Investigation of Intracellular Signal Transduction on Fetal Brain

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We analyzed developmental stage-specific protein expression in rat brain cytosol to clarify proteins which might be involved in the development of the central nervous system. About 30 proteins that are expressed specifically at the prenatal stage were isolated from the cytosol fraction by preparative electrophoresis and were subjected to amino acid sequence analysis. Although several of these fetal specific proteins, such as elongation factor-1 and HMG-1, which arc known to be related to growth and differentiation have been characterized, the structure and function of almost all other proteins are still unknown. Among these proteins, 30 kDA protein (p30) which might be related to regulatory factor of low molecular weight G protein and rat homologues of human putative HLA-DR associated proteins (PHAPI and PHAPII) were further analyzed. To obtain the cDNA encoding these proteins, polynierase chain reaction was performed using mouse 11-day embryo and mouse brain cDNA library as template and degenerated oligonucleotide primers deduced from partial amino acid sequences of these proteins. Since the specificity of used degenerated primers were low, cloning of the cDNA encoding p30 aws not successful. While, many cDNA encoding PHAPI and PIIAPII were cloned and these cDNA clones were not identical, but closely related each other. Comparison of nuelcotidesequences and deduced amino acid sequences suggest that both PIJAPI and P11 APII have many molecular spieces generated by alternative splicing.