Loss of CaMKIV signaling aggravates muscle inflammation and affects muscle regeneration

doi:10.13418/j.issn.1001-165x.2023.6.11

PDF(7477 KB)

Chinese Journal of Clinical Anatomy ›› 2023, Vol. 41 ›› Issue (6) : 692-697. DOI: 10.13418/j.issn.1001-165x.2023.6.11

Loss of CaMKIV signaling aggravates muscle inflammation and affects muscle regeneration

Wang Han¹, Li Yangyang¹, Jian Xiaoting¹, Huang Jingwen¹, Lan Haiqiang¹, Hu Jijie²*, Liao Hua¹*

Author information +

History +

Abstract

Objective To explore the effects of CaMKIV signaling on muscle inflammatory response and muscle repair post-injury. Methods CaMKIV gene deletion (CaMKIV-/-) mice were constructed by using CRISPR/Cas9 technology and identified by PCR. Cardiotoxin (CTX) induced skeletal muscle injury was performed in wild B6 and CaMKIV-/- mice. HE staining was used to observe the inflammation and repair in injured muscles. Dystrophy immunofluorescence staining was used to identify the regenerating myofibers while F4/80 staining to present infiltrated macrophage in inflamed muscle. F4/80, Ly-6C, CD206, Ki67 staining and flow cytometry skill were used to analyze the number and proportion of M1 (F4/80+Ly6C+) and M2 ((F4/80+CD206+) macrophages in the injured muscle, as well proliferation capacity of macrophages (F4/80+Ki67+). Results CaMKIV-/- mice were successfully constructed and bred. HE and fluorescence staining showed that, the absence of CaMKⅣ signaling in mice resulted in increasing intramuscular inflammation and delaying muscle repair. The number of the regenerating central nuclear muscle fibers（Dystrophin+） in CaMKⅣ-/- mice was lower than that in WT mice. The number of macrophages (F4/80+) in inflamed muscle from CaMKⅣ-/- mice was significantly higher than that of WT mice, which was dominant by M1 cells, comparing with that of WT mice. In inflamed muscle of CaMKⅣ-/- mice, the number and proportion of proliferating macrophages (F4/80+Ki67+) was significantly higher than that of control mice, but the proliferating macrophages were mainly M1 cells (Ly6C+Ki67+). Conclusions The loss of CaMKⅣ signaling leads to the aggravation of muscle inflammation, promotes the exudation and proliferation of M1 macrophages in inflamed muscle, and delays muscle repair after injury.

Key words

Muscle injury / / / CaMKⅣ / / / Macrophage / / / Inflammation / / / Proliferation

Cite this article

EndNote

Ris (Procite)

Bibtex

Download Citations

Wang Han, Li Yangyang, Jian Xiaoting, Huang Jingwen, Lan Haiqiang, Hu Jijie, Liao Hua. Loss of CaMKIV signaling aggravates muscle inflammation and affects muscle regeneration[J]. Chinese Journal of Clinical Anatomy. 2023, 41(6): 692-697 https://doi.org/10.13418/j.issn.1001-165x.2023.6.11

References

[1] Lowe DA, Warren GL, Ingalls CP, et al. Muscle function and protein metabolism after initiation of eccentric contraction-induced injury[J]. J Appl Physiol (1985),1995,79(4):1260-1270. DOI: 10.1152/jappl.1995.79.4.1260.
[2] Shi D, Gu R, Song Y, et al. Calcium/Calmodulin-Dependent Protein Kinase IV (CaMKIV) Mediates Acute Skeletal Muscle Inflammatory Response[J]. Inflammation, 2018, 41(1):199-212. DOI: 10.1007/s10753-017-0678-2.
[3] Racioppi L, Means AR. Calcium/calmodulin-dependent kinase IV in immune and inflammatory responses: novel routes for an ancient traveller[J]. Trends Immunol, 2008, 29(12):600-607. DOI: 10.1016/j.it.2008.08.005.
[4] Illario M, Giardino-Torchia ML, Sankar U, et al. Calmodulin-dependent kinase IV links Toll-like receptor 4 signaling with survival pathway of activated dendritic cells[J]. Blood, 2008, 111(2):723-731. DOI: 10.1182/blood-2007-05-091173.
[5] Tokumitsu H, Hatano N, Inuzuka H, et al. Mechanism of the generation of autonomous activity of Ca2+/calmodulin-dependent protein kinase IV[J]. J Biol Chem, 2004, 279(39):40296-40302. DOI: 10.1074/jbc.M406534200.
[6] Krebs J, Wilson A, Kisielow P. Calmodulin-dependent protein kinase IV during T-cell development[J]. Biochem Biophys Res Commun, 1997, 241(2):383-389. DOI: 10.1006/bbrc.1997.7823.
[7] Anderson KA, Means AR. Defective signaling in a subpopulation of CD4 (+) T cells in the absence of Ca(2+)/calmodulin-dependent protein kinase IV[J]. Mol Cell Biol, 2002, 22 (1):23-29. DOI: 10.1128/MCB.22.1.23-29.2002.
[8] Koga T, Kawakami A. The role of CaMK4 in immune responses[J]. Mod Rheumatol,2018, 28(2):211-214. DOI: 10.1080/14397595. 2017.1413964.
[9] Juang YT, Wang Y, Solomou EE, et al. Systemic lupus erythematosus serum IgG increases CREM binding to the IL-2 promoter and suppresses IL-2 production through CaMKIV[J]. J Clin Invest, 2005, 115(4):996-1005. DOI: 10.1172/JCI22854.
[10] Koga T, Ichinose K, Mizui M, et al. Calcium/calmodulin-dependent protein kinase IV suppresses IL-2 production and regulatory T cell activity in lupus[J]. J Immunol, 2012, 189(7):3490-3496. DOI: 10.4049/jimmunol.1201785.
[11]Chin ER. Role of Ca2+/calmodulin-dependent kinases in skeletal muscle plasticity[J]. J Appl Physiol (1985), 2005, 99(2):414-423. DOI: 10.1152/japplphysiol.00015.2005.
[12]Chin E R. The role of calcium and calcium/calmodulin-dependent kinases in skeletal muscle plasticity and mitochondrial biogenesis[J]. Proc Nutr Soc, 2004, 63(2):279-286. DOI: 10.1079/PNS2004335.
[13] Tidball JG. Inflammatory processes in muscle injury and repair[J]. Am J Physiol Regul Integr Comp Physiol, 2005, 288(2):R345-R353. DOI: 10.1152/ajpregu.00454.2004.
[14] Vitadello M, Doria A, Tarricone E, et al. Myofiber stress-response in myositis: parallel investigations on patients and experimental animal models of muscle regeneration and systemic inflammation[J]. Arthritis Res Ther, 2010, 12(2):R52. DOI: 10.1186/ar2963.
[15] Ding M, Huang T, Zhu R, et al. Immunological Behavior Analysis of Muscle Cells under IFN-gamma Stimulation in Vitro and in Vivo[J]. Anat Rec (Hoboken), 2018, 301(9):1551-1563. DOI: 10.1002/ar.23834.
[16] Arnold L, Henry A, Poron F, et al. Inflammatory monocytes recruited after skeletal muscle injury switch into antiinflammatory macrophages to support myogenesis[J]. J Exp Med, 2007,204(5):1057-1069. DOI: 10.1084/jem.20070075.
[17] Hu J, Shi D, Ding M, et al. Calmodulin-dependent signalling pathways are activated and mediate the acute inflammatory response of injured skeletal muscle[J]. J Physiol, 2019, 597(21):5161-5177. DOI: 10.1113/JP278478.
[18] Cote CH, Bouchard P, van Rooijen N, et al. Monocyte depletion increases local proliferation of macrophage subsets after skeletal muscle injury[J]. BMC Musculoskelet Disord, 2013, 14:359. DOI: 10.1186/1471-2474-14-359.
[19] Zhang J, Zhang D, Zhao J. CFNAs of RBCs affect the release of inflammatory factors through the expression of CaMKIV in macrophages[J]. Transfus Apher Sci, 2022, 61(6):103494. DOI: 10.1016/j.transci.2022.103494.
[20] Gu R, Ding M, Shi D, et al. Calcium/Calmodulin-Dependent Protein Kinase IV Mediates IFN-gamma-Induced Immune Behaviors in Skeletal Muscle Cells [J]. Cell Physiol Biochem, 2018 46(1):351-364. DOI: 10.1159/000488435.