Ingestion-activated CGRP neurons control learning but not satiety
Data files
Jul 31, 2026 version files 73.04 MB
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Figure_1.xlsx
8.71 MB
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Figure_2.xlsx
23.93 MB
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Figure_3.xlsx
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Figure_4.xlsx
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Figure_5.xlsx
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README.md
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Supplemental_Fig_1.xlsx
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Supplemental_Fig_2.xlsx
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Supplemental_Fig_3.xlsx
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Supplemental_Fig_4.xlsx
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Supplemental_Fig_5.xlsx
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Supplemental_Fig_6.xlsx
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Supplemental_Fig_7.xlsx
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Supplemental_Fig_8.xlsx
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Abstract
Calcitonin Gene-Related Peptide (CGRP) neurons in the parabrachial nucleus are critical for sickness and malaise but have also been proposed to control non-aversive (rewarding) satiation. How one cell type coordinates these opposing processes has not been explained. Here we reinvestigate the function of these cells using single-cell imaging as well as optical and chemogenetic manipulations in mice. Contrary to current models, we show that CGRP neurons do not track cumulative food consumption, and their activity is not necessary for meal termination. Instead, we identify two distinct populations of CGRP cells, one of which responds rapidly to appetitive signals during ingestion and the other of which responds slowly to aversive visceral cues. Surprisingly, the ingestion-activated CGRP neurons are important for learning about post-ingestive effects but have little effect on food consumption. This reveals two populations of CGRP neurons that are sequentially engaged during, and responsible for, distinct stages of post-ingestive learning.
Dataset DOI: 10.5061/dryad.qnk98sg03
Description of the data and file structure
This readme file provides instructions on reproducing the results presented in the research article titled "Ingestion-activated CGRP neurons control learning but not satiety," by Jarvie et al. and accepted in Neuron, 2026. We have submitted our raw data for main figure 1 (Figure_1.xlsx), main figure 2 (Figure_2.xlsx), main figure 3 (Figure_3.xlsx), main figure 4 (Figure_4.xlsx), main figure 5 (Figure_5.xlsx), and supplemental figure 1 (Supplemental_Fig_1.xlsx), supplemental figure 2 (Supplemental_Fig_2.xlsx), supplemental figure 3 (Supplemental_Fig_3.xlsx), supplemental figure 4 (Supplemental_Fig_4.xlsx), supplemental figure 5 (Supplemental_Fig_5.xlsx), supplemental figure 6 (Supplemental_Fig_6.xlsx), supplemental figure 7 (Supplemental_Fig_7.xlsx), supplemental figure 8 (Supplemental_Fig_8.xlsx).
Descriptions
Figure 1 Data
*Figure 1b
*Z-scored traces: Mean z-scored calcium activity of all neurons over time when given access to Ensure.
*Figure 1c
*Z-scored traces: Mean z-scored calcium activity of Ensure-activated neurons relative to the start of a bout of licking.
*Z-scored traces: Mean z-scored calcium activity of Ensure-activated neurons relative to the end of a bout of licking.
*Figure 1f
*Simple linear regression: Correlation between total licks for Ensure during intermittent 60-second trials and calcium activity of individual neurons.
*Figure 1g
*R2 of lick-bout state predicting neural activity of the Ensure-activated population, comparing observed activity with shuffled controls.
*Figure 1h
*Cumulative distribution function: Z-scored calcium activity selectively during bouts of licking for Ensure or air.
*Figure 1i
*Z-scored traces: Mean z-scored calcium activity of solution-activated neurons relative to the start of a bout of licking for intralipid, glucose, saline, or sucralose.
*Figure 1j
*Time to 50% peak z-score: Time in seconds to half of maximum z-scored change in activity during oral ingestion.
*Figure 1k
*Percentage: Quantification of neurons that were inhibited, activated, or non-responsive during voluntary licking for solutions.
*Figure 1l
*Mean z-score 0-10 min: Average z-scored calcium activity during oral ingestion of solutions, for both activated and inhibited neurons.
*Figure 1m
*Mean z-score normalized to ingestion (licks): Mean z-scored calcium activity selectively during bouts of licking, normalized to the number of licks in each bout.
Figure 2 Data
*Figure 2d
*Mean z-score: Average z-scored calcium activity of Ensure-activated neurons during oral ingestion and IG infusions.
*Figure 2e
*Mean z-score normalized to ingestion (licks): Average z-scored calcium activity during ingestion of Ensure before and after an IG infusion.
*Figure 2f
*Z-scored traces: Mean z-scored calcium activity of all neurons over time during ingestion of chow and high-fat diet.
*Figure 2g
*Solid food consumption (g): Food intake of chow or high-fat diet over 60 minutes.
*Figure 2i
*Percentage: Categorization of neural responses to solid food intake.
*Figure 2j
*Z-scored neural traces: Calcium activity relative to the first bite in a bout of food intake for chow and high-fat diet.
*Figure 2k
*Peak z-score: Maximum z-score of neural activity relative to the first bite in a bout of food intake.
*Figure 2l
*Time to 50% of the peak-z-score: Time in seconds to half of maximum z-scored change in activity following the first bite in a bout of food intake.
*Figure 2m
*R2 of lick-bout state predicting neural activity of the activated population, comparing observed activity with shuffled controls.
*Figure 2n
*Mean z-score: Average z-scored calcium activity during bouts, or during the interbout interval, when eating either chow to high-fat diet food.
*Figure 2o
*Percentage neurons activated in 5 minute bins: Number of neurons with elevated activity every 5 minutes during a 60-minute access period to chow or high-fat diet.
*Figure 2p
*Mean z-score over 10 minutes: Mean z-scored calcium activity of neurons activated by ingestion of solid food, looking at the first and last 10 minutes of access.
*Mean z-score over 10 minutes: Mean z-scored calcium activity of neurons with delayed activation during ingestion of solid food, looking at the first and last 10 minutes of access.
Figure 3 Data
*Figure 3b
*Z-scored traces: Mean z-scored calcium activity of activated and inhibited neurons in response to IG infusions.
*Figure 3c
*Percentages of whole population: Categorization of neural responses to IG infusions.
*Figure 3d
*Mean z-score 0-10 minutes: Average z-scored calcium activity during IG infusions.
*Figure 3f
*Response time (seconds): Time to start of neural responses relative to the start of an IG infusions.
*Figure 3g
*Inverse survival curve: Cumulative fraction of neurons that are activated by IG infusions.
*Figure 3h
*Cumulative distribution function: Mean z-scored calcium activity 0-10 minutes during IG infusions of 1 or 1.5 mL Ensure.
*Cumulative distribution function: Mean z-scored calcium activity 0-10 minutes after IG infusions of 1 or 1.5 mL Ensure.
*Figure 3j
*Preference (%): Licks for sucralose divided by licks for water and sucralose (total licks).
Figure 4 Data
*Figure 4c
*Mean z-score 0-10 min: Average z-scored calcium activity during oral ingestion of Ensure, or following a LiCl injection.
*Figure 4f
*Mean z-score 0-10 min: Average z-scored calcium activity during oral ingestion of Ensure, or following a DON injection.
*Figure 4h
*Number of cells: Number of cells in the field-of view of neural recordings from individual mice, categorized based on position of the GRIN lens.
*Figure 4i
*Percentage of neurons: The number of cells that increased calcium activity in response to different stimuli, categorized based on the position of the GRIN lens.
Figure 5 Data
*Figure 5c
*Ensure consumption (licks): Cumulative licks for Ensure during laser (closed-loop) or no laser over 60 minutes.
*Total licks: Total licks for Ensure during laser (closed-loop) or no laser over 60 minutes.
*Figure 5d
*Bout number: Total number of bouts of licking during laser (closed-loop) or no laser trials.
*Bout duration (seconds): Average length of bouts of licking during laser (closed-loop) or no laser trials.
*Meal number: Total number of meals during laser (closed-loop) or no laser trials.
*Meal duration (minutes): Average length of meals during laser (closed-loop) or no laser trials.
*Figure 5e
*Ensure consumption (licks): Cumulative licks for Ensure during laser (closed-loop) or no laser over 120 minutes.
*Total licks: Total licks for Ensure during laser (closed-loop) or no laser over 120 minutes.
*Figure 5f
*Bout number: Total number of bouts of licking during laser (closed-loop) or no laser trials.
*Bout duration (seconds): Average length of bouts of licking during laser (closed-loop) or no laser trials.
*Meal number: Total number of meals during laser (closed-loop) or no laser trials.
*Meal duration (minutes): Average length of meals during laser (closed-loop) or no laser trials.
*Figure 5g
*Preference (%): Licks for quinine divided by total licks for water and quinine.
*Figure 5i
*Total licks: Total number of licks for sucralose during training sessions.
*Figure 5j
*Preference (%): Licks for sucralose divided by total licks for water and sucralose during a two-bottle choice test across multiple trials.
*Figure 5k
*Preference (%): Licks for glucose divided by licks for water and glucose (total licks).
Supplemental Figure 1 Data
*Supplemental Figure 1b
*Mean z-scored neural activity over time: Averaged z-scored calcium activity of all neurons over time in response to oral ingestion.
*Supplemental Figure 1c
*Total licks: Oral ingestion of various solutions, reported as number of licks.
*Supplemental Figure 1e
*Mean z-score across trials: Average z-scored calcium activity normalized to amount ingested (licks) across three intermittent 60-second windows of access to Ensure at the start and end of a 60 minute session.
*Supplemental Figure 1g
*Mean z-score: Average z-scored calcium activity of activated neurons during bouts of licking for Ensure, compared to calcium activity between bouts.
*Supplemental Figure 1h
*Mean z-score: Average z-scored calcium activity of activated neurons during the interbout interval when mice are not licking, for caloric and non-caloric solutions.
*Supplemental Figure 1i
*Mean z-score: Average z-scored calcium activity of neurons that do not respond to ingestion of Ensure, specifically during the interbout intervals when mice are not licking.
*Supplemental Figure 1j
*Mean z-score: Average z-scored calcium activity of neurons activated by ingestion of caloric solutions, specifically during the interbout intervals in the elast 10-minutes of the access period.
Supplemental Figure 2 Data
*Supplemental Figure 2a
*Quantification of viral expression (load): Total GFP-positive pixels/total area sampled per nucleus from individual animals.
Supplemental Figure 3 Data
*Supplemental Figure 3a
*Percentage of neurons inhibited over time: Quantification of neurons inhibited by oral ingestion of chow or high-fat diet in consecutive 5-minute time bins.
*Supplemental Figure 3b
*Percentage neurons inhibited: Quantification of neurons inhibited by oral ingestion during the first and last 5-minutes of food access for individual animals.
*Supplemental Figure 3d
*Mean z-score over 10 minutes: Mean z-scored calcium activity of neurons inhibited by ingestion of solid food, looking at the first and last 10 minutes of access.
*Mean z-score over 10 minutes: Mean z-scored calcium activity of neurons with delayed inhibition during ingestion of solid food, looking at the first and last 10 minutes of access.
Supplemental Figure 4 Data
*Supplemental Figure 4a
*Mean z-scored neural activity over time: Averaged z-scored calcium activity of activated neurons over time in response to IG infusions of macronutrients.
*Supplemental Figure 4b
*Inverse survival curve: Cumulative fraction of neurons that are activated by IG infusions.
*Supplemental Figure 4c
*Mean z-score, 5 minute bins: Average z-scored calcium activity of activated neurons across 5-minute epochs in response to IG infusions.
*Supplemental Figure 4d
*Response time (seconds): Time to the start of inhibitory neural responses relative to the start of IG infusions.
*Supplemental Figure 4e
*Mean z-score (10 minutes): Average z-scored calcium activity of inhibited neurons in the 10 minutes following the completion of IG infusions.
Supplemental Figure 5 Data
*Supplemental Figure 5b
*Percentage of whole population: Categorization of neural responses to injection of saline, CCK, or Exendin-4.
*Supplemental Figure 5c
*Z-scored neural activity over time: Averaged z-scored calcium activity of activated neurons over time in response to injection of CCK.
*Mean z-score over 10 minutes: Average z-scored calcium activity over consecutive 10 minute time bins in response to injection of CCK.
*Supplemental Figure 5d
*Z-scored neural activity over time: Averaged z-scored calcium activity of activated neurons over time in response to injection of Exendin-4.
*Mean z-score over 10 minutes: Average z-scored calcium activity over consecutive 10 minute time bins in response to injection of Exendin-4.
Supplemental Figure 6 Data
*Supplemental Figure 6b
*Normalized weight (g): Weight of the contents of different compartments of the gastrointestinal tract following an IG infusion.
*Supplemental Figure 6c
*Tissue weight (g): Weight of the tissue of different compartments of the gastrointestinal tract.
*Supplemental Figure 6d
*Gastric emptying (volume/min): Difference in the volume infused in the stomach and what remains at the end of the IG infusion, divided by total infusion time.
*Supplemental Figure 6e
*Stomach area (cm2): Measurement of the surface area of one side of the stomach following IG infusions.
Supplemental Figure 7 Data
*Supplemental Figure 7a
*Ensure consumption (licks): Cumulative licks for Ensure during laser (continuous) or no laser over 60 minutes.
*Total licks: Total licks for Ensure during laser (continuous) or no laser over 60 minutes.
*Supplemental Figure 7b
*Bout number: Total number of bouts of licking during laser (continuous) or no laser trials.
*Bout duration (seconds): Average length of bouts of licking during laser (continuous) or no laser trials.
*Meal number: Total number of meals during laser (continuous) or no laser trials.
*Meal duration (minutes): Average length of meals during laser (continuous) or no laser trials.
*Supplemental Figure 7c
*Ensure consumption (licks): Cumulative licks for Ensure during laser (continuous) or no laser over 60 minutes.
*Total licks: Total licks for Ensure during laser (continuous) or no laser over 60 minutes.
*Supplemental Figure 7d
*Bout number: Total number of bouts of licking during laser (continuous) or no laser trials.
*Bout duration (seconds): Average length of bouts of licking during laser (continuous) or no laser trials.
*Meal number: Total number of meals during laser (continuous) or no laser trials.
*Meal duration (minutes): Average length of meals during laser (continuous) or no laser trials.
Supplemental Figure 8 Data
*Supplemental Figure 8b
*Ensure consumption (licks): Cumulative licks for Ensure following injection of saline or DCZ.
*Total licks: Total licks for Ensure following injection of saline or DCZ.
*Supplemental Figure 8c
*Bout number: Total number of bouts of licking following injection of saline or DCZ.
*Bout duration (seconds): Average length of bouts of licking following injection of saline or DCZ.
*Meal number: Total number of meals following injection of saline or DCZ.
*Meal duration (minutes): Average length of meals following injection of saline or DCZ.
*Supplemental Figure 8d
*Ensure consumption (licks): Cumulative licks for Ensure following injection of saline or DCZ.
*Total licks: Total licks for Ensure following injection of saline or DCZ.
*Supplemental Figure 8e
*Bout number: Total number of bouts of licking following injection of saline or DCZ.
*Bout duration (seconds): Average length of bouts of licking following injection of saline or DCZ.
*Meal number: Total number of meals following injection of saline or DCZ.
*Meal duration (minutes): Average length of meals following injection of saline or DCZ.
*Supplemental Figure 8f
*Food consumption (g): Cumulative food intake over time following injection of saline or DCZ.
*Supplemental Figure 8g
*Total food consumption (g): Consumption of food over 2 hours following injection of saline or DCZ.
*Supplemental Figure 8h
*Total food consumption (pellets): Number of food pellets consumed over the first 2 hours of the dark cycle following injection of saline or DCZ.
*Meal number: Total number of meals following injection of saline or DCZ.
*Meal size: Average number of pellets consumed per meal following injection of saline or DCZ.
*Supplemental Figure 8j
*Preference (%): Licks for sucralose divided by total licks for water and sucralose.
Sharing/Access information
The data were collected in a research laboratory (Knight Lab at UCSF): https://knightlab.ucsf.edu/.
Code/Software
GraphPad Prism version 10 and MATLAB 2025b were used to perform statistical tests with the source data to produce the results in the manuscript "Ingestion-activated CGRP neurons control learning but not satiety," accepted at Neuron. Custom MATLAB code can be found at (https://github.com/zackknightlabucsf/zackknightlabucsf).