Fimbria-Fornix Transection with bFGF Infusion
Objective: To analyze the anatomical distribution of basic fibroblast growth factor (bFGF) in the rat brain and its response to injury, specifically examining how chronic bFGF infusion following fimbria-fornix transection affects NGF receptors and neuronal survival in the medial septal complex
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Equipment2
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Protocol Steps
Tissue preparation and sectioning
Prepare rat brain tissue for immunohistochemical analysis
Note: Tissue from normal rat brain and lesioned animals
View evidence from paper
“Double-staining immunohistochemistry showed that bFGF immunoreactivity was localized in astrocytes, in select neuronal populations”
Double-staining immunohistochemistry
Perform double-staining immunohistochemistry to identify bFGF immunoreactivity in different cell types throughout the normal rat brain
Note: Identifies bFGF localization in astrocytes, neuronal populations, and microglial cells
View evidence from paper
“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”
Entorhinal cortex lesion
Create lesion in entorhinal cortex and examine resulting changes in bFGF immunoreactivity in hippocampus
Note: Lesion is ipsilateral; contralateral side serves as control
View evidence from paper
“The pattern of bFGF immunoreactivity in the hippocampus was examined following entorhinal cortex lesion”
Fimbria-fornix transection
Perform transection of the fimbria-fornix in experimental animals
Note: Surgical procedure to create the injury model
View evidence from paper
“After transection of the fimbria-fornix, chronic infusion of bFGF appeared to preserve NGF receptors”
Chronic bFGF infusion
Initiate chronic infusion of bFGF following fimbria-fornix transection into the medial septal complex
Note: bFGF infusion is designed to preserve NGF receptors and prevent neuronal death
View evidence from paper
“chronic infusion of bFGF appeared to preserve NGF receptors on neurons within the medial septal complex and prevent the death of medial septal neurons”
Quantification of bFGF response
Quantify changes in bFGF immunoreactivity following lesion, measuring number of bFGF astrocytes, intensity of immunoreactivity, and extracellular matrix localization
Note: Comparisons made between ipsilateral and contralateral sides
View evidence from paper
“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”
Assessment of NGF receptor preservation
Examine whether chronic bFGF infusion preserves NGF receptors on neurons in the medial septal complex
Note: Determines if bFGF enables neurons to respond to NGF
View evidence from paper
“chronic infusion of bFGF appeared to preserve NGF receptors on neurons within the medial septal complex”
Neuronal survival assessment
Evaluate medial septal neuron survival following fimbria-fornix transection with and without bFGF infusion
Note: bFGF infusion prevents death of medial septal neurons
View evidence from paper
“prevent the death of medial septal neurons”