Roles of glutamate transporter and receptors, poly (ADPribose) polymerase, and transforming growth factor-beta1 in pontosubicular neuron necrosis.

The expression of neuron-type glutamate transporters (EAAC-1), AMPA glutamate receptor subunits (GluR1 and GluR2/3), polyadenosine (5'diphosphate-ribose) polymerase (PARP), and transforming growth factor-beta1 was investigated in 20 cases of neonatal pontosubicular neuron necrosis and 12 gestational-age matched controls. Developmental immunoreactivities of EAAC-1, GluR1, and GluR2/3 appeared in the neurons of the pontine nuclei at 29 to 30 weeks' gestation in controls, and then gradually increased with age. However, these activities were decreased in the pontine nucleus of patients with pontosubicular neuron necrosis. Decreases in these immunoreactivities might indicate early degeneration of neurons. Although PARP and transforming growth factor-beta1 immunoreactivity was insignificant or very weak in the pontine nuclei at any age in controls, PARP was markedly expressed in karyorrhectic neurons of the pontine nucleus in patients with pontosubicular neuron necrosis. Transforming growth factor-beta1 immunoreactivity was observed in nonkaryorrhectic neurons of the pontine nuclei. PARP could contribute to the pathogenesis of pontosubicular neuron necrosis more than EAAC-1 or GluR1 or GluR2/3. Transforming growth factor-beta1 could play a role in the protection and repair of damaged neurons.

Dopamine transporter and nitric oxide synthase in hypoxic-ischemic brain.

Changes in dopamine transporter and neuronal nitric oxide synthase (nNOS) were investigated by immunohistochemistry in 18 cases of hypoxic-ischemic basal ganglia necrosis. Neuropil dopamine transporter immunostaining in the striatum was increased in seven cases, with relatively mild basal ganglia necrosis, and decreased in four cases, with marked basal ganglia necrosis, compared with age-matched control subjects. Correspondingly, some striatal neurons had increased immunoreactivity to dopamine transporter in the cases of increased immunostaining in the neuropil. nNOS-positive neurons did not obviously change in cases of basal ganglia necrosis within 2 days after birth and then decreased or were not detectable in cases of basal ganglia necrosis at more than 3 days after birth. The results suggest that the synthesis of dopamine transporter is up-regulated in relatively mild basal ganglia necrosis to compensate for the uptake of increased dopamine, that this compensative ability is lost in marked basal ganglia necrosis, and that nNOS-containing neurons in the striatum are relatively resistant to hypoxic ischemia. We speculate that glutamate excitotoxicity mediated by glutamate receptors 1, 2/3, and 4 and excessive dopaminergic excitatory activity may play important roles in hypoxic-ischemic basal ganglia necrosis and that nNOS does not contribute to that condition.

Developmental and age-related changes of dopamine transporter, and dopamine D1 and D2 receptors in human basal ganglia.

The developmental and age-related changes of the dopamine transporter (DAT), and the dopamine D1 and D2 receptor (D1R and D2R) subtypes were investigated in basal ganglia (BG) of human brain. DAT immunostaining was mainly observed in the neuropil, neurons, and glia of the striatum. The DAT-positive neuropil was detectable at 32 GW, a peak being reached at 9-10 years of age, with a decrease to 50-63 years of age. The developmental pattern of DAT immunoreactivity in neuron was similar to that of the neuropil. DAT-positive glia were observed in the BG at 32 GW, which increased slightly at 38-40 GW, and then did not obviously change until 6-8 months after birth. D2R-positive neurons were clearly observed at 19 GW, a peak being reached at 32 GW and 1-3 months of age in the globus pallidus and striatum, respectively, with a decrease after 9-10 years of age. D1R was expressed as early as D2R, but decreased after 6-8 months. Our results suggest that D1R and D2R expression is an intrinsic property of striatal neurons and is independent of dopaminergic innervation. D1R may play a more important role in neuronal maturation of the BG than D2R. D2R may be closely correlated with late neuronal development. The higher expression of DAT during adolescence may be related to function of the BG which learns complex behavioral patterns. The significance of the age-related decreases in DAT, D1R and D2R in the BG remains to be further investigated.

Expression of transforming growth factor-beta 1 in periventricular leukomalacia.

The expression of transforming growth factor-beta 1, which has neurotrophic effects, was investigated in 25 neonates with periventricular leukomalacia using immunohistochemistry. In controls, transforming growth factor-beta 1 immunoreactivity was not detected in the cerebral white matter or cortex. Of the 25 cases of periventricular leukomalacia, transforming growth factor-beta 1 immunoreactivity was found in 16, and was distributed mainly in the cytoplasm of astrocytes, being prominent around necrotic foci in the white matter. The immunoreactivity was negative or weak at the acute stage of periventricular leukomalacia, and increased at the subacute stage and then decreased or was absent at the chronic stage. Astrocytes that were moderately or markedly positive for transforming growth factor-beta 1 were not found before 27 weeks' gestation, but were observed after 32 weeks' gestation in the white matter of the brains of neonates with periventricular leukomalacia. Transforming growth factor-beta 1 expression tended to be more obvious in focal periventricular leukomalacia than in widespread or diffuse periventricular leukomalacia. Our results suggest that transforming growth factor-beta 1 could be involved in the delayed glial response rather than the initial glial activation, and could play a role in the inhibition and repair of injury in periventricular leukomalacia. Exogenous transforming growth factor-beta 1 could have therapeutic potential for periventricular leukomalacia.

Developmental expression of monocyte chemoattractant protein-1 in the human cerebellum and brainstem.

The developmental expression of monocyte chemoattractant protein-1 (MCP-1) in the cerebellum, medulla oblongata and pons was investigated in 26 normal human brains, ranging from 20 weeks of gestation (GW) to adulthood by means of an immunohistochemical method. Immunoreactivity to MCP-1 was observed in neurons of the cerebellum and brainstem, and was mainly distributed in the cytoplasm and dendrites. MCP-1-positive Purkinje cells appeared at 27 GW, reached a peak at 36 GW, and then decreased after 1 month of age, almost completely disappearing by 1-2 years of age. MCP-1-reactive neurons in the dentate nucleus and inferior olivary nucleus showed temporal increases similar to that of Purkinje cells. In the pons, however, MCP-1 reactivity was low in neurons of the pontine nuclei persisting from the fetal to the adult period, and was very low and short in the reticular formation and cranial nerve nuclei during the fetal and/or neonatal period. MCP-1 Western blotting of the cerebellar cortex confirmed the specificity of the immunohistochemistry. Our results suggest that MCP-1 may be related to the maturation of Purkinje cells, the dentate nucleus, the inferior olivary nucleus, and their network, promoting the growth of dendrites and synapses. Furthermore, MCP-1 may also be useful for the study of abnormal neuron development and ischemic damage.

A developmental study of the dopamine D2R receptors in the human basal ganglia and thalamus.

The development of the dopamine D2R receptors (D2R) in the human basal ganglia (BG) and thalamus was investigated in 25 normal brains by means of an immunohistochemical method and Western blotting. Immunoreactivity to D2R was detected in the cytoplasm and dendrites of small and large neurons in the BG and thalamus. D2R-positive neurons were clearly observed at 19 weeks of gestation (GW) in the globus pallidus and thalamus, and at 21 GW in the striatum. The number of D2R-positive neurons gradually increased and reached a peak at 27 GW in the globus pallidus, at 39 GW in the thalamus, and at 1 month of age in the striatum. The number of D2R-positive large neurons in the globus pallidus and small neurons in the striatum decreased after 1 year and about 10 years of age, respectively. Western blotting confirmed the specificity of the immunohistochemistry. Our results suggest that the D2R protein begins to be synthesized at an early fetal stage in the BG and thalamus, and the development of D2R is mostly consistent with neuronal maturation in the BG. D2R may play an important role in regulating the neuronal development of the BG. The decrease in D2R-positive neurons may be related to D2R post-transcriptional regulation.

Neuropathological characteristics and alteration of the dopamine D2 receptor in hypoxic-ischemic basal ganglia necrosis.

The neuropathological characteristics and alteration of the dopamine D2 receptor (D2R) were investigated in 27 cases of hypoxic-ischemic basal ganglia necrosis (BGN) by means of neuropathological and immunohistochemical methods. Perinatal hypoxic-ischemic BGN manifested neuronal karyorrhexis as well as eosinophilia, karyorrhexis being more predominant in preterm infants and eosinophilia more predominant in full-term infants. Immunoreactivity to D2R was detected in the cytoplasm and dendrites of small and large neurons in the basal ganglia, and increased with neuronal maturation during the late gestational period in normal human basal ganglia. The number of D2R-positive neurons was smaller in all cases of acute BGN than that in controls, the areas of decreased D2R-positive neurons corresponding to the damaged regions observed on HE staining. Furthermore, neurons showed high expression of D2R in a few cases of remote BGN, suggesting some plasticity as to the recovery of D2R. Thus, the neuropathological characteristics of perinatal hypoxic-ischemic BGN may be related to neuronal maturation during different developmental stages in each region, and D2R development may play a role in the basal ganglia vulnerability to hypoxic-ischemia.

Early detection of axonal and neuronal lesions in prenatal-onset periventricular leukomalacia.

The expression of beta-amyloid precursor protein (beta-APP) immunoreactivity was investigated in 16 cases of prenatal-onset periventricular leukomalacia (PVL). beta-APP positive axons were found in the early stage of prenatal PVL, which included coagulation necrosis, microglial activation, axonal swelling or astrogliosis, but were not detectable in the late stage of prenatal PVL. Furthermore, beta-APP immunoreactive neurons were also observed in the fifth layer of pyramidal neurons of the cerebral cortex, corresponding to the beta-APP positive axons in PVL. Thus, beta-APP is detected as an early sign of axonal and neuronal lesions in prenatal-onset PVL, and neuronal beta-APP in the cerebral cortex may function to repair cell damage. In addition, prenatal PVL occurred at various stages before birth.

Changes in AMPA glutamate and dopamine D2 receptors in hypoxic-ischemic basal ganglia necrosis.

Changes in the AMPA glutamate receptor subunits (GluR1, 2-3, and 4) and dopamine D2 receptor (D2R) were investigated in 16 cases of hypoxic-ischemic basal ganglia necrosis (BGN) by immunohistochemistry. Immunoreactivity to GluR1, 2-3, and 4 was observed in the cytoplasm and dendrites of small and large neurons in the basal ganglia. Neuronal immunoreactivity to GluR1, 2-3, and 4 was decreased in cases with acute BGN as compared with that in age-matched controls, the areas of decreased immunoreactivity corresponding to the damaged regions observed on hematoxylin and eosin staining. Glia in the basal ganglia exhibited immunoreactivity to GluR4 in 4 patients with acute BGN, 3 of the 4 surviving for 12 to 35 days. In addition, neuronal immunoreactivity to D2R was also decreased in cases of acute BGN, the decrease being similar to that of GluR1, 2-3, and 4. Our results suggest that excitotoxicity mediated by GluR1, 2-3, and 4 is involved in the pathogenesis of hypoxic-ischemic neuronal damage, and that GluR4 expressed in glia of the BG in the late stage of hypoxic-ischemic injury may participate in the delayed and long-term response of the glia to injury. The decrease in neuronal D2R may be related to downregulated synthesis of the D2R protein induced by the decrease in GluR1-4 in the basal ganglia.

A developmental expression of AMPA-selective glutamate receptor subunits in human basal ganglia.

The development of AMPA-selective glutamate receptors (GluR1 and GluR2-3) in human basal ganglia (BG) was investigated in 23 normal brains by means of an immunohistochemical method. Immunoreactivity to GluR1 and 2-3 was detected in the cytoplasm and dendrites of small and large neurons in the BG. GluR2-3 immunoreactivity-positive neurons were clearly observed at 23-24 gestational weeks (GW) in the globus pallidus and at 32 GW in the neostriatum, and reached a peak at 32 GW and 39 GW, respectively. GluR2-3 positive neurons in the BG began to decrease at 1-4 months of age, reaching the low level of adults by 7 months of age. The developmental pattern of GluR1 was similar to that of GluR2-3 in the BG, but the immunoreactivity to GluR1 was a little weaker than that of GluR2-3 in the neostriatum. Furthermore, GluR1 and 2-3 subunit Western blotting confirmed the specificity of the immunohistochemistry. Our results suggest that the development of GluR1 and 2-3 in the BG is consistent with neuronal development in the BG, which supports further that GluR1 and 2-3 play an important role in neuronal differentiation and maturation of the BG.