Chromosome territoriality is not random along the cell cycle and it is mainly governed by intrinsic chromosome factors and gene expression patterns. Conversely, very few studies have explored the factors that determine chromosome territoriality and its influencing factors during meiosis. In this study, we analysed chromosome positioning in murine spermatogenic cells using three-dimensionally fluorescence in situ hybridization-based methodology, which allows the analysis of the entire karyotype. The main objective of the study was to decipher chromosome positioning in a radial axis (all analysed germ-cell nuclei) and longitudinal axis (only spermatozoa) and to identify the chromosomal factors that regulate such an arrangement. Results demonstrated that the radial positioning of chromosomes during spermatogenesis was cell-type specific and influenced by chromosomal factors associated to gene activity. Chromosomes with specific features that enhance transcription (high GC content, high gene density and high numbers of predicted expressed genes) were preferentially observed in the inner part of the nucleus in virtually all cell types. Moreover, the position of the sex chromosomes was influenced by their transcriptional status, from the periphery of the nucleus when its activity was repressed (pachytene) to a more internal position when it is partially activated (spermatid). At pachytene, chromosome positioning was also influenced by chromosome size due to the bouquet formation. Longitudinal chromosome positioning in the sperm nucleus was not random either, suggesting the importance of ordered longitudinal positioning for the release and activation of the paternal genome after fertilisation.
This is a preview of subscription content, access via your institution.
Buy single article
Instant access to the full article PDF.
Tax calculation will be finalised during checkout.
Bartholdi MF (1991) Nuclear distribution of centromeres during the cell cycle of human diploid fibroblasts. J Cell Sci 99(Pt 2):255–263
Berrios S (2017) Nuclear architecture of mouse spermatocytes: chromosome topology, heterochromatin, and nucleolus. Cytogenet Genome Res. https://doi.org/10.1159/000460811
Bolzer A, Kreth G, Solovei I, Koehler D, Saracoglu K, Fauth C, Müller S, Eils R, Cremer C, Speicher MR, Cremer T (2005) Three-dimensional maps of all chromosomes in human male fibroblast nuclei and prometaphase rosettes. PLoS Biol. https://doi.org/10.1371/journal.pbio.0030157
Boyle S, Gilchrist S, Bridger JM, Mahy NL, Ellis JA, Bickmore WA (2001) The spatial organization of human chromosomes within the nuclei of normal and emerin-mutant cells. Hum Mol Genet. https://doi.org/10.1093/hmg/10.3.211
Bridger JM, Boyle S, Kill IR, Bickmore WA (2000) Re-modelling of nuclear architecture in quiescent and senescent human fibroblasts. Curr Biol. https://doi.org/10.1016/s0960-9822(00)00312-2
Chaly N, Munro SB (1996) Centromeres reposition to the nuclear periphery during L6E9 myogenesis in vitro. Exp Cell Res. https://doi.org/10.1006/excr.1996.0082
Cox M, Cox T (2008) Multidimensional scaling. In: Handbook of data visualization. Springer Handbooks Comp.Statistics. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-540-33037-0_14
Crabbe L, Cesare AJ, Kasuboski JM, Fitzpatrick JAJ, Karlseder J (2012) Human telomeres are tethered to the nuclear envelope during postmitotic nuclear assembly. Cell Rep. https://doi.org/10.1016/j.celrep.2012.11.019
Cremer T, Cremer M (2010) Chromosome territories. Cold Spring Harb Perspect Biol. https://doi.org/10.1101/cshperspect.a003889
Cremer M, Küpper K, Wagler B, Wizelman L, von Hase J, Weiland Y, Kreja L, Diebold J, Speicher MR, Cremer T (2003) Inheritance of gene density–related higher order chromatin arrangements in normal and tumor cell nuclei. J Cell Biol. https://doi.org/10.1083/jcb.200304096
Cremer M, Grasser F, Lanctôt C, Müller S, Neusser M, Zinner R, Solovei I, Cremer T (2008) Multicolor 3D fluorescence in situ hybridization for imaging interphase chromosomes. Methods Mol Biol. https://doi.org/10.1007/978-1-59745-406-3_15
Croft JA, Bridger JM, Boyle S, Perry P, Teague P, Bickmore WA (1999) Differences in the localization and morphology of chromosomes in the human nucleus. J Cell Biol. https://doi.org/10.1083/jcb.145.6.1119
da Cruz I, Rodríguez-Casuriaga R, Santiñaque FF, Farías J, Curti G, Capoano CA, Folle GA, Benavente R, Sotelo-Silveira JR, Geisinger A. (2016) Transcriptome analysis of highly purified mouse spermatogenic cell populations: gene expression signatures switch from meiotic-to postmeiotic-related processes at pachytene stage. BMC Genomics. https://doi.org/10.1186/s12864-016-2618-1
Federico C, Cantarella CD, Di Mare P, Tosi S, Saccone S (2008) The radial arrangement of the human chromosome 7 in the lymphocyte cell nucleus is associated with chromosomal band gene density. Chromosoma. https://doi.org/10.1007/s00412-008-0160-x
Ferguson M, Ward DC (1992) Cell cycle dependent chromosomal movement in pre-mitotic human T-lymphocyte nuclei. Chromosoma 101(9):557–565. https://doi.org/10.1007/BF00660315
Foster HA, Abeydeera LR, Griffin DK, Bridger JM (2005) Non-random chromosome positioning in mammalian sperm nuclei, with migration of the sex chromosomes during late spermatogenesis. J Cell Sci. https://doi.org/10.1242/jcs.02301
Fritz AJ, Stojkovic B, Ding H, Xu J, Bhattacharya S, Gaile D, Berezney R (2014) Wide-scale alterations in interchromosomal organization in breast cancer cells: defining a network of interacting chromosomes. Hum Mol Genet. https://doi.org/10.1093/hmg/ddu237
Garagna S, Zuccotti M, Thornhill A, Fernandez-Donoso R, Berrios S, Capanna E, Redi CA (2001) Alteration of nuclear architecture in male germ cells of chromosomally derived subfertile mice. J Cell Sci 114:4429–4434
Garcia-Quevedo L, Sarrate Z, Vidal F, Blanco J (2012) A sequential methodology that allows apoptotic cell sorting and FISH analysis in human testicular cells. Syst Biol Reprod Med. https://doi.org/10.3109/19396368.2012.717163
Gurevitch M, Amiel A, Ben-Zion M, Fejgin M, Bartoov B (2001) Acrocentric centromere organization within the chromocenter of the human sperm nucleus. Mol Reprod Dev. https://doi.org/10.1002/mrd.1116
Guttenbach M, Martínez-Expósito MJ, Engel W, Schmid M (1996) Interphase chromosome arrangement in Sertoli cells of adult mice. Biol Reprod. https://doi.org/10.1095/biolreprod54.5.980
Handel MA (2004) The XY body: A specialized meiotic chromatin domain. Exp Cell Res. https://doi.org/10.1016/j.yexcr.2004.03.008
Harper L, Golubovskaya I, Cande WZ (2004) A bouquet of chromosomes. J Cell Sci. https://doi.org/10.1242/jcs.01363
Hiraoka Y, Dernburg AF (2009) The SUN rises on meiotic chromosome dynamics. Dev Cell. https://doi.org/10.1016/j.devcel.2009.10.014
Kemeny S, Tatout C, Salaun G, Pebrel-Richard C, Goumy C, Ollier N, Maurin E, Pereira B, Vago P, Gouas L (2018) Spatial organization of chromosome territories in the interphase nucleus of trisomy 21 cells. Chromosoma. https://doi.org/10.1007/s00412-017-0653-6
Küpper K, Kölbl A, Biener D, Dittrich S, von Hase J, Thormeyer T, Fiegler H, Carter NP, Speicher MR, Cremer T, Cremer M (2007) Radial chromatin positioning is shaped by local gene density, not by gene expression. Chromosoma. https://doi.org/10.1007/s00412-007-0098-4
Landis JR, Koch GG (1997) The measurement of observer agreement for categorical data. Biometrics. https://doi.org/10.2307/2529310
Lefrançois P, Rockmill B, Xie P, Roeder GS, Snyder M (2016) Multiple pairwise analysis of non-homologous centromere coupling reveals preferential chromosome size-dependent interactions and a role for bouquet formation in establishing the interaction pattern. PLOS Genet. https://doi.org/10.1371/journal.pgen.1006347
Lieberman-Aiden E, van Berkum NL, Williams L, Imakaev M, Ragoczy T, Telling A, Amit I, Lajoie BR, Sabo PJ, Dorschner MO, Sandstrom R, Bernstein B, Bender MA, Groudine M, Gnirke A, Stamatoyannopoulos J, Mirny LA, Lander ES, Dekker J (2009) Comprehensive mapping of long-range interactions reveals folding principles of the human genome. Science. https://doi.org/10.1126/science.1181369
Lukásová E, Kozubek S, Kozubek M, Falk M, Amrichová J (2002) The 3D structure of human chromosomes in cell nuclei. Chromosome Res. https://doi.org/10.1023/a:1020958517788
Luo Z, Wang X, Jiang H, Wang R, Chen J, Chen Y, Xu Q, Cao J, Gong X, Wu J, Yang Y, Li W, Han C, Cheng CY, Rosenfeld MG, Sun F, Song X (2020) Reorganized 3D genome structures support transcriptional regulation in mouse spermatogenesis. IScience. https://doi.org/10.1016/j.isci.2020.101034
Meaburn KJ, Cabuy E, Bonne G, Levy N, Morris GE, Novelli G, Kill IR, Bridger JM (2007) Primary laminopathy fibroblasts display altered genome organization and apoptosis. Aging Cell. https://doi.org/10.1111/j.1474-9726.2007.00270.x
Mudrak OS, Solovjeva LV, Chagin VO (2013) Organization of chromosomes in human sperm nucleus. Hum Interphase Chromosom. New York: Springer. https://doi.org/10.1007/978-1-4614-6558-4_8
Namekawa SH, Park PJ, Zhang LF, Shima JE, McCarrey JR, Griswold MD, Lee JT (2006) Postmeiotic sex chromatin in the male germline of mice. Curr Biol. https://doi.org/10.1016/j.cub.2006.01.066
Sarrate Z, Solé M, Vidal F, Anton E, Blanco J (2018) Chromosome positioning and male infertility: it comes with the territory. J Assist Reprod Genet. https://doi.org/10.1007/s10815-018-1313-3
Schindelin J, Arganda-Carreras I, Frise E, Kaynig V, Longair M, Pietzsch T, Preibisch S, Rueden C, Saalfeld S, Schmid B, Tinevez JY, White DJ, Hartenstein V, Eliceiri K, Tomancak P, Cardona A (2013) Fiji-an Open Source platform for biological image analysis. Nat Methods. https://doi.org/10.1038/nmeth.2019
Sim J, Wright CC (2005) The kappa statistic in reliability studies: use, interpretation, and sample size requirements. Phys Ther 85(3):257–268
Solari AJ (1974) The behavior of the XY pair in mammals. Int Rev Cytol. https://doi.org/10.1016/s0074-7696(08)60928-6
Sun HB, Shen J, Yokota H (2000) Size-dependent positioning of human chromosomes in interphase nuclei. Biophys J. https://doi.org/10.1016/S0006-3495(00)76282-5
Tagawa Y, Nanashima A, Yasutake T, Hatano K, Nishizawa-Takano JE, Ayabe H (1997) Differences in spatial localization and chromatin pattern during different phases of cell cycle between normal and cancer cells. Cytometry 27(4):327–335
Tanabe H, Habermann FA, Solovei I, Cremer M, Cremer T (2002) Non-random radial arrangements of interphase chromosome territories: evolutionary considerations and functional implications. Mutat Res. https://doi.org/10.1016/S0027-5107(02)00077-5
Tanaka H, Pereira LA, Nozaki M, Tsuchida J, Sawada K, Mori H, Nishimune Y (1998) A germ cell-specific nuclear antigen recognized by a monoclonal antibody raised against mouse testicular germ cells. Int J Androl. https://doi.org/10.1046/j.1365-2605.1998.00080.x
Turner JMA (2007) Meiotic sex chromosome inactivation. Develop. https://doi.org/10.1242/dev.000018
Vara C, Paytuví-Gallart A, Cuartero Y, Le Dily F, Garcia F, Salvà-Castro J, Gómez-H L, Julià E, Moutinho C, Aiese Cigliano R, Sanseverino W, Fornas O, Pendás AM, Heyn H, Waters PD, Marti-Renom MA, Ruiz-Herrera A (2019) Three-dimensional genomic structure and cohesin occupancy correlate with transcriptional activity during spermatogenesis. Cell Rep. https://doi.org/10.1016/j.celrep.2019.06.037
Vergés L, Blanco J, Valero O, Vidal F, Sarrate Z (2014) Chromosome size, morphology, and gene density determine bivalent positioning in metaphase I human spermatocytes. Fertil Steril. https://doi.org/10.1016/j.fertnstert.2013.11.013
Vourc’h C, Taruscio D, Boyle AL, Ward DC (1993) Cell cycle-dependent distribution of telomeres, centromeres, and chromosome-specific subsatellite domains in the interphase nucleus of mouse lymphocytes. Exp Cell Res. https://doi.org/10.1006/excr.1993.1068
Werner RJ, Schultz BM, Huhn JM, Jelinek J, Madzo J, Engel N (2017) Sex chromosomes drive gene expression and regulatory dimorphisms in mouse embryonic stem cells. Biol Sex Differ. https://doi.org/10.1186/s13293-017-0150-x
Wright SJ (1999) Sperm nuclear activation during fertilization. Curr Top Dev Biol. https://doi.org/10.1016/S0070-2153(08)60328-2
Zalenskaya IA, Bradbury EM, Zalensky AO (2000) Chromatin structure of telomere domain in human sperm. Biochem Biophys Res Commun. https://doi.org/10.1006/bbrc.2000.3917
Zalensky A, Zalenskaya I (2007) Organization of chromosomes in spermatozoa: an additional layer of epigenetic information? Biochem Soc Trans. https://doi.org/10.1042/BST0350609
Zickler D, Kleckner N (1998) The leptotene-zygotene transition of meiosis. Annu Rev Genet. https://doi.org/10.1146/annurev.genet.32.1.619
Zickler D, Kleckner N (2015) Recombination, pairing, and synapsis of homologs during meiosis. Cold Spring Harb Perspect Biol. https://doi.org/10.1101/cshperspect.a016626
We thank Dr. Ignasi Roig for his advice in optimising the technique of immunofluorescence and cell stage identification. We thank M.A Handel from Jackson Laboratories for providing us the H1T antibody.
This study was supported by CF-180034 (UAB), DPI2015-65286-R/SAF2016-77165-P/RTI2018-095209 (MINECO), 2017-SGR-1624 and CERCA (Generalitat de Catalunya). Mireia Solé is the recipient of a grant from UAB (PIF/2015). Debora Gil is a Serra Hunter Fellow.
The Ethics Committee on Animal and Human Experimentation (CEEAH) of the Autonomous University of Barcelona declares that according to the Spanish regulation RD53/2013, the use of biological material in the term used in this study does not require formal consent by the Committee.
Consent to participate
Consent for publication
Conflict of interest
The authors declare no competing interests.
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Below is the link to the electronic supplementary material.
About this article
Cite this article
Solé, M., Blanco, J., Gil, D. et al. Chromosomal positioning in spermatogenic cells is influenced by chromosomal factors associated with gene activity, bouquet formation and meiotic sex chromosome inactivation . Chromosoma 130, 163–175 (2021). https://doi.org/10.1007/s00412-021-00761-0
- Chromosome territories
- Three-dimensional analysis