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Loss of NCB5OR in the cerebellum disturbs iron pathways, potentiates behavioral abnormalities, and exacerbates harmaline-induced tremor in mice

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Abstract

Iron dyshomeostasis has been implicated in many diseases, including a number of neurological conditions. Cytosolic NADH cytochrome b5 oxidoreductase (NCB5OR) is ubiquitously expressed in animal tissues and is capable of reducing ferric iron in vitro. We previously reported that global gene ablation of NCB5OR resulted in early-onset diabetes and altered iron homeostasis in mice. To further investigate the specific effects of NCB5OR deficiency on neural tissue without contributions from known phenotypes, we generated a conditional knockout (CKO) mouse that lacks NCB5OR only in the cerebellum and midbrain. Assessment of molecular markers in the cerebellum of CKO mice revealed changes in pathways associated with cellular and mitochondrial iron homeostasis. 59Fe pulse-feeding experiments revealed cerebellum-specific increased or decreased uptake of iron by 7 and 16 weeks of age, respectively. Additionally, we characterized behavioral changes associated with loss of NCB5OR in the cerebellum and midbrain in the context of dietary iron deprivation-evoked generalized iron deficiency. Locomotor activity was reduced and complex motor task execution was altered in CKO mice treated with an iron deficient diet. A sucrose preference test revealed that the reward response was intact in CKO mice, but that iron deficient diet consumption altered sucrose preference in all mice. Detailed gait analysis revealed locomotor changes in CKO mice associated with dysfunctional proprioception and locomotor activation independent of dietary iron deficiency. Finally, we demonstrate that loss of NCB5OR in the cerebellum and midbrain exacerbated harmaline-induced tremor activity. Our findings suggest an essential role for NCB5OR in maintaining both iron homeostasis and the proper functioning of various locomotor pathways in the mouse cerebellum and midbrain.

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Notes

  1. Wang WF, et al., NCB5OR deficiency causes anemia, iron dyshomeostasis and increased susceptibility to mitochondrial dysfunction. Manuscript in preparation;

  2. Wang WF, et al., Monogenic NCB5OR diabetes is a result of impaired iron homeostasis and mitochondrial dysfunction in beta-cells. Manuscript in preparation;

  3. Low-Fe is used in lieu of low-iron for figures.

  4. Stroh MA, et al., NCB5OR deficiency in the cerebellum and midbrain affects fasted feeding behavior, thirst response, and voluntary exercise in mice. Publication in process.

Abbreviations

NCB5OR:

NADH Cytochrome b5 Oxidoreductase

CKO:

Conditional knockout

WT:

Wild-type

MPP:

Motion power percentage

VTA:

Ventral tegmental area

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Acknowledgments

Authors thank Dr. John Stanford (University of Kansas Medical Center, KUMC) for providing critiques and comments for the manuscript. Authors acknowledge Dr. WenFang Wang (KUMC) for preparing the NCB5OR-floxed line for crossing and Dr. Alexandra Joyner at Memorial Sloan-Kettering Cancer Center for providing the En1-cre driver. This study was supported by the KUMC School of Health Professions research funds (H.Z.) and a Ruth L. Kirschstein National Research Service Award (T32 HD057850, PI: R. Nudo) supporting M.A.S. Access to the KUMC Rodent Behavior Facility and shared equipment core facilities was provided by the Kansas Intellectual and Developmental Disabilities Research Center (P30 HD02528).

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Authors and Affiliations

  1. Landon Center on Aging, University of Kansas Medical Center, 3901 Rainbow Blvd., MSN 1005, Kansas City, KS, 66160, USA

    Matthew A. Stroh

  2. Department of Biochemistry and Molecular Biology, University of Kansas Medical Center, 3901 Rainbow Blvd., MSN 3030, Kansas City, KS, 66160, USA

    Matthew A. Stroh, Russell H. Swerdlow & Hao Zhu

  3. Neuroscience Graduate Program, University of Kansas Medical Center, 3901 Rainbow Blvd., MSN 3038, Kansas City, KS, 66160, USA

    Matthew A. Stroh, Russell H. Swerdlow & Hao Zhu

  4. Kansas Intellectual and Developmental Disabilities Research Center, University of Kansas Medical Center, 3901 Rainbow Blvd., MSN 3051, Kansas City, KS, 66160, USA

    Michelle K. Winter & Kenneth E. McCarson

  5. Department of Neurology, University of Kansas Medical Center, 3599 Rainbow Blvd., MSN 2012, Kansas City, KS, 66160, USA

    Russell H. Swerdlow

  6. Department of Pharmacology, Toxicology and Therapeutics, University of Kansas Medical Center, 3901 Rainbow Blvd., MSN 1018, Kansas City, KS, 66160, USA

    Kenneth E. McCarson

  7. Department of Clinical Laboratory Sciences, University of Kansas Medical Center, 3901 Rainbow Blvd., MSN 4048G-Eaton, Kansas City, KS, 66160, USA

    Hao Zhu

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  1. Matthew A. Stroh
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Correspondence to Hao Zhu.

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Stroh, M.A., Winter, M.K., Swerdlow, R.H. et al. Loss of NCB5OR in the cerebellum disturbs iron pathways, potentiates behavioral abnormalities, and exacerbates harmaline-induced tremor in mice. Metab Brain Dis 31, 951–964 (2016). https://doi.org/10.1007/s11011-016-9834-x

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