Skip to main content
Article thumbnail
Location of Repository

Prenatal Hypoxic-Ischemic Insult Changes the Distribution and Number of NADPH-Diaphorase Cells in the Cerebellum

By Tiago Savignon, Everton Costa, Frank Tenorio, Alex C. Manhães and Penha C. Barradas


Astrogliosis, oligodendroglial death and motor deficits have been observed in the offspring of female rats that had their uterine arteries clamped at the 18th gestational day. Since nitric oxide has important roles in several inflammatory and developmental events, here we evaluated NADPH-diaphorase (NADPH-d) distribution in the cerebellum of rats submitted to this hypoxia-ischemia (HI) model. At postnatal (P) day 9, Purkinje cells of SHAM and non-manipulated (NM) animals showed NADPH-d+ labeling both in the cell body and dendritic arborization in folia 1 to 8, while HI animals presented a weaker labeling in both cellular structures. NADPH-d+ labeling in the molecular (ML), and in both the external and internal granular layer, was unaffected by HI at this age. At P23, labeling in Purkinje cells was absent in all three groups. Ectopic NADPH-d+ cells in the ML of folia 1 to 4 and folium 10 were present exclusively in HI animals. This labeling pattern was maintained up to P90 in folium 10. In the cerebellar white matter (WM), at P9 and P23, microglial (ED1+) NADPH-d+ cells, were observed in all groups. At P23, only HI animals presented NADPH-d labeling in the cell body and processes of reactive astrocytes (GFAP+). At P9 and P23, the number of NADPH-d+ cells in the WM was higher in HI animals than in SHAM and NM ones. At P45 and at P90 no NADPH-d+ cells were observed in the WM of the three groups. Our results indicate that HI insults lead to long-lasting alterations in nitric oxide synthase expression in the cerebellum. Such alterations in cerebellar differentiation might explain, at least in part, the motor deficits that are commonly observed in this model

Topics: Research Article
Publisher: Public Library of Science
OAI identifier:
Provided by: PubMed Central
Download PDF:
Sorry, we are unable to provide the full text but you may find it at the following location(s):
  • http://www.pubmedcentral.nih.g... (external link)
  • Suggested articles


    1. (2002b) Nitric oxide-mediated expression of Bax protein and DNA fragmentation during hypoxia in neuronal nuclei from newborn piglets.
    2. (2001). A nitric oxide donor induces neurogenesis and reduces functional deficits after stroke in rats.
    3. (1971). A theory of cerebelar function.
    4. (1969). A theory of cerebellar cortex.
    5. (1999). Almeida A
    6. (1993). Altered distribution of nicotinamide-adenine dinucleotide phosphate-diaphorase cells in frontal lobe of schizophrenics implies disturbances of cortical development.
    7. (2005). Apoptosis and necrosis in developing cerebellum and brainstem induced after focal cerebral hypoxic-ischemic injury.
    8. (1990). Apparent hydroxyl radical production by peroxynitrite: implications for endothelial injury from nitric oxide and superoxide.
    9. (1969). Autoradiographic and histological studies of postnatal neurogenesis. 3. Dating the time of production and onset of differentiation of cerebellar microneurons in rats.
    10. (2009). Brain injury in premature infants: a complex amalgam of destructive and developmental disturbances.
    11. (2011). Cerebellar abnormalities following hypoxia alone compared to hypoxic-ischemic forebrain injury in the developing rat brain.
    12. (2005). Cerebellar injury in the extremely premature infant: newly recognized but relatively common outcome.
    13. (1994). Cerebellar injury induces NOS in Purkinje cells and cerebellar afferent neurons.
    14. Contestabile A (2010) Role of Nitric Oxide in Cerebellar Development and Function: Focus on Granule Neurons. Cerebellum DOI:
    15. (1994). Correlation of nitric oxide synthase expression with changing patterns of axonal projections in the developing visual system.
    16. (2002). Correlation of VEGF and angiopoietin expression with disruption of bloodbrain barrier and angiogenesis after focal cerebral ischemia.
    17. (1997). Delayed reduction of ischemic brain injury and neurological deficits in mice lacking the inducible nitric oxide synthase gene.
    18. (1988). Demonstration and biochemical characterization of rat brain NADPH-dependent diaphorase.
    19. (1988). Development of macroglial cells in rat cerebellum II. An in situ immunohistochemical study of oligodendroglial lineage from precursor to mature myelinating cell.
    20. (2005). Developmental changes induced by graded prenatal systemic hypoxic-ischemic insults in rats.
    21. (1995). Developmental changes of nitric oxide synthase in the rat superior colliculus.
    22. (2004). Developmental lag in superoxide dismutases relative to other antioxidant enzymes in premyelinated human telencephalic white matter.
    23. (1999). Developmental neuropathology and impact of perinatal brain damage. III: gray matter lesions of the neocortex.
    24. (1997). Developmental neuropathology and impact of perinatal brain damage.II: white matter lesions of the neocortex.
    25. (2003). Differences in nitric oxide production: a comparison of retinal ganglion cells and retinal glial cells cultured under hypoxic conditions.
    26. (1992). DNA damage and mutation in human cells exposed to nitric oxide in vitro.
    27. (2009). Does inflammation after stroke affect the developing brain differently than adult brain?
    28. (2006). Early response of neurons and glial cells to hypoxia in the retina.
    29. (2001). Endogenous activation of group-I metabotropic glutamate receptors is required for differentiation and survival of cerebellar Purkinje cells.
    30. (1999). Endogenous nitric oxide synthesis: biological functions and pathophysiology.
    31. (1994). Expression of mouse brain soluble guanylyl cyclase and NO synthase during ontogeny.
    32. (1998). Expression of neuronal nitric oxide synthase during embryonic development of the rat cerebral cortex.
    33. (1998). Eye enucleation alters intracellular distribution of NO synthase in the superior colliculus.
    34. (2006). Free radicals, mitochondria, and hypoxia ischemia in the developing brain.
    35. (1970). How the cerebellum may be used.
    36. (1996). Induction of NADPH-diaphorase/nitric oxide synthase in the brainstem trigeminal system resulting from cerebellar lesions.
    37. (2009). Intrinsic versus extrinsic determinants during the development of Purkinje cell dentrites.
    38. (2004). Maternal hypoxia during pregnancy induces fetal neurodevelopmental brain damage: partial protection by magnesium sulfate.
    39. (2003). Mitochondrial nitric oxide synthase: role in pathophysiology.
    40. (1996). Morphology of NADPH-diaforase positive cells in the retinoceptive layers of the developing rat superior colliculus.
    41. (1993). NADPH-diaphorase histochemistry in the postnatal mouse cerebellum suggests specific developmental functions for nitric oxide.
    42. (1997). NADPH-diaphorase reactivity is reduced in the molecular layer but increased in the granular layer of primate cerebellum after prolonged anaesthesia.
    43. (2003). Neonatal cytokines and cerebral palsy in very preterm infants.
    44. (1993). Neuronal growth cone collapse and inhibition of protein fatty acylation by nitric oxide. Nature 366: 562–565. NADPH-d Distribution after a Prenatal HI
    45. (1991). Neuronal NADPH diaphorase is a nitric oxide synthase.
    46. (2007). Nitric oxide and peroxynitrite inhealth and disease.
    47. (1996). Nitric oxide inhibits creatine kinase and regulates rat heart contractile reserves.
    48. (2000). Nitric oxide synthase expression in ischemic rat retinas.
    49. (2004). Nitric oxide-mediated activation of extracellular signal-regulated kinase (ERK) and c-jun N-terminal kinase (JNK) during hypoxia in cerebral cortical nuclei of newborn piglets.
    50. (2002). Nitric oxide-mediated Ca2+/calmodulin-dependent protein kinase IV activity during hypoxia in neuronal nuclei from newborn piglets.
    51. (1994). Nitric oxyde synthase expression reveals compartments of cerebellar granule cells and suggests a role for mossy fibers in their development.
    52. (2006). Nuclear Ca(++)-influx, Ca (++)/calmodulin-dependent protein kinase IV activity and CREB protein phosphorylation during post-hypoxic reoxygenation in neuronal nuclei of newborn piglets: the role of nitric oxide.
    53. (1993). Parasagittal patches in the granular layer of the developing and adult rat cerebellum as demonstrated by NADPHdiaphorase histochemistry.
    54. (2004). Perinatal hypoxia/ischemia damages and depletes progenitors from the mouse subventricular zone.
    55. (2002). Phosphorylation of cAMP response element binding (CREB) protein during hypoxia in cerebral cortex of newborn piglets and the effect of nitric oxide synthase inhibition.
    56. (1972). Postnatal Development of the Cerebellar Cortex in the Rat I. The Eternal Germinal Layer and the Transitional Molecular Layer.
    57. (2010). Postnatal erythropoietin treatment mitigates neural cell loss after systemic prenatal hypoxic-ischemic injury: Laboratory investigation.
    58. (2001). Predominant role of endothelial nitric oxide synthase in vascular endothelial growth factor induced angiogenesis and vascular permeability.
    59. (1996). Prenatal development of NADPHdiaphorase-reactive neurons in human frontal cortex.
    60. (1991). Presence of nitric oxide synthase activity in the neurons of the rat embryonal cerebrum.
    61. (1993). Preterm birth and cerebral palsy: is tumor necrosis factor the missing link?
    62. (2006). Redox regulation of heat shock protein expression by signaling in volving nitric oxide and carbon monoxide: relevance to brain aging, neurodegenerative disorders and longevity.
    63. (2006). Regional metabolic status of the E–18 rat fetal brain following transient hypoxia/ ischemia.
    64. (1996). Role for nitric oxide in the development of the ferret retinogeniculate projection.
    65. (1994). Roles of endogenous nitric oxide in cerebellar cortical development in slice cultures.
    66. (2003). Signal molecules and receptors in the differential development of cerebellum lobules. Acute effects of cisplatin on nitric oxide and glutamate systems in Purkinje cell population.
    67. (1994). Stamataki A
    68. (1997). Synaptic plasticity, place cells and spatial memory: study with second generation knockouts.
    69. (1994). Temporal profile of ischemic tissue damage, neutrophil response, and vascular plugging following permanent and transient (2H) middle cerebral artery occlusion in the rat.
    70. (1993). The maternal-fetal interaction: some controversies and solutions.
    71. (1990). The NO hypothesis: possible effects of a short-lived, rapidly diffusible signal in the development and function of the nervous system.
    72. (2000). The regulatory connection between the activity of granule cell NMDA receptors and dendritic differentiation of cerebellar Purkinje cells.
    73. (1997). The vulnerability of the fetal sheep brain to hypoxemia at mid-gestation.
    74. (1994). Transient nitric oxide synthase neurons in embryonic cerebral cortical plate, sensory ganglia and olfactory epithelium.
    75. (2000). VEGF enhances angiogenesis and promotes blood-brain barrier leakage in the ischemic brain.

    To submit an update or takedown request for this paper, please submit an Update/Correction/Removal Request.