabbit monoclonal | Cell Signaling Technology | RRID:AB_2129575 | IF 1:800 |
| Antibody | phospho-CREB | Cell Signaling Technology | RRID:AB_2561044 | IF 1:800 |
| Commercial assay or kit | Insulin elisa | Crystal Chem | RRID:AB_2783626 | |
| Commercial assay or kit | Leptin elisa | Cayman Chemical | Cat # 10007609 | |
| Commercial assay or kit | Corticosterone elisa | Cayman Chemical | Cat # 501320 | |
| Commercial assay or kit | Glucose meter | Bayer | One touch ultra 2 | |
| Commercial assay or kit | Ketone monitor | Abbott | Precision xtra | |
| Peptide, recombinant protein | Ghrelin | Phoenix Pharmaceuticals, inc | Cat # 031–31 | ICV 1 ug |
| Software, algorithm | Prism 8 | Graphpad | RRID:SCR_002798 | |
| Software, algorithm | FIJI | FIJI | RRID:SCR_002285 | |
| Other | DAPI stain | Southern Biotech | Cat # 0100–20 | |
| Other | Opto M4 Activity Monitor | Columbus Instruments |
