Co-expression of vesicular glutamate transporters (VGLUT1 and VGLUT2) and their association with synaptic-like microvesicles in rat pinealocytes

Riyo Morimoto, Mitsuko Hayashi, Shouki Yatsushiro, Masato Otsuka, Akitsugu Yamamoto, Yoshinori Moriyama

Research output: Contribution to journalArticle

45 Citations (Scopus)

Abstract

A vesicular glutamate transporter (VGLUT) is responsible for the accumulation of L-glutamate in synaptic vesicles in glutamatergic neurons. Two isoforms, VGLUT1 and VGLUT2, have been identified, which are complementarily expressed in these neurons. Mammalian pinealocytes, endocrine cells for melatonin, are also glutamatergic in nature, accumulate L-glutamate in synaptic-like microvesicles (SLMVs), and secrete it through exocytosis. Although the storage of L-glutamate in SLMVs is mediated through a VGLUT, the molecular nature of the transporter is less understood. We recently observed that VGLUT2 is expressed in pinealocytes. In the present study, we show that pinealocytes also express VGLUT1. RT-PCR and northern blot analyses indicated expression of the VGLUT1 gene in pineal gland. Western blotting with specific antibodies against VGLUT1 indicated the presence of VGLUT1 in pineal gland. Indirect immunofluorescence microscopy with a section of pineal gland and cultured cells indicated that VGLUT1 and VGLUT2 are co-localized with process terminal regions of pinealocytes. Furthermore, immunoelectronmicroscopy as well as subcellular fractionation studies revealed that both VGLUT1 and VGLUT2 are specifically associated with SLMVs. These results indicate that both VGLUTs are responsible for storage of L-glutamate in SLMVs in pinealocytes. Pinealocytes are the first exception as to complementary expression of VGLUT1 and VGLUT2.

Original languageEnglish
Pages (from-to)382-391
Number of pages10
JournalJournal of Neurochemistry
Volume84
Issue number2
DOIs
Publication statusPublished - Jan 2003

Fingerprint

Vesicular Glutamate Transport Proteins
Pineal Gland
Rats
Glutamic Acid
Neurons
Cells
Endocrine Cells
Synaptic Vesicles
Exocytosis
Melatonin
Indirect Fluorescent Antibody Technique
Fractionation
Fluorescence Microscopy
Northern Blotting
Cultured Cells
Microscopic examination
Protein Isoforms
Genes
Western Blotting
Gene Expression

Keywords

  • Pineal gland
  • Pinealocytes
  • Synaptic-like microvesicle
  • Vesicular glutamate transporter 1
  • Vesicular glutamate transporter 2

ASJC Scopus subject areas

  • Biochemistry
  • Cellular and Molecular Neuroscience

Cite this

Co-expression of vesicular glutamate transporters (VGLUT1 and VGLUT2) and their association with synaptic-like microvesicles in rat pinealocytes. / Morimoto, Riyo; Hayashi, Mitsuko; Yatsushiro, Shouki; Otsuka, Masato; Yamamoto, Akitsugu; Moriyama, Yoshinori.

In: Journal of Neurochemistry, Vol. 84, No. 2, 01.2003, p. 382-391.

Research output: Contribution to journalArticle

Morimoto, Riyo ; Hayashi, Mitsuko ; Yatsushiro, Shouki ; Otsuka, Masato ; Yamamoto, Akitsugu ; Moriyama, Yoshinori. / Co-expression of vesicular glutamate transporters (VGLUT1 and VGLUT2) and their association with synaptic-like microvesicles in rat pinealocytes. In: Journal of Neurochemistry. 2003 ; Vol. 84, No. 2. pp. 382-391.
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