The present study was aimed to investigate the role of cerebrospinal fluid-contacting nucleus (CSF-CN) neurons in modulation of inflammatory pain and underlying mechanism. The inflammatory pain model was made by subcutaneous injection of the complete Freund's adjuvant (CFA) into the left hind paw of rats. The phosphorylation level of PKC (p-PKC) was examined by Western blot. Thermal withdrawal latency (TWL) of the rats was measured to assess inflammatory pain. The results showed that, compared with the sham controls, the inflammatory pain model rats showed shortened TWL on day 1, 3, and 7 after CFA injection, as well as increased level of p-PKC in CSF-CN neurons at 24 h after CFA injection. The administration of GF109203X, a PKC inhibitor, into lateral ventricle decreased the level of p-PKC protein expression and increased TWL in the model rats. These results suggest that blocking the PKC pathway in CSF-CN neurons may be an effective way to reduce or eliminate the inflammatory pain.
Animals ; Freund's Adjuvant ; Inflammation ; enzymology ; Neurons ; enzymology ; Pain ; enzymology ; Phosphorylation ; Protein Kinase C ; cerebrospinal fluid ; chemistry ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo study the role of neuronal nitric oxide synthase (nNOS) in aged rats' hippocampal delayed neuronal death (DND) following brain ischemia.

METHODSModels of incomplete brain ischemia were induced by clipping common carotid artery. A total of 46 aged SD rats were divided into 8 groups: normal control group (Group A, n=5), sham-operation group (Group B, n=5), reperfusion 1, 6, 12, 24, 48, and 96 hours groups after brain ischemia for 30 minutes (Group C, D, E, F, G, and H, n=6/group). The expression of nNOS was examined by immunohistochemistry and neuronal ultrastructural changes were observed by the transmission electron microscopy (TEM) at different time points after reperfusion.

RESULTSImmunohistochemistry showed that nNOS expression in the hippocampal neurons was high in Group E, low expression in Group D, moderate expression in Group F and G. There was nearly no expression of nNOS in Group A, B, C, and H. Ultrastructure of hippocampal neurons was damaged more severely in reperfusion over 24 hours groups.

CONCLUSIONSNitric oxide (NO) may be one of the important factors in inducing DND after ischemia/reperfusion.

Animals ; Apoptosis ; Brain Ischemia ; enzymology ; Female ; Hippocampus ; enzymology ; pathology ; Immunohistochemistry ; Male ; Microscopy, Electron ; Neurons ; enzymology ; Nitric Oxide Synthase ; metabolism ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; enzymology

Chronic spinal cord compression is the common clinical prognosis with various outcomes, but the affecting factors and mechanisms still remain unexplored. The structure and function of neurovascular unit manifest great significance in the central nervous system diseases. This paper discusses matrix metalloproteinase (MMP) impact on the stability of the neural vascular unit, by directly decomposing extracellular matrix, inducing the glial cell migration, activating angiogenesis, regulating function of blood spinal cord barrier, and put forward the MMP may be the key points in regulation of spinal cord neurovascular unit structure and function change to affect the outcome of chronic oppressive cervical spinal cord.
Cell Movement ; Humans ; Matrix Metalloproteinases ; physiology ; Nerve Compression Syndromes ; diagnosis ; enzymology ; Neurons ; cytology ; Prognosis ; Spinal Cord Injuries ; diagnosis ; enzymology

OBJECTIVETo establish a simple, reproducible, and practical mechanical injury model of hippocampal neurons of Sprague-Dawley rats in vitro.

METHODSHippocampal neurons isolated from 1-2-day old rats were cultured in vitro. Mild, moderate and severe mechanical injuries were delivered to the neurons by syringe needle tearing, respectively. The control neurons were treated identically with the exception of trauma. Cell damage was assessed by measuring the Propidium Iodide (PI) uptaking at different time points (0.5, 1, 6, 12 and 24 hours) after injury. The concentration of neuron specific enolase was also measured at some time points.

RESULTSPathological examination showed that degeneration, degradation and necrosis occurred in the injured cultured neurons. Compared with the control group, the ratio of PI-positive cells in the injured groups increased significantly after 30 minutes of injury (P<0.05). More severe the damage was, more PI-positive neurons were detected. Compared with the control group, the concentration of neuron specific enolase in the injured culture increased significantly after 1 hour of injury (P<0.05).

CONCLUSIONSThe established model of hippocampal neuron injury in vitro can be repeated easily and can simulate the damage mechanism of traumatic brain injury, which can be used in the future research of traumatic brain injury.

Analysis of Variance ; Animals ; Brain Injuries ; enzymology ; pathology ; Equipment Design ; Hippocampus ; enzymology ; injuries ; In Vitro Techniques ; Neurons ; enzymology ; pathology ; Phosphopyruvate Hydratase ; biosynthesis ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Reproducibility of Results

The serine/threonine p21-activated kinases (PAKs), as main effectors of the Rho GTPases Cdc42 and Rac, represent a group of important molecular switches linking the complex cytoskeletal networks to broad neural activity. PAKs show wide expression in the brain, but they differ in specific cell types, brain regions, and developmental stages. PAKs play an essential and differential role in controlling neural cytoskeletal remodeling and are related to the development and fate of neurons as well as the structural and functional plasticity of dendritic spines. PAK-mediated actin signaling and interacting functional networks represent a common pathway frequently affected in multiple neurodevelopmental and neurodegenerative disorders. Considering specific small-molecule agonists and inhibitors for PAKs have been developed in cancer treatment, comprehensive knowledge about the role of PAKs in neural cytoskeletal remodeling will promote our understanding of the complex mechanisms underlying neurological diseases, which may also represent potential therapeutic targets of these diseases.
Animals ; Cytoskeleton/genetics* ; Humans ; Nervous System Diseases/genetics* ; Neurons/enzymology* ; Signal Transduction ; p21-Activated Kinases/metabolism*

The expression of both constitutive and inducible forms of nitric oxide synthase (NOS) was investigated by immunohistochemical staining of formalin-fixed paraffin-embedded sections in normal and Listeria monocytogenes-infected brains of goats. In normal control goats, a small number of neurons showed immunoreactivity of both iNOS and nNOS, and the number of iNOS-positive neurons was higher than the number of nNOS-positive neurons. In natural listeriosis, listeria antigens were easily immunostained in the inflammatory cells of microabscesses. In this lesion, the immunoreactivity of iNOS in neurons was more intense than the control, but nNOS was not. In microabscesses, nNOS was weakly visualized in macrophages and neutrophils, while iNOS was expressed in macrophages, but not in neutrophils. These findings suggest that normal caprine brain cells, including neurons, constitutively express iNOS and nNOS, and the expressions of these molecules is increased in Listeria monocytogenes infections. Furthermore, inflammatory cells, including macrophages, expressing both nNOS and iNOS may play important roles in the pathogenesis of bacterial meningoencephalitis in goat.
Animals ; Brain/cytology/*enzymology ; Glial Fibrillary Acidic Protein/analysis ; Goat Diseases/*enzymology ; Goats ; Immunohistochemistry ; Listeria Infections/enzymology/*veterinary ; Neurons/*enzymology ; Nitric Oxide Synthase/*analysis ; Nitric Oxide Synthase Type I ; Nitric Oxide Synthase Type II

OBJECTIVETo explore action mechanism of acupuncture and moxibustion for Alzheimer's disease (AD) to provide evidence for prevention and treatment with acupuncture and moxibustion on AD in clinic.

METHODSEighty SPF-grade male Wistar rats, (200 +/- 20) g, were randomly divided into a normal group, a sham-operation group, a model group and a treatment group, 20 cases in each one. The model was duplicated with injection of Abeta1-42 in rats' hippocampus. Expect the treatment group, the rest groups were treated with regular feeding after respective intervention. The treatment group was treated with acupuncture and moxibustion at "Baihui" (GV 20) and "Shenshu" (BL 23), once a day, seven days as a treatment course and totally for two courses. There was one day of interval between the courses. The immunohistochemistry and quantitative RT-PCR methods were applied to test level of Abeta-binding alcohol dehydrogense (ABAD) and cytochrome oxidase IV (COX IV) in hippocampal neurons mitochondria.

RESULTSAcupuncture and moxibustion could reduce effectively level of ABAD and improve activity of COX IV in hippocampal neurons mitochondria in the treatment group, which has statistical significance compared with that in the model group (P < 0.01) and no statistical significance compared with that in the normal group and sham-operation group (P > 0.05). This indicated that acupuncture and moxibustion could effectively suppress overexpression of ABAD, improve activity of COX IV and reduce leak of reactive oxygen species, which could improve metabolic disturbance of mitochondria energy to achieve the goal of prevention and treatment of AD.

CONCLUSIONThe prevention and treatment of AD with acupuncture and moxibustion could be related with suppressing overexpression of ABAD and improving activity of COX IV in hippocampal neurons mitochondria to improve mitochondria energy metabolism.

3-Hydroxyacyl CoA Dehydrogenases ; genetics ; metabolism ; Acupuncture Therapy ; Alzheimer Disease ; enzymology ; metabolism ; therapy ; Animals ; Electron Transport Complex IV ; genetics ; metabolism ; Energy Metabolism ; Hippocampus ; cytology ; enzymology ; metabolism ; Humans ; Male ; Mitochondria ; enzymology ; metabolism ; Moxibustion ; Neurons ; enzymology ; metabolism ; Rats ; Rats, Wistar

OBJECTIVE: To study the changes and actions of neuronal constructive nitric oxide synthase(ncNOS) in heroin drug abuse. METHODS: The expression of ncNOS and ncNOS mRNA in neurons of cerebral cortex, periaqueductal gray matter and the ventral tegmental area was observed by immunohistochemistry, in situ hybridization and image analysis technique after heroin dependence and spontaneous withdrawal in rats. RESULTS: The quantity of ncNOS and ncNOS mRNA rised clearly and the number of ncNOS and ncNOS mRNA positive cells increased greatly in heroin dependence and withdrawal. The changes of ncNOS and ncNOS mRNA in spontaneous withdrawal were more clear than ones of dependence. Heroin dependence and withdrawal led to alterations in ncNOS and ncNOS mRNA expression in important regions implicated in the physical tolerance and dependence. CONCLUSION The ncNOS plays an important role in heroin dependence and withdrawal. The ncNOS immunohistochemical changes observed in the present study might be useful for the forensic pathological diagnosis of heroin drug abuse.
Animals ; Cerebral Cortex/enzymology* ; Forensic Medicine ; Heroin Dependence/enzymology* ; Immunohistochemistry ; Male ; Neurons/enzymology* ; Nitric Oxide Synthase/metabolism* ; Nitric Oxide Synthase Type I ; RNA, Messenger/metabolism* ; Rats ; Rats, Sprague-Dawley ; Substance Withdrawal Syndrome/enzymology*

Axotomy of the vagal motor neurons by cervical vagotomy induces NADPH diaphorase staining due to increased nitric oxide synthase expression in both the rat dorsal motor nucleus and nucleus ambiguous; furthermore, cerical vagotomy leads to cell death of the dorsal motor nucleus cells. Subdiaphragmatic vagotomy axotomizes the vagal motor cells further from the brainstem than cervical vagotomy, and cuts the fibers running only to the abdominal viscera. Here we report that subdiaphragmatic vagotomy is sufficient to induce NADPH diaphorase staining in the dorsal motor nucleus but does not induce staining in the nucleus ambiguus. Because the neurons of the dorsal motor nucleus do not undergo cell death after subdiaphragmatic vagotomy and are able to re-enervate the gut, the increased nitric oxide synthase expression after distal axotomy may be related more to regeneration than degeneration.
Animal ; Fourth Ventricle/physiology* ; Fourth Ventricle/enzymology* ; Fourth Ventricle/cytology ; Male ; Motor Neurons/enzymology ; NADPH Dehydrogenase/metabolism* ; Rats ; Rats, Sprague-Dawley ; Vagotomy/methods* ; Vagus Nerve/physiology*

Group totalling 55 young rabbits (both sexes), whose right optic nerves had been severed intraorbitally, were fed for 1 week, 2 weeks, 4 weeks and 8 weeks respectively. The retina of the left eye was used as a control and that of the right eye for the experiment. The histochemical changes of cholinesterase, acid phosphatase and ribonucleic acid in the reitna after to severance of the optic nerve were observed for 8 weeks after section. In the retina of the young rabbit, whose visual connection to the central nervous system was blocked, there was a decreasing specific cholinesterase activity beginning at the 4th week after the section of it. By the 8th week, the enzyme activity in the perikaryon of the ganglion cell and the inner plexiform layer was considerably decreased. Acid phosphatase activity in the young rabbit's retina peaked at the 2nd week, but decreaseed below normal after the 4th week. This rapid decline of acid phosphatase activity was characteristic in the experimental retinae and was in contrast to the rather slow alteration of enzymatic activity in neurons undergoing wallerian degeneration. Pyroninophilic granules contained in neural cytoplasm of the retina were affected by the surgical blocking of the visual connection with the central nervous system. By the 4th week the granules had partially disappeared from the perikaryon of the ganglion cell and from the inner nuclear layer. Consequently, as the result of histochemical studies, firstly it is postulated that the gradual decline of specific cholinesterase activity in the rabbit's retina was closely related to the intraorbital blocking of the optic nerve, and secondly, that the typical degeneration of the ganglion cell in the ganglion cell layer (which was associated with a partial disappearance of the ganglion cell) was related to the changes in the acid phosphatase activity and alteration of the pyroninophilic granules in the retina following optic nerve transection.
Acid Phosphatase/metabolism* ; Animal ; Cholinesterases/metabolism* ; Histocytochemistry ; Nerve Degeneration ; Neurons/enzymology ; Optic Nerve/surgery* ; Rabbits ; Retina/enzymology* ; Substances: ; Cholinesterases ; Acid Phosphatase