Preliminary notes the Swedish Medical Research Council @%78-16X02183-12). from the Swedish Societv for Medical Research aid from the Medical Faculty, University of Linkiiping.

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Specificity of NOR staining in Vi& faba References 1. Wilkinson, P C, Clin exp immunol25 (1976) 355.

2. Bjiirksten, B, Casann, R & Quie, P G, Infect immunol 14 (1976) 315. 3. Wilkinson, P C, Immunology 33 (1977)407. 4. Dahlgren, C, Kihlstriim, E, Magnusson, K-E, Stendahl, 0 & Tagesson, C, Exp cell res 108 (1977) 175. 5. Woodin, A M & Harris, A, Exp cell res 77 (1973) 41. 6. Woodin, A M, Poole, A R & Dunn, G A, Exp cell res 94 (1975) 292. 7. Shanbhag, V & Johansson, G, Biochem biophys res commun 61 (1974) 1141. 8. StjemstrGm, I, Magnusson, K-E, Stendahl, 0 & Tagesson, C, Infect immunol 18 (1978) 261. 9. B@yum, A, Stand j clin invest 21 (1968) suppl. 97,31. 10. Hed, J, FEMS microbial lett 1 (1977) 357. II. Oliver, J M, Zurica, R B & Berlin, R D, Nature 253 (1975)471. 12. Ward, H A & Fothergill, J E, Fluorescent protein tracing (ed R C Naim) 4th edn, p. 27. Churchill Livingstone, Edinburgh (1976). 13. Nelson, R D, Quie, P G & Simmons, R L, Immunoloav 115(1975) 1650. 14. StendaG: 0, M‘olin,‘L & Dahlgren, C, J clin invest 62 (1978) 214. 15. Repo, H, Leukocyte migration agarose test for the assessment of human neutroohil chemotaxis. Thesis, University of Helsinki, I%nland (1976). 16. Dahlgren, C, Edebo, L & Munktell, G, Movement, metabolism and bactericidal mechanisms of phagocytes (ed F Rossi, P L Patriarca & D komeoj p. 41. Piccin medical books, Padua (1977). 17. Dahlgren, C, Hed, J, Magnusson, K-E & Sundqvist, T, Stand j immunol. In press. 18. $n, W S & Mukherjee, C, Am j pathol 1 (1973) 19. Mandell, B F, Spilberg, I & Lichtman, J, J immunol 118(1977) 1375. 20. Hawley, H P & Gordon, G B, Lab invest 34 (1976) 216. 21. Tanford, C, The hydrophobic effect. John Wiley & Sons, New York (1973). 22. Ryan, G B, Borysenko, J 2 & Karnovsky, M J, J cell biol 62 (1974) 35I Received November 10, 1978 Revised version received January 5, 1979 Accepted January 19, 1979

I. SCHUBERT, M. ANASTASSOVA-KRISTEVA’ fiir Genetik und Kuland R. RIEGER. Zentralinstitut turpjlanzenforschung der Aka&emie der schaften der DDR, 4325 Gatersleben, DDR

Wissen-

Silver staining of the nucleolus organizing regions (NORs) in Vicia faba has been demonstrated and its specificity compared with “N-banding” in plant material is discussed.

Summary.

On the basis of silver staining, NOR (nucleolus organizing region) staining was obtained in Viciafaba by adapting the method of Goodpasture & Bloom [l] as simplified by Varley [2]; air-dried squash preparations of pectinase-digested root tips were incubated for 12.5 h at 65°C in a AgN03 (Merck) solution (5 g/5 ml, i.e., double the concentration used by Varley), then the slides were rinsed in distilled water, air-dried again, dipped in xylol, and mounted in cedax. Both the interphase nuclei and the metaphase chromosomes were stained yellow. In interphase cells the nucleoli and in metaphase chromosomes the secondary constrictions became exclusively stained, dark brown (fig. 1). In a small number of interphase nuclei, some additional darkly stained spots appeared for which no corresponding bands in metaphase chromosomes were found. By means of in situ hybridization with radioactively labelled 18 and 25s rRNA the nucleolus organizing secondary constriction was previously found to be the only recognizable site of these rRNA genes in Viciafuba [3,4]. The result described here supports, for plant material, the original interpretation of NOR staining by Goodpasture & Bloom [I], who suggested that silver staining of the ’ Permanent address: Institute of Morphology, Bulgarian Academy of Sciences, Sofia 1113,Bulgaria. Exp

Cell

Res IZO (1979)

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Preliminary notes

I. (a) Metaphase cell with silver-stained NORs in the secondary constrictions (arrows) of both third chromosomes. The karyotype used (termed ACB) differs from the normal karyotype of Vicia fuba by the homozygous combination of a pericentric inversion in the fifth chromosome (=B), and two reciprocal translocations, one of them between the long arms of chro-

Fig.

Exp Cd/ Res 120 (1979)

mosomes I and VI (=C), and one between the short arm of chromosome I and the long arm of chromosome III (=A). By the latter the NOR (secondary constriction) was transposed from the first to the third chromosome. For further description of this karyotype see [lo]; (b) interphase nucleus with darkly stained nucleoli.

Preliminary notes NOR might be due to some non-histone protein specific for rDNA loci (probably only those which are active in transcription [5,

61). Gerlach [7] observed bands in cereals after application of a method developed and termed “N-banding” by Funaki et al. [8], not only at the sites of NORs but also in some other chromosomal regions. With the other bands appearing, sometimes even the NORs remained unstained. Since the technique by Funaki et al. [8] as well as by Gerlach [7] are modified Giemsa techniques the “additional bands” [8] exactly correspond to the bands described in Vicia after performance of a variety of Giemsa staining techniques (for review see [9]). Some of the dark bands, inclusive those found in the secondary constriction, after Giemsa techniques in Vicia faba become manifest with different frequencies [9]. This frequency of manifestation may vary due to modifications of the technique used (Dobel, unpublished). Keeping this in mind, it seems that NOR staining obtained by the silver staining method is indeed very specific for NORs and the “unspecific” bands obtained with modified Giemsa techniques represent a phenomenon different from original NOR staining. (Similar conclusions were drawn by Faust & Vogel [ 1l] concerning the specificity of “N-banding” in animal cells.) For that reason silver staining seems to be a more useful tool for the detection of NORs (and their transcriptive activity?) in plant tissues than “N-banding”. We are grateful to Miss A. Scheer for skilful technical assistance.

References 1. Goodpasture, C & Bloom, S E, Chromosoma 53 (1975) 37. 2. Varley, J M, Chromosoma 61 (1977) 207.

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3. Scheuermann, W & Knllmann, M, Exp cell res 90 (1975) 463. 4. Schubert, I, BPumlein, H & Wobus, U, Biol Zb197 (1978) 129. 5. Miller, D A, Dev, V G, Tantravahi, R & Miller, 0 J, Exp cell res 101 (1976) 235. 6. Howell, W M, Chromosoma 62 (1977) 361. 7. Gerlach, W L, Chromosoma 62 (1977) 49. 8. Funaki, K, Matsui, S & Sasaki, M, Chromosoma 49 (1975) 357. 9. Dobel, P, Schubert, I & Rieger, R, Chromosoma 69 (1978) 193. 10. Michaelis, A & Rieger, R, Chromosoma 35 (1971) 1. 1I. Faust, J & Vogel, W, Nature 249 (1974) 352. Received November 10, 1978 Revised version received January 3, 1979 Accepted January 8, 1979

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Ciliogenesis in tissue-cultured cells by the increased density of cell population Y. MORI,’ H. AKED0,2 Y. TANIGAKI,l K. TANAKA’ and M. OKADA,’ ‘Department of Experimental Pathology, and 2Department of Biochemistry, The Center for Adult Diseases, I-3-3, Nakamichi, Higashinari-ku, Osuka, 537, Japan

Cilia developed on the surfaces of cells of an established line derived from rat liver (LI 10) as the population of the cultures increased. Ultrastructural examination by the scanning electron microscope (SEM) showed that a cilium appeared on a cell surface when cells were cultured more than 5 days after reaching the confluent stage. The essential components of the cilium were a basal body and central and peripheral librils displaying 9+2 or 9+0 doublet arrangement in the cross section. The cilium was not observed during the time when the cells were growing.

Summary.

It has been reported that phenotypic expressions of cultured cells are often modified when the density of cell population changes during culture [l-4]. Williams et al. examined differences in cellular ultrastructures and in the activities of several cellular enzymes among pre-confluent, confluent and post-confluent culture cells originating from rat liver, and suggested that the cells E,rp Cell Res I20 (1979)

Specificity of NOR staining in Vicia faba.

Preliminary notes the Swedish Medical Research Council @%78-16X02183-12). from the Swedish Societv for Medical Research aid from the Medical Faculty,...
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