Galanin and cholecystokinin in cultured magnocellular neurons isolated from adult rat supraoptic nuclei: a correlative light and scanning electron microscopical study.
Cultured magnocellular neurons, isolated from adult rat supraoptic nuclei, were characterized by immunocytochemistry, using the avidin-biotin-peroxidase complex and antisera to vasopressin, oxytocin, galanin and cholecystokinin. Light microscope examination of the immunostained cultures revealed the presence of vasopressin- and oxytocin-like immunoreactivity, as well as neurons containing either galanin- or cholecystokinin-like immunoreactivity. In contrast, no significant galanin- or cholecystokinin-like immunoreactivity could be observed in freshly dispersed cells. Correlative scanning electron microscopical observations in the secondary electron imaging mode revealed that the stained neurons appeared significantly brighter than the unstained structures. Complementary observations with toad brain sections (preoptic area), immunostained for galanin, led to the same result. Considering previous results, it is suggested that the presence of galanin- and cholecystokinin-like immunoreactivity in the cultured neurons and its virtual absence in freshly dispersed cells is indicating a participation of these peptides in the regenerative processes taking place during culture. It is further concluded that the avidin-biotin-peroxidase method is suitable for correlative light and scanning electron microscopical studies of smooth surfaces and cultured cells.
Sanchez
A
A
Facultad de Ciencias Veterinarias, Universidad de Buenos Aires, Argentina.
Bilinski
M
M
Gonzalez Nicolini
V
V
Villar
M J
MJ
Tramezzani
J H
JH
D000818:Animals; D002478:Cells, Cultured; D002766:Cholecystokinin / Q000032:analysis*; D019004:Galanin / Q000032:analysis*; D007124:Immunoenzyme Techniques; D008297:Male; D008853:Microscopy; D008855:Microscopy, Electron, Scanning; D009474:Neurons / Q000737:chemistry* / Q000166:cytology / Q000648:ultrastructure; D010121:Oxytocin / Q000032:analysis; D051381:Rats; D013495:Supraoptic Nucleus / Q000737:chemistry* / Q000166:cytology / Q000648:ultrastructure; D014667:Vasopressins / Q000032:analysis
D014667:Vasopressins; D010121:Oxytocin; D019004:Galanin; D002766:Cholecystokinin