Ultrastructural immunolocalization of rat oxytocin-neurophysin in transgenic mice expressing the rat oxytocin gene.
Cell-specific expression of the rat oxytocin (OT)-neurophysin transgene in mice was achieved using a construct containing both OT and vasopressin genes (Young III, W.S., Reynolds, K., Shepard, E.A., Gainer, H. and Castel, M., Cell-specific expression of the rat oxytocin gene in transgenic mice, J. Neuroendocrinol., 2 (1990) 1-9). The present study describes the distribution of the protein products of these genes in various regions of the cell, and determines whether the transgenic rat and endogenous mouse OT-neurophysins are colocalized within the same neurosecretory granules. Two monoclonal antibodies against OT-neurophysins were used: PS38 which can react with both rat and mouse OT-neurophysin (pan-specific), and PS67 which is specific for rat OT-neurophysin only. Various approaches to double immunolabeling at the ultrastructural level were employed; these included: (1) pre-embedding immunoperoxidase followed by post-embedding immunogold; (2) post-embedding immunolabeling using gold particles of different sizes; and (3) labeling of consecutive ultrathin sections with different antibodies. Results from each of these approaches showed that both in the transgenic mouse and in the rat (used as control), immunocytochemical labeling for both PS38 and PS67 occurred in the same OT-ergic neurosecretory granules. In the control mouse, only PS38 elicited labeling. Hence, it may be concluded that the protein and peptide products of the transgene and the endogenous gene for OT-neurophysin are being processed similarly in the cell and finally concentrated together in the same neurosecretory granules.
Belenky
M
M
Department of Experimental Zoology, Silberman Institute of Life Sciences, Hebrew University of Jerusalem, Israel.
Castel
M
M
Young
W S
WS
Gainer
H
H
Cohen
S
S
D000818:Animals; D000911:Antibodies, Monoclonal; D003594:Cytoplasmic Granules / Q000648:ultrastructure*; D004721:Endoplasmic Reticulum / Q000648:ultrastructure; D007124:Immunoenzyme Techniques; D051379:Mice; D008815:Mice, Inbred Strains; D008822:Mice, Transgenic; D016253:Microscopy, Immunoelectron; D009481:Neurophysins / Q000032:analysis*; D010121:Oxytocin / Q000032:analysis* / Q000235:genetics*; D010286:Paraventricular Hypothalamic Nucleus / Q000502:physiology / Q000648:ultrastructure*; D011401:Promoter Regions, Genetic; D051381:Rats
D000911:Antibodies, Monoclonal; D009481:Neurophysins; D010121:Oxytocin