Serotonin transporter and receptor expression in osteocytic MLO-Y4 cells

M. Bliziotes, A. Eshleman, B. Burt-Pichat, X. W. Zhang, J. Hashimoto, K. Wiren, C. Chenu

Research output: Contribution to journalArticle

90 Scopus citations

Abstract

Neurotransmitter regulation of bone metabolism has been a subject of increasing interest and investigation. We reported previously that osteoblastic cells express a functional serotonin (5-HT) signal transduction system, with mechanisms for responding to and regulating uptake of 5-HT. The clonal murine osteocytic cell line, MLO-Y4, demonstrates expression of the serotonin transporter (5-HTT), and the 5-HT1A, and 5-HT2A receptors by real-time RT-PCR and immunoblot analysis. Immunohistochemistry using antibodies for the 5-HTT, and the 5-HT1A and 5-HT2A receptors reveals expression of all three proteins in both osteoblasts and osteocytes in rat tibia. 5-HTT binding sites were demonstrated in the MLO-Y4 cells with nanomolar affinity for the stable cocaine analog [125I]RTI-55. Imipramine and fluoxetine, antagonists with specificity for 5-HTT, show the highest potency to antagonize [125I]RTI-55 binding in the MLO-Y4 cells. GBR-12935, a relatively selective dopamine transporter antagonist, had a much lower potency, as did desipramine, a selective norepinephrine transporter antagonist. The maximal [3H]5-HT uptake rate in MLO-Y4 cells was 2.85 pmol/15 min/well, with a Km value of 290 nM. Imipramine and fluoxetine inhibited specific [3H]5-HT uptake with IC50 values in the nanomolar range. 5-HT rapidly stimulated PGE2 release from MLO-Y4 cells; the EC50 for 5-HT was 0.1 μM, with a 3-fold increase seen at 60 min. The rate-limiting enzyme for serotonin synthesis, tryptophan hydroxylase, is expressed in MLO-Y4 cells as well as osteoblastic MC3T3-E1 cells. Thus, osteocytes, as well as osteoblasts, are capable of 5-HT synthesis, and express functional receptor and transporter components of the 5-HT signal transduction system.

Original languageEnglish (US)
Pages (from-to)1313-1321
Number of pages9
JournalBone
Volume39
Issue number6
DOIs
StatePublished - Dec 1 2006

    Fingerprint

Keywords

  • Bone
  • Neurotransmitter
  • Osteoblast
  • Osteocyte
  • Transporter

ASJC Scopus subject areas

  • Endocrinology, Diabetes and Metabolism
  • Physiology
  • Histology

Cite this

Bliziotes, M., Eshleman, A., Burt-Pichat, B., Zhang, X. W., Hashimoto, J., Wiren, K., & Chenu, C. (2006). Serotonin transporter and receptor expression in osteocytic MLO-Y4 cells. Bone, 39(6), 1313-1321. https://doi.org/10.1016/j.bone.2006.06.009