Arginine-vasopressin mediates counter-regulatory glucagon release and is diminished in type 1 diabetes
- Division of Endocrinology, Diabetes, and Metabolism, Beth Israel Deaconess Medical Center, United States
- Program in Neuroscience, Harvard Medical School, United States
- Oxford Centre for Diabetes, Endocrinology and Metabolism, Radcliffe Department of Medicine, University of Oxford, United Kingdom
- Section for Cell Biology and Physiology, Department of Biology, University of Copenhagen, Denmark
- Metabolic Research Unit, Institute of Neuroscience and Physiology, Sahlgrenska Academy at University of Gothenburg, Sweden
- Alberta Diabetes Institute, Li Ka Shing Centre for Health Research Innovation, Canada
- Department of Clinical Biochemistry, John Radcliffe, Oxford NHS Trust, United Kingdom
- Section of Cell Biology and Functional Genomics, Department of Metabolism, Digestion and Reproduction, Imperial College London, United Kingdom
- Center for Clinical Metabolic Research, Gentofte Hospital, Denmark
- Department of Clinical Pharmacology, Bispebjerg Hospital, University of Copenhagen, Denmark
- Department of Clinical Medicine, Faculty of Health and Medical Sciences, University of Copenhagen, Denmark
- Steno Diabetes Center Copenhagen, Denmark
- Novo Nordisk Foundation Center for Basic Metabolic Research, Faculty of Health and Medical Sciences, University of Copenhagen, Denmark
- Department of Computer Science, University of Oxford, United Kingdom
Figures
Figure 1 with 2 supplements
Insulin-induced hypoglycemia enhances population activity of AVP neurons in the supraoptic nucleus (SON), driving glucagon secretion AAV-DIO-hM3Dq-mCherry was injected bilaterally into the SON of Avpires-Cre/+ mice.
CNO (3 mg/kg) or vehicle was injected i.p. In the same cohort (different trial), LY2409021 (5 mg/kg) or SSR149415 (30 mg/kg) was injected (i.p.) 30 min prior to CNO. See Figure 1—figure supplement 1.…
Figure 1—figure supplement 1
Effects of CNO in animals expressing mCherry in supraoptic nucleus (SON) AVP neurons (a) AAV-DIO-hM3Dq-mCherry was injected into the SON of Avpires-Cre/+ mice.
Immunostaining for AVP. See Figure 1. (b) Staining for mCherry; same brain sample as (a). (c) Merge of (a) and (b). (d) Patch-clamp recording from mCherry+ neuron in the SON in response to CNO. …
Figure 1—figure supplement 2
Simultaneous continuous glucose monitoring (CGM) and in vivo fiber photometry of AVP neurons Grouped analysis of the glucose value at which the GCaMP6 signal crosses >2 SD from baseline, >3 SD from baseline and first exhibits a peak.
Six trials, two mice. One-way repeated measures ANOVA (Sidak), p<0.05=*. AVP, arginine-vasopressin.
Figure 2 with 2 supplements
AVP increases glucagon release and alpha-cell activity ex vivo, in situ, and in vivo.
(a) Glucagon secretion from the perfused mouse pancreas in response AVP (10 nM). All data are represented as mean ± SEM. n=5 mice. Extracellular glucose of 3 mM. (b) Islets from GcgCre/+:GCaMP3/+ …
Figure 2—figure supplement 1
Expression of the vasopressin 1b receptor in mouse and human.
(a) mRNA expression of Avpr family in mouse heart, kidney, islets and adrenal glands. Samples from n=3 wild-type mice, each run in triplicate. Calculated with the Pfaffl method, using Actb as the …
Figure 2—figure supplement 2
AVP and beta-cell function.
(a) Glucagon secretion (% content) from isolated mouse islets in response to AVP. One-way ANOVA (p<0.05=*). n=5–6 wild-type mice per condition. AVP was applied in high (15 mM glucose). (b) Insulin …
Figure 3
AVP increases action potential firing, Ca2+ activity, and intracellular DAG in alpha-cells in intact islets.
(a) Membrane potential (Vm) recording (perforated patch-clamp) of an alpha-cell in response to 100 pM AVP. (b) Frequency-response curve for varying concentrations of AVP (17 alpha-cells, 10 Gcg-GCaMP…
Figure 4
The vasopressin 1b receptor mediates hypoglycemia-induced glucagon secretion.
(a) Blood glucose during an ITT (0.75 U/kg; injection at 30 min). 30 min prior to the commencement of the ITT (t=0 min), either the V1bR antagonist SSR149415 (30 mg/kg) or vehicle was administered …
Figure 5 with 2 supplements
Insulin-induced AVP secretion is mediated by A1/C1 neurons.
(a) Upper: AAV-DIO-ChR2-mCherry was injected into the VLM of ThCre/+ mice or AvpGFP/+::ThCre/+ mice, targeting A1/C1 neurons. Lower: CRACM. Excitatory post-synaptic currents (EPSCs) were recorded in …
Figure 5—figure supplement 1
Viral tracing of A1/C1 terminals.
(a) Injection of a Cre-dependent viral vector containing the light-gated ion channel Channelrhodopsin-2 (AAV-DIO-ChR2-mCherry) into A1/C1 neurons of ThCre/+ mice. Top left: mCherry expression in …
Figure 5—figure supplement 2
c-Fos expression in A1/C1 neurons during an ITT.
(a) Left: c-Fos expression 30 min following i.p. saline injection. Right: Same animal, but merge of Th and c-Fos. Note the Th+ immunoreactive neurons in the A1/C1 region (white box). (b) Left: c-Fos …
Figure 6
Insulin-induced hypoglycemia evokes copeptin and glucagon secretion in human participants.
(a) Blood glucose was clamped at euglycemia (Eug) and followed during insulin-induced hypoglycemia (Hypo). n=10 healthy human subjects. The clamp was initiated at time 0 min and terminated at 60 …
Figure 7
Insulin-induced copeptin and glucagon secretion is diminished in people with type 1 diabetes.
(a) Hypoglycemia was induced by an insulin infusion in patients with T1D (n=10) and non-diabetic individuals (Controls; n=10). The insulin infusion was initiated at 0 min. (b) Plasma glucagon during …
Tables
Table 1
Participant characteristics for Figure 7.
| Christensen et al., 2011 | Christensen et al., 2015 | P | |
|---|---|---|---|
| Cohort | Control (n=10) | T1D (n=10) | |
| Age (years) | 23±1 | 26±1 | 0.06 |
| BMI (kg/m2) | 23±0.5 | 24±0.5 | 0.1744 |
| HbA1c (%) | 5.5±0.1 | 7.3±0.2 | <0.0001 |
Additional files
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Transparent reporting form
- https://cdn.elifesciences.org/articles/72919/elife-72919-transrepform1-v2.docx
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Source data 1
Data for figures.
- https://cdn.elifesciences.org/articles/72919/elife-72919-supp1-v2.xlsx
