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Citation
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HERO ID
1515400
Reference Type
Journal Article
Title
Quantitative immunocytochemistry of tyrosine hydroxylase in rat brain. I. Development of a computer assisted method using the peroxidase-antiperoxidase technique
Author(s)
Benno, RH; Tucker, LW; Joh, TH; Reis, DJ
Year
1982
Is Peer Reviewed?
Yes
Journal
Brain Research
ISSN:
0006-8993
EISSN:
1872-6240
Publisher
Elsevier
Volume
246
Issue
2
Page Numbers
225-236
Language
English
PMID
6181853
URL
http://www.sciencedirect.com/science/article/pii/0006899382911702
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Abstract
We sought to develop a quantitative immunocytochemical procedure using the peroxidase-antiperoxidase (PAP) technique to analyze amounts of neurotransmitter biosynthetic enzyme tyrosine hydroxylase (TH) within the nucleus locus coeruleus (LC) of rat brain. Rats were perfused with 4% paraformaldehyde. The brains were embedded in paraffin, sectioned at 5 micrometers in the sagittal plane and immunocytochemically stained with antibodies to bovine adrenal TH. Staining intensity, measured by a TV image analysis system was reproducible within +/- 5%. Reaction conditions required so that the intensity of the PAP reaction product was directly and linearly related to the amount of TH enzyme protein in tissue were obtained by reacting tissues with saturating concentrations (2.5 mM) of diaminobenzidine (DAB) substrate, a constant dilution of antibody, and incubation time adjusted so that the darkest elements in tissue were below saturation. Variations in staining intensity of serial sections through the LC were found to be insignificant when compared to variations in staining between animals. In order to increase the amount of immunoreactive TH in the LC, groups of rats were treated with reserpine. Immunocytochemical and biochemical analyses were performed in parallel groups of control and reserpine animals allowed to survive 1-3 days following a single injection (10 mg/kg s.c.) of reserpine. A close correlation was found to exist between the amount of TH enzyme protein determined biochemically and the density of staining for TH. The maximal increase in TH measured immunocytochemically was 2.2-fold which was about 80% of the maximal induction determined biochemically. We conclude that the PAP method can be used for quantitative immunocytochemistry of brain TH providing that optimal reaction conditions are established.
Keywords
quantitative immunocytochemistry; tyrosine hydroxylase
Tags
IRIS
•
Formaldehyde [archived]
Nervous system effects
Found
Database search results
PubMed
Screened
Title/abstract
Related to use in methodology
Reproductive and Developmental Effects
Screened
Title/abstract
Methodology/therapeutics
Retroactive RIS import
Pre2013
Merged Litsearch Results 100912
Merged LitSearch Results ToxNet 101012
Merged LitSearch Additions 86 Reviews SCREEN
PubMed Search 100912
2013
HCHON tox Ref Identification 022713
2015
FA DevRepro 072115
Methodology/Therapeutics-Population Criteria
•
IRIS Formaldehyde (Inhalation) [Final 2024]
Literature Indexing
PubMed
Literature Identification
Nervous System Effects
Excluded
Reproductive and Developmental Effects
Excluded
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