Quantitative analysis of long-term survival and neuritogenesis in vitro: cochleovestibular ganglion of the chick embryo in BDNF, NT-3, NT-4/5, and insulin

Experimental Neurology
B H Sokolowski

Abstract

The dynamics of survival and growth were examined for cochleovestibular ganglion (CVG) cells maintained in long-term cultures. CVG cells were explanted from chick embryos after 90 h of incubation into a defined-medium containing BDNF, NT-3, or NT-4/5 and an insulin, transferrin, selenium, and progesterone supplement. Explant survival and neuritogenesis was measured for 23 to 24 days in vitro. All three neurotrophins prolonged CVG survival in a dose-dependent manner although insulin acted as a cofactor. In 0.872 microM insulin-containing medium the ED50 for BDNF and NT-3 was 100 pg/ml, whereas the ED50 for NT-4/5 was 600-1200 pg/ml. However, at later ages in vitro, survival decreased with concentrations of BDNF greater than 2 ng/ml. In insulin-free medium, concentrations of 5-200 ng/ml of BDNF or 30-200 ng/ml of NT-4/5 maintained the survival of explants at a rate that was equivalent to or less than the survival rate of cultures treated with insulin but not with neurotrophin. In contrast, NT-3-treated explants in insulin-free medium did not survive the duration of the experiment. Dose-dependent effects of BDNF and NT-3 on explant neuritogenesis were reflected as an initial delay in outgrowth, whereas NT-4/5 had no effect. Insuli...Continue Reading

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Citations

Oct 7, 2004·Hearing Research·Isabel Varela-NietoYolanda Leon
Oct 19, 1999·The Journal of Comparative Neurology·S L CochranE W Rubel
Sep 15, 2004·Acta Neuropathologica·Kazuhiro SugimotoSoroku Yagihashi
Jul 4, 2013·Developmental Neurobiology·Janet AlderSmita Thakker-Varia

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