Heterogeneity of tau protein and mRNA expression during the development of cerebellar granule cell neurons in vitro.

Tau microtubule-associated proteins constitute a group of developmentally regulated neuronal proteins which promote microtubule polymerization and stabilization and hence have important implications during neuronal morphogenesis. We have examined the expression of tau mRNA and protein levels during the differentiation of cerebellar granule neurons over a period of 3 weeks in vitro. Oligonucleotide probes directed towards either immature or mature forms of tau mRNA were detected by in situ hybridization. Such experiments demonstrated that the time interval between 1 and 4 days in vitro represents a developmental epoch in the regulation of tau mRNA whereby the dominant immature tau messages were gradually replaced by mature mRNAs. Analysis of the profile of the various tau isoforms showed further developmental regulation with the transient rise in immature tau variants followed by the appearance of mature isoforms in older cultures. The increase in tau heterogeneity during granule neuron differentiation was enhanced by and could be attributed to intensive post-translational phosphorylation. Dephosphorylation of cell cultures demonstrated that the majority of tau was phosphorylated and that such a modification had profound affects on the localization of tau within developing neurons by immunocytochemistry. This study describes the profile of tau protein and mRNA levels expressed by differentiating cerebellar granule neurons in vitro and clearly demonstrates that tau is developmentally regulated and that important changes in tau expression occur at a time when processes are consolidating their first contacts.