Expressions of iron transport related proteins in the spinal cord of amyotrophic lateral sclerosis transgenic mice

ZHANG Ya-Wen; GAO Ying; SUN Han-Cong; ZHANG Hao-Yun; WANG Feng-Bin

doi:10.16098/j.issn.0529-1356.2021.02.001

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Acta Anatomica Sinica ›› 2021, Vol. 52 ›› Issue (2) : 161-167. DOI: 10.16098/j.issn.0529-1356.2021.02.001

Neurobiology

Expressions of iron transport related proteins in the spinal cord of amyotrophic lateral sclerosis transgenic mice

ZHANG Ya-wen^1,5 GAO Ying^2,5 SUN Han-cong^3,5 ZHANG Hao-yun^4,5* WANG Feng-bin^1*

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Abstract

Objective To investigate the relationship between the expressions of iron transport related proteins and the dysregulation of iron homeostasis in the spinal cord of amyotrophic lateral sclerosis(ALS) transgenic mice. Methods The hSOD1G93A transgenic mice (ALS mice) and littermate wild-type mice (WT mice) were selected to separate the spinal cord at day 70, day 95, and day 122 after birth, 9 mice per time point and per group. Western blotting was used to detect the expressions of iron transporter divalent metal transporter-1 (DMT1), ferroportin 1 (FPN1) and regulatory protein iron regulatory protein 1(IRP1) in the spinal cord. Double immunofluorescence labeling was used to detect the co-localization of cells in the ventral horn of lumbar spinal cord. Results Western blotting results showed that compared with WT mice the expressions of DMT1 protein were down-regulated with the disease progression from day 70 to day 122 (P<0.05, P<0.01); FPN1 protein was transiently up-regulated at day 70 (P<0.05), and decline expressions were observed at day 95 and day 122 (P<0.01); IRP1 was down-regulated at day 95 and day 122 (P<0.01). Double immunofluorescence labeling revealed that at day 70, DMT1 co-expressed mainly with β-tubulin Ⅲ both in WT and ALS mice lumbar spinal cord. Compared with the WT group, the DMT1 immunoreactivity in the neurons of the ventral horn lumbar spinal cord of ALS mice was elevated at day 95, while the FPN1 fluorescence intensity was weak. With the disease progression, the co-localization expression of DMT1, FPN1 with reactive glial cells increased. With the disease progresses, the expression of IRP1 decreased. Conclusion With the progression of ALS, iron influx increases and iron outflux decreases in neurons at the early-symptomatic stage of ALS, the activity of iron transport in reactive glial cells is enhanced, which participates in local iron homeostasis imbalance and progressive loss of motor neurons in ventral horn of spinal cord. Decreased expression of IRP1 partly participates in the regulation of local iron metabolism.

Key words

Amyotrophic lateral sclerosis / Iron homeostasis / Divalent metal transporter-1 / Ferroportin 1 / Iron regulatory protein / Western blotting / Mouse

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ZHANG Ya-wen GAO Ying SUN Han-cong ZHANG Hao-yun WANG Feng-bin. Expressions of iron transport related proteins in the spinal cord of amyotrophic lateral sclerosis transgenic mice[J]. Acta Anatomica Sinica. 2021, 52(2): 161-167 https://doi.org/10.16098/j.issn.0529-1356.2021.02.001

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