Christopher D. Breder

Intravenous lipopolysaccharide induces cyclooxygenase 2-like immunoreactivity in rat brain perivascular microglia and meningeal macrophages

Production of prostaglandins is a critical step in transducing immune stimuli into central nervous system (CNS) responses, but the cellular source of prostaglandins responsible for CNS signalling is unknown. Cyclooxygenase catalyzes the rate‐limiting step in the synthesis of prostaglandins and exists in two isoforms. Regulation of the inducible isoform, cyclooxygenase 2, is thought to play a key role in the brains response to acute inflammatory stimuli. In this paper, we report that intravenous lipopolysaccharide (LPS or endotoxin) induces cyclooxygenase 2‐like immunoreactivity in cells closely associated with brain blood vessels and in cells in the meninges. Neuronal staining was not noticeably altered or induced in any brain region by endotoxin challenge. Furthermore, many of the cells also were stained with a perivascular microglial/macrophage‐specific antibody, indicating that intravenous LPS induces cyclooxygenase in perivascular microglia along blood vessels and in meningeal macrophages at the edge of the brain. These findings suggest that perivascular microglia and meningeal macrophages throughout the brain may be the cellular source of prostaglandins following systemic immune challenge. We hypothesize that distinct components of the CNS response to immune system activation may be mediated by prostaglandins produced at specific intracranial sites such as the preoptic area (altered sleep and thermoregulation), medulla (adrenal corticosteroid response), and cerebral cortex (headache and encephalopathy). J. Comp. Neurol. 381:119‐129, 1997.

Expression of inducible cyclooxygenase mRNA in the mouse brain after systemic administration of bacterial lipopolysaccharide

Cyclooxygenase (COX) is an enzyme involved in the biosynthesis of prostaglandins and is one of the principle targets of non-steroidal anti-inflammatory drugs. Two isoforms of this enzyme are known to exist in the brain; one of these (type 1 COX or COX1) is constitutively expressed, whereas the other form of the enzyme, which is inducible, has been called type 2 COX (COX2). We have used systemic administration of bacterial lipopolysaccharide (LPS) as a model of the acute phase response to study the expression of COX2 in the murine CNS. We observed COX2 expression in neurons of several regions of the normal murine telencephalon. Robust expression of COX2 mRNA was induced in perivascular cells between 45 min and 6 h after LPS injection. The role of prostaglandins produced by these perivascular cells in the cerebral components of the acute phase response remains to be elucidated.

Endogenous pyrogens in the CNS: Role in the febrile response

The febrile reaction is an integrated endocrine, autonomic and behavioral response, coordinated by the hypothalamus, that includes certain components of the stress response, such as elevated corticosteroid secretion. It is produced by the actions of circulating cytokines, such as interleukin-1 (IL-1) and tumor necrosis factor (TNF), on the organum vasculosum of the lamina terminalis (OVLT), resulting in the secretion of prostaglandin E2, which initiates a variety of responses, including elevation of body temperature and corticosteroid secretion. Although circulating cytokines apparently do not enter the brain, injections of IL-1 or TNF well within the blood-brain barrier produce identical effects. We have examined the localization of possible central sources of cytokines and prostaglandins, using immunohistochemistry, immunoblotting and enzyme assay. Our data indicate that in the brain cyclooxygenase, the key enzyme in the synthesis of prostaglandins, is found in neurons in the OVLT, but is also made by neurons in many sensory and visceral regulatory systems. We present evidence also that IL-1 beta in the human brain and TNF alpha in the mouse may be present in the central nervous system as neuromodulators that are important for producing the autonomic, endocrine and behavioral components of the febrile reaction. We propose a sequence of events in the febrile reaction involving: (1) action of circulating cytokines on cyclooxygenase containing neurons within the OVLT to produce local prostaglandin secretion; (2) local diffusion of prostaglandin E2 into the preoptic and anterior hypothalamic areas; (3) action of prostaglandin E2 on cytokine containing neurons in the preoptic and anterior hypothalamic areas; and (4) release of cytokines from neuronal terminals at distal sites involved in producing the autonomic, endocrine and behavioral components of the febrile reaction.

Long‐term elevation of cyclooxygenase‐2, but not lipoxygenase, in regions synaptically distant from spreading depression

Eicosanoids, produced from arachidonic acid by cyclooxygenases (COXs) and lipoxygenases (LIPOXs), are involved in numerous brain processes. To explore if brief and noninjurious stimuli chronically alter expression of these enzymes, we examined the induction of COX‐2 and LIPOX expression following unilateral neocortical spreading depression (SD). Expression was examined over time and in regions not experiencing SD (hippocampus) but synaptically connected to the site of stimulation (cortex). One hundred six male Wistar rats had SD induced via microinjection of 0.5 M KCl (0.5 M NaCl for sham) into left parietal cortex every 9 minutes for 1 or 3 hours. One hour before SD some animals received dexamethasone (Dex), mepacrine (Mep), indomethacin (Indo), nordihydroguaiaretic acid (Ndga), phenylephrine (Pe), sodium nitroprusside (Snp) with Pe, or Nω‐nitro‐L‐arginine methyl ester (Lnam). Animals survived for 0, 3, or 6 hours, or 1, 2, 3, 7, 14, 21, or 28 days. Brains were processed immunohistochemically for COX‐2 and LIPOX, and the optical density (OD) of the left and right cortex, dentate gyrus (DG), CA3, and CA1 immunoreactivity (IR) were measured. Induction was expressed as the log of left divided by right side OD for each region. COX‐2 IR in the left cortex was elevated rapidly and was sustained for 21 days following SD. COX‐2 IR was also elevated in the ipsilateral hippocampus not experiencing SD, with the rank order of induction as follows: DG > CA3 > CA1. Dex, Snp, and/or Pe significantly reduced the induction of COX‐2. No changes in LIPOX IR were observed. These results show that long‐term changes in COX‐2 expression are induced by SD and these changes decrease with synaptic distance. Benign stimuli increase COX‐2 expression and thus may influence brain function for extended periods and at distant locations.