The Journal of Experimental Medicine
Fluorescence In Vivo Endomicroscopy
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Published online 9 February 2004 doi:10.1084/jem.20031802
Rockefeller University Press, 0022-1007 $8.00
JEM, Volume 199, Number 4, 483-489
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Developmental Separation of V(D)J Recombinase Expression and Initiation of IgH Recombination in B Lineage Progenitors In Vivo

Lisa Borghesi and Rachel M. Gerstein

Molecular Genetics and Microbiology, University of Massachusetts Medical School (UMMS), Worcester, MA 01655

Address correspondence to Rachel M. Gerstein, University of Massachusetts Medical School, Molecular Genetics and Microbiology, 55 Lake Ave. North, S5-714, Worcester, MA 01655. Phone: (508) 856-1044; Fax: (508) 856-5920; email: rachel.gerstein{at}umassmed.edu

In B lineage progenitors, V(D)J recombination occurs only during distinct stages of development and is restricted to immunoglobulin loci. This process is thought to be controlled by both regulated expression of the V(D)J recombinase and by limited accessibility of target loci to the recombinase complex. However, it is unknown whether these two processes occur concomitantly in developing B lineage progenitors or whether these events are temporally distinct and, therefore, potentially independently regulated. To distinguish between these possibilities, we developed a transgenic V(D)J recombination substrate that is not governed by the same chromatin remodeling constraints as endogenous immunoglobulin heavy chain (IgH) loci and examined the requirements for V(D)J recombination to initiate in early B lineage progenitors. We find that single B lineage precursors express an active V(D)J recombinase in vivo before the stage when IgH rearrangements are frequently detectable. Our results indicate that the onset of recombinase activity and the initiation of IgH recombination are developmentally distinct events in the B lineage.

Key Words: B lymphopoiesis • V(D)J recombination • chromatin • immunoglobulin • heavy chain • hematopoiesis

Abbreviations used in this paper: GFP, green fluorescence protein; IgH, Ig heavy chain; RAG, recombinase-activating gene; RSS, recombination signal sequence.


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