A Comparative Evaluation of the Strengths and Potential Caveats of the Microglial Inducible CreER Mouse Models

Overview

Journal Cell Rep

Publisher Cell Press

Specialties Biology
Cell Biology
Molecular Biology

Date 2024 Jan 13

PMID 38217856

Authors

Alicia M Bedolla

Gabriel L McKinsey

Kierra Ware

Nicolas Santander

Thomas D Arnold

Yu Luo

Affiliations

Soon will be listed here.

Abstract

The recent proliferation of new Cre and CreER recombinase lines provides researchers with a diverse toolkit to study microglial gene function. To determine how best to apply these lines in studies of microglial gene function, a thorough and detailed comparison of their properties is needed. Here, we examined four different microglial CreER lines (Cx3cr1, Cx3cr1, P2ry12, and Tmem119), focusing on (1) recombination specificity, (2) leakiness (the degree of tamoxifen-independent recombination in microglia and other cells), (3) the efficiency of tamoxifen-induced recombination, (4) extraneural recombination (the degree of recombination in cells outside of the CNS, particularly myelo/monocyte lineages), and (5) off-target effects in the context of neonatal brain development. We identify important caveats and strengths for these lines, which will provide broad significance for researchers interested in performing conditional gene deletion in microglia. We also provide data emphasizing the potential of these lines for injury models that result in the recruitment of splenic immune cells.

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References

Cheng X, Li Y, Huang Y, Feng X, Feng G, Xiong Z . Pulse labeling and long-term tracing of newborn neurons in the adult subgranular zone. Cell Res. 2010; 21(2):338-49. PMC: 3193432. DOI: 10.1038/cr.2010.141. View

Yona S, Kim K, Wolf Y, Mildner A, Varol D, Breker M . Fate mapping reveals origins and dynamics of monocytes and tissue macrophages under homeostasis. Immunity. 2013; 38(1):79-91. PMC: 3908543. DOI: 10.1016/j.immuni.2012.12.001. View

Kim J, Kolesnikov M, Peled-Hajaj S, Scheyltjens I, Xia Y, Trzebanski S . A Binary Cre Transgenic Approach Dissects Microglia and CNS Border-Associated Macrophages. Immunity. 2020; 54(1):176-190.e7. DOI: 10.1016/j.immuni.2020.11.007. View

Mastorakos P, Mihelson N, Luby M, Burks S, Johnson K, Hsia A . Temporally distinct myeloid cell responses mediate damage and repair after cerebrovascular injury. Nat Neurosci. 2021; 24(2):245-258. PMC: 7854523. DOI: 10.1038/s41593-020-00773-6. View

Jin Y, Raviv N, Barnett A, Bambakidis N, Filichia E, Luo Y . The shh signaling pathway is upregulated in multiple cell types in cortical ischemia and influences the outcome of stroke in an animal model. PLoS One. 2015; 10(4):e0124657. PMC: 4415811. DOI: 10.1371/journal.pone.0124657. View

Lagace D, Whitman M, Noonan M, Ables J, DeCarolis N, Arguello A . Dynamic contribution of nestin-expressing stem cells to adult neurogenesis. J Neurosci. 2007; 27(46):12623-9. PMC: 3718551. DOI: 10.1523/JNEUROSCI.3812-07.2007. View

Li X, Du Z, Chen M, Chen J, Liang Z, Ding X . The effects of tamoxifen on mouse behavior. Genes Brain Behav. 2019; 19(4):e12620. DOI: 10.1111/gbb.12620. View

Van Hove H, Pombo Antunes A, De Vlaminck K, Scheyltjens I, Van Ginderachter J, Movahedi K . Identifying the variables that drive tamoxifen-independent CreERT2 recombination: Implications for microglial fate mapping and gene deletions. Eur J Immunol. 2019; 50(3):459-463. DOI: 10.1002/eji.201948162. View

Ajami B, Bennett J, Krieger C, Tetzlaff W, Rossi F . Local self-renewal can sustain CNS microglia maintenance and function throughout adult life. Nat Neurosci. 2007; 10(12):1538-43. DOI: 10.1038/nn2014. View

10.

Wieghofer P, Knobeloch K, Prinz M . Genetic targeting of microglia. Glia. 2014; 63(1):1-22. DOI: 10.1002/glia.22727. View

11.

Pezoldt J, Wiechers C, Erhard F, Rand U, Bulat T, Beckstette M . Single-cell transcriptional profiling of splenic fibroblasts reveals subset-specific innate immune signatures in homeostasis and during viral infection. Commun Biol. 2021; 4(1):1355. PMC: 8640036. DOI: 10.1038/s42003-021-02882-9. View

12.

Wong K, Noubade R, Manzanillo P, Ota N, Foreman O, Hackney J . Mice deficient in NRROS show abnormal microglial development and neurological disorders. Nat Immunol. 2017; 18(6):633-641. DOI: 10.1038/ni.3743. View

13.

McKinsey G, Lizama C, Keown-Lang A, Niu A, Santander N, Larpthaveesarp A . A new genetic strategy for targeting microglia in development and disease. Elife. 2020; 9. PMC: 7375817. DOI: 10.7554/eLife.54590. View

14.

Fernandez-Chacon M, Casquero-Garcia V, Luo W, Lunella F, Ferreira Rocha S, Del Olmo-Cabrera S . iSuRe-Cre is a genetic tool to reliably induce and report Cre-dependent genetic modifications. Nat Commun. 2019; 10(1):2262. PMC: 6531465. DOI: 10.1038/s41467-019-10239-4. View

15.

Zhang Z, Park J, Ahn I, Diamante G, Sivakumar N, Arneson D . Estrogen receptor alpha in the brain mediates tamoxifen-induced changes in physiology in mice. Elife. 2021; 10. PMC: 7924955. DOI: 10.7554/eLife.63333. View

16.

Carroll J, Race B, Williams K, Striebel J, Chesebro B . Innate immune responses after stimulation with Toll-like receptor agonists in ex vivo microglial cultures and an in vivo model using mice with reduced microglia. J Neuroinflammation. 2021; 18(1):194. PMC: 8419892. DOI: 10.1186/s12974-021-02240-w. View

17.

Fonseca M, Chu S, Hernandez M, Fang M, Modarresi L, Selvan P . Cell-specific deletion of C1qa identifies microglia as the dominant source of C1q in mouse brain. J Neuroinflammation. 2017; 14(1):48. PMC: 5340039. DOI: 10.1186/s12974-017-0814-9. View

18.

Paolicelli R, Bolasco G, Pagani F, Maggi L, Scianni M, Panzanelli P . Synaptic pruning by microglia is necessary for normal brain development. Science. 2011; 333(6048):1456-8. DOI: 10.1126/science.1202529. View

19.

Jin Y, Barnett A, Zhang Y, Yu X, Luo Y . Poststroke Sonic Hedgehog Agonist Treatment Improves Functional Recovery by Enhancing Neurogenesis and Angiogenesis. Stroke. 2017; 48(6):1636-1645. PMC: 5667564. DOI: 10.1161/STROKEAHA.117.016650. View

20.

Chen H, Sun Y, Chen C, Kuo Y, Kuan I, Tiger Li Z . Fate mapping via CCR2-CreER mice reveals monocyte-to-microglia transition in development and neonatal stroke. Sci Adv. 2020; 6(35):eabb2119. PMC: 7449686. DOI: 10.1126/sciadv.abb2119. View