Biochemical end points can be applied on tissue samples collected during or after all of the studies. Effects of transgene, surgical manipulation and/or compound treatment can be studied on three levels: transcription (when DNA is transcripted to mRNA), translation (when mRNA is processed into protein), and activity (how the active protein / enzyme function is modified).
Charles River provides several tools to study these different levels of biomarker expression:
Synaptophysin mRNA is decreased significantly in aged rat cortex.
Validated immunohistochemical end points include:
Immunoreactivity of myelin basic protein, a marker for white matter, in rat spinal cord
CD68 immunoreactive monocytes/macrophages in rat spinal cord after injury
Aβ immunoreactive plaques can be found in 9-month-old CVN mouse hippocampus, but not in wild-type littermates.
Tyrosine hydroxylase immunoreactive neurons in rat substantia nigra
Iba-1 immunoreactive microglia in R6/2 mouse striatum
Expression of different C-terminal APP-fragments can be analyzed from AD mouse tissue samples by Western blotting.
Validated ELISA end points include
- Aβ1-40 (soluble and insoluble)
- Aβ 1-42 (soluble and insoluble)
Increased amount of Aβ1-42 can be detected in plasma, CSF and tissue samples of Tg2576 mice already at 5 months of age. Gamma-secretase inhibitor DAPT significantly decreases the amount of Aβ1-42.
Histology end points include:
- Cresyl Fast Violet
- Fluoro-Jade B
- Hematoxylin & Eosin
- Luxol Fast Blue
- Perls (iron)
Histological staining of rat brain with CFV, close-up on hippocampus
Histological staining of rat spinal cord with Giemsa
Fluoro-Jade staining can be used for identification of degenerating neurons
Hematoxylin-Eosin stain can be applied to evaluation of brain tissue damage.
IN VITRO SPECTROSCOPY
Using the Muromachi microwave brain fixation system the short-lived brain metabolites and neurotransmitters can be detected using in vitro spectroscopy.
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