Quantitation of tyrosine hydroxylase and neuropeptide Y immunoreactivity in single rat sympathetic neurons: effects of preganglionic nerve activity.
Using computerised densitometry to measure immunocytochemical reaction product in a model system, we established conditions that produced a linear relationship between the logarithm of antigen concentration and the measured intensity of staining. We then applied the densitometric technique to assess the changes in tyrosine hydroxylase (TH) and neuropeptide tyrosine (NPY) within sympathetic neurons of rat superior cervical ganglion following chronic decentralization and following reserpine treatment. One week after surgical or pharmacological decentralization, there was appreciable reduction of neuronal levels of both TH and NPY. However, there remained considerable variation in the immunoreactivities of individual cells. Three days of treatment with reserpine elevated TH levels but substantially reduced NPY. Both these effects were prevented by prior decentralization of the ganglia. No differences were seen between normotensive and the Otago strain of genetically hypertensive rats, either in basal TH or NPY immunoreactivities or in responses to the maneuvers performed. Comparison of our findings with previous biochemical data indicate that densitometric immunocytochemistry provides an accurate index of neuronally localised antigen concentrations but also allows analysis of interneuronal differences that are not otherwise apparent.
Department of Physiology, University of Melbourne, Parkville, Victoria, Australia.
C J
CJ
Gurusinghe
P J
PJ
Harris
D F
DF
Abbott
A
A
Messina
C
C
Bell
D000818:Animals; D005724:Ganglia / Q000502:physiology; D007150:Immunohistochemistry; D009474:Neurons / Q000378:metabolism*; D009478:Neuropeptide Y / Q000276:immunology*; D009638:Norepinephrine / Q000096:biosynthesis; D051381:Rats; D011919:Rats, Inbred Strains; D012110:Reserpine / Q000494:pharmacology; D013564:Sympathetic Nervous System / Q000276:immunology / Q000378:metabolism*; D014446:Tyrosine 3-Monooxygenase / Q000032:analysis*
D009478:Neuropeptide Y; D012110:Reserpine; D014446:Tyrosine 3-Monooxygenase; D009638:Norepinephrine