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Adrenomedullin-2/Intermedin Induces cAMP Accumulation in Dissociated Rat Spinal Cord Cells: Evidence for the Existence of a Distinct Class of Sites of Action

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Abstract

Adrenomedullin-2/intermedin is structurally related to the calcitonin family of peptides, which includes calcitonin gene-related peptide (CGRP), adrenomedullin, and amylin. We recently reported that CGRP and adrenomedullin act through distinct receptors to induce cyclic adenosine monophosphate (cAMP) accumulation in dispersed cells from embryonic rat spinal cord. Here, we investigated the apparent affinity and efficacy of adrenomedullin-2/intermedin for these receptors. Adrenomedullin-2/intermedin competed with [125I]-CGRP for binding to specific embryonic spinal cord cells with a pIC50 of 9.73 ± 0.06. Interestingly, adrenomedullin-2/intermedin competed for specific [125I]-adrenomedullin binding in a biphasic manner with pIC50 of 9.03 ± 0.22 and 6.45 ± 0.24, respectively. Cellular levels of cAMP were increased by adrenomedullin-2/intermedin (pEC50 7.84 ± 0.08) when cells were exposed to this peptide for 10 min at 37°C. This effect was partially inhibited by the non-peptide antagonist BIBN4096BS (pA2 6.56 ± 0.12), the adrenomedullin antagonist hAM22–52 (pA2 6.36 ± 0.30), and the adrenomedullin/CGRP antagonist CGRP8–37 (pA2 7.24 ± 0.60). More interestingly, a highly significant effect of adrenomedullin-2/intermedin on cAMP accumulation (pEC50 7.3 ± 0.14) was still observed even in the presence of a mixture of saturating concentrations of BIBN4096BS, hAM22–52, and the amylin antagonist AC187. Taken together, these data provide evidence for the possible existence of a distinct class of receptor sites for adrenomedullin-2/intermedin in embryonic rat spinal cord cells.

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References

  • Dennis, T., Fournier, A., Cadieux, A., et al. (1990). hCGRP8–37, a calcitonin gene-related peptide antagonist revealing calcitonin gene-related peptide receptor heterogeneity in brain and periphery. Journal of Pharmacology and Experimental Therapeutics, 254, 123–128.

    PubMed  CAS  Google Scholar 

  • Doods, H., Hallermayer, G., Wu, D., et al. (2000). Pharmacological profile of BIBN4096BS, the first selective small molecule CGRP antagonist. British Journal of Pharmacology, 129, 420–423.

    Article  PubMed  CAS  Google Scholar 

  • Goltzman, D., & Mitchell, J. (1985). Interaction of calcitonin and calcitonin gene-related peptide at receptor sites in target tissues. Science, 227, 1343–1345.

    Article  PubMed  CAS  Google Scholar 

  • Hashimoto, H., Hyodo, S., Kawasaki, M., et al. (2007). Adrenomedullin 2 (AM2)/intermedin is a more potent activator of hypothalamic oxytocin-secreting neurons than AM possibly through an unidentified receptor in rats. Peptides, 28, 1104–1112.

    Article  PubMed  CAS  Google Scholar 

  • Hay, D. L., Christopoulos, G., Christopoulos, A., Poyner, D. R., & Sexton, P. M. (2005). Pharmacological discrimination of calcitonin receptor: receptor activity-modifying protein complexes. Molecular Pharmacology, 67, 1655–1665.

    Article  PubMed  CAS  Google Scholar 

  • Hay, D. L., Conner, A. C., Howitt, S. G., Smith, D. M., & Poyner, D. R. (2004). The pharmacology of adrenomedullin receptors and their relationship to CGRP receptors. Journal of Molecular Neuroscience, 22, 105–113.

    Article  PubMed  Google Scholar 

  • Hay, D. L., Howitt, S. G., Conner, A. C., Schindler, M., Smith, D. M., & Poyner, D. R. (2003). CL/RAMP2 and CL/RAMP3 produce pharmacologically distinct adrenomedullin receptors: a comparison of effects of adrenomedullin22–52, CGRP8–37 and BIBN4096BS. British Journal of Pharmacology, 140, 477–486.

    Article  PubMed  CAS  Google Scholar 

  • Mimeault, M., Fournier, A., Dumont, Y., St-Pierre, S., & Quirion, R. (1991). Comparative affinities and antagonistic potencies of various human calcitonin gene-related peptide fragments on calcitonin gene-related peptide receptors in brain and periphery. Journal of Pharmacology and Experimental Therapeutics, 258, 1084–1090.

    PubMed  CAS  Google Scholar 

  • Morimoto, R., Satoh, F., Murakami, O., et al. (2007). Expression of adrenomedullin2/intermedin in human brain, heart, and kidney. Peptides, 28, 1095–1103.

    Article  PubMed  CAS  Google Scholar 

  • Owji, A. A., Gardiner, J. V., Upton, P. D., et al. (1996). Characterisation and molecular identification of adrenomedullin binding sites in the rat spinal cord: a comparison with calcitonin gene-related peptide receptors. Journal of Neurochemistry, 67, 2172–2179.

    Article  PubMed  CAS  Google Scholar 

  • Poyner, D. R., Andrew, D. P., Brown, D., Bose, C., & Hanley, M. R. (1992). Pharmacological characterization of a receptor for calcitonin gene-related peptide on rat, L6 myocytes. British Journal of Pharmacology, 105, 441–447.

    PubMed  CAS  Google Scholar 

  • Poyner, D. R., Sexton, P. M., Marshall, I., et al. (2002). International Union of Pharmacology. XXXII. The mammalian calcitonin gene-related peptides, adrenomedullin, amylin, and calcitonin receptors. Pharmacological Reviews, 54, 233–246.

    Article  PubMed  CAS  Google Scholar 

  • Roh, J., Chang, C. L., Bhalla, A., Klein, C., & Hsu, S. Y. (2004). Intermedin is a calcitonin/calcitonin gene-related peptide family peptide acting through the calcitonin receptor-like receptor/receptor activity-modifying protein receptor complexes. Journal of Biological Chemistry, 279, 7264–7274.

    Article  PubMed  CAS  Google Scholar 

  • Smith, D. M., Coppock, H. A., Withers, D. J., et al. (2002). Adrenomedullin: receptor and signal transduction. Biochemical Society Transactions, 30, 432–437.

    Article  PubMed  CAS  Google Scholar 

  • Takei, Y., Inoue, K., Ogoshi, M., Kawahara, T., Bannai, H., & Miyano, S. (2004). Identification of novel adrenomedullin in mammals: a potent cardiovascular and renal regulator. FEBS Letters, 556, 53–58.

    Article  PubMed  CAS  Google Scholar 

  • Takhshid, M. A., Poyner, D. R., Chabot, J. G., et al. (2006). Characterization and effects on cAMP accumulation of adrenomedullin and calcitonin gene-related peptide (CGRP) receptors in dissociated rat spinal cord cell culture. British Journal of Pharmacology, 148, 459–468.

    Article  PubMed  CAS  Google Scholar 

  • Taylor, M. M., Bagley, S. L., & Samson, W. K. (2005). Intermedin/adrenomedullin-2 acts within central nervous system to elevate blood pressure and inhibit food and water intake. American Journal of Physiology. Regulatory Integrative and Comparative Physiology, 288, R919–R927.

    CAS  Google Scholar 

  • Taylor, M. M., Bagley, S. L., & Samson, W. K. (2006). Intermedin/adrenomedullin-2 inhibits growth hormone release from cultured, primary anterior pituitary cells. Endocrinology, 147, 859–864.

    Article  PubMed  CAS  Google Scholar 

  • van Rossum, D., Hanisch, U. K., & Quirion, R. (1997). Neuroanatomical localization, pharmacological characterization and functions of CGRP, related peptides and their receptors. Neuroscience and Biobehavioral Reviews, 21, 649–678.

    Article  PubMed  Google Scholar 

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Acknowledgement

This research was supported by a Canadian Institutes of Health Research grant to RQ.

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Authors and Affiliations

  1. Department of Biochemistry, Shiraz University of Medical Sciences, Shiraz, Fars, Iran

    Ali Akbar Owji

  2. Douglas Mental Health University Institute, Department of Psychiatry, McGill University, 6875 LaSalle Boulevard, Verdun, Montréal, QC, H4H 1R3, Canada

    Jean-Guy Chabot, Yvan Dumont & Rémi Quirion

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  1. Ali Akbar Owji
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  2. Jean-Guy Chabot
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  3. Yvan Dumont
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  4. Rémi Quirion
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Correspondence to Ali Akbar Owji or Rémi Quirion.

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Owji, A.A., Chabot, JG., Dumont, Y. et al. Adrenomedullin-2/Intermedin Induces cAMP Accumulation in Dissociated Rat Spinal Cord Cells: Evidence for the Existence of a Distinct Class of Sites of Action. J Mol Neurosci 35, 355–361 (2008). https://doi.org/10.1007/s12031-008-9062-x

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  • DOI: https://doi.org/10.1007/s12031-008-9062-x

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