Accuri C6 Flow Cytometer System
Originally published online as doi:10.1189/jlb.0708405 on February 13, 2009

Published online before print February 13, 2009
This Article
Right arrow Full Text Free
Right arrow Full Text (PDF) Free
Right arrow Table 1
Right arrow All Versions of this Article:
jlb.0708405v1
85/5/751    most recent
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Hall, C.
Right arrow Articles by Crosier, P.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Hall, C.
Right arrow Articles by Crosier, P.
(Journal of Leukocyte Biology. 2009;85:751-765.)
© 2009 Society for Leukocyte Biology

Transgenic zebrafish reporter lines reveal conserved Toll-like receptor signaling potential in embryonic myeloid leukocytes and adult immune cell lineages

Chris Hall*, Maria Vega Flores*, Annie Chien*, Alan Davidson{dagger}, Kathryn Crosier* and Phil Crosier*,1

* Department of Molecular Medicine and Pathology, School of Medical Sciences, The University of Auckland, Auckland, New Zealand; and
{dagger} Center for Regenerative Medicine, Massachusetts General Hospital, Boston, Massachusetts, USA

1 Correspondence: Department of Molecular Medicine and Pathology, School of Medical Sciences, The University of Auckland, Private Bag 92019, Auckland 1142, New Zealand. E-mail: ps.crosier{at}auckland.ac.nz

ABSTRACT

The immune response of a host to an invading pathogen is dependent on the capacity of its immune cell compartment to recognize highly conserved pathogen components using an ancient class of pattern recognition receptors known as Toll-like receptors (TLRs). Initiation of TLR-mediated signaling results in the induction of proinflammatory cytokines that help govern the scale and duration of any ensuing response. Specificity for TLR signaling is, in part, a result of the differential recruitment of intracellular adaptor molecules. Of these, MyD88 is required for the majority of TLR signaling. Zebrafish have been shown to possess TLRs and adaptor molecules throughout early development, including MyD88, strongly suggesting conservation of this ancient defense mechanism. However, information about which embryonic cells/tissues possess this conserved signaling potential is lacking. To help define which embryonic cells, in particular, those of the innate immune system, have the potential for MyD88-dependent, TLR-mediated signaling, we generated transgenic reporter lines using regulatory elements of the myd88 gene to drive the fluorescent reporters enhanced GFP and Discosoma red fluorescent protein 2 within live zebrafish. These lines possess fluorescently marked cells/tissues consistent with endogenous myd88 expression, including a subset of myeloid leukocytes. These innate immune cells were confirmed to express other TLR adaptors including Mal, trif, and Sarm. Live wound-healing and infection assays validated the potential of these myd88-expressing leukocytes to participate in immune responses. These lines will provide a valuable resource for further resolving the contribution of MyD88 to early vertebrate immunity.

Key Words: MyD88 • TLR • immunity • live imaging • neutrophils • macrophages