Vesicular monoamine transporter-1 (VMAT-1) mRNA and immunoreactive proteins in mouse brain

Karen M. Ashe, Wan Ling Chiu, Ahmed M. Khalifa, Antoine N. Nicolas, Bonnie L. Brown, Randall R. De Martino, Clayton P. Alexander, Christopher T. Waggener, Krista Fischer-Stenger, Jennifer K. Stewart

Research output: Contribution to journalArticle

7 Scopus citations

Abstract

OBJECTIVE: Vesicular monoamine transporter 1 (VMAT-1) mRNA and protein were examined (1) to determine whether adult mouse brain expresses full-length VMAT-1 mRNA that can be translated to functional transporter protein and (2) to compare immunoreactive VMAT-1 proteins in brain and adrenal. METHODS: VMAT-1 mRNA was detected in mouse brain with RT-PCR. The cDNA was sequenced, cloned into an expression vector, transfected into COS-1 cells, and cell protein was assayed for VMAT-1 activity. Immunoreactive proteins were examined on western blots probed with four different antibodies to VMAT-1. RESULTS: Sequencing confirmed identity of the entire coding sequences of VMAT-1 cDNA from mouse medulla oblongata/pons and adrenal to a Gen-Bank reference sequence. Transfection of the brain cDNA into COS-1 cells resulted in transporter activity that was blocked by the VMAT inhibitor reserpine and a proton ionophore, but not by tetrabenazine, which has a high affinity for VMAT-2. Antibodies to either the C- or N- terminus of VMAT-1 detected two proteins (73 and 55 kD) in transfected COS-1 cells. The C-terminal antibodies detected both proteins in extracts of mouse medulla/pons, cortex, hypothalamus, and cerebellum but only the 73 kD protein and higher molecular weight immunoreactive proteins in mouse adrenal and rat PC12 cells, which are positive controls for rodent VMAT-1. CONCLUSIONS: These findings demonstrate that a functional VMAT-1 mRNA coding sequence is expressed in mouse brain and suggest processing of VMAT-1 protein differs in mouse adrenal and brain.

Original languageEnglish (US)
Pages (from-to)253-258
Number of pages6
JournalNeuroendocrinology Letters
Volume32
Issue number3
StatePublished - Aug 29 2011

Keywords

  • Brain
  • Schizophrenia
  • Solute carrier family 18 member 1
  • VMAT-1
  • Vesicular monoamine transporter

ASJC Scopus subject areas

  • Endocrinology, Diabetes and Metabolism
  • Endocrinology
  • Neurology
  • Endocrine and Autonomic Systems
  • Clinical Neurology
  • Psychiatry and Mental health

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    Ashe, K. M., Chiu, W. L., Khalifa, A. M., Nicolas, A. N., Brown, B. L., De Martino, R. R., Alexander, C. P., Waggener, C. T., Fischer-Stenger, K., & Stewart, J. K. (2011). Vesicular monoamine transporter-1 (VMAT-1) mRNA and immunoreactive proteins in mouse brain. Neuroendocrinology Letters, 32(3), 253-258.