Chronic postnatal chemogenetic activation of forebrain excitatory neurons evokes persistent changes in mood behavior
- Department of Biological Sciences, Tata Institute of Fundamental Research, India
- Centre for Cellular and Molecular Biology, India
- Neuroscience Unit, Jawaharlal Nehru Centre for Advanced Scientific Research, India
Figures
Figure 1 with 4 supplements
Selective expression and activation of hM3Dq DREADD in CamKIIα-positive forebrain excitatory neurons in CamKIIα-tTA::TetO-hM3Dq bigenic mice during the postnatal window.
(A) Shown is a schematic of the experimental strategy for the generation of the bigenic CamKIIα-tTA::TetO-hM3Dq mouse line to selectively drive the expression of the hM3Dq DREADD in CamKIIα-positive …
Figure 1—figure supplement 1
Selective expression of the HA-tagged hM3Dq DREADD in CamKIIα-positive forebrain excitatory neurons in adult CamKIIα-tTA::TetO-hM3Dq bigenic mice.
(A) Shown is a schematic of the experimental strategy for the generation of the bigenic CamKIIα-tTA::TetO-hM3Dq mouse line to selectively drive the expression of the HA-tagged hM3Dq DREADD in …
Figure 1—figure supplement 2
Enhanced p-ERK/ERK expression following chronic postnatal hM3Dq DREADD activation in CamKIIα-positive forebrain excitatory neurons.
(A) Experimental paradigm to assess the influence of chronic PNCNO-mediated hM3Dq DREADD activation of CamKIIα-positive forebrain excitatory neurons on p-ERK/ERK expression. CamKIIα-tTA::TetO-hM3Dq …
Figure 1—figure supplement 3
Spontaneous network activity and intrinsic excitability following chronic postnatal hM3Dq DREADD activation in CamKIIα-positive forebrain excitatory neurons.
(A) Shown is the experimental paradigm to assess the effects of chronic CNO-mediated hM3Dq DREADD activation of CamKIIα-positive forebrain excitatory neurons using whole-cell patch clamp. CamKIIα-tTA…
Figure 1—figure supplement 4
Distribution of spontaneous network events following chronic postnatal hM3Dq DREADD activation in CamKIIα-positive forebrain excitatory neurons.
(A) Shown is an experimental paradigm to assess the effects of chronic CNO-mediated hM3Dq DREADD activation of CamKIIα-positive forebrain excitatory neurons using whole-cell patch clamp. CamKIIα-tTA:…
Figure 2 with 7 supplements
Chronic chemogenetic activation of CamKIIα-positive forebrain excitatory neurons during the early postnatal window results in a long-lasting increase in anxiety- and despair-like behavior in adult male mice.
(A) Shown is a schematic of the experimental paradigm to induce chronic CNO-mediated hM3Dq DREADD activation in CamKIIα-positive forebrain excitatory neurons using bigenic CamKIIα-tTA::TetO-hM3Dq …
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Figure 2—source data 1
Source data for Figure 2.
- https://cdn.elifesciences.org/articles/56171/elife-56171-fig2-data1-v2.xlsx
Figure 2—figure supplement 1
Chronic chemogenetic activation of CamKIIα-positive forebrain excitatory neurons during the early postnatal window does not alter weight during CNO administration and in adulthood.
(A) Shown is a schematic of the experimental paradigm used to determine the influence of chronic CNO-mediated hM3Dq DREADD activation of CamKIIα-positive forebrain excitatory neurons in …
Figure 2—figure supplement 2
Chronic chemogenetic activation of CamKIIα-positive forebrain excitatory neurons during the early postnatal window does not alter the developmental emergence of reflex behaviors.
(A) Shown is a schematic of the experimental paradigm used to determine the influence of chronic CNO-mediated hM3Dq DREADD activation of CamKIIα-positive forebrain excitatory neurons in …
Figure 2—figure supplement 3
Chronic chemogenetic activation of CamKIIα-positive forebrain excitatory neurons during the early postnatal window results in a long-lasting increase in anxiety-like behavior in adult female mice.
(A) Shown is a schematic of the experimental paradigm to induce chronic CNO-mediated hM3Dq DREADD activation in CamKIIα-positive forebrain excitatory neurons using bigenic CamKIIα-tTA::TetO-hM3Dq …
Figure 2—figure supplement 4
Chronic chemogenetic activation of CamKIIα-positive forebrain excitatory neurons during the early postnatal window does not alter repetitive behavior in adult male mice.
(A) Shown is a schematic of the experimental paradigm to induce chronic CNO-mediated hM3Dq DREADD activation in CamKIIα-positive forebrain excitatory neurons using bigenic CamKIIα-tTA::TetO-hM3Dq …
Figure 2—figure supplement 5
Chronic CNO administration during the early postnatal window does not influence anxiety- and despair-like behavior in genotype-control, adult male mice.
(A) Shown is a schematic of the experimental paradigm to assess the influence of chronic CNO administration in genotype-control mice. Mouse pups, single-positive for either CamKIIα-tTA or TetO-hM3Dq …
Figure 2—figure supplement 6
Chronic CNO administration during the early postnatal window does not influence anxiety- and despair-like behavior in C57BL/6J adult male mice.
(A) Shown is a schematic of the experimental paradigm to assess the influence of chronic CNO administration in the background strain for the bigenic CamKIIα-tTA::TetO-hM3Dq, namely the C57BL/6J …
Figure 2—figure supplement 7
Chronic chemogenetic activation of CamKIIα-positive forebrain excitatory neurons during the early postnatal window using the DREADD agonist Compound 21 (C21) results in a long-lasting increase in anxiety-like behavior in adult male mice.
(A) Shown is a schematic of the experimental paradigm to induce chronic C21-mediated hM3Dq DREADD activation in CamKIIα-positive forebrain excitatory neurons using bigenic CamKIIα-tTA::TetO-hM3Dq …
Figure 3 with 2 supplements
Chronic chemogenetic activation of CamKIIα-positive forebrain excitatory neurons during the juvenile window or in adulthood does not evoke any long-lasting changes in anxiety- and despair-like behavior in male mice.
(A) Shown is a schematic of the experimental paradigm to induce chronic CNO-mediated hM3Dq DREADD activation in CamKIIα-positive forebrain excitatory neurons using bigenic CamKIIα-tTA::TetO-hM3Dq …
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Figure 3—source data 1
Source data for Figure 3.
- https://cdn.elifesciences.org/articles/56171/elife-56171-fig3-data1-v2.xlsx
Figure 3—figure supplement 1
Chronic chemogenetic activation of CamKIIα-positive forebrain excitatory neurons during the juvenile window or in adulthood does not evoke any long-lasting changes in anxiety-like behavior in male mice.
(A) Shown is a schematic of the experimental paradigm to induce chronic CNO-mediated hM3Dq DREADD activation in CamKIIα-positive forebrain excitatory neurons using bigenic CamKIIα-tTA::TetO-hM3Dq …
Figure 3—figure supplement 2
Chronic chemogenetic activation of CamKIIα-positive forebrain excitatory neurons in adulthood does not evoke any long-lasting changes in anxiety-like behavior in male mice during or soon after cessation of CNO treatment.
(A) Shown is a schematic of the experimental paradigm to induce chronic CNO-mediated hM3Dq DREADD activation in CamKIIα-positive forebrain excitatory neurons using bigenic CamKIIα-tTA::TetO-hM3Dq …
Figure 4 with 2 supplements
Chronic chemogenetic activation of CamKIIα-positive forebrain excitatory neurons during the early postnatal window evokes impaired sensorimotor gating in adulthood.
(A) Shown is a schematic of the experimental paradigm to induce chronic CNO-mediated hM3Dq DREADD activation in CamKIIα-positive forebrain excitatory neurons using bigenic CamKIIα-tTA::TetO-hM3Dq …
Figure 4—figure supplement 1
Chronic chemogenetic activation of CamKIIα-positive forebrain excitatory neurons during the juvenile window does not alter sensorimotor gating behavior.
(A) Shown is a schematic of the experimental paradigm to induce chronic CNO-mediated hM3Dq DREADD activation in CamKIIα-positive forebrain excitatory neurons using bigenic CamKIIα-tTA::TetO-hM3Dq …
Figure 4—figure supplement 2
Chronic chemogenetic activation of CamKIIα-positive forebrain excitatory neurons in adulthood does not alter sensorimotor gating behavior.
(A) Shown is a schematic of the experimental paradigm to induce chronic CNO-mediated hM3Dq DREADD activation in CamKIIα-positive forebrain excitatory neurons using bigenic CamKIIα-tTA::TetO-hM3Dq …
Figure 5 with 3 supplements
Chronic chemogenetic activation of CamKIIα-positive forebrain excitatory neurons during the early postnatal window results in long-lasting alterations in neurotransmitter cycling flux and neuronal metabolic rate in hippocampus and cortex.
(A) Shown is a schematic of the experimental paradigm to induce chronic CNO-mediated hM3Dq DREADD activation in CamKIIα-positive forebrain excitatory neurons using bigenic CamKIIα-tTA::TetO-hM3Dq …
Figure 5—figure supplement 1
Schematic of 13C labeling of various metabolites from [1,6-13C2]glucose in a three-compartment metabolic model [1,6-13C2]glucose is converted to PyruvateC3 via glycolysis and subsequently enters the TCA cycle.
Metabolism of [1,6-13C2]glucose in glutamatergic and GABAergic neurons produces GluC4, which is then decarboxylated to GABAC2 by GAD enzyme in GABAergic neurons. The glutamate–glutamine and …
Figure 5—figure supplement 2
Representative 1H-[13C]-NMR spectra from the cortex of vehicle and PNCNO-treated CamKIIα-tTA::TetO-hM3Dq adult male mice.
(A) Shown is a schematic of the experimental paradigm to induce chronic CNO-mediated hM3Dq DREADD activation in CamKIIα-positive forebrain excitatory neurons using bigenic CamKIIα-tTA::TetO-hM3Dq …
Figure 5—figure supplement 3
Influence of chronic chemogenetic activation of CamKIIα-positive forebrain excitatory neurons during the early postnatal window on the levels of other 13C-labeled metabolites in the hippocampus and cortex in adult male mice.
(A) Shown is a schematic of the experimental paradigm to induce chronic CNO-mediated hM3Dq DREADD activation in CamKIIα-positive forebrain excitatory neurons using bigenic CamKIIα-tTA::TetO-hM3Dq …
Figure 6
Chronic chemogenetic activation of CamKIIα-positive forebrain excitatory neurons during the early postnatal window results in a long-lasting decline in neuronal activity-related gene expression, and c-Fos immunopositive cell numbers, in the adult hippocampus.
(A) Shown is a schematic of the experimental paradigm to induce chronic CNO-mediated hM3Dq DREADD activation in CamKIIα-positive forebrain excitatory neurons using bigenic CamKIIα-tTA::TetO-hM3Dq …
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Figure 6—source data 1
List of qPCR primers.
- https://cdn.elifesciences.org/articles/56171/elife-56171-fig6-data1-v2.xlsx
Figure 7 with 2 supplements
Chronic chemogenetic activation of CamKIIα-positive forebrain excitatory neurons during the early postnatal window alters excitatory and inhibitory spontaneous currents in the hippocampi of adult male mice.
(A) Shown is a schematic of the experimental paradigm to induce chronic CNO-mediated hM3Dq DREADD activation in CamKIIα-positive forebrain excitatory neurons using bigenic CamKIIα-tTA::TetO-hM3Dq …
Figure 7—figure supplement 1
Chronic chemogenetic activation of CamKIIα-positive forebrain excitatory neurons during the early postnatal window does not change intrinsic excitability but alters spontaneous network activity in the hippocampi of adult male mice.
(A) Shown is a schematic of the experimental paradigm to induce chronic CNO-mediated hM3Dq DREADD activation in CamKIIα-positive forebrain excitatory neurons using bigenic CamKIIα-tTA::TetO-hM3Dq …
Figure 7—figure supplement 2
Effect of chronic chemogenetic activation of CamKIIα-positive forebrain excitatory neurons during the early postnatal window on the distribution of spontaneous network events in hippocampi of adult male mice.
(A) Shown is a schematic of the experimental paradigm to induce chronic CNO-mediated hM3Dq DREADD activation in CamKIIα-positive forebrain excitatory neurons using bigenic CamKIIα-tTA::TetO-hM3Dq …
Tables
Table 1
Effects of chronic postnatal hM3Dq DREADD activation in CamKIIα-positive forebrain excitatory neurons on intrinsic membrane properties.
| Postnatal Day 7 | CA1 pyramidal neurons | |||||
|---|---|---|---|---|---|---|
| Intrinsic Properties | ||||||
| Group | RMP (mV) | Input resistance (MΩ) | τ (ms) | Sag (mV) | Sag (%) | Accomodation index |
| Vehicle | −55.96 ± 1.923 | 310.2 ± 39.43 | 34.77 ± 4.62 | −10.09 ± 2.65 | 10.15 ± 2.42 | 0.32 ± 0.06 |
| PNCNO | −51.66 ± 1.151$ | 352.3 ± 35.23 | 33.62 ± 3.65 | −8.99 ± 1.66 | 9.10 ± 1.22 | 0.48 ± 0.09 |
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CamKIIα-tTA::TetO-hM3Dq bigenic mouse pups were fed either CNO (1 mg/kg) or vehicle from P2 to P7, and whole-cell patch clamp was performed at P7 to determine intrinsic membrane properties. No significant effect was noted for input resistance, membrane time constant (τ), sag voltage, percent sag and accommodation index across treatment groups. We noted a trend toward an increase in resting membrane potential (RMP) in hippocampi derived from the PNCNO-treated mouse pups as compared to vehicle-treated controls. $p=0.06 as compared to vehicle-treated controls (n = 6 cells for vehicle; n = 9 cells for PNCNO) using the two-tailed, unpaired Student’s t-test.
Table 2
Influence of chronic chemogenetic activation of CamKIIα-positive forebrain excitatory neurons during the early postnatal window on the total levels of metabolites relative to [2-13C]glycine in the hippocampus and cortex in adult male mice.
| [1,6-13C]glucose Infusion | ||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Concentration of brain metabolites determined relative to [2-13C]glycine (µmol/g) | ||||||||||||
| Brain Region | Group | Glu | GABA | Gln | Asp | NAA | Ala | Lac | Ino | Tau | Cho | Cre |
| Hippocampus | Vehicle | 13.5 ± 0.3 | 3.6 ± 0.2 | 5.0 ± 0.1 | 2.3 ± 0.1 | 8.9 ± 0.1 | 0.7 ± 0.1 | 3.0 ± 0.6 | 7.3 ± 0.2 | 8.6 ± 0.2 | 2.3 ± 0.1 | 13.5 ± 0.2 |
| PNCNO | 13.7 ± 0.3 | 3.7 ± 0.2 | 4.8 ± 0.1 | 2.5 ± 0.1 | 9.0 ± 0.1 | 0.6 ± 0.1* | 2.3 ± 0.2 | 7.5 ± 0.2 | 8.5 ± 0.2 | 2.3 ± 0.1 | 13.5 ± 0.3 | |
| Cortex | Vehicle | 13.5 ± 0.3 | 3.4 ± 0.1 | 4.7 ± 0.1 | 2.9 ± 0.1 | 9.5 ± 0.4 | 0.7 ± 0.1 | 7.0 ± 1.3 | 6.0 ± 0.2 | 9.1 ± 0.1 | 2.0 ± 0.1 | 12.9 ± 0.1 |
| PNCNO | 14.0 ± 0.2 | 3.3 ± 0.1 | 4.5 ± 0.1 | 3.2 ± 0.1$ | 9.6 ± 0.4 | 0.6 ± 0.1 | 5.3 ± 0.5 | 6.0 ± 0.1 | 9.1 ± 0.2 | 2.1 ± 0.1 | 12.9 ± 0.2 | |
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Chronic CNO-mediated hM3Dq DREADD activation in CamKIIα-positive forebrain excitatory neurons was achieved using bigenic CamKIIα-tTA::TetO-hM3Dq mouse pups that were fed CNO (PNCNO; 1 mg/kg) or vehicle from P2 to P14 and then left undisturbed for 3 months prior to metabolic analysis performed in adulthood on male mice and NMR spectroscopy on the hippocampus and cortex was performed to acquire 1H-[12C + 13C] spectra. The concentration of metabolites was determined relative to [2-13C]glycine. Glu: Glutamate; GABA: γ-aminobutyric acid; Gln: Glutamine; Asp: Aspartate; NAA: N-acetylaspartate; Suc: Succinate; Ala: Alanine; Lac: Lactose; Ino; Inositol; Tau: Taurine; Cho: Choline; Cre: Creatine. Results are expressed as the mean ± S.E.M (n = 7 per group). *p<0.05, $p=0.06; as compared to vehicle-treated controls using the two-tailed, unpaired Student’s t-test.
Table 3
Effect of chronic chemogenetic activation of CamKIIα-positive forebrain excitatory neurons during the early postnatal window on intrinsic membrane properties in adulthood.
| Adults | CA1 pyramidal neurons | |||||
|---|---|---|---|---|---|---|
| Intrinsic properties | ||||||
| Group | RMP (mV) | Input resistance (MΩ) | τ (ms) | Sag (mV) | Sag (%) | Accomodation index |
| Vehicle | −62.3 ± 0.655 | 195.1 ± 10.89 | 18.87 ± 1.574 | −4.282 ± 0.288 | 4.956 ± 0.317 | 0.375 ± 0.054 |
| PNCNO | −62.81 ± 0.682 | 180.2 ± 10.46 | 18.66 ± 1.269 | −3.603 ± 0.397 | 4.173 ± 0.420 | 0.392 ± 0.046 |
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Chronic CNO-mediated hM3Dq DREADD activation of CamKIIα-positive forebrain excitatory neurons was performed using bigenic CamKIIα-tTA::TetO-hM3Dq mouse pups that were fed CNO (PNCNO; 1 mg/kg) or vehicle from P2 to P14 and then left undisturbed for 3 months prior to electrophysiological analysis, in acute hippocampal slices derived from adult male mice. Whole-cell patch clamp was performed to determine intrinsic properties in the somata of CA1 pyramidal neurons. RMP: Resting membrane potential, τ: Membrane time constant. Results are expressed as the mean ± S.E.M. (n = 18 cells for vehicle; n = 25 cells for PNCNO).
Additional files
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Source data 1
Summary of statistical analysis.
- https://cdn.elifesciences.org/articles/56171/elife-56171-data1-v2.xlsx
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Transparent reporting form
- https://cdn.elifesciences.org/articles/56171/elife-56171-transrepform-v2.pdf
