Radioimmunoassay of met-enkephalin in microdissected areas of paraformaldehyde-fixed rat brain
The effects were studied of various sample preparation procedures on rat brain met-enkephalin content, measured by radioimmunoassay. Whole brain met-enkephalin content of rats killed by decapitation followed by immediate tissue freezing was similar to that of rats killed by microwave irradiation and to those of rats anesthetized with pentobarbital or halothane before killing, whether previously perfused with paraformaldehyde or not. In contrast, a decrease (up to 80%) in met-enkephalin concentrations was observed when brain samples were frozen and thawed to mimic the procedure utilized in the ''punch'' technique for analysis of discrete brain nuclei. This decrease was totally prevented by paraformaldehyde perfusion of the brain prior to sacrifice. Brain perfusion did not alter the amount of immunoassayable met-enkephalin extracted from tissue or its profile after Sephadex chromatography. Paraformaldehyde perfusion results in better morphological tissue preservation and facilitates the ''punch'' dissecting technique. Paraformaldehyde perfusion may be the procedure of choice for the measurement of neuropeptides in specific brain nuclei dissected by the ''punch'' technique.
- Research Organization:
- National Inst. of Mental Health, Bethesda, MD
- OSTI ID:
- 6007745
- Journal Information:
- Life Sci.; (United States), Vol. 34:9
- Country of Publication:
- United States
- Language:
- English
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ENKEPHALINS
RADIOIMMUNOASSAY
BIOLOGICAL LOCALIZATION
BRAIN
CYTOLOGICAL TECHNIQUES
FORMALDEHYDE
MEASURING METHODS
RATS
ALDEHYDES
ANIMALS
AUTONOMIC NERVOUS SYSTEM AGENTS
BODY
CENTRAL NERVOUS SYSTEM
DRUGS
ENDORPHINS
ISOTOPE APPLICATIONS
MAMMALS
NERVOUS SYSTEM
NEUROREGULATORS
ORGANIC COMPOUNDS
ORGANS
PEPTIDES
POLYPEPTIDES
PROTEINS
RADIOASSAY
RODENTS
TRACER TECHNIQUES
VERTEBRATES
550201* - Biochemistry- Tracer Techniques