In situ hybridization and immunohistochemical localization of heme oxygenase-2 mRNA and protein in normal rat brain: Differential distribution of isozyme 1 and 2

https://doi.org/10.1016/1044-7431(92)90068-DGet rights and content

Abstract

Heme oxygenase isozymes, HO-1 (HSP32) and HO-2, stereospecifically bind and degrade the potent prooxidant, the heme molecule, and convert it to the effective antioxidant, biliverdin, and the potential cellular messenger, carbon monoxide. In the present study we have examined the pattern of expression of the two HO-2 transcripts and protein in normal rat brain by in situ hybridization and immunochemical analysis, respectively. We have found by Northern blot analysis that HO-2 isozyme is by far the most prevalent form in the brain. Analysis of HO-2 1.3- and 1.9-kb mRNAs by in situ hybridization histochemistry showed that these transcripts are abundantly expressed in many neuronal and nonneuronal cell populations in forebrain, diencephalon, cerebellum, and brain stem regions. Furthermore, the pattern of expression of HO-2 transcripts, as detected by oligonucleotide probes, is in good agreement with that of immunoreactive protein detected by immunohistochemical analysis. Impressive levels of HO-2 transcripts and immunoreactive protein were observed in Purkinje cells of cerebellum, red nucleus, superior and inferior colliculus, nucleus of the trapezoid body, cochlear neurons, and facial nucleus of brain stem. Furthermore, in certain select brain cell populations the pattern of expression of HO-1- and HO-2-immunoreactive proteins overlapped. We suggest that the high levels of heme degradation activity and the localization of HO-2 transcripts and protein in the brain may reflect the functions of this enzyme in processes such as production of cellular messenger, regulation of the activity of heme-dependent enzymes catalyzing intracellular signaling molecule synthesis, and production of antioxidants.

References (51)

  • W.K. McCoubrey et al.

    Human heme oxygenase-2: Characterization and expression of a full length cDNA and evidence suggesting that the two HO-2 tran scripts may differ by choice of polyadenylation signal

    Arch. Biochem. Biophys

    (1992)
  • G.M. Trakshel et al.

    Resolution of the rat brain heme oxygenase activity: Absence of a detectable amount of the inducible form (HO-1)

    Arch. Biochem. Biophys

    (1988)
  • G.M. Trakshel et al.

    Multiplicity of heme oxygenase isozymes-HO-1 and HO-2 are different molecular species in rat and rabbit

    J. Biol. Chem

    (1989)
  • O.H. Lowry et al.

    Protein measurement with the Folin phenol reagent

    J. Biol. Chem

    (1951)
  • A.J. Minty et al.

    Mouse actin messenger RNA's

    J. Biol. Chem

    (1981)
  • J. Utz et al.

    Carbon monoxide relaxes ileal smooth muscle through activation of guanylate cyclase

    Biochem. Pharmacol

    (1991)
  • S.R. Vincent et al.

    Neurons that say NO

    Trends Neurosci

    (1992)
  • S. Shibahara et al.

    Transcriptional control of the rat heme oxygenase by heat shock

    J. Biol. Chem

    (1987)
  • S. Taketani et al.

    The human 32-kDa stress protein induced by exposure to arsenite and cadmium ions is heme oxygenase

    FEBS Lett

    (1989)
  • S. Keyse et al.

    Heme oxygenase is the major 32-kDa stress protein induced in human skin fibroblasts by UVA radiation, hydrogen peroxide and sodium arsenite

  • M. Bienz et al.

    Mechanisms of heat-shock gene activation in higher eukaryotes

    Adv. Genet

    (1987)
  • I.R. Brown

    Induction of heat shock (stress) genes in the mammalian brain by hyperthermia and other traumatic events: A current perspective

    J. Neurosci. Res

    (1990)
  • I.R. Brown et al.

    Expression of heat shock genes (hsp70) in the mammalian brain: Distinguishing constitutively expressed and hyperthermia-inducible mRNA species

    J. Neurosci. Res

    (1990)
  • T.D. Ingolia et al.

    Drosophila gene related to the major heat shock-induced gene is transcribed at normal temperatures and not induced by heat shock

  • P. Stocker et al.

    Antioxidant activity of albumin bound bilirubin

  • Cited by (194)

    • Upregulation of hemeoxygenase enzymes HO-1 and HO-2 following ischemia-reperfusion injury in connection with experimental cardiac arrest and cardiopulmonary resuscitation: Neuroprotective effects of methylene blue

      2021, Progress in Brain Research
      Citation Excerpt :

      For HO-2 ELISA Kit (#MBS2506874; My BioSource, San Diego, CA, USA) was used. The ELISA kit sensitivity of HO-1 was 0.937 ng/mL to 60 ng/mL and HO-2 detection range was 0.375 ng/mL to 40 ng/mL (Ewing and Maines, 1992; Kitchin et al., 2001). Small tissue pieces of porcine brain from cerebral cortex, hippocampus and cerebellum were dissected immediately after removal and weighed immediately on a preweighed filter paper on an electronic analytical balance (sensitivity 0.1 mg; Mettler Toledo AB, Stockholm, Sweden).

    • Heme oxygenase-1 and chronic hypoxia

      2012, Respiratory Physiology and Neurobiology
      Citation Excerpt :

      HO-1, an inducible isoform of the enzyme, has been shown to increase under various conditions including heat shock or oxidative stress. HO-2 is found in discrete neuronal populations throughout the brain closely paralleling the distribution of guanylyl cyclase (Ewing and Maines, 1992; Maines et al., 1993; Mazza et al., 2001; Verma et al., 1993) as well as in peripheral autonomic ganglia including the petrosal, superior cervical, nodose and myenteric ganglia (Zakhary et al., 1996). A role for HO mediated production of CO has been proposed to be important for many brain processes (for a review see Ryter et al., 2006).

    View all citing articles on Scopus
    View full text