Autoradiographic measurement of tyrosine hydroxylase immunoreactivity in rat brain using enhanced chemiluminescence: a general method for combined quantitative and cytochemical detection

• Published in: Brain research Vol. 638; no. 1; pp. 352 - 356
• Main Authors: Norman, Andrew B., Khosla, Uday M., Klug, Jennifer M., Thompson, Herbert R.
• Format: Journal Article
• Language: English
• Published: London Elsevier B.V 28.02.1994; Amsterdam Elsevier; New York, NY
• Subjects: 3,3'-Diaminobenzidine; 6-Hydroxydopamine lesion; Animals; Antibodies, Monoclonal; Autoradiography; Autoradiography - methods; Biochemistry and metabolism; Biological and medical sciences; Brain - cytology; Brain - enzymology; Brain - pathology; Central nervous system; Fundamental and applied biological sciences. Psychology; Immunoenzyme Techniques; Immunohistochemistry - methods; Luminescent Measurements; Male; Monoclonal antibody; Oxidopamine - toxicity; Quantitative immunocytochemistry; Rats; Rats, Sprague-Dawley; Striatum; Substantia nigra; Tyrosine 3-Monooxygenase - analysis; Vertebrates: nervous system and sense organs; Striatum; Autoradiography; 6-Hydroxydopamine lesion; Monoclonal antibody; Substantia nigra; Quantitative immunocytochemistry; Vertebrata; Mammalia; Rat; Enzyme; Rodentia; Central nervous system; Chemiluminescence; Oxidoreductases; Lesion; Tyrosine 3-monooxygenase; Brain (vertebrata)
• ISSN: 0006-8993; 1872-6240
• DOI: 10.1016/0006-8993(94)90671-8

Autoradiographic measurements of peroxidase labelled tyrosine immunoreactivity in brain sections from rats with unilateral 6-hydroxydopamine lesions were obtained using enhanced chemiluminescence. The autoradiograms required only 5–60 s exposure and displayed a good sinnal-to-noise ratio. The autoradiograms were quantitated using densitometry and by comparison with a standard curve. Following acquisition of the autoradiograms the sections were stained with the chromogen diaminobenzidine (DAB). Distribution of the DAB closely followed the grain densities observed in the autoradiograms on both the lesioned and unlesioned side of the brain. This method represents a simple, rapid and inexpensive technique for performing both quantitative autoradiography and cytochemical studies in individual tissue sections of any perosidase labllled immunoreactivity.