Inhibition of nNOS and iNOS following Hypoxia-Ischaemia Improves Long-Term Outcome but Does Not Influence the Inflammatory Response in the Neonatal Rat Brain

Tweel, Evelyn R.W. van den; Peeters-Scholte, Cacha; Bel, Frank van; Heijnen, Cobi J.; Groenendaal, Floris

doi:10.1159/000069044

Cited by 36 publications

(32 citation statements)

References 37 publications

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“…20 Eight-micron-thick coronal sections were stained with anti-GRK2, horseradish peroxidase-conjugated goat anti-rabbit antibody (Biotin), and Vectastain-ABC (Vector Laboratories).…”

Section: Immunohistochemistrymentioning

confidence: 99%

Hypoxia/Ischemia Modulates G Protein–Coupled Receptor Kinase 2 and β-Arrestin-1 Levels in the Neonatal Rat Brain

Lombardi

Tweel

Kavelaars

et al. 2004

Stroke

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Background and Purpose-Neurotransmitters, neuropeptides, chemokines, and many other molecules signal through G protein-coupled receptors (GPCRs). GPCR kinases (GRKs) and ␤-arrestins play a crucial role in regulating the responsiveness of multiple GPCRs. Reduced expression of GRK and ␤-arrestins leads to supersensitization of GPCRs and will thereby increase the response to neuropeptides and neurotransmitters. We analyzed GRK and ␤-arrestin expression after cerebral hypoxia/ischemia (HI). Materials and Methods-Twelve-day-old rat pups were exposed to 90 minutes of hypoxia (fraction of inspired oxygen [FiO 2 ] 0.08) after ligation of the right carotid artery, a procedure that induces unilateral damage in the right hemisphere. At 6, 12, 24, and 48 hours after HI, the left (hypoxic) and right (hypoxic/ischemic) hemispheres were analyzed for GRK and ␤-arrestin protein and mRNA expression by Western blotting and real-time polymerase chain reaction, respectively. In addition, we analyzed GRK2 expression in the hippocampus by immunohistochemistry. Results-HI downregulated GRK2 protein expression in both hemispheres at 24 to 48 hours after HI, and the effect was more pronounced in the ipsilateral hemisphere. HI induced no global change in GRK6 protein expression. However, GRK2 was markedly decreased in the hippocampal region of the ipsilateral hemisphere that will be severely damaged after HI. No changes in global mRNA levels for GRK2 were detected. In contrast, HI increased ␤-arrestin-1 protein expression as well as mRNA levels at 6 to 12 hours after HI. Conclusions-Neonatal HI-induced brain damage is associated with specific changes in the GPCR desensitization machinery. We hypothesize that these changes result in supersensitization of multiple GPCRs and might therefore contribute to HI-induced brain damage.

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“…20 Eight-micron-thick coronal sections were stained with anti-GRK2, horseradish peroxidase-conjugated goat anti-rabbit antibody (Biotin), and Vectastain-ABC (Vector Laboratories).…”

Section: Immunohistochemistrymentioning

confidence: 99%

Hypoxia/Ischemia Modulates G Protein–Coupled Receptor Kinase 2 and β-Arrestin-1 Levels in the Neonatal Rat Brain

Lombardi

Tweel

Kavelaars

et al. 2004

Stroke

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“…However, measurement of the areas of the left and right hemisphere indicated that 2-iminobiotin is only neuroprotective at a dose of 30 mg/kg per day. In previous studies, neuropathological scores, area measurements, or weight measurements of the ipsilateral and contralateral hemispheres were used to quantify the extent of neuroprotection in the Vannucci-Rice rat model (Bagenholm et al, 1996;Hagberg et al, 1997;Taylor et al, 2002;van den Tweel et al, 2002;Vannucci et al, 1998;Yuan et al, 2001). In the present study, two different methods of quantification of the cerebral injury resulted in slightly different results.…”

Section: Discussionmentioning

confidence: 74%

“…Next, we focused on the short-term effects of 2-iminobiotin after hypoxia-ischemia and reperfusion, using the optimal dose of 30 mg/kg per day. The presence of HSP70 in brain tissue has been suggested to be an early marker of injury (Ferriero et al, 1990;van den Tweel et al, 2002). Therefore, to investigate whether reduced HSP70 expression can be used as an early indicator of neuroprotection, we determined the effects of 2-iminobiotin administration on HSP70 expression at 24 hours after hypoxiaischemia.…”

Section: Discussionmentioning

confidence: 99%

“…Activation of nitric oxide synthase (NOS) contributes to hypoxia-ischemia-induced cerebral injury in the perinatal period (Hamada et al, 1994;Ikeno et al, 2000;Ishida et al, 2001;Tsuji et al, 2000;van den Tweel et al, 2002). Inhibition of both neuronal NOS (nNOS) and inducible NOS (iNOS) using 2-iminobiotin resulted in short-term neuroprotection, whereas simultaneous inhibition of all three isoforms of NOS, that is, nNOS, iNOS, and endothelial NOS (eNOS), which regulates cerebral blood flow, did not (Groenendaal et al, 1999;Marks et al, 1996;Peeters-Scholte et al, 2002).…”

Section: Introductionmentioning

confidence: 99%

“…If so, we wanted to determine the optimal dose of 2-iminobiotin using histology. It has been demonstrated in the same model of hypoxia-ischemia that cerebral levels of HSP70 protein may serve as an early marker of brain injury (Ferriero et al, 1990;van den Tweel et al, 2002). Therefore, HSP70 expression was determined 24 hours after the insult as an indicator of early neuroprotection.…”

Section: Introductionmentioning

confidence: 99%

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Long-Term Neuroprotection with 2-Iminobiotin, An Inhibitor of Neuronal and Inducible Nitric Oxide Synthase, after Cerebral Hypoxia-Ischemia in Neonatal Rats

Tweel

Bel

Kavelaars

et al. 2005

J Cereb Blood Flow Metab

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The short-and long-term neuroprotective effects of 2-iminobiotin, a selective inhibitor of neuronal and inducible nitric oxide synthase, were studied in 12-day-old rats following hypoxia-ischemia. Hypoxia-ischemia was induced by occlusion of the right carotid artery followed by 90 minutes of hypoxia (FiO 2 0.08). Immediately on reoxygenation, 12 and 24 hours later the rats were treated with vehicle or 2-iminobiotin at a dose of 5.5, 10, 30, or 60 mg/kg per day. Histologic analysis of brain damage was performed at 6 weeks after hypoxia-ischemia. To assess early changes of cerebral tissue, levels of HSP70, nitrotyrosine, and cytochrome c were determined 24 hours after reoxygenation. Significant neuroprotection was obtained using a dose of 30 mg/kg per day of 2-iminobiotin. Levels of HSP70 were increased in the ipsilateral hemisphere in both groups (Po0.05), but the increase was significantly (Po0.05) less in the rats receiving the optimal dose of 2-iminobiotin (30 mg/kg per day). Hypoxia-ischemia did not lead to increased levels of nitrotyrosine, nor did 2-iminobiotin influence levels of nitrotyrosine. In contrast, hypoxia-ischemia induced an increase in cytochrome c level that was prevented by 2-iminobiotin. In conclusion, 2-iminobiotin administered after hypoxia-ischemia provides long-term neuroprotection. This neuroprotection is obtained by mechanisms other than a reduction of nitrotyrosine formation in proteins.

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Nitric oxide system alteration at spinal cord as a result of perinatal asphyxia is involved in behavioral disabilities: Hypothermia as preventive treatment

Dorfman

Rey-Funes

Bayona

et al. 2008

J of Neuroscience Research

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Perinatal asphyxia (PA) is able to induce sequelae such as spinal spasticity. Previously, we demonstrated hypothermia as a neuroprotective treatment against cell degeneration triggered by increased nitric oxide (NO) release. Because spinal motoneurons are implicated in spasticity, our aim was to analyze the involvement of NO system at cervical and lumbar motoneurons after PA as well as the application of hypothermia as treatment. PA was performed by immersion of both uterine horns containing full-term fetuses in a water bath at 37 degrees C for 19 or 20 min (PA19 or PA20) or at 15 degrees C for 20 min (hypothermia during PA-HYP). Some randomly chosen PA20 rats were immediately exposed for 5 min over grain ice (hypothermia after PA-HPA). Full-term vaginally delivered rats were used as control (CTL). We analyzed NO synthase (NOS) activity, expression and localization by nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-d) reactivity, inducible and neuronal NOS (iNOS and nNOS) by immunohistochemistry, and protein nitrotyrosilation state. We observed an increased NOS activity at cervical spinal cord of 60-day-old PA20 rats, with increased NADPH-d, iNOS, and nitrotyrosine expression in cervical motoneurons and increased NADPH-d in neurons of layer X. Lumbar neurons were not altered. Hypothermia was able to maintain CTL values. Also, we observed decreased forelimb motor potency in the PA20 group, which could be attributed to changes at cervical motoneurons. This study shows that PA can induce spasticity produced by alterations in the NO system of the cervical spinal cord. Moreover, this situation can be prevented by perinatal hypothermia.

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Inhibition of nNOS and iNOS following Hypoxia-Ischaemia Improves Long-Term Outcome but Does Not Influence the Inflammatory Response in the Neonatal Rat Brain

Cited by 36 publications

References 37 publications

Hypoxia/Ischemia Modulates G Protein–Coupled Receptor Kinase 2 and β-Arrestin-1 Levels in the Neonatal Rat Brain

Hypoxia/Ischemia Modulates G Protein–Coupled Receptor Kinase 2 and β-Arrestin-1 Levels in the Neonatal Rat Brain

Long-Term Neuroprotection with 2-Iminobiotin, An Inhibitor of Neuronal and Inducible Nitric Oxide Synthase, after Cerebral Hypoxia-Ischemia in Neonatal Rats

Nitric oxide system alteration at spinal cord as a result of perinatal asphyxia is involved in behavioral disabilities: Hypothermia as preventive treatment

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