Chromosome Research

, Volume 15, Issue 6, pp 755–765 | Cite as

Nucleolus size variation during meiosis and NOR activity of a B chromosome in the grasshopper Eyprepocnemis plorans

  • María Teruel
  • Josefa Cabrero
  • Francisco Perfectti
  • Juan Pedro M. CamachoEmail author


The number of nucleoli and nucleolar area were measured in meiotic cells from males of the grasshopper Eyprepocnemis plorans collected in three natural populations. Number of nucleoli per cell showed no significant correlation among cells in different meiotic stages, but there was strong positive correlation for nucleolar area between leptotene and interkinesis cells in individuals from distant populations (Salobreña in Spain, and Smir in Morocco). No correlation was, however, observed for both parameters between the meiotic stages analysed in individuals from the population of Torrox (Spain). The number of nucleoli at leptotene was about double the number at interkinesis, as expected from the double ploidy level at leptotene and the corresponding double number of rDNA clusters. Leptotene nucleolar area, however, was about fourfold that in interkinesis, presumably due to higher requirements for ribosome biogenesis in meiosis I than meiosis II. In Torrox, diplotene cells showed a lower number of nucleoli but larger nucleolar area than in leptotene cells, suggesting an increase in nucleolus size during prophase I. Significant differences were found among populations for nucleolar area but not for number of nucleoli, the smallest nucleolar area being observed in Torrox, which is the population harbouring the most parasitic B chromosome variant. No clear effects on nucleolar area or number of nucleoli were associated with the B-chromosome number. However, B-chromosome effects on the nucleolar area were apparent in the Torrox population when data were analysed with respect to a B-chromosome odd–even pattern in leptotene and interkinesis cells. However, in diplotene cells no odd–even pattern was observed for both nucleolar parameters, suggesting that the increase in nucleolar size from leptotene to diplotene dilutes the leptotene odd–even pattern. The rDNA distally located in the B chromosome was associated with a nucleolus in 6.5% out of the 247 diplotene cells analysed. The implications of these findings are discussed in the context of B chromosomes as stress-causing genome parasites and the nucleolus as a sensor of stress.

Key words

B chromosomes Eyprepocnemis plorans meiosis nucleolus rRNA 


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  1. Andersen JS, Lam YW, Leung AK et al. (2005) Nucleolar proteome dynamics. Nature 433: 77–83.PubMedCrossRefGoogle Scholar
  2. Bakkali M, Camacho JPM (2004) The B chromosome polymorphism of the grasshopper Eyprepocnemis plorans in North Africa. III. Mutation rate of B chromosomes. Heredity 92: 428–433.PubMedCrossRefGoogle Scholar
  3. Bakkali M, Cabrero J, López-León MD, Perfectti F, Camacho JPM (1999) The B chromosome polymorphism of the grasshopper Eyprepocnemis plorans in North Africa. I. B variants and frequency. Heredity 83: 428–434.PubMedCrossRefGoogle Scholar
  4. Bakkali M, Cabrero J, López-León MD, Perfectti F, Camacho JPM (2001) Population differences in the expression of nucleolus organizer regions in the grasshopper Eyprepocnemis plorans. Protoplasma 217: 185–190.PubMedCrossRefGoogle Scholar
  5. Bakkali M, Perfectti F, Camacho JPM (2002) The B-chromosome polymorphism of the grasshopper Eyprepocnemis plorans in North Africa. II. Parasitic and neutralized B1 chromosomes. Heredity 88: 14–18.PubMedCrossRefGoogle Scholar
  6. Busch H (1997) Nucleolar and nucleolonemal proteins of cancer cells. J Tumor Marker Oncol 12: 5–68.Google Scholar
  7. Busch H, Daskal Y, Gyorkey F, Smetana K (1979) Silver staining of nucleolar granules in tumor cells. Cancer Res 39: 857–863.PubMedGoogle Scholar
  8. Cabrero J, Alché JD, Camacho JPM (1987) Effects of B chromosome on the activity of nucleolus organizer regions in the grasshopper Eyprepocnemis plorans: activation of a latent nucleolus organizer region on a B chromosome fused to an autosome. Genome 29: 116–121.Google Scholar
  9. Cabrero J, López-León MD, Bakkali M, Camacho JPM (1999) Common origin of B chromosome variants in the grasshopper Eyprepocnemis plorans. Heredity 83: 435–439.PubMedCrossRefGoogle Scholar
  10. Camacho JPM (2005) B Chromosomes. In: Gregory TR, ed., The Evolution of the Genome. San Diego: Elsevier, pp. 223–286.Google Scholar
  11. Camacho JPM, Sharbel TF, Beukeboom LW (2000) B chromosome evolution. Phil Trans R Soc Lond B 355: 163–178.CrossRefGoogle Scholar
  12. Camacho JPM, Shaw MW, López–León MD, Pardo MC, Cabrero J (1997) Population dynamics of a selfish B chromosome neutralized by the standard genome in the grasshopper Eyprepocnemis plorans. Am Nat 149: 1030–1050.CrossRefPubMedGoogle Scholar
  13. Camacho JPM, Perfectti F, Teruel M, López-León MD, Cabrero J (2004) The odd–even effect in mitotically unstable B chromosomes in grasshoppers. Cytogenet Genome Res 106: 325–331.PubMedCrossRefGoogle Scholar
  14. Caspersson T (1950) Cell Growth and Cell Function, a Cytochemical Study. New York: WW Norton.Google Scholar
  15. Derenzini M (2000) The AgNORs. Micron 31: 117–120.PubMedCrossRefGoogle Scholar
  16. Derenzini M, Farabegoli F, Trerè D (1992) Relationship between interphase AgNOR distribution and nucleolar size in cancer cells. J Mol Histol 24: 951–956.CrossRefGoogle Scholar
  17. Derenzini M, Sirri V, Trerè D (1994) Nucleolar organizer regions in tumour cells. Cancer J 7: 71–77.Google Scholar
  18. Derenzini M, Trerè D, Pession A, Montanaro L, Sirri V, Ochs RL (1998) Nucleolar function and size in cancer cells. Am J Pathol 152: 1291–1297.PubMedGoogle Scholar
  19. Derenzini M, Trerè D, Pession A, Govoni M, Sirri V, Chieco P (2000) Nucleolar size indicates the rapidity of cell proliferation in cancer tissues. J Pathol 191: 181–186.PubMedCrossRefGoogle Scholar
  20. Guo M, Davis D, Birchler JA (1996) Dosage effects on gene expression in a maize ploidy series. Genetics 142: 1349–1355.PubMedGoogle Scholar
  21. Herbener GH, Bendayan M (1998) A correlated morphometric and cytochemical study on hepatocyte nucleolar size and RNA distribution during vitellogenesis. Histochem J 20: 194–200.CrossRefGoogle Scholar
  22. Hudson LA, Ciborowski JJH (1996) Teratogenic and genotoxic responses of larval Chironomus salinarius group (Diptera: Chironomidae) to contaminated sediment. Environ Toxicol Chem 15: 1375–1381.CrossRefGoogle Scholar
  23. Lagerstedt S (1949) Cytological studies on the protein metabolism of the liver in the rat. Acta Anat Suppl 9: 1–140.Google Scholar
  24. López-León MD, Cabrero J, Pardo MC, Viseras E, Camacho JPM, Santos JL (1993) Generating high variability of B chromosomes in Eyprepocnemis plorans (grasshopper). Heredity 71: 352–362.Google Scholar
  25. López-León MD, Neves N, Schwarzacher T, Heslop-Harrison JS, Hewitt GM, Camacho JPM (1994) Possible origin of a B chromosome deduced from its DNA composition using double FISH technique. Chromosome Res 2: 87–92.PubMedCrossRefGoogle Scholar
  26. López-León MD, Cabrero J, Camacho JPM (1995) Changes in NOR activity pattern in the presence of supernumerary heterochromatin in the grasshopper Eyprepocnemis plorans. Genome 38: 68–74.CrossRefPubMedGoogle Scholar
  27. McClintock B (1934) The relationship of a particular chromosomal element to the development of the nucleoli in Zea mays. Z Zellforch Mikrosk Anat 21: 294–328.CrossRefGoogle Scholar
  28. Mosgoeller W (2004) Nucleolar ultrastructure in vertebrates. In: Olson MOJ, ed., The Nucleolus. New York: Kluwer, pp. 10–20.Google Scholar
  29. Nakamoto K, Ito A, Watabe K et al. (2001) Increased expression of a nucleolar Nop5/Sik family member in metastatic melanoma cells: evidence for its role in nucleolar sizing and function. Am J Pathol 159: 1363–1374.PubMedGoogle Scholar
  30. Okabe Y, Nakamura S, Okumura H et al. (1991) The relation of argyrophilic proteins of nucleolar organizer regions (AgNORs) to the proportions of Ki–67 or DNA polymerase á-reacting cells in non-Hodgkin's lymphomas. Anticancer Res 11: 2031–2035.PubMedGoogle Scholar
  31. Olson MOJ (2004) Sensing cellular stress: another new function for the nucleolus? Sci STKE 2004: e10.CrossRefGoogle Scholar
  32. Pikaard CS (2002) Transcription and tyranny in the nucleolus: the organization, activation, dominance and repression of ribosomal RNA genes. In: Somerville CR & Meyerowitz EM, eds., The Arabidopsis Book. Rockville: American Society of Plant Biologists, pp. 1–23.Google Scholar
  33. Rasband WS (1997) ImageJ, U. S. National Institutes of Health, Bethesda, Maryland, USA,, 1997–2006.
  34. Rubbi CP, Milner J (2003) Disruption of the nucleolus mediates stabilisation of p53 in response to DNA damage and other stresses. EMBO J 22: 6068–6077.PubMedCrossRefGoogle Scholar
  35. Rufas JS, Iturra P, de Souza W, Esponda P (1982) Simple silver staining procedure for the localization of nucleolus and nucleolar organizer under light and electron microscopy. Arch Biol 93: 267–274.Google Scholar
  36. Schmid M, Löser C, Schmidtke J, Engel W (1982) Evolutionary conservation of a common pattern of activity of nucleolus organizers during spermatogenesis in vertebrates. Chromosoma 86: 149–179.PubMedCrossRefGoogle Scholar
  37. Schmid M, Müller H, Stasch S, Engel W (1983) Silver staining of nucleolus organizer regions during human spermatogenesis. Hum Genet 64: 363–370.PubMedCrossRefGoogle Scholar
  38. Shubert I, Künzel G (1990) Position dependent NOR activity in barley. Chromosoma 99: 352–359.CrossRefGoogle Scholar
  39. Sumner AT (2003) Chromosomes: Organization and Function. Oxford: Blackwell.Google Scholar
  40. Thiele J, Fischer R (1993) Bone marrow tissue and proliferation markers: results and general problems. Virchows Arch A Pathol Anat Histopathol 423: 409–416.PubMedCrossRefGoogle Scholar
  41. Trerè D (2000) AgNOR staining and quantification. Micron 31: 127–131.PubMedCrossRefGoogle Scholar
  42. Tsai RYL, McKay RDG (2005) A multistep, GTP-driven mechanism controlling the dynamic cycling of nucleostemin. J Cell Biol 168: 179–184.PubMedCrossRefGoogle Scholar
  43. Tuma RS (2005) A GTP signal to the nucleolus. J Cell Biol 168: 172.Google Scholar
  44. Wachtler F, Stahl A (1993) The nucleolus: a structural and functional interpretation. Micron 24: 473–505.CrossRefGoogle Scholar
  45. Wannemacher RW (1972) Ribosomal ribonucleic acid synthesis and function as influenced by amino acid supply and stress. Biochem J 129: 5P–6P.Google Scholar
  46. Yezerinac SM, Lougheed SC, Handford P (1992) Measurement error and morphometric studies- Statistical power and observer experience. Syst Biol 41: 471–482.CrossRefGoogle Scholar
  47. Zurita S, Cabrero J, López-León MD, Camacho JPM (1998) Polymorphism regeneration for a neutralized selfish B chromosome. Evolution 52: 274–277.CrossRefGoogle Scholar

Copyright information

© Springer 2007

Authors and Affiliations

  • María Teruel
    • 1
  • Josefa Cabrero
    • 1
  • Francisco Perfectti
    • 1
  • Juan Pedro M. Camacho
    • 1
    Email author
  1. 1.Departamento de GenéticaUniversidad de GranadaGranadaSpain

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