Skip to main content
SpringerLink
Log in

Correlation of meiotic events in testis sections and microspreads of mouse spermatocytes relative to the mid-pachytene checkpoint

  • Research Article
  • Published:
Chromosoma Aims and scope Submit manuscript

Abstract

In mouse, asynaptic meiotic mutants arrest at Testis Epithelial Stage IV. This arrest is 4.5 days after homologous chromosomes begin to synapse and approximately 2.5 days after synapsis is usually completed. To correlate cytological events with meiotic progression in testis and to determine which meiotic events are normally completed by Stage IV, we induced spermatogenic arrest by placing mice on a vitamin A deficient diet. Subsequent injection of retinoic acid and a return to a normal diet resulted in resumption of spermatogenesis with all spermatocytes proceeding through meiosis in a highly synchronous cohort. Between Days 11 and 16 post-injection we prepared one testis for immunocytological and the other for histological evaluation, then used antibodies to SCP3 and either RPA, or MLH1 to follow quantitative changes in synapsis and recombination. RPA was found at sites along the synaptonemal complex as soon as homologs synapsed, and most, but not all, RPA disappeared by Stage IV. MLH1 foci appeared between Stage II and IV and remained through Stage VII, the end point of the study. The data suggest that the earliest the mid-pachytene checkpoint can be activated is Epithelial Stage IV, but that activities monitored by the checkpoint may not be completed by this time.

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

Access this article

Log in via an institution

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

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1a–c
Fig. 2a–f
Fig. 3
Fig. 4

Similar content being viewed by others

References

  • Anderson LK, Reeves A, Webb LM, Ashley T (1999) Distribution of crossovers on mouse chromosomes using immunofluorescent localization of MLH1 protein. Genetics 151:1569–1579

    CAS  PubMed  Google Scholar 

  • Baker SM, Plug AW, Prolla TA, Bronner CE, Harris AC, Yao X, Christie D-M, Monell C, Arnheim N, Bradley A, Ashley T, Liskay RM (1996) Involvement of mouse Mlh1 in DNA mismatch repair and meiotic crossing over. Nat Genet 13:336–342

    CAS  PubMed  Google Scholar 

  • van Beek ME, Meistrich ML (1991) Stage synchronized seminiferous epithelium in rats after manipulation of retinol levels. Biol Reprod 45:235–244

    Google Scholar 

  • Bloom SE, Goodpasture C (1976) An improved technique for selective silver staining of nucleolar organizer regions in human chromosomes. Hum Genet 34:199–206

    CAS  PubMed  Google Scholar 

  • de Boer P, Searl AG, van der Hoeven FA, de Rooij DG, Beechey CV (1986) Male pachytene pairing in single and double translocation heterozygotes and spermatogenic impairment in the mouse. Chromosoma 93:326–336

    CAS  PubMed  Google Scholar 

  • Davis TW, Wilson-Van Patten C, Meyers M, Kunugi KA, Cuthill S, Reznikoff C, Garces C, Boland CR, Kinsella TJ, Fishel R, Boothman DA (1998) Defective expression of the DNA mismatch repair protein, MLH1, alters G2-M cell cycle checkpoint arrest following ionizing radiation. Cancer Res 58:767–778

    CAS  PubMed  Google Scholar 

  • Eaker S, Cobb J, Pyle A, Handel MA (2002) Meiotic prophase abnormalities and metaphase cell death in MLH1-deficient mouse spermatocytes; insights into the regulation of spermatogenic progress. Dev Biol 249:85–95

    Google Scholar 

  • Edelmann W, Cohen PE, Kneitz B, Winand N, Lia M, Heyer J, Kolodner R, Pollard JW, Kucherlapati R (1999) Mammalian MutS homologue 5 is required for chromosome pairing in meiosis. Nat Genet 21:123–127

    Article  CAS  PubMed  Google Scholar 

  • Froenicke L, Anderson LK, Wienberg J, Ashley T (2002) Integrating genetic recombination and meiotic chromosome structure in male mice. Am J Hum Genet 71:1353–1368

    Article  CAS  PubMed  Google Scholar 

  • Gaemers IC, van Pelt AM, van der Saag PT, de Rooij DG (1996) All-trans-4-oxo-retinoic acid: a potent inducer of in vivo proliferation of growth-arrested A spermatogonia in the vitamin A-deficient mouse testis. Endocrinology 137:479–485

    Article  CAS  PubMed  Google Scholar 

  • Goetz P, Chandley AC, Speed RM (1984) Morphological and temporal sequence of meiotic prophase development at puberty in male mouse. J Cell Sci 65:249–263

    CAS  PubMed  Google Scholar 

  • Huang HF, Hembree WC (1979) Spermatogenic response to vitamin A in vitamin A deficient rats. Biol Reprod 21:891–904

    CAS  PubMed  Google Scholar 

  • Koehler KE, Cherry JP, Lynn A, Hunt PA, Hassold TJ (2004) Genetic control of mammalian meiotic recombination I. Variation in exchange frequencies among males from inbred mouse strains. Genetics 166:1199–1214

    CAS  PubMed  Google Scholar 

  • Lipkin SM, Moens PB, Wang V, Lenzi M, Shanmugarajah D, Gilgeous A, Thomas J, Cheng J, Touchman JW, Green ED, Schwartzberg P, Collins FS, Cohen PE (2002) Meiotic arrest and aneuploidy in MLH3-deficient mice. Nat Genet 4:385–390

    Google Scholar 

  • Moens PB, Heyting C, Dietrich AJJ, van Raamsdonk W, Chen Q (1987) Synaptonemal complex antigen localization and conservation. J Cell Biol 105:93–103

    Article  CAS  PubMed  Google Scholar 

  • Moens PB, Kolas NK, Tarsounas M, Marcon E, Cohen PE, Spyropoulos B (2002) The time course and chromosomal localization of recombination-related proteins at meiosis in the mouse are compatible with models that can resolve the early DNA–DNA interactions without reciprocal recombination. J Cell Sci 115:1611–1622

    CAS  PubMed  Google Scholar 

  • Morales CGM, Griswold MD (1987) Retinol induced stage synchronization in seminiferous tubules of the rat. Endocrinology 121:432–434

    CAS  PubMed  Google Scholar 

  • Moses MJ (1980) New cytogenetic studies on mammalian meiosis. In: Serio M, Martini L (eds) Animal models in human reproduction. Raven, New York, pp 169–190

  • Moses MJ, Poorman PA (1981) Synaptonemal complex analysis of mouse chromosome rearrangements. II. Synaptic adjustment in a tandem duplication. Chromosoma 81:519–535

    CAS  PubMed  Google Scholar 

  • Moses MJ, Slatton GH, Gambling TM, Starmer CF (1977) Synaptonemal complex karyotyping in spermatocytes of the Chinese hamster (Cricetulus griseus). III. Quantitative evaluation. Chromosoma 60:345–375

    CAS  PubMed  Google Scholar 

  • Moses MJ, Poorman PA, Roderick TH, Davisson MT (1982a) Synaptonemal complex analysis of mouse chromosome rearrangements. II. Synaptic adjustment in a tandem duplication. Chromosoma 81:519–535

    Google Scholar 

  • Moses MJ, Poorman PA, Roderick TH, Davisson MT (1982b) Synaptonemal complex analysis of mouse chromosome rearrangements. IV. Synapsis and synaptic adjustment in two paracentric inversions. Chromosoma 84:457–474

    CAS  PubMed  Google Scholar 

  • Oakberg EF (1956) Duration of spermatogenesis in the mouse and timing of stages of the cycle in the seminiferous epithelium. Am J Anat 99:507–516

    CAS  PubMed  Google Scholar 

  • Peters AHFM, Plug AW, van Vugt MJ, de Boer P (1997) A drying-down technique for spreading of mammalian meiocytes from the male and female germ line. Chromosome Res 5:66–71

    Article  CAS  PubMed  Google Scholar 

  • van Pelt AM, de Rooij DG (1990) Synchronization of the seminiferous epithelium after vitamin A replacement in vitamin A-deficient mice. Biol Reprod 43:363–367

    PubMed  Google Scholar 

  • Pittman DL, Cobb J, Schimenti KJ, Wilson LA, Cooper DM, Brignull E, Handel MA, Schimenti JC (1998) Meiotic prophase arrest with failure of chromosome synapsis in mice deficient for Dmc1, a germline-specific RecA homolog. Mol Cell 1:697–705

    Article  CAS  PubMed  Google Scholar 

  • Plug AW, Clairmont CA, Sapi E, Ashley T, Sweasy JB (1997a) Evidence for a role for DNA polymerase β in mammalian meiosis. Proc Natl Acad Sci USA 94:1327–1331

    Article  CAS  PubMed  Google Scholar 

  • Plug AW, Peters AHFM, Xu Y, Keegan KS, Hoekstra MF, Baltimore D, de Boer P, Ashley T (1997b) ATM and RPA in meiotic chromosome synapsis and recombination. Nat Genet 17:457–461

    CAS  PubMed  Google Scholar 

  • Plug AW, Peters AHFM, van Breuklen B, Keegan KS, Hoekstra M, de Boer P, Ashley T (1998) Changes in protein composition of meiotic nodules during mammalian meiosis. J Cell Sci 111:413–423

    CAS  PubMed  Google Scholar 

  • Reeves A (2001) Micromeasure: a new computer program for the collection and analysis of cytogenetic data. Genome 44:439–443

    Article  CAS  PubMed  Google Scholar 

  • Russell LD, Ettlin RA, Hikim APS, Clegg ED (1990) Histological and histopathological evaluation of the testes. Cache River, Clearwater

  • Shiloh Y (2001) ATM and ATR: network in cellular responses to DNA damage. Curr Opin Genet Dev 11:71–77

    Article  CAS  PubMed  Google Scholar 

  • Shimada K, Pasero P, Gasser SM (2002) ORC and the intra-S-phase checkpoint: a threshold regulates Rad53p activation in S phase. Genes Dev 16:3236–3252

    Article  CAS  PubMed  Google Scholar 

  • Van der Meer Y, Cattanach BC, de Rooij DG (1993) The radiosensitivity of spermatogonial stem cells in C3H/101 F1 hybrid mice. Mutat Res 290:201–210

    PubMed  Google Scholar 

  • de Vries SS, Baart EB, Dekker M, Siezen A, de Rooij DG, de Boer P, te Riele H (1999) Mouse MutS-like protein MSH5 is required for proper chromosome synapsis in male and female meiosis. Genes Dev 13:523–531

    PubMed  Google Scholar 

  • Walpita D, Plug AW, Neff N, German J, Ashley T (1999) Bloom’s syndrome protein (BLM) co-localizes with RPA in meiotic prophase nuclei of mammalian spermatocytes. Proc Natl Acad Sci USA 96:5622–5627

    Article  CAS  PubMed  Google Scholar 

  • Wold MS (1997) Replication protein A: a heterotrimeric, single-strand DNA-binding protein required for eukaryotic DNA metabolism. Annu Rev Biochem 66:61–92

    Article  CAS  PubMed  Google Scholar 

  • Yoshida K, Kondoh G, Matsuda Y, Habu T, Nishimune Y, Marita T (1998) The mouse RecA-like gene Dmc1 is required for homologous chromosome synapsis during meiosis. Mol Cell 1:707–718

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We thank Anita Tellier for the fluorescence photography. We also thank Drs. Lorrie Anderson and Paula Cohen for their comments and suggestions on the manuscript. This work was supported by NIH grant RO1-HD39384.

Author information

Authors and Affiliations

  1. Department of Genetics, School of Medicine, Yale University, 333 Cedar St, New Haven, CT 06520, USA

    Terry Ashley, Ann P. Gaeth & Adelle M. Hack

  2. Departments of Endocrinology, Faculty of Biology, Utrecht University and of Cell Biology, University Medical Center Utrecht, Utrecht, The Netherlands

    Laura B. Creemers & Dirk G. de Rooij

Authors
  1. Terry Ashley
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Ann P. Gaeth
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Laura B. Creemers
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Adelle M. Hack
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Dirk G. de Rooij
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Terry Ashley.

Additional information

Communicated by T. Hassold

Rights and permissions

Reprints and permissions

About this article

Cite this article

Ashley, T., Gaeth, A.P., Creemers, L.B. et al. Correlation of meiotic events in testis sections and microspreads of mouse spermatocytes relative to the mid-pachytene checkpoint. Chromosoma 113, 126–136 (2004). https://doi.org/10.1007/s00412-004-0293-5

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00412-004-0293-5

Keywords

Search

Navigation