Source Paper
Regulation of BDNF and trkB mRNA in rat brain by chronic electroconvulsive seizure and antidepressant drug treatments
M Nibuya, S Morinobu, RS Duman
Journal of Neuroscience • 1995
Source Paper
M Nibuya, S Morinobu, RS Duman
Journal of Neuroscience • 1995
The influence of chronic electroconvulsive seizure (ECS) or antidepressant drug treatments on expression of brain-derived neurotrophic factor (BDNF) and its receptor, trkB, was examined by in situ hybridization and Northern blot. In frontal cortex, acute ECS increased BDNF mRNA approximately twofold, an effect significantly augmented by a prior course of chronic ECS treatment (10 d). In the hippocampus, the influence of chronic ECS varied between the major subfields. In the dentate gyrus granule cell layer, chronic ECS decreased the acute induction of BDNF and trkB mRNA by approximately 50%, but prolonged their expression: levels remained elevated two- to threefold 18 hr later after the last chronic ECS treatment, but returned to control 18 hr after acute ECS. In CA3 and CA1 pyramidal cell layers, chronic ECS significantly elevated the acute induction of BDNF, and tended to prolong the expression of BDNF and trkB mRNA. A similar effect was observed in layer 2 of the piriform cortex, where chronic ECS significantly increased the acute induction and prolonged the expression of BDNF and trkB mRNA. Chronic (21 d), but not acute (1 d), administration of several different antidepressant drugs, including tranylcypromine, sertraline, desipramine, or mianserin, significantly increased BDNF mRNA and all but mianserin increased trkB mRNA in hippocampus. In contrast, chronic administration of nonantidepressant psychotropic drugs, including morphine, cocaine, or haloperidol, did not increase levels of BDNF mRNA. Furthermore, chronic administration of ECS or antidepressant drugs completely blocked the down-regulation of BDNF mRNA in the hippocampus in response to restraint stress. The enhanced induction and prolonged expression of BDNF in response to chronic ECS and antidepressant drug treatments could promote neuronal survival, and protect neurons from the damaging effects of stress.
Objective: Assessment of BDNF mRNA down-regulation in response to restraint stress and its prevention by ECS or antidepressant treatments
This is a Restraint Stress Paradigm protocol using rat as the model organism. The procedure involves 8 procedural steps, 3 equipment items, 7 materials. Extracted from a 1995 paper published in Journal of Neuroscience.
Model and subjects
rat • Not specified • unknown • Not specified • Not specified
Study window
~3 week study window
Core workflow
Chronic ECS Treatment • Acute ECS Administration • Chronic Antidepressant Drug Administration
Primary readouts
Key equipment and reagents
Verified items
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Administer chronic electroconvulsive seizure treatment to rats
Note: This is the chronic ECS treatment condition
“an effect significantly augmented by a prior course of chronic ECS treatment (10 d)”
Administer acute electroconvulsive seizure treatment following chronic ECS treatment or as standalone acute treatment
Note: Can be administered after chronic ECS or as acute-only control
“In frontal cortex, acute ECS increased BDNF mRNA approximately twofold”
Administer antidepressant drugs chronically to rats. Drugs tested include tranylcypromine, sertraline, desipramine, or mianserin
Note: Chronic administration only; acute administration (1 day) did not increase BDNF mRNA
“Chronic (21 d), but not acute (1 d), administration of several different antidepressant drugs, including tranylcypromine, sertraline, desipramine, or mianserin, significantly increased BDNF mRNA”
Administer non-antidepressant psychotropic drugs chronically as control conditions. Drugs tested include morphine, cocaine, or haloperidol
Note: These drugs serve as negative controls and did not increase BDNF mRNA levels
“chronic administration of nonantidepressant psychotropic drugs, including morphine, cocaine, or haloperidol, did not increase levels of BDNF mRNA”
Apply restraint stress to rats to induce down-regulation of BDNF mRNA in hippocampus
Note: Restraint stress causes BDNF mRNA down-regulation which is prevented by prior ECS or antidepressant treatment
“chronic administration of ECS or antidepressant drugs completely blocked the down-regulation of BDNF mRNA in the hippocampus in response to restraint stress”
Collect brain tissue and analyze BDNF and trkB mRNA expression using in situ hybridization and Northern blot
Note: Acute ECS increased BDNF mRNA approximately twofold in frontal cortex
“In frontal cortex, acute ECS increased BDNF mRNA approximately twofold”
Collect brain tissue 18 hours after the last chronic ECS treatment to assess prolonged BDNF and trkB mRNA expression
Note: In dentate gyrus, levels remained elevated two- to threefold 18 hr after last chronic ECS but returned to control 18 hr after acute ECS
“levels remained elevated two- to threefold 18 hr later after the last chronic ECS treatment, but returned to control 18 hr after acute ECS”
Analyze BDNF and trkB mRNA expression in specific brain regions: frontal cortex, hippocampus (dentate gyrus, CA3, CA1), and piriform cortex layer 2
Note: Different brain regions showed different patterns of BDNF and trkB mRNA regulation
“In the hippocampus, the influence of chronic ECS varied between the major subfields”
This section explains what the experiment is doing, which readouts matter, what the data artifacts usually look like, and how the analysis should flow from raw capture to reported result.
Assessment of BDNF mRNA down-regulation in response to restraint stress and its prevention by ECS or antidepressant treatments
Objective
Assessment of BDNF mRNA down-regulation in response to restraint stress and its prevention by ECS or antidepressant treatments
Subjects
From paperrat • Not specified • unknown • Not specified • Not specified
Cohort notes
From paperNot specified
Chronic ECS Treatment (10 days)
Acute ECS Administration (Single acute treatment)
Chronic Antidepressant Drug Administration (21 days)
Non-antidepressant Psychotropic Drug Administration (Control) (Chronic administration)
BDNF mRNA expression levels measured by in situ hybridization
From paperNot specified in methods section
Artifact type
Band images paired with densitometry summaries
Comparison focus
Compare normalized protein expression between experimental groups
TrkB mRNA expression levels measured by in situ hybridization
From paperNot specified in methods section
Artifact type
Band images paired with densitometry summaries
Comparison focus
Compare normalized protein expression between experimental groups
BDNF mRNA expression levels measured by Northern blot
From paperNot specified in methods section
Artifact type
Band images paired with densitometry summaries
Comparison focus
Compare normalized protein expression between experimental groups
TrkB mRNA expression levels measured by Northern blot
From paperNot specified in methods section
Artifact type
Band images paired with densitometry summaries
Comparison focus
Compare normalized protein expression between experimental groups
BDNF mRNA expression levels measured by in situ hybridization
From paperRaw artifact
Membrane or gel image with visible bands for target and control proteins
Processed artifact
Band quantification and normalized densitometry values
Final reported form
Relative expression values or fold-change comparisons across groups
TrkB mRNA expression levels measured by in situ hybridization
From paperRaw artifact
Membrane or gel image with visible bands for target and control proteins
Processed artifact
Band quantification and normalized densitometry values
Final reported form
Relative expression values or fold-change comparisons across groups
BDNF mRNA expression levels measured by Northern blot
From paperRaw artifact
Membrane or gel image with visible bands for target and control proteins
Processed artifact
Band quantification and normalized densitometry values
Final reported form
Relative expression values or fold-change comparisons across groups
TrkB mRNA expression levels measured by Northern blot
From paperRaw artifact
Membrane or gel image with visible bands for target and control proteins
Processed artifact
Band quantification and normalized densitometry values
Final reported form
Relative expression values or fold-change comparisons across groups
Acquisition
Collect the raw assay or blot output and retain identifiers for each sample and experimental group.
Preprocessing / cleaning
Not specified in methods section
Scoring or quantification
Quantify the primary readouts for this experiment: BDNF mRNA expression levels measured by in situ hybridization; TrkB mRNA expression levels measured by in situ hybridization; BDNF mRNA expression levels measured by Northern blot; TrkB mRNA expression levels measured by Northern blot.
Normalization
Normalize expression or signal values against the stated control or loading reference before comparing groups.
Statistical comparison
Statistical method not yet structured for this page.
Reporting output
Report representative outputs alongside summary comparisons for BDNF mRNA expression levels measured by in situ hybridization, TrkB mRNA expression levels measured by in situ hybridization, BDNF mRNA expression levels measured by Northern blot, TrkB mRNA expression levels measured by Northern blot.
Source links and direct wording from the methods section for validation and deeper review.
Citation
M Nibuya et al. (1995). Regulation of BDNF and trkB mRNA in rat brain by chronic electroconvulsive seizure and antidepressant drug treatments. Journal of Neuroscience
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