Spatiotemporal dynamics of the CD11c+ microglial population in the mouse brain and spinal cord from developmental to adult stages

doi:10.1186/s13041-024-01098-2

. 2024 May 18;17(1):24.

doi: 10.1186/s13041-024-01098-2.

Spatiotemporal dynamics of the CD11c⁺ microglial population in the mouse brain and spinal cord from developmental to adult stages

Kohei Nomaki¹, Risako Fujikawa¹, Takahiro Masuda², Makoto Tsuda^{3

4}

Affiliations

¹ Department of Molecular and System Pharmacology, Graduate School of Pharmaceutical Sciences, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka, 812-8582, Japan.
² Division of Molecular Neuroimmunology, Medical Institute of Bioregulation, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka, 812-8582, Japan.
³ Department of Molecular and System Pharmacology, Graduate School of Pharmaceutical Sciences, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka, 812-8582, Japan. tsuda@phar.kyushu-u.ac.jp.
⁴ Kyushu University Institute for Advanced Study, 744 Motooka Nishi-ku, Fukuoka, 819-0395, Japan. tsuda@phar.kyushu-u.ac.jp.

PMID: 38762724
PMCID: PMC11102220
DOI: 10.1186/s13041-024-01098-2

Spatiotemporal dynamics of the CD11c⁺ microglial population in the mouse brain and spinal cord from developmental to adult stages

Kohei Nomaki et al. Mol Brain. 2024.

. 2024 May 18;17(1):24.

doi: 10.1186/s13041-024-01098-2.

Authors

Kohei Nomaki¹, Risako Fujikawa¹, Takahiro Masuda², Makoto Tsuda^{3

4}

Affiliations

¹ Department of Molecular and System Pharmacology, Graduate School of Pharmaceutical Sciences, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka, 812-8582, Japan.
² Division of Molecular Neuroimmunology, Medical Institute of Bioregulation, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka, 812-8582, Japan.
³ Department of Molecular and System Pharmacology, Graduate School of Pharmaceutical Sciences, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka, 812-8582, Japan. tsuda@phar.kyushu-u.ac.jp.
⁴ Kyushu University Institute for Advanced Study, 744 Motooka Nishi-ku, Fukuoka, 819-0395, Japan. tsuda@phar.kyushu-u.ac.jp.

PMID: 38762724
PMCID: PMC11102220
DOI: 10.1186/s13041-024-01098-2

Abstract

CD11c-positive (CD11c⁺) microglia have attracted considerable attention because of their potential implications in central nervous system (CNS) development, homeostasis, and disease. However, the spatiotemporal dynamics of the proportion of CD11c⁺ microglia in individual CNS regions are poorly understood. Here, we investigated the proportion of CD11c⁺ microglia in six CNS regions (forebrain, olfactory bulb, diencephalon/midbrain, cerebellum, pons/medulla, and spinal cord) from the developmental to adult stages by flow cytometry and immunohistochemical analyses using a CD11c reporter transgenic mouse line, Itgax-Venus. We found that the proportion of CD11c⁺ microglia in total microglia varied between CNS regions during postnatal development. Specifically, the proportion was high in the olfactory bulb and cerebellum at postnatal day P(4) and P7, respectively, and approximately half of the total microglia were CD11c⁺. The proportion declined sharply in all regions to P14, and the low percentage persisted over P56. In the spinal cord, the proportion of CD11c⁺ microglia was also high at P4 and declined to P14, but increased again at P21 and thereafter. Interestingly, the distribution pattern of CD11c⁺ microglia in the spinal cord markedly changed from gray matter at P4 to white matter at P21. Collectively, our findings reveal the differences in the spatiotemporal dynamics of the proportion of CD11c⁺ microglia among CNS regions from early development to adult stages in normal mice. These findings improve our understanding of the nature of microglial heterogeneity and its dynamics in the CNS.

Keywords: Adult; Brain; CD11c+ microglia; Mouse; Pre/postnatal development; Spatiotemporal dynamics; Spinal cord.

PubMed Disclaimer

Conflict of interest statement

The authors declare that they have no competing interests.

Figures

**Fig. 1**
CD11c⁺ cells in the brain of *Itgax-Venus* mice at P4. (A) Representative immunofluorescence images of Venus⁺ (CD11c⁺) cells in the whole brain of *Itgax-Venus* mice at P4. Each brain region for quantitative analyses in further experiments is indicated by dashed lines. (B) Immunolabeling of Venus⁺ cells (green) with IBA1 (red) in the forebrain. Scale bars, 1000 μm (A) and 50 μm (B)

**Fig. 2**
Flow cytometry analysis of the proportion of CD11c⁺ microglia in the forebrain. (A) Gating strategy for the CD11c⁺ and CD11c^neg microglia (CD11b⁺ CD45^low CD206^neg cells with and without Venus fluorescence in red and blue squares, respectively) in the forebrain of *Itgax-Venus* mice at P4. (B) qPCR analysis of *P2ry12*, *Igf1*, *Clec7a*, and *Trem2* mRNA expression in FACS-isolated CD11c^neg and CD11c⁺ microglia from the forebrain at P4 (n = 5 mice). Values represent the relative ratio of the mRNA levels (normalized to *Actb* mRNA) of the CD11c^neg microglia group. (C) Temporal analysis of the proportion of CD11c⁺ microglia in the forebrain from P0 to P56 (n = 4–8 mice for each time point tested). The proportion of CD11c⁺ microglia is indicated as the percentage of Venus⁺ cells in the total microglia (CD11b⁺ CD45^low CD206^neg cells). The proportion of CD11c^high microglia (the upper half of CD11c⁺ microglia in the scattered plot) to CD11c⁺ microglia at P4 and P21 was also analyzed. Data are shown as means ± SEM. **P < 0.01, ***P < 0.001, and ****P < 0.0001

**Fig. 3**
Spatiotemporal analysis of CD11c⁺ microglia in other brain regions. Flow cytometric analysis of the percentage of CD11c⁺ (upper panels) and CD11c^high (lower panels) microglia per total microglia (CD11b⁺ CD45^low CD206^neg cells) at P4, P7, and P21 (n = 4 mice at each time point). Data are shown as means ± SEM. **P < 0.01, ***P < 0.001, and ****P < 0.0001

**Fig. 4**
CD11c⁺ microglia in the prenatal mouse brain. (A) Representative immunofluorescence images of Venus⁺ (CD11c⁺; green) cells in the whole brain of *Itgax-Venus* mice at E14.5. The parenchymal brain region was indicated by a dashed line. Venus⁺ cells (green) were colocalized with IBA1 (red). Scale bars, 1000 μm (left panel) and 100 μm (right panel). (B) Flow cytometry analysis of the percentage of CD11c⁺ microglia per total microglia (CD11b⁺ CD45^low CD206^neg cells) at E12.5, E14.5, and E16.5 (n = 6–8 mice for each time point tested). Data are shown as means ± SEM. **P < 0.01, ***P < 0.001, and ****P < 0.0001

**Fig. 5**
Temporal analysis of CD11c⁺ microglia in the mouse spinal cord. (A) Representative immunofluorescence images of Venus⁺ (CD11c⁺; green) cells in the spinal cord of *Itgax-Venus* mice at E12.5 and E14.5. Venus⁺ cells (green) were colocalized with IBA1 (red) in the spinal cord. Scale bars, 500 μm. (B) Representative scattered plot of CD11c^neg and CD11c⁺ microglia (blue and red square, respectively) in the spinal cord of *Itgax-Venus* mice at P4. (C) qPCR analysis of *P2ry12*, *Igf1*, *Clec7a*, and *Trem2* mRNA in FACS-isolated CD11c^neg and CD11c⁺ microglia in the spinal cord at P4 (n = 7 mice). Values represent the relative ratio of the mRNA levels (normalized to *Actb* mRNA) of CD11c^neg microglia group. (D) Temporal analysis of the percentage of CD11c⁺ and CD11c^high microglia per total microglia (CD11b⁺ CD45^low CD206^neg cells) during development and adult (n = 4–10 mice for each time point tested). Data are shown as means ± SEM. ***P < 0.001, and ****P < 0.0001

**Fig. 6**
Spatial changes in spinal CD11c⁺ microglia during development. (A) Representative immunofluorescence images of Venus⁺ (CD11c⁺; green) cells in the spinal cord of *Itgax-Venus* mice at P4, P14, P7, P21, P28, and P56. GM regions are indicated by dashed lines. (B) High magnification images of the areas (indicated by white arrowheads in panel A). Dashed lines indicate the boundary between WM and GM. Scale bars, 1000 μm (A) and 200 μm (B)

See this image and copyright information in PMC

References

1. Li Q, Barres BA. Microglia and macrophages in brain homeostasis and disease. Nat Rev Immunol. 2018;18:225–42. doi: 10.1038/nri.2017.125. - DOI - PubMed
1. Prinz M, Jung S, Priller J. Microglia biology: one century of evolving concepts. Cell. 2019;179:292–311. doi: 10.1016/j.cell.2019.08.053. - DOI - PubMed
1. Keren-Shaul H, Spinrad A, Weiner A, Matcovitch-Natan O, Dvir-Szternfeld R, Ulland TK, David E, Baruch K, Lara-Astaiso D, Toth B, et al. A unique microglia type associated with restricting development of alzheimer’s disease. Cell. 2017;169:1276–e12901217. doi: 10.1016/j.cell.2017.05.018. - DOI - PubMed
1. Masuda T, Sankowski R, Staszewski O, Bottcher C, Amann L, Sagar; Scheiwe C, Nessler S, Kunz P, van Loo G, et al. Spatial and temporal heterogeneity of mouse and human microglia at single-cell resolution. Nature. 2019;566:388–92. doi: 10.1038/s41586-019-0924-x. - DOI - PubMed
1. Masuda T, Sankowski R, Staszewski O, Prinz M. Microglia heterogeneity in the single-cell era. Cell Rep. 2020;30:1271–81. doi: 10.1016/j.celrep.2020.01.010. - DOI - PubMed

MeSH terms

Actions
Actions
Actions
Actions
Actions
Actions
Actions
Actions
Actions
Actions
Actions
Actions
Actions

Grants and funding

LinkOut - more resources

Full Text Sources
- BioMed Central
- PubMed Central
Research Materials
- NCI CPTC Antibody Characterization Program

[1] Li Q, Barres BA. Microglia and macrophages in brain homeostasis and disease. Nat Rev Immunol. 2018;18:225–42. doi: 10.1038/nri.2017.125. - DOI - PubMed

[2] Li Q, Barres BA. Microglia and macrophages in brain homeostasis and disease. Nat Rev Immunol. 2018;18:225–42. doi: 10.1038/nri.2017.125. - DOI - PubMed

[3] Prinz M, Jung S, Priller J. Microglia biology: one century of evolving concepts. Cell. 2019;179:292–311. doi: 10.1016/j.cell.2019.08.053. - DOI - PubMed

[4] Prinz M, Jung S, Priller J. Microglia biology: one century of evolving concepts. Cell. 2019;179:292–311. doi: 10.1016/j.cell.2019.08.053. - DOI - PubMed

[5] Keren-Shaul H, Spinrad A, Weiner A, Matcovitch-Natan O, Dvir-Szternfeld R, Ulland TK, David E, Baruch K, Lara-Astaiso D, Toth B, et al. A unique microglia type associated with restricting development of alzheimer’s disease. Cell. 2017;169:1276–e12901217. doi: 10.1016/j.cell.2017.05.018. - DOI - PubMed

[6] Keren-Shaul H, Spinrad A, Weiner A, Matcovitch-Natan O, Dvir-Szternfeld R, Ulland TK, David E, Baruch K, Lara-Astaiso D, Toth B, et al. A unique microglia type associated with restricting development of alzheimer’s disease. Cell. 2017;169:1276–e12901217. doi: 10.1016/j.cell.2017.05.018. - DOI - PubMed

[7] Masuda T, Sankowski R, Staszewski O, Bottcher C, Amann L, Sagar; Scheiwe C, Nessler S, Kunz P, van Loo G, et al. Spatial and temporal heterogeneity of mouse and human microglia at single-cell resolution. Nature. 2019;566:388–92. doi: 10.1038/s41586-019-0924-x. - DOI - PubMed

[8] Masuda T, Sankowski R, Staszewski O, Bottcher C, Amann L, Sagar; Scheiwe C, Nessler S, Kunz P, van Loo G, et al. Spatial and temporal heterogeneity of mouse and human microglia at single-cell resolution. Nature. 2019;566:388–92. doi: 10.1038/s41586-019-0924-x. - DOI - PubMed

[9] Masuda T, Sankowski R, Staszewski O, Prinz M. Microglia heterogeneity in the single-cell era. Cell Rep. 2020;30:1271–81. doi: 10.1016/j.celrep.2020.01.010. - DOI - PubMed

[10] Masuda T, Sankowski R, Staszewski O, Prinz M. Microglia heterogeneity in the single-cell era. Cell Rep. 2020;30:1271–81. doi: 10.1016/j.celrep.2020.01.010. - DOI - PubMed

Save citation to file

Email citation

Add to Collections

Add to My Bibliography

Your saved search

Create a file for external citation management software

Your RSS Feed

Spatiotemporal dynamics of the CD11c⁺ microglial population in the mouse brain and spinal cord from developmental to adult stages

Affiliations

Spatiotemporal dynamics of the CD11c⁺ microglial population in the mouse brain and spinal cord from developmental to adult stages

Authors

Affiliations

Abstract

Conflict of interest statement

Figures

Similar articles

References

MeSH terms

Grants and funding

LinkOut - more resources

Full Text Sources

Research Materials

Abstract

Conflict of interest statement

Figures

Similar articles

References

MeSH terms

Related information

Grants and funding

LinkOut - more resources

Full Text Sources

Research Materials