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Effects of hypoxia and intracellular iron chelation on hypoxia-inducible factor-1α and -1β in the rat carotid body and glomus cells

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Abstract

The present investigation provides for the first time, unambiguous information on the occurrence of hypoxia-inducible factors (HIF-1α and HIF-1β proteins) in normoxia (Nx) and their interaction with hypoxia (Hx) and intracellular Fe2+ chelation in the rat carotid body (CB) glomus cells. HIF-1α bound to HIF-1β translocated into the nucleus is identified on the basis of immunohistochemistry and immunofluorescence. In Nx, a weak expression of HIF-1α was observed in CB glomus cells. However, exposure of CB and glomus cells to Hx (Po2≃7 Torr) and Nx with ciclopirox olamine (CPX, 5 μM) for 1 h showed a significant (P<0.001) increase in HIF-1α protein. The CBs and glomus cells exposed to Nx, Hx, and Nx with CPX showed a constant level of HIF-1β protein expression. HIF-1α subunit is continuously synthesized and degraded under normoxic conditions, while it accumulates rapidly following exposure to low oxygen tensions. Hydroxylation of HIF-1α by prolyl hydroxylase for proteasomal degradation was dependent on iron, 2-oxoglutarate, and oxygen concentration. The intracellular iron that acts as a cofactor for prolyl hydroxylase activity belongs to the labile iron pool and can be easily chelated. Thus, chelation of intracellular labile iron by CPX in Nx significantly increased HIF-1α in CB glomus cells. Thus, the results are consistent with the hypothesis that HIF-1α which is present in the glomus cells translocates to the nucleus during exposure to Hx and to CPX in Nx.

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Acknowledgements

The authors are thankful to Dr. L. Kubin, Department of Animal Biology, for his excellent guidance in designing the experimental protocol, and to Mrs. Kathy Notarfrancesco, Institute for Environmental Medicine, Juli Burns, Department of Pathology, and Maggie Grimes, Department of Physiology, University of Pennsylvania for their skillful technical assistance. S.M.B. is a recipient of an NRSA fellowship award. This work was supported by NIH training grants T-32-HL07027 and in part by ONR N-00014-01-0948, R01-HL50180, and R37-HL43413.

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  1. Department of Physiology, B-400 Richards Building, University of Pennsylvania School of Medicine, 3700 Hamilton Walk, Philadelphia, PA 19104-6085, USA

    Santhosh M. Baby, Arijit Roy, Anil M. Mokashi & Sukhamay Lahiri

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  1. Santhosh M. Baby
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  2. Arijit Roy
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  4. Sukhamay Lahiri
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Correspondence to Sukhamay Lahiri.

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Baby, S.M., Roy, A., Mokashi, A.M. et al. Effects of hypoxia and intracellular iron chelation on hypoxia-inducible factor-1α and -1β in the rat carotid body and glomus cells. Histochem Cell Biol 120, 343–352 (2003). https://doi.org/10.1007/s00418-003-0588-2

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