Abstract
The ultrastructural localization of rDNA and rRNA within the nucleolus of human spermatids was investigated by in situ hybridization at steps 1 and 2. Two different digoxigenin-labeled human probes from the rRNA transcription unit were used. Identification of hybrids was performed with immunogold techniques. Comparative observations in the Sertoli cell nucleolus as controls revealed that rDNA was predominantly visualized in the threads of the dense fibrillar component, while rRNA was detected over both the fibrillar component and the granular component. Whithin the nucleolus of round spermatids in the same sections of seminiferous tubules, rDNA labeling was localized over the spherical or stranded dense fibrillar components. rRNA labeling was found not only over these components but also in the adjacent nucleoplasm rich in ribonucleoprotein particles. These results are consistent with the view that the round spermatid nucleolus is transcriptionally active.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Altman LG, Schneider BG, Papermaster DS (1984) Rapid embedding of tissues in Lowicryl K4 M for immunoelectron microscopy. J Histochem Cytochem 32:1217–1223
D'Agostino A, Geremia R, Monesi V (1978) Post-meiotic gene activity in spermatogenesis of the mouse. Cell Differentiation 7:175–183
Dadoune JP, Alfonsi MF (1989) Nuclear changes during spermiogenesis in man and the monkey. In: Motta PM (ed) Developments in ultrastructure of reproduction. Liss, New York, pp 165–170
Dadoune JP, Alfonsi MF, Fain-Maurel MA (1991) Cytochemical variations in the nucleolus during spermiogenesis in man and monkey. Cell Tissue Res 264:167–173
Fakan S, Hernandez-Verdun D (1986) The nucleolus and the nucleolar organizer regions. Biol Cell 56:189–206
Financsek I, Mizumoto K, Mishima Y, Muramatsu M (1982) Human ribosomal RNA gene: nucleotide sequence of the transcription initiation region and comparison of three mammalian genes. Proc Natl Acad Sci USA 79:3092–3096
Goessens G (1984) Nucleolar structure. Int Rev Cyt 87:107–158
Grootegoed JA, Grollé-Hey AH, Rommerto FFG, Van der Moless HJ (1977) Ribonucleic acid synthesis in vitro in primary spermatocytes isolated from rat testis. Biochem J 168:23–31
Hecht NB (1990) Gene expression during spermiogenesis. In: Hamilton DW, Waites GMH (eds) Cellular and molecular events in spermiogenesis. Cambridge University Press, pp 265–283
Kierszenbaum AL, Tres LL (1974) Nucleolar and perichromosomal RNA synthesis during meiotic prophase in the mouse testis. J Cell Biol 60:39–53
Puvion-Dutilleul F, Bachellerie JP, Puvion E (1991) Nucleolar organization of HeLa cells as studied by in situ hybridization. Chromosoma 100:395–409
Scheer U, Benavente R (1990) Functional and dynamic aspects of the mammalian nucleolus. Bioessays 12:14–21
Schwarzacher H, Wachtler F (1987) Nucleolus organizer regions and nucleoli: cytological findings. Chromosomes Today 9:252–260
Stahl A, Wachtler F, Hartung M, Devictor M, Schöfer C, Mosgöller W, Lanversin A de, Fouet C, Schwarzacher HG (1991) Nucleoli, nucleolar chromosomes and ribosomal genes in the human spermatocyte. Chromosoma 101:231–244
Sylvester JE, Whiteman DA, Podolsky R, Pozsgay JM, Respess J, Schmickel RD (1986) The human ribosomal RNA genes: structure and organization of the complete repeating unit. Hum Genet 73:193–198
Thiry M (1988) Immunoelectron microscope localization of bromodeoxyuridine incorporated into DNA of Ehrlich tumor cell nucleoli. Exp Cell Res 179:204–213
Thiry M (1991) DNase I-senstive sites within the nuclear architecture visualized by immunoelectron microscopy. DNA Cell Biol 10:169–180
Thiry M (1992) Ultrastructural detection of DNA within the nucleolus by sensitive molecular immunocytochemistry. Exp Cell Res 200:135–144
Thiry M, Thiry-Blaise L (1989) In situ hybridization at the electron microscope level:an improved method for precise localization of ribosomal DNA and RNA. Eur J Cell Biol 50:235–243
Thiry M, Thiry-Blaise L (1991) Locating transcribed and nontranscribed rDNA spacer sequences within the nucleolus by in situ hybridization and immunoelectron microscopy. Nucleic Acids Res 19:11–15
Thiry M, Scheer U, Goessens G (1988) Localization of DNA within Ehrlich tumour cell nucleoli by immunoelectron microscopy. Biol Cell 63:27–34
Thiry M, Ploton D, Menager M, Goessens G (1993) Ultrastructural distribution of DNA within the nucleolus of various animal cell lines or tissues revealed by terminal deoxynucleotidyl transferase. Cell Tissue Res 271:33–45
Wachtler F, Mosgöller W, Schwarzacher H (1990) Electron microscopic in situ hybridization and autoradiography: localization and transcription of rDNA in human lymphocyte nucleoli. Exp Cell Res 187:346–348
Wachtler F, Schofer C, Mosgöller W, Weipoltshammer K, Schwarzacher HG, Guichaoua M, Hartung M, Stahl A, Bergé-Lefranc JL, Gonzalez I, Sylvester J (1992) Human ribosomal RNA gene repeats are localized in the dense fibrillar component of nucleoli: light and electron microscopic in situ hybridization in human Sertoli cells. Exp Cell Res 198:135–143
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Dadoune, JP., Siffroi, JP. & Alfonsi, MF. Ultrastructural localization of rDNA and rRNA by in situ hybridization in the nucleolus of human spermatids. Cell Tissue Res 278, 611–616 (1994). https://doi.org/10.1007/BF00331381
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00331381