Study of nuclear membrane differentiation between germ and vegetative nuclei in early development of Paramecium.

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1. Isolated macronuclei of Paramecium caudatum were injected into mice, and hybridomas bet ween the spleen cells and mieroma cells (NS-1) were obtained by cell fusion. The hybridoma was screened by indirect immunofluorescence microscopy and cloned by limiting dilution. Monoclonal antibody (MAM-1) against macronuclear membrane of P. caudatum was obtained from cell-fee medium of the hybridoma cells.2. Molecular weight of the MAM-1 antigen was ca. 70 kDa by immunoblotting.3. Indirect immunofluorescence microscopy showed that the antigens appeared in 4 out of 8 postzygotic nuclei immediately after the 4 nuclei differentiated morphologically into the macronuclear anlagen, and the antigens were constitutionally present in the membrane through the cell cycle. The antigens were not detected in the micronucleus. The results indicate that the nuclear membranes differentiate between the two kinds of nuclei in early development.4. Immunofluorescence microscopy on species-specificity of the cross reactivity of the antibody showed that all strains of P. caudatum and P. jenningsi examined showed the flurescence in the macronuclear membranes but not in P. bursaria and Tetrahymena thermophila and T. pyriformis examined.