Source Paper
Basic FGF in adult rat brain: cellular distribution and response to entorhinal lesion and fimbria-fornix transection
F Gomez-Pinilla, JW Lee, CW Cotman
Journal of Neuroscience • 1992
Source Paper
F Gomez-Pinilla, JW Lee, CW Cotman
Journal of Neuroscience • 1992
Basic fibroblast growth factor (bFGF) is a potent trophic factor for neurons and astrocytes and recently has been implicated in the pathology of Alzheimer's disease. In order to better understand the role of bFGF in normal brain function and during pathology, we have analyzed its anatomical distribution and its response to injury in the CNS. Double-staining immunohistochemistry showed that bFGF immunoreactivity was localized in astrocytes, in select neuronal populations, and occasionally in microglial cells throughout the normal rat brain. Neuronal populations that showed bFGF immunoreactivity included septohippocampal nucleus, cingulate cortex, subfield CA2 of the hippocampus, cerebellar Purkinje cells, cerebellar deep nuclei, facial nerve nucleus, and the motor and spinal subdivisions of the trigeminal nucleus and facial nerve nucleus. The pattern of bFGF immunoreactivity in the hippocampus was examined following entorhinal cortex lesion, or fimbria-fornix transection. After entorhinal cortex lesion, bFGF immunoreactivity increased in the outer molecular layer of the dentate gyrus ipsilateral to the lesion. The lesion effect on bFGF immunoreactivity was expressed as an increase in the number of bFGF astrocytes, as an increase in the intensity of bFGF immunoreactivity within astrocytes, and as an increase of bFGF immunoreactivity in the surrounding extracellular matrix, relative to the contralateral side. The time course and pattern of reorganization paralleled the sprouting of septal cholinergic terminals in response to the same type of lesion, suggesting that bFGF may play an important role in lesion-induced plasticity. After transection of the fimbria-fornix, chronic infusion of bFGF appeared to preserve NGF receptors on neurons within the medial septal complex and, as previously reported, prevent the death of medial septal neurons. Therefore, it appears that bFGF infusion, which has been shown to increase the synthesis of NGF by astrocytes (Yoshida and Gage, 1991), also helps enable neurons to respond to NGF. This suggests that after injury bFGF may participate in a cascade of neurotrophic events, directly and indirectly facilitating neuronal repair and/or promoting neuronal survival.
Objective: Analyze anatomical distribution of basic fibroblast growth factor (bFGF) immunoreactivity in the rat brain and examine its response to entorhinal cortex lesion and fimbria-fornix transection
This is a Entorhinal Cortex Lesion with bFGF Analysis protocol using rat as the model organism. The procedure involves 9 procedural steps, 4 equipment items, 2 materials. Extracted from a 1992 paper published in Journal of Neuroscience.
Model and subjects
rat • not specified • unknown • adult • not specified
Study window
Estimated timing pending
Core workflow
Tissue preparation and sectioning • Double-staining immunohistochemistry • Map bFGF distribution in normal brain
Primary readouts
Key equipment and reagents
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Prepare rat brain tissue for immunohistochemical analysis
Note: Tissue from normal adult rats and lesioned animals
“Double-staining immunohistochemistry showed that bFGF immunoreactivity was localized in astrocytes, in select neuronal populations”
Perform double-staining immunohistochemistry to detect bFGF immunoreactivity and identify cell types
Note: Allows simultaneous identification of bFGF in astrocytes, neurons, and microglial cells
“Double-staining immunohistochemistry showed that bFGF immunoreactivity was localized in astrocytes, in select neuronal populations, and occasionally in microglial cells”
Document anatomical distribution of bFGF immunoreactivity throughout normal rat brain
Note: Identify neuronal populations and brain regions with bFGF immunoreactivity
“Neuronal populations that showed bFGF immunoreactivity included septohippocampal nucleus, cingulate cortex, subfield CA2 of the hippocampus, cerebellar Purkinje cells”
Create lesion of the entorhinal cortex in experimental animals
Note: Lesion is unilateral; contralateral side serves as control
“The pattern of bFGF immunoreactivity in the hippocampus was examined following entorhinal cortex lesion”
Examine changes in bFGF immunoreactivity in hippocampus following entorhinal cortex lesion
Note: Compare ipsilateral (lesioned side) to contralateral (control) side
“After entorhinal cortex lesion, bFGF immunoreactivity increased in the outer molecular layer of the dentate gyrus ipsilateral to the lesion”
Measure increases in bFGF immunoreactivity as number of bFGF astrocytes, intensity within astrocytes, and extracellular matrix immunoreactivity
Note: Compare lesioned side to contralateral control side
“The lesion effect on bFGF immunoreactivity was expressed as an increase in the number of bFGF astrocytes, as an increase in the intensity of bFGF immunoreactivity within astrocytes”
Create transection of the fimbria-fornix in separate cohort of experimental animals
Note: Separate experimental group from entorhinal cortex lesion animals
“After transection of the fimbria-fornix, chronic infusion of bFGF appeared to preserve NGF receptors”
Administer chronic infusion of bFGF following fimbria-fornix transection
Note: bFGF infusion preserves NGF receptors and prevents medial septal neuron death
“After transection of the fimbria-fornix, chronic infusion of bFGF appeared to preserve NGF receptors on neurons within the medial septal complex”
Examine NGF receptor expression on neurons in medial septal complex following bFGF infusion
Note: Assess neuronal survival in medial septal complex
“chronic infusion of bFGF appeared to preserve NGF receptors on neurons within the medial septal complex and prevent the death of medial septal neurons”
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.
Analyze anatomical distribution of basic fibroblast growth factor (bFGF) immunoreactivity in the rat brain and examine its response to entorhinal cortex lesion and fimbria-fornix transection
Objective
Analyze anatomical distribution of basic fibroblast growth factor (bFGF) immunoreactivity in the rat brain and examine its response to entorhinal cortex lesion and fimbria-fornix transection
Subjects
From paperrat • not specified • unknown • adult • not specified
Cohort notes
From paperNormal rat brain tissue examined; lesion studies performed on separate cohorts
Tissue preparation and sectioning (not specified)
Double-staining immunohistochemistry (not specified)
Map bFGF distribution in normal brain (not specified)
Perform entorhinal cortex lesion (not specified)
BFGF immunoreactivity localization in astrocytes, neurons, and microglial cells
From paperComparison of bFGF immunoreactivity between ipsilateral (lesioned) and contralateral (control) sides; analysis of time course and pattern of reorganization; assessment of NGF receptor preservation and neuronal survival
Artifact type
Representative image panels with region or marker comparisons
Comparison focus
Compare staining intensity, structure, or cell counts across matched conditions
Anatomical distribution of bFGF in normal rat brain
From paperComparison of bFGF immunoreactivity between ipsilateral (lesioned) and contralateral (control) sides; analysis of time course and pattern of reorganization; assessment of NGF receptor preservation and neuronal survival
Artifact type
Representative image panels with region or marker comparisons
Comparison focus
Compare staining intensity, structure, or cell counts across matched conditions
Number of bFGF-immunoreactive astrocytes in dentate gyrus following entorhinal cortex lesion
From paperComparison of bFGF immunoreactivity between ipsilateral (lesioned) and contralateral (control) sides; analysis of time course and pattern of reorganization; assessment of NGF receptor preservation and neuronal survival
Artifact type
Representative image panels with region or marker comparisons
Comparison focus
Compare staining intensity, structure, or cell counts across matched conditions
Intensity of bFGF immunoreactivity within astrocytes
From paperComparison of bFGF immunoreactivity between ipsilateral (lesioned) and contralateral (control) sides; analysis of time course and pattern of reorganization; assessment of NGF receptor preservation and neuronal survival
Artifact type
Representative image panels with region or marker comparisons
Comparison focus
Compare staining intensity, structure, or cell counts across matched conditions
BFGF immunoreactivity localization in astrocytes, neurons, and microglial cells
From paperRaw artifact
Field or section images captured from matched samples
Processed artifact
Selected representative panels with quantified intensity, counts, or area measurements
Final reported form
Per-group imaging summaries with representative figures and quantified endpoints
Anatomical distribution of bFGF in normal rat brain
From paperRaw artifact
Field or section images captured from matched samples
Processed artifact
Selected representative panels with quantified intensity, counts, or area measurements
Final reported form
Per-group imaging summaries with representative figures and quantified endpoints
Number of bFGF-immunoreactive astrocytes in dentate gyrus following entorhinal cortex lesion
From paperRaw artifact
Field or section images captured from matched samples
Processed artifact
Selected representative panels with quantified intensity, counts, or area measurements
Final reported form
Per-group imaging summaries with representative figures and quantified endpoints
Intensity of bFGF immunoreactivity within astrocytes
From paperRaw artifact
Field or section images captured from matched samples
Processed artifact
Selected representative panels with quantified intensity, counts, or area measurements
Final reported form
Per-group imaging summaries with representative figures and quantified endpoints
Acquisition
Capture matched images from the relevant tissue region using the same acquisition settings across samples.
Preprocessing / cleaning
Comparison of bFGF immunoreactivity between ipsilateral (lesioned) and contralateral (control) sides; analysis of time course and pattern of reorganization; assessment of NGF receptor preservation and neuronal survival
Scoring or quantification
Quantify the primary readouts for this experiment: BFGF immunoreactivity localization in astrocytes, neurons, and microglial cells; Anatomical distribution of bFGF in normal rat brain; Number of bFGF-immunoreactive astrocytes in dentate gyrus following entorhinal cortex lesion; Intensity of bFGF immunoreactivity within astrocytes.
Normalization
Normalize image-derived measurements against the matched acquisition or segmentation rules before comparing groups.
Statistical comparison
Statistical method not yet structured for this page.
Reporting output
Report representative outputs alongside summary comparisons for BFGF immunoreactivity localization in astrocytes, neurons, and microglial cells, Anatomical distribution of bFGF in normal rat brain, Number of bFGF-immunoreactive astrocytes in dentate gyrus following entorhinal cortex lesion, Intensity of bFGF immunoreactivity within astrocytes.
Source links and direct wording from the methods section for validation and deeper review.
Citation
F Gomez-Pinilla et al. (1992). Basic FGF in adult rat brain: cellular distribution and response to entorhinal lesion and fimbria-fornix transection. Journal of Neuroscience
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