Localization of endogenous biotin-containing proteins in mouse Bergmann glial cells.
A peroxidase-conjugated avidin-biotin complex was used to detect endogenous biotin-containing proteins in mouse cerebellum. By this method, Bergmann glial cells were found to be strongly labelled in the adult mouse cerebellum. Developmentally, cells in the granular layer, probably astrocytes, appeared to be labelled around postnatal 10-day (P10). Their labelling decreased after P20, although the positive-labelling remained in the Bergmann glial cells up to the adult stage. The findings were confirmed by using a Alexa Fluor 488-conjugated streptavidin technique. The labelling was not affected by routine hydrogen peroxide treatment, but it was eliminated by avidin-biotin blocking. By another transblot method, the reactive proteins in the mouse cerebellum were found to be 120 kDa (the strongest one) and 75 kDa. For electron microscopy, a gold-conjugated anti-biotin antibody was immunoreacted to the mitochondria of Bergmann glial cells. These results suggest that endogenous biotin-containing proteins are abundant in the Bergmann glial cells. Therefore, the avidin-biotin complex method is useful for detecting Bergmann glial cells, probably because of the difference of biotin metabolism in the cerebellar glial cells.
Yagi
Takashi
T
Department of Anatomy, Faculty of Medicine, University of Yamanashi, Tamaho, Yamanashi 409-3898, Japan.
Terada
Nobuo
N
Baba
Takeshi
T
Ohno
Shinichi
S
D000818:Animals; D000831:Animals, Newborn; D001360:Avidin / Q000032:analysis / Q000378:metabolism; D001710:Biotin / Q000032:analysis / Q000378:metabolism*; D015153:Blotting, Western; D002525:Cerebellar Cortex / Q000737:chemistry / Q000166:cytology*; D007124:Immunoenzyme Techniques; D051379:Mice; D008810:Mice, Inbred C57BL; D016253:Microscopy, Immunoelectron; D008928:Mitochondria / Q000737:chemistry / Q000378:metabolism / Q000648:ultrastructure; D009457:Neuroglia / Q000737:chemistry / Q000378:metabolism* / Q000648:ultrastructure
D001360:Avidin; D001710:Biotin