Induction and quantitation of the RI cAMP-binding protein in clonal mouse neuroblastoma cell lines: evidence that the increase in RI is not linked to neurite outgrowth.

This is a study of the regulation of expression of the RI cAMP-binding protein in mouse neuroblastoma cells as it relates to neurotransmitter phenotype and neurite outgrowth. Dibutyryl cAMP was used to promote differentiation of the cholinergic NS-20, the adrenergic N1E-115, the neurotransmitter-inactive N-18, and the neurite-minus N1A-103 mouse neuroblastoma cells. The amount of the RI cAMP-binding protein was quantitated by photoaffinity labeling of the 47,000-dalton RI protein with 8-N3-[32P]cAMP and by Western blot, ELISA, and immunocytochemistry. Our results showed that dibutyryl cAMP induced the RI cAMP-binding protein by three to fivefold in each of the four neuroblastoma cell lines examined. The increased expression of the RI cAMP-binding protein was not linked to neurite outgrowth, a parameter of morphological differentiation in the neuroblastoma cells. Thus, the RI cAMP-binding protein can be induced in the neurite-minus N1A-103 neuroblastoma round cells; further, 8-bromo-cAMP effected neurite outgrowth without inducing the RI cAMP-binding protein in the neurite-positive cell lines. Indirect immunocytochemistry of RI showed a cytoplasmic localization with little evidence of nuclear staining. The increase in RI cAMP-binding protein coincided with an increase in the cAMP-phosphodiesterase and a decrease in cAMP-dependent phosphotransferase activity in the mouse neuroblastoma cells.(ABSTRACT TRUNCATED AT 250 WORDS)