Abstract
Immunohistochemical techniques were used to examine the distribution of prostaglandin H synthase (PGHS)-2 and neuronal nitric oxide synthase (nNOS) in piglet brain. Samples from parietal cortex, hippocampus, and cerebellum were immersion fixed in 10% formalin, sectioned at 50 μm, and immunostained using specific antibodies against PGHS-2 and nNOS. Immunoreactivity for PGHS-2 was extensive throughout the areas examined. For example, PGHS-2 immunoreactive cells were present in all layers of the cortex, but were particularly dense among neurons in layers II/III, V, and VI. In addition, glial cells associated with microvessels in white matter showed PGHS-2 immunoreactivity. In contrast, nNOS immunoreactive neurons were limited in number and widely dispersed across all layers of the cortex and thus did not form a definable pattern. In the hippocampus, heavy PGHS-2 immunoreactivity was present in neurons and glial cells in the subgranular region, stratum radiatum, adjacent to the hippocampal sulcus, and in CA1 and CA3 pyramidal cells. Immunostaining for nNOS displayed a different pattern from PGHS-2 in the hippocampus, and was mainly localized to the granule cell layer of the dentate gyrus and the mossy fiber layer. In the cerebellum, PGHS-2 immunoreactivity was heavily represented in the Bergmann glia and to a lesser extent in cells of the granular layer, whereas nNOS was detected only in Basket cells. There are four conclusions from this study. First, PGHS-2 immunoreactivity is widely represented in the cerebral cortex, hippocampus, and cerebellum of neonatal pigs. Second, glia cells as well as neurons can show immunoreactivity for PGHS-2. And third, the distribution of nNOS is different from PGHS-2 immunoreactivity in the cerebral cortex, hippocampus, and cerebellum.
Similar content being viewed by others
Log in or create a free account to read this content
Gain free access to this article, as well as selected content from this journal and more on nature.com
or
Abbreviations
- PGHS:
-
prostaglandin H synthase
- NOS:
-
nitric oxide synthase
- PGH2:
-
prostaglandin H2
- NO:
-
nitric oxide
- NGS:
-
normal goat serum
- GFAP:
-
glial fibrillary acidic protein
References
DeWitt DL 1991 Prostaglandin endoperoxide synthase: regulation of enzyme expression. Biochim Biophys Acta 1083: 121–134.
Smith WL 1992 Prostanoid biosynthesis and mechanisms of action. Am J Physiol 263:F181–F191.
Wu KK 1996 Cyclooxygenase 2 induction: molecular mechanism and pathophysiologic roles. J Lab Clin Med 128: 242–245.
Yamagata K, Adreasson KT, Kaufmann WE, Barnes CA, Worley PF 1993 Expression of a mitogen-inducible cyclooxygenase in brain neurons: regulation by synaptic activity and glucocorticoids. Neuron 11: 371–386.
Breder CD, DeWitt D, Kraig RP 1995 Characterization of inducible cyclooxygenase in rat brain. J Comp Neurol 355: 296–315.
Peri KG, Hardy P, Li DY, Varma DR, Chemtob S 1995 Prostaglandin G/H synthase-2 is a major contributor of brain prostaglandins in the newborn. J Biol Chem 270: 24615–24620.
Thore CR, Nam MJ Busija DW 1996 Immunofluorescent localization of constitutive and inducible prostaglandin H synthase in ovine astroglia. J Comp Neurol 367: 1–9.
Busija DW, Thore C, Beasley T, Bari F 1996 Induction of cyclooxygenase-2 following anoxic stress in piglet cerebral arteries. Microcirculation 3: 379–386.
Busija DW 1997 Eicosanoids and cerebrovascular control. In: Welch KMA, Caplan LR, Reis DJ, Siesjö BK, Weir B (eds) Primer on Cerebrovascular Diseases. Academic Press, New York, PP 93–96.
Pourcyrous M, Leffler CW, Bada HS, Korones SB, Busija DW 1993 Brain superoxide anion generation in asphyxiated piglets and the effect of indomethacin at therapeutic dose. Pediatr Res 34: 366–369.
Beasley TC, Bari F, Thore C, Thrikawala N, Louis TM, Busija DW 1998 Cerebral ischemia/reperfusion increases endothelial nitric oxide synthase levels via an indomethacin sensitive mechanism. J Cereb Blood Flow Metab 18: 88–96.
Vannuci RC 1993 Mechanisms of perinatal hypoxic-ischemic brain damage. Semin Perinatol 17: 330–337.
Breder CD, Smith WL, Raz A, Masferrer J, Seibert K, Needleman P, Saper CB 1992 Distribution and characterization of cyclooxygenase immunoreactivity in the ovine brain. J Comp Neurol 322: 409–438.
Beckman JS, Beckman TW, Chen J, Marshall PA, Freeman BA 1990 Apparent hydroxyl radical production by peroxynitrite: implications for endothelial injury from nitric oxide and superoxide. Proc Natl Acad Sci USA 87: 1620–1624.
Beckman JS 1991 The double-edged role of nitric oxide in brain function and superoxide-mediated injury. J Dev Physiol 15: 53–59.
Busija DW, Thore C 1997 Modulation of prostaglandin production by nitric oxide in astroglia. Prostaglandins Leukot Essent Fatty Acids 56: 355–359.
Hedqvist P 1977 Basic mechanisms of prostaglandin action on autonomic neurotransmission. Annu Rev Pharmacol Toxicol 17: 259–273.
Ueno R, Narumiya S, Ogorochi T, Nakayama T, Ishikawa Y, Hayaishi O 1982 Role of prostaglandin D2 in the hypothermia of rats caused by bacterial lipopolysaccharide. Proc Natl Acad Sci USA 79: 6093–6097.
Kimura H, Okamoto K, Sakai Y 1985 Modulatory effects of prostaglandins D2, E2 and F2α on the postsynaptic actions of inhibitory and excitatory amino acids in cerebellar Purkinje cell dendrites in vitro. Brain Res 330: 235–244.
Stitt J 1986 Prostaglandin E as the neural mediator of the febrile response. Yale J Biol Med 59: 137–149.
Hayaishi O 1988 Sleep-wake regulation by prostaglandins D2 and E2. J Biol Chem 263: 14593–14596.
Shimizu T, Wolfe LS 1990 Arachidonic acid cascade and signal transduction. J Neurochem 55: 1–15.
Busija DW 1996 Nervous control of the cerebral circulation. In: T. Bennett and S. M. Gardiner (eds) Nervous Control of the Circulation. Harwood Academic Publishers, Amsterdam, PP 177–206.
Bari F, Louis TM, Busija DW 1997 Kainate-induced cerebrovascular dilation is resistant to ischemia in piglets. Stroke 28: 1272–1277.
Tsubokura SY, Watanabe H, Ehara K, Imamura K, Saugimoto O, Kagamiyama H, Yamamoto S, Hayaishi O 1991 Localization of prostaglandin endoperoxide synthase in neurons and glia of the monkey brain. Brain Res 543: 15–24.
Kaufmann WE, Worley PF, Pegg J, Bremer M, Isakson P 1996 COX-2, a synaptically induced enzyme, is expressed by excitatory neurons at postsynaptic sites in rat cerebral cortex. Proc Natl Acad Sci USA 93: 2317–2321.
Adams J, Collaço-Moraes Y, de Belleroche J 1996 Cyclooxygenase-2 induction in cerebral cortex: an intracellular response to synaptic excitation. J Neurochem 66: 6–13.
Smith WL, Gutekunst DI, Lyons RH Jr 1980 Immunocytochemical localization of the prostaglandin-forming cyclooxygenase in the cerebellar cortex. Prostaglandins 19: 61–69.
Bredt DS, Synder SH 1994 Transient nitric oxide synthase neurons in embryonic cerebral cortical plate, sensory ganglia, and olfactory epithelium. Neuron 13: 301–313.
Kugler P, Drenckhahn D 1996 Astrocyte and Bergmann glia as an important site of nitric oxide synthase I. Glia 16: 165–173.
Busija DW Leffler CW 1989 Dilator effects of amino acid neurotransmitters on piglet pial arterioles. Am J Physiol 257:H1200–H1203.
Meng W, Tobin J, Busija DW 1995 Glutamate-induced cerebral vasodilation is mediated by nitric oxide through NMDA receptors. Stroke 26: 857–863.
Salvemini D, Misko TP, Masferrer JL, Seibert K, Currie MG, Needleman P 1993 Nitric oxide activates cyclooxygenase enzymes. Proc Natl Acad Sci USA 90: 7240–7244.
Stadler J, Harbrecht BG, DiSilivi M, Curran RD, Jordan ML, Simmons RL, Billiar TR 1993 Endogenous nitric oxide inhibits the synthesis of cyclooxygenase products and interleukin-6 by rat Kupffer cells. J Leuk Biol 53: 165–172.
Curtis JF, Reddy NG, Mason RP, Kalyanaraman B, Eling TE 1996 Nitric oxide: a prostaglandin H synthase 1 and 2 reducing cosubstrate that does not stimulate cyclooxygenase activity or prostaglandin H synthase expression in murine macrophages. Arch Biochem Biophys 335: 369–376.
Acknowledgements
The authors thank Dr. A. W. Ford-Hutchinson of the Merck Frosst Center for Therapeutic Research, Pointe Claire, CA) for providing the PGHS-2 antibody, and Nancy Busija for editing the manuscript.
Author information
Authors and Affiliations
Additional information
Supported by National Institutes of Health Grants HL-30260, HL-46558, and HL-50587 and by the Hungarian Ministry of Education Grant FKFP 0713/1997.
Rights and permissions
About this article
Cite this article
Dégì, R., Bari, F., Beasley, T. et al. Regional Distribution of Prostaglandin H Synthase-2 and Neuronal Nitric Oxide Synthase in Piglet Brain. Pediatr Res 43, 683–689 (1998). https://doi.org/10.1203/00006450-199805000-00018
Received:
Accepted:
Issue date:
DOI: https://doi.org/10.1203/00006450-199805000-00018
