Skip to main content
Advertisement

Main menu

  • Home
  • Articles
    • Accepted manuscripts
    • Issue in progress
    • Latest complete issue
    • Issue archive
    • Archive by article type
    • Special issues
    • Subject collections
    • Cell Scientists to Watch
    • First Person
    • Sign up for alerts
  • About us
    • About JCS
    • Editors and Board
    • Editor biographies
    • Travelling Fellowships
    • Grants and funding
    • Journal Meetings
    • Workshops
    • The Company of Biologists
    • Journal news
  • For authors
    • Submit a manuscript
    • Aims and scope
    • Presubmission enquiries
    • Fast-track manuscripts
    • Article types
    • Manuscript preparation
    • Cover suggestions
    • Editorial process
    • Promoting your paper
    • Open Access
    • JCS Prize
    • Manuscript transfer network
    • Biology Open transfer
  • Journal info
    • Journal policies
    • Rights and permissions
    • Media policies
    • Reviewer guide
    • Sign up for alerts
  • Contacts
    • Contact JCS
    • Subscriptions
    • Advertising
    • Feedback
  • COB
    • About The Company of Biologists
    • Development
    • Journal of Cell Science
    • Journal of Experimental Biology
    • Disease Models & Mechanisms
    • Biology Open

User menu

  • Log in

Search

  • Advanced search
Journal of Cell Science
  • COB
    • About The Company of Biologists
    • Development
    • Journal of Cell Science
    • Journal of Experimental Biology
    • Disease Models & Mechanisms
    • Biology Open

supporting biologistsinspiring biology

Journal of Cell Science

  • Log in
Advanced search

RSS   Twitter  Facebook   YouTube  

  • Home
  • Articles
    • Accepted manuscripts
    • Issue in progress
    • Latest complete issue
    • Issue archive
    • Archive by article type
    • Special issues
    • Subject collections
    • Cell Scientists to Watch
    • First Person
    • Sign up for alerts
  • About us
    • About JCS
    • Editors and Board
    • Editor biographies
    • Travelling Fellowships
    • Grants and funding
    • Journal Meetings
    • Workshops
    • The Company of Biologists
    • Journal news
  • For authors
    • Submit a manuscript
    • Aims and scope
    • Presubmission enquiries
    • Fast-track manuscripts
    • Article types
    • Manuscript preparation
    • Cover suggestions
    • Editorial process
    • Promoting your paper
    • Open Access
    • JCS Prize
    • Manuscript transfer network
    • Biology Open transfer
  • Journal info
    • Journal policies
    • Rights and permissions
    • Media policies
    • Reviewer guide
    • Sign up for alerts
  • Contacts
    • Contact JCS
    • Subscriptions
    • Advertising
    • Feedback
Research Article
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
Peter B. Moens, Nadine K. Kolas, Madalena Tarsounas, Edyta Marcon, Paula E. Cohen, Barbara Spyropoulos
Journal of Cell Science 2002 115: 1611-1622;
Peter B. Moens
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Nadine K. Kolas
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Madalena Tarsounas
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Edyta Marcon
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Paula E. Cohen
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Barbara Spyropoulos
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • Article
  • Figures & tables
  • Info & metrics
  • PDF
Loading

Article Figures & Tables

Figures

  •   Fig. 7.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Fig. 7.

    Definition of a rat recombination nodule, RN, in a shadow-cast preparation of an SC. (A) The SC consists of two lateral elements, le, and a median central element. (B) Occasionally RPA can be detected at these RNs. Mouse RNs are somewhat smaller and less distinct and are therefore visualized with PTA in the other illustrations. The width of the SC is about 200 nm.

  •   Fig. 1.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Fig. 1.

    Time course and interactions of SC-associated RAD51/DMC1, RPA and BLM protein in mouse spermatocytes. (A,B) RAD51/DMC1 foci initiate prior to detection of RPA. To the left of the white line is a spermatocyte early zygotene nucleus with mostly unpaired centromeres (red), which has about 200 RAD51/DMC1 foci (yellow). A nucleus in a later stage of meiotic prophase to the right of the line has fewer, about 90, foci. The same two nuclei stained for RPA in Fig. 1B show that the early prophase nucleus on the left has few and indistinct RPA foci, whereas the pachytene nucleus to the right of the line has an abundance of RPA foci. (C,D) The shift from RAD51/DMC1 to RPA foci is a developmental progression for the nucleus as a whole, but details seem to be regulated at the level of the individual bivalent. SC#1 and 2 have acquired a full complement of RPA foci (1D) and have lost most RAD51/DMC1 foci (1C), whereas SC#3 still has RAD51/DMC1 foci but few and indistinct RPA foci. (E) The replacement of RAD51/DMC1 by RPA is demonstrated from the relative amounts of the two proteins in individual foci based on electron microscopy of two types of immunogold grains as in Fig. 2. The data from nine of 40 SCs are presented in a bar graph; the top portion is the percentage of foci with only RPA antigen, the middle portion represents the foci with both antigens, and the lower part is the percentage of pure RAD51/DMC1 foci. Each bar represents one SC, and the SCs were sorted according to the prophase stage of the nucleus and by the previously documented decline in RAD51/DMC1 foci from zygotene to pachytene (details in Materials and Methods). The composition of the protein complexes change at successively later stages of meiotic prophase. (F) The line graph shows the number of fluorescent foci at progressively later stages of prophase. RAD51/DMC1 peaks at leptotene. Somewhat later, RPA reaches its maximum and later still, BLM does. These three antigens frequently are present together in individual foci. MLH1 appears at late prophase and is present in low numbers. Each data point is one nucleus, and the staging of the nuclei is described in the Materials and Methods.

  •   Fig. 2.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Fig. 2.

    Immunoelectron microscopy of RAD51/DMC1 (10 nm gold grains) and RPA (5 nm gold grains) in association with chromosome cores/SCs in an early and later meiotic prophase spermatocyte. (A) At early meiotic prophase, there is an abundance of RAD51/DMC1 foci but relatively few RPA foci in association with partially paired zygotene chromosome cores and SCs. (B; insert) At a later stage, pachytene, there are numerous RPA foci and very few RAD51/DMC1 foci. The RPA foci are concentrated on central element-associated nodules, TN. The width of the SC is about 200 nm. (C) At early pachytene, there are occasionally a few unpaired centromeric ends (15 nm gold grains). One of these `laggards' is shown to have RPA associated with a single, unpaired chromosome core, indicating that RPA is not always associated with RAD51/DMC1 protein. The image further demonstrates the presence of mixed foci containing RAD51/DMC1 and RPA antigen (mix). Incidental is the observation that RPA gold grains occur as a small loop (arrow) in this and other electron micrographs. The width of the SC is about 200 nm.

  •   Fig. 4.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Fig. 4.

    Colocation of RPA and BLM protein in pachytene spermatocytes. (A,B) The same nucleus immunostained for BLM (A) and for RPA (B). (C) The BLM image of (A) is made into a negative and superimposed on the positive RPA image of (B). Nearly all BLM foci colocate with RPA foci. The relative contributions of the two proteins to the individual foci are semi-quantifiable by the size and the intensity of the immunofluorescent signal, and this is somewhat more efficient than multiple colour combinations. (D-G) Electron micrographs of the mix of RPA 5 nm immunogold grains and 10 nm BLM immunogold grains at phosphotungstic acid-stained SCs and nodules. (H) With our antibodies, there are fewer 10 nm BLM gold grains than there are 5 nm RPA gold grains at the nodules.

  •   Fig. 3.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Fig. 3.

    RPA at the pseudoautosomal region, PAR, of the X-Y chromosomes. (A) Chromosome cores and centromeres visualized with fluorescent tags. The X and Y chromosomes have a short paired region, PAR. (B) The same chromosome visualized with anti-RPA antibody. The autosomal SCs and the unpaired X-chromosome core have numerous RPA foci. The PA region frequently has the brightest RPA focus or foci. Scale: the SCs are about 15 mm in length. (C; insert) At early pachytene there is extensive non-homologous synapsis between the distal portions of the X and Y chromosomes. RPA is present in the most distal portion, presumably the actual PA region. (D; insert) In the rat, the late RNs are better defined than in the mouse, and the Figure illustrates a group of 5 nm RPA gold grains in association with a nodule.

  •   Fig. 5.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Fig. 5.

    The association of MSH4p with RPA and RAD51/DMC1p. (A) The images of the red MSH4 foci and the green RPA foci are slightly offset to better demonstrate the colocation of the two types of foci. The short white lines indicate the direction of the offset and examples of colocating foci. The large bright green foci are the centromeres. (B) The colocation is also evident with immune electron microscopy. The 10 nm gold particles mark RPA antigen and the 5 nm grains mark the MSH4 antigen. The grains are associated with a transitional nodule at the central region of the SC. (C) The association of RAD51/DMC1 with MSH4 foci is less pronounced, and evidence from counts of five complete nuclei with some 150 foci each suggest that at most 10% of the foci possibly overlap and that, in general, the MSH4 foci appear after the Rad51/DMC1 foci have declined in numbers. The X chromosome at the base of (C) has, as usual, prolonged presence of RAD51/DMC1 foci. (D,E) At the EM level MSH4 foci (5 nm gold) are mostly separate from RAD51/DMC1 foci (10 nm gold). (E) The MSH4 antigen is associated with the transitional nodule, which has no evidence of RAD51/DMC1 antigen in this double-labeled preparation.

  •   Fig. 6.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Fig. 6.

    MLH1, RPA and BLM foci at meiotic prophase. (A) MLH foci (orange) appear in association with the SCs (green) when there are only a very few RPA foci (blue) left and there is no obvious colocation at this time. (B) BLM foci (red) are still fairly abundant when the MLH1 foci (green) are present in their nearly full complement of 20-30 foci. There does not appear to be colocation of BLM with MLH1. (C,E) Evidence that MLH1 antigen, which is marked by 5 nm gold grains, locates to the EM-defined RNs. The RNs are the dark-stained (PTA) bodies in association with the SCs. In the nuclei that are positive for MLH1, each SC has one or two immunogold labeled RNs. (D) The bar graph illustrates the decline in immunofluorescent BLM foci, whereas the numbers of MLH1 foci remain relatively constant at late pachytene. (F,G) A precocious diplotene bivalent that was generated by a 2 hour okadaic acid treatment of a pachytene bivalent demonstrates the presence of an MLH1-labeled RN at the site of a presumptive chiasma/crossover.

Previous ArticleNext Article
Back to top
Previous ArticleNext Article

This Issue

 Download PDF

Email

Thank you for your interest in spreading the word on Journal of Cell Science.

NOTE: We only request your email address so that the person you are recommending the page to knows that you wanted them to see it, and that it is not junk mail. We do not capture any email address.

Enter multiple addresses on separate lines or separate them with commas.
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
(Your Name) has sent you a message from Journal of Cell Science
(Your Name) thought you would like to see the Journal of Cell Science web site.
CAPTCHA
This question is for testing whether or not you are a human visitor and to prevent automated spam submissions.
Share
Research Article
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
Peter B. Moens, Nadine K. Kolas, Madalena Tarsounas, Edyta Marcon, Paula E. Cohen, Barbara Spyropoulos
Journal of Cell Science 2002 115: 1611-1622;
del.icio.us logo Digg logo Reddit logo Twitter logo CiteULike logo Facebook logo Google logo Mendeley logo
Citation Tools
Research Article
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
Peter B. Moens, Nadine K. Kolas, Madalena Tarsounas, Edyta Marcon, Paula E. Cohen, Barbara Spyropoulos
Journal of Cell Science 2002 115: 1611-1622;

Citation Manager Formats

  • BibTeX
  • Bookends
  • EasyBib
  • EndNote (tagged)
  • EndNote 8 (xml)
  • Medlars
  • Mendeley
  • Papers
  • RefWorks Tagged
  • Ref Manager
  • RIS
  • Zotero
Alerts

Please log in to add an alert for this article.

Sign in to email alerts with your email address

Article navigation

  • Top
  • Article
    • Summary
    • Introduction
    • Materials and Methods
    • Results
    • Discussion
    • Acknowledgements
    • References
  • Figures & tables
  • Info & metrics
  • PDF

Related articles

Cited by...

More in this TOC section

  • Hydrostatic pressure prevents chondrocyte differentiation through heterochromatin remodeling
  • PLEKHG4B enables actin cytoskeletal remodeling during epithelial cell–cell junction formation
  • Matrix stiffness regulates α-TAT1-mediated acetylation of α-tubulin and promotes silica-induced epithelial–mesenchymal transition via DNA damage
Show more RESEARCH ARTICLE

Similar articles

Other journals from The Company of Biologists

Development

Journal of Experimental Biology

Disease Models & Mechanisms

Biology Open

Advertisement

2020 at The Company of Biologists

Despite the challenges of 2020, we were able to bring a number of long-term projects and new ventures to fruition. While we look forward to a new year, join us as we reflect on the triumphs of the last 12 months.


Mole – The Corona Files

"This is not going to go away, 'like a miracle.' We have to do magic. And I know we can."

Mole continues to offer his wise words to researchers on how to manage during the COVID-19 pandemic.


Cell scientist to watch – Christine Faulkner

In an interview, Christine Faulkner talks about where her interest in plant science began, how she found the transition between Australia and the UK, and shares her thoughts on virtual conferences.


Read & Publish participation extends worldwide

“The clear advantages are rapid and efficient exposure and easy access to my article around the world. I believe it is great to have this publishing option in fast-growing fields in biomedical research.”

Dr Jaceques Behmoaras (Imperial College London) shares his experience of publishing Open Access as part of our growing Read & Publish initiative. We now have over 60 institutions in 12 countries taking part – find out more and view our full list of participating institutions.


JCS and COVID-19

For more information on measures Journal of Cell Science is taking to support the community during the COVID-19 pandemic, please see here.

If you have any questions or concerns, please do not hestiate to contact the Editorial Office.

Articles

  • Accepted manuscripts
  • Issue in progress
  • Latest complete issue
  • Issue archive
  • Archive by article type
  • Special issues
  • Subject collections
  • Interviews
  • Sign up for alerts

About us

  • About Journal of Cell Science
  • Editors and Board
  • Editor biographies
  • Travelling Fellowships
  • Grants and funding
  • Journal Meetings
  • Workshops
  • The Company of Biologists

For Authors

  • Submit a manuscript
  • Aims and scope
  • Presubmission enquiries
  • Fast-track manuscripts
  • Article types
  • Manuscript preparation
  • Cover suggestions
  • Editorial process
  • Promoting your paper
  • Open Access
  • JCS Prize
  • Manuscript transfer network
  • Biology Open transfer

Journal Info

  • Journal policies
  • Rights and permissions
  • Media policies
  • Reviewer guide
  • Sign up for alerts

Contacts

  • Contact JCS
  • Subscriptions
  • Advertising
  • Feedback

Twitter   YouTube   LinkedIn

© 2021   The Company of Biologists Ltd   Registered Charity 277992