Skip to main content
Log in

Silver staining of Drosophila polytene chromosomes and the effect of hyaluronidase and lysozyme pretreatment

  • Published:
Genetica Aims and scope Submit manuscript

Abstract

The Ag-AS technique was used for staining the polytene chromosomes of D. melanogaster and D. lummei. Bands were stained dark reddish-brown, interbands light yellow. A toromere was heavily stained on the sixth chromosome of D. lummei. The staining intensity of nucleoli was lower than that of chromosomes. During a prolonged staining ectopic threads and the nonhomogeneous structure of nucleoli were revealed. Pretreatment with RNase caused slight changes in the silver staining pattern of chromosomes; pretreatment with DNase did not result in any visible changes, while after preincubation with proteolytic enzymes chromosome morphology was destroyed. Hyaluronidase and lysozyme removed the silver-reducing components from chromosomes without destroying the general chromosome structure. Each of these two enzymes acts specifically: hyaluronidase affects the morphology of chromosomes, but not nucleoli and bands at heat shock puffs, whereas the action of lysozyme is probably evenly distributed between chromosomes and nucleoli.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Black, M. M. & Ansley, H. R., 1964. Histone staining with ammoniacal silver. Science 143: 693–695.

    Google Scholar 

  • Bloom, S. E., Shalit, P. & Bacon, L. D., 1978. Chromosomal localization of nucleolus organizer in chicken. Genetics 88: 13.

    Google Scholar 

  • Bourgeois, C. A., Hernandez-Verdun, D., Hubert, J. & Bouteille, M., 1979. Silver staining of NORs in electron microscopy. Expl Cell Res. 123: 449–452.

    Google Scholar 

  • Buys, C. H. C. M. & Osinga, J., 1980. Abundance of protein-bound sulfhydryl and disulfide groups at chromosomal nucleolus organizing regions. A cytochemical study on the selective silver staining of NORs. Chromosoma 77: 1–11.

    Google Scholar 

  • Byarugaba, W., 1978. A pepsin-revealed material possibly related to chromosomal banding. Chromosoma 67: 275–283.

    Google Scholar 

  • Conn, P. M. & O'Malley, B. W., 1975. Lysozyme (muramidase) in isolated chick oviduct nuclei. Biochem. biophys. Res. Communs 64: 740–746.

    Google Scholar 

  • Czaker, R., 1978. Silver staining in transcriptionally active NORs of meiotic and mitotic cells in Achaeta domesticus (L.) (Orthoptera). Chromosoma 68: 187–193.

    Google Scholar 

  • Dresser, M. E. & Moses, M. J., 1980. Synaptonemal complex karyotyping in spermatocytes of the Chinese hamster (Cricetus griseus). IV. Light and electron microscopy of synapsis and nucleolar development by silver staining. Chromosoma 76: 1–22.

    Google Scholar 

  • Evgen'ev, M. B., Stepanova, N. G., Levin, A. V. & Shilov, A. A., The structure of toromere in Drosophila lummei Hackman and its role in chromosome behavior. Molec. Biol. (Russ.) (in press).

  • Goodpasture, C. & Bloom, S. E., 1975. Visualization of nucleolar organizer regions in mammalian chromosomes using silver staining. Chromosoma, 53: 37–50.

    Google Scholar 

  • Hausmann, I., Gebauer, J., Bihl, L. & Crimm, T., 1978. Onset of nucleolus organizer activity in early mouse embryogenesis and evidence for its regulation. Expl Cell Res. 114: 263–268.

    Google Scholar 

  • Heneen, W. K., 1979. Silver staining of nucleolar organizers and nucleoli during mitosis and interphase of mammalian cells. Genetica 50: 31–39.

    Google Scholar 

  • Hofgartner, F. J., Krone, W. & Kamiesh, J., 1979. Correlated inhibition of ribosomal RNA synthesis and silver staining by actinomicin D. Hum. Genet. 47: 329–333.

    Google Scholar 

  • Howell, M. W., 1977. Vizualization of ribosomal gene activity: silver stains proteins associated with rRNA transcribed from oocyte chromosomes. Chromosoma 62: 361–367.

    Google Scholar 

  • Howell, M. W. & Denton, T. E., 1976. Negative silver staining in A-T and satellite DNA-rich regions of human chromosomes. Chromosoma 57: 165–169.

    Google Scholar 

  • Howell, M. W. & Hsu, T. C., 1979. Chromosome core structure revealed by silver staining. Chromosoma 73: 61–66.

    Google Scholar 

  • Hozier, J. C., Adolph, K. W., & Furcht, L. T., 1979. Binding of lectins to human mitotic chromosomes and interphase nuclei. J. Cell Biol. 83: 160a.

  • Khachaturov, E. N., Barsky, V. E., Galkina, I. G., Stonova, N. S., & Maksimaldo, Yu. B., 1975. Cytochemical study of the substrate responsible for differential staining of human chromosomes along the length. Izv. AN SSSR, ser. biol. N 6: 873–875.

    Google Scholar 

  • Martin-DeLeon, P. A., Flemming, M. E., & Petrosky, D. E., 1978. Patterns of silver staining in cells of six-day blastocyst and kidney fibroblast of the domestic rabbit. Chromosoma 67: 245–252.

    Google Scholar 

  • Matsui, S., Goyanes, V. & Sandberg, A. A., 1978. Non-histone chromosomal proteins (NHCP) associated with nucleolarorganizers (NOR) of metaphase chromosomes. J. Cell Biol. 79: 116a.

  • Miki, B. L. A., Gurd, J. W., & Brown, I., 1978. Heterogeneity of monucleosomes. J. Cell Biol. 79: 116a.

  • Moutschen, J., 1976. Fine structure of chromosomes as revealed by fluorescence analysis. Progr. biophys. molec. Biol. 31: 39–66.

    Google Scholar 

  • Nardi, I., DeLucchini, S., Barsacchi-Pilone, G. & Andronico, F., 1978. Chromosome location of the ribosomal RNA genes in Triturus vulgaris meridionalis (Amphibia, Urodela). IV. Comparison between in situ hybridization with 3H 18S+28S rRNA and AS-SAT staining. Chromosoma 70: 91–99.

    Google Scholar 

  • Olert, J., Sawatzki, G., Kling, H. & Gebauer, J., 1979. Cytological and histochemical studies on the mechanism of the selective silver staining of nucleolus organizer regions (NORs). Histochemistry 60: 91–99.

    Google Scholar 

  • Pelliccia, P., DeCapos, A., Belloni, G., Rocchi, A. & Ferraro, M., 1978. Localization of silver staining in interphase, prophase and metaphase lymphocytes. Expl Cell Res. 115: 439–441.

    Google Scholar 

  • Plagens, U., 1978. Effects of salt-treatment on manually isolated polytene chromosomes from Chironomus tentans. Chromosome 68: 1–19.

    Google Scholar 

  • Puchtler, H. & Waldrop, F. S., 1978. Silver impregnation methods for reticulum fibers and reticulin: a re-investigation of their origins and specificity. Histochemistry 57: 177–187.

    Google Scholar 

  • Rizzo, W. B. & Bustin, M., 1977. Lectins as probes of chromatin structure. Binding of concanavalin A to purified rat liver chromatin. J. biol. Chem. 252: 7062–7067.

    Google Scholar 

  • Schmid, M., 1978. Chromosome banding in Amphibia. I. Constitutive heterochromatin and nucleolus organizer regions in Bufo and Hyla. Chromosoma 66: 361–388.

    Google Scholar 

  • Schubert, I., Anastassova-Kristeva, M. & Rieger, R., 1979. Specificity of NOR staining in Vicia faba. Expl Cell Res. 120: 433–435.

    Google Scholar 

  • Schwarzacher, H. G., Mikelsaar, A.-V. & Schnedl, W., 1978. The nature of the Ag-staining of nucleolus organizing regions: electron and light microscopic studies on human cells in interphase, mitosis, and meiosis. Cytogenet. Cell Genet. 20: 24–39.

    Google Scholar 

  • Stocker, A. J., 1978. Correspondence of silver banding with rRNA hybridization sites in polytene chromosomes of Rhynchosciara hollaenderi. Expl Cell Res. 114: 429–434.

    Google Scholar 

  • Stocker, A. J., Fresquez, C. & Leutzions, G., 1978. Banding studies on the polytene chromosomes of Rhynchosciara hollaenderi. Chromosoma 68: 337–356.

    Google Scholar 

  • Tantravahi, R., Miller, D. A., Dev, V. G. & Miller, O. G., 1976. Detection of nucleolus organizer regions in chromosomes of human, chimpanzee, gorilla, orangutan and gibbon. Chromosoma 56: 15–27.

    Google Scholar 

  • Varley, J. M. & Morgan, G. T., 1977. Silver staining of the lampbrush chromosomes of Triturus cristatus carnifex. Chromosoma 67: 233–244.

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Birstein, V.J. Silver staining of Drosophila polytene chromosomes and the effect of hyaluronidase and lysozyme pretreatment. Genetica 56, 167–173 (1981). https://doi.org/10.1007/BF00057556

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00057556

Keywords

Navigation