Cellular localization of lipocalin-type prostaglandin D synthase (β-trace) in the central nervous system of the adult rat

M. A. Howard, I. O. Volkov, R. Mirsky, P. C. Garell, M. D. Noh, M. Granner, H. Damasio, Mitchell Steinschneider, R. A. Reale, J. E. Hind, J. F. Brugge

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Abstract

We applied high-resolution laser-scanning microscopy, electron microscopy, and non-radioactive in situ hybridization histochemistry to determine the cellular and intracellular localization of lipocalin-type prostaglandin D synthase, the major brain-derived protein component of cerebrospinal fluid, and its mRNA in leptomeninges, choroid plexus, and parenchyma of the adult rat brain. Both immunoreactivity and mRNA for prostaglandin D synthase were located in arachnoid barrier cells, arachnoid trabecular cells, and arachnoid pia mater cells. Furthermore, meningeal macrophages and perivascular microglial cells, identified by use of ED2 antibody, were immunopositive for prostaglandin D synthase. In the arachnoid trabecular cells, the immunoreactivity for prostaglandin D synthase was located in the nuclear envelope, Golgi apparatus, and secretory vesicles, indicating the active production and secretion of prostaglandin D synthase. In the meningeal macrophages, prostaglandin D synthase was not found around the nucleus but in lysosomes in the cytoplasm, pointing to an uptake of the protein from the cerebrospinal fluid. Furthermore, the existence of meningeal cyclooxygenase (COX)-1 and COX-2 was investigated by Western blot, Northern blot, and reverse transcriptase - polymerase chain reaction (RT-PCR), and the colocalization of COX-2 and prostaglandin D synthase was demonstrated in virtually all cells of the leptomeninges, choroid plexus epithelial cells, and perivascular microglial cells, suggesting that these cells synthesize prostaglandin D2 actively. Alternatively, oligodendrocytes showed prostaglandin D synthase immunoreactivity without detectable COX-2. The localization of lipocalin-type prostaglandin D synthase in meningeal cells and its colocalization with COX-2 provide evidence for its function as a prostaglandin D2-producing enzyme. (C) 2000 Wiley-Liss, Inc.

Original languageEnglish (US)
Pages (from-to)62-78
Number of pages17
JournalJournal of Comparative Neurology
Volume428
Issue number1
DOIs
StatePublished - Dec 4 2000

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prostaglandin R2 D-isomerase
Lipocalins
Central Nervous System
Arachnoid
Cyclooxygenase 2
Cerebrospinal Fluid Proteins
Prostaglandin D2
Choroid Plexus
Pia Mater
Macrophages
Cyclooxygenase 1
Messenger RNA
Nuclear Envelope
Oligodendroglia
Secretory Vesicles
Brain
Golgi Apparatus
Lysosomes
Reverse Transcriptase Polymerase Chain Reaction
Confocal Microscopy

Keywords

  • Arachnoid membrane
  • COX
  • Electron microscopy
  • Immunohistochemistry
  • In situ hybridization
  • Perivascular microglial cells

ASJC Scopus subject areas

  • Neuroscience(all)

Cite this

Cellular localization of lipocalin-type prostaglandin D synthase (β-trace) in the central nervous system of the adult rat. / Howard, M. A.; Volkov, I. O.; Mirsky, R.; Garell, P. C.; Noh, M. D.; Granner, M.; Damasio, H.; Steinschneider, Mitchell; Reale, R. A.; Hind, J. E.; Brugge, J. F.

In: Journal of Comparative Neurology, Vol. 428, No. 1, 04.12.2000, p. 62-78.

Research output: Contribution to journalArticle

Howard, M. A. ; Volkov, I. O. ; Mirsky, R. ; Garell, P. C. ; Noh, M. D. ; Granner, M. ; Damasio, H. ; Steinschneider, Mitchell ; Reale, R. A. ; Hind, J. E. ; Brugge, J. F. / Cellular localization of lipocalin-type prostaglandin D synthase (β-trace) in the central nervous system of the adult rat. In: Journal of Comparative Neurology. 2000 ; Vol. 428, No. 1. pp. 62-78.
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AU - Mirsky, R.

AU - Garell, P. C.

AU - Noh, M. D.

AU - Granner, M.

AU - Damasio, H.

AU - Steinschneider, Mitchell

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N2 - We applied high-resolution laser-scanning microscopy, electron microscopy, and non-radioactive in situ hybridization histochemistry to determine the cellular and intracellular localization of lipocalin-type prostaglandin D synthase, the major brain-derived protein component of cerebrospinal fluid, and its mRNA in leptomeninges, choroid plexus, and parenchyma of the adult rat brain. Both immunoreactivity and mRNA for prostaglandin D synthase were located in arachnoid barrier cells, arachnoid trabecular cells, and arachnoid pia mater cells. Furthermore, meningeal macrophages and perivascular microglial cells, identified by use of ED2 antibody, were immunopositive for prostaglandin D synthase. In the arachnoid trabecular cells, the immunoreactivity for prostaglandin D synthase was located in the nuclear envelope, Golgi apparatus, and secretory vesicles, indicating the active production and secretion of prostaglandin D synthase. In the meningeal macrophages, prostaglandin D synthase was not found around the nucleus but in lysosomes in the cytoplasm, pointing to an uptake of the protein from the cerebrospinal fluid. Furthermore, the existence of meningeal cyclooxygenase (COX)-1 and COX-2 was investigated by Western blot, Northern blot, and reverse transcriptase - polymerase chain reaction (RT-PCR), and the colocalization of COX-2 and prostaglandin D synthase was demonstrated in virtually all cells of the leptomeninges, choroid plexus epithelial cells, and perivascular microglial cells, suggesting that these cells synthesize prostaglandin D2 actively. Alternatively, oligodendrocytes showed prostaglandin D synthase immunoreactivity without detectable COX-2. The localization of lipocalin-type prostaglandin D synthase in meningeal cells and its colocalization with COX-2 provide evidence for its function as a prostaglandin D2-producing enzyme. (C) 2000 Wiley-Liss, Inc.

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