Differential immunolabeling for electron microscopy of diverse peptidergic neurons.
We describe a simple and reliable method for differential immunolabeling of pre- and post-synaptic signal peptides at the ultrastructural level. Hypothalamic tissues of rats, including the suprachiasmatic nucleus, were cut on a Vibratome. Visualization of the immunolabeling of somatostatin (SRIH) and vasoactive intestinal polypeptide (VIP) was performed with avidin-biotin-peroxidase-diaminobenzidine (DAB). The end product of the DAB to VIP was further silver-intensified in a physical processing using silver nitrate, and the silver grains were finally substituted for gold. DAB-labeled SRIH fibers synapse on gold-labeled VIP perikarya and dendrites in the suprachiasmatic nucleus.
D000818:Animals; D006651:Histocytochemistry / Q000379:methods*; D007031:Hypothalamus / Q000166:cytology; D007124:Immunoenzyme Techniques; D008854:Microscopy, Electron; D009474:Neurons / Q000648:ultrastructure*; D051381:Rats; D011919:Rats, Inbred Strains; D013004:Somatostatin / Q000032:analysis*; D013493:Suprachiasmatic Nucleus / Q000166:cytology; D014660:Vasoactive Intestinal Peptide / Q000032:analysis*
D014660:Vasoactive Intestinal Peptide; D013004:Somatostatin