Regulator of calcineurin 1 (RCAN1) can be induced by an intracellular calcium increase and oxidative stress, which are characteristic features of temporal lobe epilepsy. Thus, we investigated the spatiotemporal expression and cellular localization of RCAN1 protein and mRNA in the mouse hippocampus after pilocarpine-induced status epilepticus (SE). Male C57BL/6 mice were given pilocarpine hydrochloride (280 mg/kg, i.p.) and allowed to develop 2 h of SE. Then the animals were given diazepam (10 mg/kg, i.p.) to stop the seizures and sacrificed at 1, 3, 7, 14, or 28 day after SE. Cresyl violet staining showed that pilocarpine-induced SE resulted in cell death in the CA1 and CA3 subfields of the hippocampus from 3 day after SE. RCAN1 immunoreactivity showed that RCAN1 was mainly expressed in neurons in the shammanipulated hippocampi. At 1 day after SE, RCAN1 expression became detected in hippocampal neuropils. However, RCAN1 signals were markedly enhanced in cells with stellate morphology at 3 and 7 day after SE, which were confirmed to be reactive astrocytes, but not microglia by double immunofluorescence. In addition, real-time reverse transcriptase–polymerase chain reaction showed a significant upregulation of RCAN1 isoform 4 (RCAN1-4) mRNA in the SE-induced hippocampi. Finally, in situ hybridization with immunohistochemistry revealed astrocytic expression of RCAN1-4 after SE. These results demonstrate astrocytic upregulation of RCAN1 and RCAN1-4 in the mouse hippocampus in the acute and subacute phases of epileptogenesis, providing foundational information for the potential role of RCAN1 in reactive astrocytes during epileptogenesis.
Animals ; Astrocytes ; Calcineurin ; Calcium ; Cell Death ; Diazepam ; Epilepsy ; Epilepsy, Temporal Lobe ; Fluorescent Antibody Technique ; Hippocampus ; Humans ; Immunohistochemistry ; In Situ Hybridization ; Male ; Mice ; Microglia ; Neurons ; Neuropil ; Oxidative Stress ; Pilocarpine ; RNA, Messenger ; Seizures ; Status Epilepticus ; Up-Regulation ; Viola

Chronic traumatic encephalopathy (CTE) is a distinct neurodegenerative disease that associated with repetitive head trauma. CTE is neuropathologically defined by the perivascular accumulation of abnormally phosphorylated tau protein in the depths of the sulci in the cerebral cortices. In advanced CTE, hyperphosphorylated tau protein deposits are found in widespread regions of brain, however the mechanisms of the progressive neurodegeneration in CTE are not fully understood. In order to identify which proteomic signatures are associated with CTE, we prepared RIPA-soluble fractions and performed quantitative proteomic analysis of postmortem brain tissue from individuals neuropathologically diagnosed with CTE. We found that axonal guidance signaling pathwayrelated proteins were most significantly decreased in CTE. Immunohistochemistry and Western blot analysis showed that axonal signaling pathway-related proteins were down regulated in neurons and oligodendrocytes and neuron-specific cytoskeletal proteins such as TUBB3 and CFL1 were reduced in the neuropils and cell body in CTE. Moreover, oligodendrocyte-specific proteins such as MAG and TUBB4 were decreased in the neuropils in both gray matter and white matter in CTE, which correlated with the degree of axonal injury and degeneration. Our findings indicate that deregulation of axonal guidance proteins in neurons and oligodendrocytes is associated with the neuropathology in CTE. Together, altered axonal guidance proteins may be potential pathological markers for CTE.
Axons ; Blotting, Western ; Brain Injury, Chronic ; Brain ; Cell Body ; Cerebral Cortex ; Craniocerebral Trauma ; Cytoskeletal Proteins ; Gray Matter ; Humans ; Immunohistochemistry ; Neurodegenerative Diseases ; Neurons ; Neuropathology ; Neuropil ; Oligodendroglia ; tau Proteins ; White Matter

The circling mice with tmie gene mutation are known as an animal deafness model, which showed hyperactive circling movement. Recently, the reinvestigation of circling mouse was performed to check the inner ear pathology as a main lesion of early hearing loss. In this trial, the inner ear organs were not so damaged to cause the hearing deficit of circling (cir/cir) mouse at 18 postnatal day (P18) though auditory brainstem response data indicated hearing loss of cir/cir mice at P18. Thus, another mechanism may be correlated with the early hearing loss of cir/cir mice at P18. Hearing loss in the early life can disrupt the ascending and descending information to inferior colliculus (IC) as integration site. There were many reports that hearing loss could result in the changes in Ca²⁺ concentration by either cochlear ablation or genetic defect. However, little was known to be reported about the correlation between the pathology of IC and Ca²⁺ changes in circling mice. Therefore, the present study investigated the distribution of calcium-binding proteins (CaBPs), calbindin-D28k, parvalbumin, and calretinin immunoreactivity (IR) in the IC to compare among wild-type (+/+), heterozygous (+/cir), and homozygous (cir/cir) mice by immunohistochemistry. The decreases of CaBPs IR in cir/cir were statistically significant in the neurons as well as neuropil of IC. Thus, this study proposed overall distributional alteration of CaBPs IR in the IC caused by early hearing defect and might be helpful to elucidate the pathology of central auditory disorder related with Ca²⁺ metabolism.
Animals ; Calbindin 1* ; Calbindin 2* ; Calcium-Binding Proteins ; Deafness ; Ear, Inner ; Evoked Potentials, Auditory, Brain Stem ; Hearing ; Hearing Loss ; Immunohistochemistry ; Inferior Colliculi* ; Metabolism ; Mice* ; Neurons ; Neuropil ; Parvalbumins ; Pathology

Here we present an autopsy case of chronic traumatic encephalopathy (CTE) in a 36-year-old man. He had a history of febrile seizures at the age of four and was severely demented at age 10 when he was admitted to a mental hospital. He had suffered repetitive self-harm, such as frequent banging of the head on the wall in his hospital record, but he had no clear history between the ages of four and ten. Autopsy revealed global cerebral atrophy, including the basal ganglia, thalamus, hippocampus, amygdala, mammilary bodies and lateral geniculate bodies. This case showed typical pathological features of CTE. Phosphorylated tau (p-tau)-positive neurofibrillary tangles (NFTs) and neuropil threads (NT) we are widely distributed in the brain, especially in the depth of the cerebral sulci. NFT and NT were also found in the basal ganglia, thalamus, amygdala and brainstem. Scanty β-amyloid deposits were found in the motor and sensory cortices, but α-synuclein was completely negative in the brain. This example showed that CTE can occur in young ages and that even children can experience CTE dementia.
Adult ; Amygdala ; Atrophy ; Autopsy* ; Basal Ganglia ; Brain ; Brain Injuries ; Brain Injury, Chronic* ; Brain Stem ; Child* ; Dementia ; Geniculate Bodies ; Head ; Hippocampus ; Hospital Records ; Hospitals, Psychiatric ; Humans ; Neurofibrillary Tangles ; Neuropil Threads ; Pathology ; Seizures, Febrile ; Thalamus

OBJECTIVETo investigate the clinicopathologic features and 19q13.42 gene changes in embryonal tumors with multilayered rosettes (ETMR).

METHODSImmunohistochemistry and fluorescence in situ hybridization (FISH) were performed in three ETMRs.

RESULTSThe average age of the patients were 34 months. Imaging revealed huge masses with inhomogeneous enhancement and two cases showed cystic lesions. Follow-up data showed 14 and 38 months survival in two children, the third had a recurrence 4 months after operation. Morphologically, the tumor was mainly composed of dense small primitive neuroepithelial cells in patchy or multilayer rosettes within a background of advanced neuronal differentiation, containing neurocytes, ganglion cells, and neuropil-like background. Immunohistochemical staining showed the neuronal marker, synaptophysin, was positive in differentiated areas. Nestin as a neural stem cell marker was immunoreactive in the primitive neuroepithelial cells including multilayered rosettes. Neurons with positive expression of NeuN were observed occasionally. Ki-67 index was up to 40%-80% in the undifferentiated cells and rosettes, but was only 1%-3% in the differentiated areas. CD99 was positive in perivascular papillary pattern areas in one case. 19q13.42 amplification was detected in more than 30% of tumor cells in all cases.

CONCLUSIONSETMR is a unique entity with distinctive clinical and pathological features. Chromosome 19q13.42 abnormality is valuable for confirming the diagnosis and for further treatment research.

Antigens, Nuclear ; genetics ; Child, Preschool ; Chromosomes, Human, Pair 19 ; genetics ; Genetic Testing ; Humans ; Immunohistochemistry ; In Situ Hybridization, Fluorescence ; Neoplasm Recurrence, Local ; Neoplasms, Germ Cell and Embryonal ; genetics ; pathology ; Nerve Tissue Proteins ; genetics ; Neuropil ; pathology ; Synaptophysin ; genetics

Ischemic stroke is the most common type of stroke in humans. The purpose of this study was to evaluate the diagnostic value of magnetic resonance imaging (MRI) in a canine model of stroke. Ischemic stroke was induced by using prepared autologous thrombus. The dogs were placed in lateral recumbency on the operation table and the cervical area of each dog was sterilized by using alcohol. After making a cervical incision, the common carotid artery and internal carotid artery (a branch of the common carotid artery that supplies an anterior part of the brain) were exposed. A 200 microL injection of the autologous thrombus prepared 24 hr prior to surgery was delivered with a 20 gauge venous catheter through an internal carotid artery. After successful delivery of the autologous thrombus, the venous catheter was removed, and the cervical incision was sutured. Neurologic signs including generalized seizures, tetraparesis, and altered mental status, were observed in all 3 dogs after induction of ischemic stroke and the signs manifested immediately after awakening from anesthesia. T1- and T2-weighted images and fluid-attenuated inversion recovery (FLAIR) images of the brain were acquired 1 day before and 1 day after surgery. On the day following ischemic stroke induction, MRI revealed multifocal lesions in the cerebral cortex and subcortex such as T1 hypointensity, T2 hyperintensity, FLAIR hyperintensity, and diffusion-weighted hyperintensity in all 3 dogs. Upon postmortem examination, ischemic lesions were found to be consistent with the MRI findings and they were unstained with 2% triphenyltetrazolium chloride. Histologic features of the earliest neuronal changes such as cytoplasmic eosinophilia with pyknotic nuclei were identified. Neuropil spongiosis and perivascular cuffing were also prominently observed at the infarcted area. The present study demonstrated the features of MRI and histopathologic findings in canine ischemic stroke models.
Anesthesia ; Animals ; Autopsy ; Brain ; Carotid Artery, Common ; Carotid Artery, Internal ; Catheters ; Cerebral Cortex ; Cytoplasm ; Dogs* ; Eosinophilia ; Equipment and Supplies ; Humans ; Magnetic Resonance Imaging* ; Neurologic Manifestations ; Neurons ; Neuropil ; Operating Tables ; Seizures ; Stroke* ; Thrombosis*

Neuroblastoma is a malignant tumor of primordial neural crest origin. It usually develops along the sympathetic nervous system, such as the adrenal glands or paramedian sympathetic chain and metastasizes to the liver most frequently. However, a primary hepatic neuroblastoma has not been reported yet. Here, we report a case of 29-year-old woman who presented with a solitary hepatic mass. Grossly, the mass was large, creamy, rubbery firm, and showed focal hemorrhage and central cavitation. Microscopically, the tumor cells were arranged in small nests of spindle to ovoid cells with abundant neuropil. The neuroblastic nature of the tumor was confirmed by immunohistochemistry and electron microscopy. No extrahepatic mass was found, despite a thorough systemic survey such as chest and abdominopelvic computed tomography (CT) scans and a whole body positron emission tomography-CT study. To the best of our knowledge, this is the first report of a bona fide primary hepatic neuroblastoma.
Adrenal Glands ; Adult ; Electrons ; Female ; Hemorrhage ; Humans ; Immunohistochemistry ; Liver ; Liver Neoplasms ; Microscopy, Electron ; Neural Crest ; Neuroblastoma ; Neuropil ; Sympathetic Nervous System ; Thorax

The major defining pathological hallmarks of Alzheimer's disease (AD) are the accumulations of Abeta in senile plaques and hyperphosphorylated tau in neurofibrillary tangles and neuropil threads. Recent studies indicate that rather than these insoluble lesions, the soluble Abeta oligomers and hyperphosphorylated tau are the toxic agents of AD pathology. Such pathological protein species are accompanied by cytoskeletal changes, mitochondrial dysfunction, Ca2+ dysregulation, and oxidative stress. In this review, we discuss how the binding of Abeta to various integrins, defects in downstream focal adhesion signaling, and activation of cofilin can impact mitochondrial dysfunction, cytoskeletal changes, and tau pathology induced by Abeta oligomers. Such pathological consequences can also feedback to further activate cofilin to promote cofilin pathology. We also suggest that the mechanism of Abeta generation by the endocytosis of APP is mechanistically linked with perturbations in integrin-based focal adhesion signaling, as APP, LRP, and beta-integrins are physically associated with each other.
Alzheimer Disease ; Amyloid ; Cytoskeleton ; Endocytosis ; Focal Adhesions ; Integrins ; Mitochondria ; Neurofibrillary Tangles ; Neuropil Threads ; Oxidative Stress ; Plaque, Amyloid

Aged ; Aged, 80 and over ; Alzheimer Disease ; metabolism ; pathology ; Brain ; metabolism ; Dentate Gyrus ; metabolism ; Dynamin I ; metabolism ; Hippocampus ; metabolism ; Humans ; Monomeric Clathrin Assembly Proteins ; metabolism ; Neuropil ; metabolism ; Synaptophysin ; metabolism

Channels formed by the transient receptor potential (TRP) family of proteins have a variety of physiological functions. In the present study, we examined the localization of canonical transient receptor potential channels (TRPCs) in rat cerebellum. Twelve adult (4~6 month old) Sprague-Dawley rats were examined in this study. We performed immunohistochemistry using specific antibodies against TRPCs to investigate the detailed and comparative distribution of six TRPCs in rat cerebellum. There was a high density of TRPC1, TRPC3, TRPC4, TRPC5 and TRPC7, with a much lower density of TRPC6 in the rat cerebellar cortex. The somatodendritic Purkinje cell areas were intensely stained with antiTRPC3, TRPC4, TRPC5 or TRPC7 antibodies, whereas the staining intensity of TRPC6 was relatively low in the Purkinje cell bodies. In the cerebellar nuclei, the cell bodies of cerebellar output neurons showed moderate TRPC1, TRPC3, TRPC5 and TRPC7 immunoreactivities, while TRPC6 immunoreactivity was observed in the surrounding neuropil. This study showing the differential localizations of TRPC channels in the cerebellum may provide useful data for the future investigations on the structural and functional properties of TRPCs.
Adult ; Animals ; Antibodies ; Cerebellar Cortex ; Cerebellar Nuclei ; Cerebellum ; Humans ; Immunohistochemistry ; Neurons ; Neuropil ; Proteins ; Rats ; Rats, Sprague-Dawley ; Transient Receptor Potential Channels