This study investigated the effects of Rehmanniae Radix Praeparata on striatal neuronal apoptosis in rats with attention deficit hyperactivity disorder(ADHD) based on the B-cell lymphoma-2(Bcl-2)/Bcl-2-associated X protein(Bax)/caspase-3 signaling pathway. Twenty-four 3-week-old male spontaneously hypertensive rats(SHR) were randomly divided into a model group, a methylphenidate group(2 mg·kg~(-1)·d~(-1)), and a Rehmanniae Radix Praeparata group(2.4 mg·kg~(-1)·d~(-1)). Age-matched male Wistar Kyoto(WKY) rats were used as the normal control group, with 8 rats in each group. The rats were administered by gavage for 28 days. Body weight and food intake were recorded for each group. The open field test and elevated plus maze test were used to assess hyperactivity and impulsive behaviors. Nissl staining was used to detect changes in striatal neurons and Nissl bodies. Terminal deoxynucleotidyl transferase dUTP nick end labeling(TUNEL) fluorescence staining was used to detect striatal cell apoptosis. Western blot was employed to detect the expression levels of Bcl-2, Bax, and caspase-3 proteins in the striatum. The results showed that compared with the model group, Rehmanniae Radix Praeparata significantly reduced the total movement distance, average movement speed, and central area residence time in the open field test, and significantly reduced the ratio of open arm entries, open arm stay time, and head dipping in the elevated plus maze test. Furthermore, it increased the number of Nissl bodies in striatal neurons, significantly downregulated the apoptosis index, significantly increased Bcl-2 protein expression and the Bcl-2/Bax ratio, and reduced Bax and caspase-3 protein expression. In conclusion, Rehmanniae Radix Praeparata can reduce hyperactivity and impulsive behaviors in ADHD rats. Its mechanism may be related to the regulation of the Bcl-2/Bax/caspase-3 signaling pathway in the striatum, enhancing the anti-apoptotic capacity of striatal neurons.
Animals ; Male ; Apoptosis/drug effects* ; Rats ; Drugs, Chinese Herbal/administration & dosage* ; Caspase 3/genetics* ; Proto-Oncogene Proteins c-bcl-2/genetics* ; bcl-2-Associated X Protein/genetics* ; Rehmannia/chemistry* ; Attention Deficit Disorder with Hyperactivity/physiopathology* ; Signal Transduction/drug effects* ; Neurons/cytology* ; Rats, Inbred SHR ; Rats, Inbred WKY ; Humans ; Corpus Striatum/cytology* ; Plant Extracts

The striatum and globus pallidus are principal nuclei of the basal ganglia. Nissl- and acetylcholinesterase-stained sections of the tree shrew brain showed the neuroanatomical features of the caudate nucleus (Cd), internal capsule (ic), putamen (Pu), accumbens, internal globus pallidus, and external globus pallidus. The ic separated the dorsal striatum into the Cd and Pu in the tree shrew, but not in rats and mice. In addition, computer-based 3D images allowed a better understanding of the position and orientation of these structures. These data provided a large-scale atlas of the striatum and globus pallidus in the coronal, sagittal, and horizontal planes, the first detailed distribution of parvalbumin-immunoreactive cells in the tree shrew, and the differences in morphological characteristics and density of parvalbumin-immunoreactive neurons between tree shrew and rat. Our findings support the tree shrew as a potential model for human striatal disorders.
Acetylcholinesterase ; metabolism ; Animals ; Brain Mapping ; Corpus Striatum ; anatomy & histology ; cytology ; metabolism ; Globus Pallidus ; anatomy & histology ; cytology ; metabolism ; Imaging, Three-Dimensional ; Male ; Mice ; Mice, Inbred C57BL ; Models, Neurological ; Neurons ; metabolism ; Parvalbumins ; metabolism ; Rats ; Rats, Sprague-Dawley ; Statistics, Nonparametric ; Tupaiidae ; anatomy & histology

OBJECTIVE: To explore the status of apoptosis in human striatum derived neural stem cells (NSCs) aft er accutase dissociation and passage. METHODS: The NSCs were isolated from fetuses obtained through spontaneous abortion at 13- 18 weeks of pregnancy, which formed neurospheres in vitro. At passages of 3-5, the neurospheres were disassociated into single cell by accutase digestion and then passaged. At 1, 24 and 72 h after passage, the apoptosis of NSCs was measured by several methods, including active caspase-3 or TUNEL staining for fixed cells, Annexin V, Hoechst or PI staining for live cells. RESULTS: At all of the 3 time points, the staining of TUNEL and active caspase-3 overlapped perfectly. The apoptosis rate of NSCs increased significantly from 20%-25% at 1 h to 75%-80% at 24 h after passage (P<0.01). At 72 h, the apoptosis rate was significantly decreased as compared to that at 24 h time point because of the self-renewal and proliferation of survived NSCs (P<0.01). CONCLUSION Many cells in the neurospheres formed by human striatum-derived NSCs underwent apoptosis soon after accutase disassociation. For NSCs cultured in vitro, anti-apoptosis treatments might be a good method to increase the self-renewal and the proliferation of NSCs.
Apoptosis ; Caspase 3 ; metabolism ; Cells, Cultured ; Collagenases ; chemistry ; Corpus Striatum ; cytology ; Female ; Humans ; In Situ Nick-End Labeling ; Neural Stem Cells ; cytology ; Peptide Hydrolases ; chemistry ; Pregnancy

Neuroprotective effect of scorpion venom on Parkinson's disease (PD) has already been reported. The present study was aimed to investigate whether scorpion venom heat resistant peptide (SVHRP) could attenuate ultrastructural abnormalities in mitochondria and oxidative stress in midbrain neurons of early-stage PD model. The early-stage PD model was established by injecting 6-hydroxydopamine (6-OHDA) (20 μg/3 μL normal saline with 0.1% ascorbic acid) into the striatum of Sprague Dawley (SD) rats unilaterally. The rats were intraperitoneally administered with SVHRP (0.05 mg/kg per day) or vehicle (saline) for 1 week. Two weeks after 6-OHDA treatment, the rats received behavior tests for validation of model. Three weeks after 6-OHDA injection, the immunoreactivity of dopaminergic neurons were detected by immunohistochemistry staining, and the ultrastructure of neuronal mitochondria in midbrain was observed by electron microscope. In the meantime, the activities of monoamine oxidase-B (MAO-B), superoxide dismutase (SOD) and content of malondialdehyde (MDA) in the mitochondria of the midbrain neurons, as well as the inhibitory ability of hydroxyl free radical and the antioxidant ability in the serum, were measured by corresponding kits. The results showed that 6-OHDA reduced the optical density of dopaminergic neurons, induced damage of mitochondrial ultrastructure of midbrain neurons, decreased SOD activity, increased MAO-B activity and MDA content, and reduced the antioxidant ability of the serum. SVHRP significantly reversed the previous harmful effects of 6-OHDA in early-stage PD model. These findings indicate that SVHRP may contribute to neuroprotection by preventing biochemical and ultrastructure damage changes which occur during early-stage PD.
Animals ; Antioxidants ; metabolism ; Corpus Striatum ; Disease Models, Animal ; Dopaminergic Neurons ; drug effects ; Malondialdehyde ; metabolism ; Mesencephalon ; cytology ; Mitochondria ; metabolism ; ultrastructure ; Neuroprotective Agents ; pharmacology ; Oxidative Stress ; Oxidopamine ; Parkinson Disease ; drug therapy ; Peptides ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Scorpion Venoms ; pharmacology ; Superoxide Dismutase ; metabolism

OBJECTIVETo investigate the effects of dopamine (DA) and metabolite in striatum of Parkinson's disease (PD) rats treated by bone mesenchymal stem cells (MSCs) modified by plasmid pIRESneo-EGFP-BDNF.

METHODpIRESneo-EGFP-BDNF was transfected to MSCs with electroporation. The rat models of PD were set up by 6-OHDA and then divided into four groups randomly, which were Sham group, PD group, BDNF group. The rotating behavior of rat models induced by apomorphine (APO) intraperitoneally which transplanting bone MSCs or MSCs modified by plasmid pIRESneo-EGFP-BDNF through cerebral lateral ventricle after 2, 4 and 8 weeks. The levels of DA, homovanillic acid (HVA), dihydroxy-phenylacetic acid (DOPAC) were measured by high performance liquid chromatography (HPLC) in striatum of each group.

RESULTSThe rotation numbers (r/min) of MSCs group or BDNF group in the 2nd, 4th and 8th week after transplanting were significantly decreased compared with that of PD group (P < 0.05). Those of BDNF group were specially significant compared with those of MSCs group (P < 0.05). The levels of DA, HVA, DOPAC and the ratios of DA/HVA, DA/DOPAC in stratum after PD rats intervened by transplanting cells through cerebral lateral ventricle after eight weeks were increased significantly in BDNF group or MSCs group while compared with PD group, especially in BDNF group.

CONCLUSIONThe behavior of rat with PD was improved significantly by increasing the levels of DA and decreasing metabolic rate of DA in striatum while transplanting BDNF modified bone MSCs through cerebral lateral ventricle.

Animals ; Brain-Derived Neurotrophic Factor ; genetics ; Corpus Striatum ; metabolism ; Disease Models, Animal ; Dopamine ; metabolism ; Genetic Engineering ; Male ; Mesenchymal Stem Cell Transplantation ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Parkinson Disease ; metabolism ; Plasmids ; Rats ; Rats, Sprague-Dawley

Parkinson's disease (PD) is a progressive neurodegenerative movement disorder characterized by severe loss of substantia nigra dopamine (DA) neurons. The target region of substantia nigra DA neurons is the dorsal striatum. According to the classic model, activation of DA receptors on striatal medium spiny neurons (MSNs) modulates their intrinsic excitability. Activation of D1 receptors makes MSNs in the direct "Go" pathway more excitable, whereas activation of D2 receptors makes MSNs in the indirect "NoGo" pathway less excitable. Therefore increased DA increases the responsiveness of the Go pathway while decreases the responsiveness of the NoGo pathway. Both mechanisms increase motor output. Conversely, diminished DA will favor the inhibitory NoGo pathway. Therefore, DA has direct, "on-line" effect on motor performance. However, in addition to modulating the intrinsic excitability of MSNs "on-line", DA also modulates corticostriatal plasticity, therefore could potentially produce cumulative and long-lasting changes in corticostriatal throughput. Studies in my lab suggest that DA blockade leads to both direct motor performance impairment and D2 receptor dependent NoGo learning ("learned" motor inhibition) that gradually deteriorates motor performance. NoGo learning is experience dependent and task specific. It is different from blocked learning since NoGo learning impairs future performance even after DA is restored. More recent data from my lab suggest that NoGo learning in the absence of DA arises from increased LTP at the indirect pathway corticostriatal synapses and contributes significantly to PD-like motor symptoms. Our data and hypotheses suggest a novel therapeutic strategy for PD that targets directly signaling molecules for corticostriatal plasticity (e.g. the cAMP pathway and downstream signaling molecules) and prevents aberrant plasticity under conditions of DA denervation.
Corpus Striatum ; cytology ; Dopamine ; physiology ; Dopaminergic Neurons ; pathology ; Humans ; Neuronal Plasticity ; Parkinson Disease ; physiopathology ; Receptors, Dopamine D1 ; physiology ; Receptors, Dopamine D2 ; physiology ; Substantia Nigra ; pathology

This study examined the neuroprotective effect of cluster of differentiation molecule 200 (CD200) against methamphetamine (METH)-induced neurotoxicity. In the in vitro experiment, neuron-microglia cultures were treated with METH (20 μmol/L), METH (20 μmol/L)+CD200-Fc (10 μg/mL) or CD200-Fc (10 μg/mL). Those untreated served as control. Microglia activation expressed as the ratio of MHC-II/CD11b was assessed by flow cytometry. The cytokines (IL-1β, TNF-α) secreted by activated microglia were detected by enzyme-linked immunosorbent assay (ELISA). In the in vivo experiment, 40 SD rats were divided into control, METH, METH+CD200-Fc and CD200-Fc groups at random. Rats were intraperitoneally injected with METH (15 mg/kg 8 times at 12 h interval) in METH group, with METH (administered as the same dose and time as the METH group) and CD200-Fc (1 mg/kg at day 0, 2, 4 after METH injection) in METH+CD200-Fc group, with CD200-Fc (1 mg/kg injected as the same time as the METH+CD200-Fc group) or with physiological saline solution in the control group. The level of striatal dopamine (DA) in rats was measured by high-performance liquid chromatography (HPLC). The microglial cells were immunohistochemically detected for the expression of Iba-1, a marker for microglial activation. The results showed that METH could increase the microglia activation in the neuron-microglia cultures and elevate the secretion of IL-1β and TNF-α, which could be attenuated by CD200-Fc. Moreover, CD200-Fc could partially reverse the striatal DA depletion induced by METH and reduce the number of activated microglia, i.e. Iba-1-positive cells. It was concluded that CD200 may have neuroprotective effects against METH-induced neurotoxicity by inhibiting microglial activation and reversing DA depletion in striatum.
Animals ; Animals, Newborn ; Antigens, CD ; administration & dosage ; Cells, Cultured ; Coculture Techniques ; Corpus Striatum ; cytology ; drug effects ; immunology ; Cytokines ; immunology ; Dopamine ; immunology ; Drug Interactions ; Male ; Methamphetamine ; toxicity ; Microglia ; drug effects ; immunology ; Neurons ; metabolism ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo construct a morphine tolerance model in primarily cultured striatal neurons, and screen the differentially expressed genes in this model using suppression subtractive hybridization (SSH).

METHODSSbtracted cDNA libraries were constructed using SSH from normal primarily cultured striatal neurons and long-term morphine treated striatal neurons (10-5 mol/L for 72 hours). To check reliability of the cell culture model, RT-PCR was performed to detect the cAMP-responsive element-binding protein (CREB) mRNA expression. The subtracted clones were prescreened by PCR. The clones containing inserted fragments from forward libraries were sequenced and submitted to GenBank for homology analysis. And the expression levels of genes of interest were confirmed by RT-PCR. Results CREB mRNA expression showed a significant increase in morphine treated striatal neurons (62.85 ± 1.98) compared with normal striatal neurons (28.43 ± 1.46, P < 0.01). Thirty-six clones containing inserted fragments were randomly chosen for sequence analysis. And the 36 clones showed homology with 19 known genes and 2 novel genes. The expression of 2 novel genes, mitochondrial carrier homolog 1 (Mtch1; 96.81 ± 2.04 vs. 44.20 ± 1.31, P < 0.01) and thymoma viral proto-oncogene 1 (Akt1; 122.10 ± 2.17 vs. 50.11 ± 2.01, P < 0.01), showed a significant increase in morphine-treated striatal neurons compared with normal striatal neurons.

CONCLUSIONSA reliable differential cDNA library of striatal neurons treated with long-term morphine is constructed. Mtch1 and Akt1 might be the candidate genes for the development of morphine tolerance.

Analgesics, Opioid ; pharmacology ; Animals ; Cells, Cultured ; Corpus Striatum ; cytology ; Drug Tolerance ; physiology ; Gene Expression Profiling ; Gene Library ; Molecular Sequence Data ; Morphine ; pharmacology ; Neurons ; cytology ; drug effects ; Nucleic Acid Hybridization ; methods ; Rats ; Rats, Wistar ; Reverse Transcriptase Polymerase Chain Reaction ; methods

OBJECTIVETo investigate the cell proliferation and differentiation in the developing brain of mouse.

METHODSC57/BL6 mice were divided into 3 groups at random. Bromodeoxyuridine (BrdU) was injected into the brains in different development periods once a day for 7 d. The brains were retrieved 4 weeks after the last BrdU injection. Immunohistochemical and immunofluorescent studies were carried out for detecting cell proliferation (BrdU) and cell differentiation (NeuN, APC, Iba1, and S100beta), respectively.

RESULTSThe number of BrdU labeled cells decreased significantly with the development of the brain. Cell proliferation was prominent in the cortex and striatum. A small portion of BrdU and NeuN double labeled cells could be detected in the cortex at the early stage of development, and in the striatum and CA of the hippocampus in all groups. The majority of BrdU labeled cells were neuroglia, and the number of neuroglia cells decreased dramatically with brain maturation. Neurogenesis is the major cytogenesis in the dentate gyrus.

CONCLUSIONThese results demonstrated that cell proliferation, differentiation and survival were age and brain region related.

Animals ; Animals, Newborn ; Brain ; cytology ; growth & development ; Bromodeoxyuridine ; Cell Count ; Cell Differentiation ; physiology ; Cell Proliferation ; Cerebral Cortex ; cytology ; growth & development ; Corpus Striatum ; cytology ; growth & development ; Fluorescent Antibody Technique ; Hippocampus ; cytology ; growth & development ; Male ; Mice ; Mice, Inbred C57BL ; Nerve Tissue Proteins ; metabolism ; Neuroglia ; cytology ; physiology ; Neurons ; cytology ; physiology ; Nuclear Proteins ; metabolism

OBJECTIVETo explore the mechanism of acupuncture in treatment of Parkinson's disease.

METHODSFifty Wistar rats were randomly divided into a normal control group, a sham-operation control group, a model group, a Fengfu-Taichong group and a Shuanggu Yitong group. The hemilateral rotation Parkinson's disease model was prepared by micro-injection of 6-hydroxyl-dopamine into the right striatum. Effects of acupuncture at "Fengfu" (GV 16), "Taichong" (LR 3) hy Shoanggu Yitong needling method on proliferation and differentiation of nerve stem cells in the Parkinson's disease model rat were compared. Proliferation and numher changes of the nerve stem cells in the suhstantia nigra and the striatum were investigated with immunohistochemical method, and the number of nerve stem cells transforming neurons were observed with immunohistochemical double-labeling method.

RESULTSIn the two acupuncture groups, the proliferative cells in the substantia nigra and thestriatum of the destroyed side increased significantly (P < 0.01), and the number of nerve stem cells and the number of transformed neurons in the destroyed side of the substantia nigra and the striatum increased significantly only in the Shuanggu Yitong group (P < 0.01).

CONCLUSIONShuanggu Yitong needling method can induce proliferation and differentiation of nerve stem cells in the substantia nigra and the striatum regions in the Parkinson's disease model rat.

Acupuncture Therapy ; methods ; Animals ; Bromodeoxyuridine ; metabolism ; Cell Differentiation ; Cell Proliferation ; Corpus Striatum ; pathology ; Male ; Neurons ; cytology ; Parkinson Disease ; therapy ; Rats ; Rats, Wistar ; Stem Cells ; cytology ; Substantia Nigra ; pathology