Involvement of 14-nm Filament-Forming Protein and Tubulin in Gametic Pronuclear Behavior during Conjugation in Tetrahymena

We have studied in detail the immunofluorescence localizations of Tetrahymena 14-nm filament-forming protein (49-kDa protein) in relation to tubulin in conjugating wild-type Tetrahymena thermophila (B strain) pairs and in pairs between B strain and star strains with defective micronuclei. The results suggest that germ nuclear behavior during conjugation may involve the following cytoskeletal structures: (1) during meiosis, microtubule structures are involved in micronuclear elongation and meiotic division; (2) at the postmeiotic stage, 49-kDa protein network structures that are formed independently of the existence of pronuclei are involved in the selection and the survival of one of four meiotic products; (3) during the third prezygotic division, gametic pronuclear transfer, and zygote formation, a cytoskeletal structure in which the 49-kDa protein colocalizes with microtubules and which is dependent on the existence of a normal gametic pronucleus is involved in gametic pronuclear behavior, and (4) during the postzygotic divisions, the microtubules are involved in nuclear behavior.     

Ciliary Membranes and Mating Substances in Paramecium caudatum

SYNOPSIS Cilia detached from mating reactive cells of Paramecium caudatum were fractionated for the purpose of identifying the structural component bearing mating substances. Purified axoneme fractions had no mating reactivity. The membrane fraction obtained by dialyzing against a solution of Tris-EDTA (0.1 m m EDTA, 1 m m Tris-HCI, pH 7.6) and 0.6 m KCI, and then by centrifuging over 40% (w/v) sucrose was strongly reactive. No mating reactivity was detected in the soluble fractions containing axonemal and matrix proteins. The results indicate that the mating substances in active from are localized only on the ciliary membranes.     

ELECTRON TRANSPORT SYSTEMS OF RHIZOBIUM GROWN IN NODULES AND IN LABORATORY MEDIUM

The electon transport systems of Rhizobium japonicum were studied,comparing cells harvested from effective nodules with thosefrom artificial culture. Participation of the cytochrome systemwas confirmed in both forms of cells. Absorption peaks of thecytochromes of cultured cells were a, b, c type, resemblingthose of Bacillus subtilis, yeast and mammalian tissue. Cytochromea could not be detected in the absorption spectrum of symbioticcells, although the CO binding difference spectrum showed apeak at about 438 mµ, which can be attributed to a componenta₃ or a₁. CO difference spectrum also showed a shoulder at about416 mµ. Cells cultivated under the insufficient supply of oxygen showedthe cytochrome absorption spectrum closely resembled that ofsymbiotic cells. Diaphorase activity was lower in symbioticcells. These results are considered to be due to the insufficientsupply of oxygen within nodule tissue. Succinate oxidation bythe symbiotic cell paniculate was shown to be carbon monooxideresistant. NADH₂ oxidation by the supernatant fraction of symbioticcells was accelerated by flavin mononucleotide, 2, 6-dichiorophenolindophenol, methylene blue and vitamin K₃. ¹Present address: Faculty of Agriculture, Tôhoku University,Sendai. ²Present address: Central Agricultural Experiment Station, Kitamoto.