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ORlGINS OF NEUROPEPTIDE Y-LIKE IMMUNOREACTIVE FIBERS IN THE LATERAL AND BASOLATERAL AMYGDALOID NUCLEI OF THE RAT

Gaoge SUN ; Yamano MARIKO ; Tohyama MASAYA ; Qunyuan XU

Chinese Journal of Neuroanatomy.2000;16(4):347-352,58.

The origin of neuropeptide Y(NPY)-like immunoreactive(IR)fibers in the lateral and basolateral amygdaloid nuclei was examined by using indirect immunofluorescence and retrograde tracing method in the rat.1njection of a retrograde tracer.fluorogold(FG),into the lateral and basolateral amygdaloid nuclei,labeled many neurons in the ipsilateral anterior amygdaloid area,and simultaneous treatment with antiserum against NPY stained some of these neurons.Destruction of the anterior amygdaloid area caused an ipsilateral decrease of NPY-IR fibers in the lateral and basolateral amygdaloid nuclei.These findings indicate that NPY-IR neurons in the anterior amygdaloid area project ipsilaterally to the lateral and basolateral amygdaloid nuclei.In addition,the present study also shows that NPY-IR neurons located in the lateral and basolateral amygdaloid nuclei are intrinsic to the amygdaloid complex,since after destruction of the anterior amygdaloid area,some of NPY-IR fibers still can be found in lateral and baso lateral nuclei,and transection of the two major amygdalofugal system,stria terminalis and ventral amygdalofugal pathway,failed to cause the accumulation of NPY-like immunoreactivity in the axons proximal to the section.

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THE EFFECTS OF HYPERTHERMIA ON THE NEUROEPITHELIAL CELLS IN PRIMARY CULTURE

Yingjun GUAN ; Yingmao GAO ; Zhaodong JUAN ; Shouxun WANG ; Rujiang LI

Chinese Journal of Neuroanatomy.2000;16(4):-.

The animal experiment studies have demonstrated that hyperthermia is a strong teratogen to many kinds of animal with high incidence of neural tube defects(NTD).Epidermiological investigation showed that hyperthermia was closely related to the acencephaly and excenphaly in human being,but little was known about the distinct mechanism of NTD induced by hyperthermia.In order to study the developmental mechanism of NTD induced by hyperthermia,we observed the effects of hyperthermia on the neuroepithelial cells in primary culture.The neural tubes of the hamster embryos on 10 d after fertilization were obtained.the neuroepithelia were dissociated and then seeded at the density of 1×106 cells per well.The cells were divided into experimental and control groups randomly.The experimental groups were exposed to 42 C for 20 min,whereas groups exposed to 37 Cserved as control.After treatment of hyperthermia,the cells were incubated continuously at 37 C in a 95%air/5%CO2 humidified incubator,the culture was terminated after different intervals.Phase-contrast microscopy.scanning electron microscopy.transmission electron microscopy,MTT assay,agarose gel electrophoresis analysis,TUNEL detection,immunocytochemistryand image analysis were made.The results of the experimental groups indicated that the floating cells and apoptotic cells increased in number,the neurites of the cells were shortened even disappeared,the survival cells decreased in number,the ultrastructure of the cells demonstrated distinct abnormal changes,the function of mitochondria was impaired,the expressions of both bcl-2 and bax were abnormal.The above results suggest that hyperthermia may induce apoptosis of neuroepithelial cells,duringwhich bcl-2 and bax genes may play important regulatory roles.

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COEXISTENCE OF ADRENOCORTICOTROPIN AND SECRETOGRANIN ⅡIN THE RAT PITUITARY GLAND AFTER BRAIN INJURY

Chunmei WANG ; Xiaofeng HUANG ; Jiaji YANG ; Mei HUANG ; Xiangdang LI ; Dan CHEN

Chinese Journal of Neuroanatomy.2000;16(4):337-340,55.

The localization of adrenocorticotropin(ACTH),secretogranin Ⅱ in rat pituitary gland Was studied by using specific antisera in conjunction with double immunofluorescence staining techniques.Stainings with secretogranin Ⅱ serum showed intense cytoplasmic reaction in the anterior lobe of the pituitary gland.Double immunofluorescence staining was used to identify the cells that expressed secretogranin Ⅱ and ACTH.Confocal laser scanning microscopy revealed that ACTH existed simultaneouslywith secretogranin Ⅱ in quite a number of cells.The results indicate that secretogranin Ⅱ is expressed in corticotrophs of rat pituitary gland,its physiological role may be associated with the regulation of optimal pH in the secretory vesicles for the cleavage of ACTH from its precursor.

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NEURONAL MIGRATION AND SYNTHESIS OF PEPTIDIC NEUROTRANSMITTERS IN CO-CULTURES OF NODOSE GANGLIA AND CARDIAC MYOCYTES

Huaxiang LIU ; Zhenzhong LI ; Xingfu LI ; Fei HUANG ; Xingyi LI ; Yonggang LI ; Jianjun LIU

Chinese Journal of Neuroanatomy.2005;21(6):636-640.

To investigate the neuronal migration and synthesis of peptidic neurotransmitters in vitro, neuromuscular co-cultures of nodose ganglia and cardiac muscle cells of Wistar rats were established. The living cells of co-cultures were observed with an inverted contrast microscope at different culture age. At 72 h and 96 h of culture age, the neuromuscular co-cultures were examined by Holmes' reduced silver staining technique for observing the neuronal migration. Immunohistochemical staining method was used for investigating the synthesis of peptidic neurotransmitters- substance P (SP) and calcitonin gene-related peptide (CGRP). The neuronal morphorlogical maturation in neuromuscular co-cultures seems at 72 h of culture age in terms of the observation of living cells and Holmes' reduced silver staining technique. SP- and CGRP-immunoreactive neurons were observed in neuromuscular co-cultures at 96 h of culture age but not 72 h. The results indicate that the neuronal morphological maturation can not represent the maturation of neurotransmitter synthesis. The synthesis of neurotransmitters is not maturated in the neuromuscular co-cultures until 96 h of the culture age.

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EFFECTS OF INSULIN-LIKE GROWTH FACTOR 1 ON β-AMYLOID INDUCED NEURON APOPTOSIS AND TAU PHOSPHORYLATION

Changhong XING ; Ying PENG ; Zuoping XIE

Chinese Journal of Neuroanatomy.2005;21(6):569-575.

The aim of this study is to elucidate the protective and anti-apoptotic effects of insulin-like growth factor 1 ( IGF-1 ) against β-amyloid (Aβ) and investigate the effect of IGF-1 on Aβ-induced tau phosphorylation. Cell viability was measured using the MTT (3-(4,5-dimethylthiazolyl-2 )-2,5-diphenyltetrazolium bromide) assay, early apoptosis and late apoptosis/necrosis were analyzed by flow cytometry using Annexin V-FITC and propidium iodide (PI) double staining, and morphology was examined by Hoechst 33342 staining. Tau phosphorylation was detected using AT8 immunostaining. Preincubation of cultured rat hippocampal neurons with IGF-1 for 24 h prevented cytotoxicity induced by Aβ25-35 for 48 h. The MTT value significantly increased from 54.51% to 61.8% of the control group, and the percentage of Hoechst 33342-positive cells decreased from 30.77% to 22.81%. Incubation with Aβ25-35 for 48 h caused a marked increase in the percentages of Annexin V-FITC single-labeled cells (Annexin V +/PI-) and Annexin V/PI double-stained cells (Annexin V +/PI + ) (3.41% and 19.47% , respectively), which were significantly decreased by pretreatment with 100 ng/ml of IGF-1 for 24 h (to 2.98% and 15.16% , respectively). Aβ25-35 treatment increased tau phosphorylation and AT8 positive cells were 41.84%. This effect could be inhibited by different concentrations of IGF-1. Our findings showed that IGF-1 protected against Aβ-induced cytotoxicity, decreased the percentage of early and late apoptosis/necrosis cells, and inhibited tau phosphorylation, which may be the cellular mechanisms for its neuroprotective action.

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EFFECTS OF CORTICOTROPIN RELEASING FACTOR ON LEARNING AND MEMORY IN THE NUCLEUS BASALIS OF MEYNERT IN RATS

Boning YANG ; Guohe TAN ; Li WEI ; Ling LAN

Chinese Journal of Neuroanatomy.2005;21(5):503-506.

The present study was performed to explore the role of corticotropin releasing factor (CRF) in the nucleus basalis of Meynert (NBM) in spatial learning and memory of rats. The latency, distance and swimming path to find the platform were measured by Morris water maze after intra-NBM injections of 0, 0.01, 0.1 and 0.4 nmol of CRF. Intra-NBM injections of 0.1 or 0.4 nmol of CRF induced significant increase of the latency for spatial learning and memory, and there were no significant changes in the swimming speed in Morris water maze test. The results suggest that CRF plays an inhibitory role in spatial learning and memory consolidation in the NBM of rats.

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pCREB IN GLUTAMATE CONTAINING NEURONS GREATLY UPREGULATED IN RAT AMYGDALA AFTER A STRESS BY FORCED SWIMMING

Lu GAO ; Ruixi LI ; Jie WANG ; Zhongliang DING ; Yuwen PENG

Chinese Journal of Neuroanatomy.2005;21(5):463-470.

Amygdala (AM) plays crucial roles in emotional learning, memory and behavior. These functions of AM are carried out by three main subnuclei (lateral nucleus, basolateral nucleus and central nucleus) in AM and closely related with a transcription factor, cAMP- responsive element binding protein (CREB) in the neurons of the AM. CREB can be phosphorylated (pCREB) in many kinds of neuronal processes to regulate the synthesis of proteins for the formation of memory processes. In order to identify what neuronal types express pCREB and how the pCREB levels changed at different time intervals after an emotional stress stimulation, the present study is designed to investigate pCREB-, glutamate (Glu)- and parvalbumin (PV)- immunoreactive (IR) profiles in AM and the levels of pCREB in AM after a stress of forced swimming (FS). The results showed that the pCREB expressed in the Glu-IR neurons but not in the PV-IR neurons, and the expression level of the pCREB increased dramatically after the stress. The present results suggested that pCREB modulates the emotional processes through the Glu-IR neurons and that the pCREB greatly upregulated to response to the emotional stimuli.

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EFFECTS OF NERVE INJURY ON THE EXPRESSION OF Trk RECEPTOR PROTEINS LOCALIZED ON THE TRIGEMINAL MESENCEPHALIC NEURONS

Fuxing ZHANG ; Yulin DONG ; Feng GUO ; Youwang PANG ; Jinlian LI

Chinese Journal of Neuroanatomy.2005;21(6):625-630.

Immunofluorescence histochemistry combined with retrograde tracing technique was employed to observe the effects of masseteric nerve transection on the expression of Trk ( tropomyosin-related kinase) receptor proteins, namely TrkA, TrkB and TrkC in the trigeminal mesencephalic nucleus ( Me5 ) of the rat. At 7 and 14 days following transection of masseteric nerve through which Fluorogold (FG) was applied to identify the Me5 neurons innervating masseter, brain sections were immunohistochemically processed to detect the three Trk isoforms in FG-labeled Me5 neurons. With the percentage of double-labeled neurons to the total number of FG-labeled neurons as the index,we demonstrated ( 1 ) a significant increase in the percentage of TrkA-immunoreactive (IR) Me5 neurons at both 7 and 14 days after nerve transection, (2) no significant, but gradual, increase in the percentage of TrkB-IR Me5 neruons with longer survival time post transection and ( 3 ) little change of TrkC expression. The current findings indicate that axotomy differently affected the expression of the individual Trk receptors and these expression patterns may reflect an adaptation of the Me5 neurons to the peripheral nerve injury.

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HIGH CONCENTRATION OF IRON IN CEREBROSPINAL FLUID OF RATS INDUCES ALZHEIMER-LIKE BEHAVIORAL AND PATHOLOGICAL CHANGES

Lin LI ; Jianliang WU ; Zheng JIN ; Yan DOU

Chinese Journal of Neuroanatomy.2005;21(3):252-258.

In order to estimate the relationship between iron and the Alzheimer's disease, the behavioral and pathological changes were observed by Morris water maze and immunohistochemical staining respectively after injecting FeC13 into brain ventricle of rats. The apoptosis was tested by flow cytometry and the electron microscopy was used to observe ultrastructural changes. There were significant differences in escape latency of time and distance between normal animals and iron treated rats. Percentage and fluorescence intension (FI) of AnnexinV FITC loaded cells undergoing apoptosis were higher in iron treated rats compared with normal animals. Fawn-coloured products of β amyloid protein were interspersedly distributed in extensive areas of cerebral cortex and hippocampus. Under electron microscope, vacuolate degeneration of neuronal processes with mitochondria degeneration and accumulation of microtubule near vacuolar nucleus were observed in iron treated rats. These results suggest that a local higher concentration of iron in brain may induce Alzheimer-like impairment of intelligence and pathological changes.

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ENDOMORPHIN-1 IS MORE POTENT THAN ENDOMORPHIN-2 IN INHIBITION OF SYNAPTIC TRANSMISSION IN SUBSTANTIA GELATINOSA OF ADULT RAT SPINAL CORD

Dongni LENG ; Yupeng FENG ; Yunqing LI

Chinese Journal of Neuroanatomy.2005;21(3):269-275.

Effects of endomorphin-1 (EM-1) and endomorphin-2 (EM-2) on synaptic transmission were investigated on neurons in substantia gelatinosa (SG) of the spinal dorsal horn by whole-cell voltage clamp recording. Both EM-1 (1 μmol/L) and EM-2 (1 μmol/L)remarkably reduced the frequency but not the amplitude of miniature excitatory postsynaptic currents (mEPSCs) and miniature inhibitory postsynaptic currents (mIPSCs). These effects were antagonized by 3-funaltrexamine ( β-FNA, 10 μmol/L), a selective μ-opioid receptor antagonist. Noticeably, EM-1 showed higher potency in decreasing the frequency of mEPSCs and mIPSCs than that of EM-2. These results indicate that EMs suppress both excitatory and inhibitory synaptic transmission by activating presynaptic μ-opioid receptors in the SG and EM-1, compared with EM-2, might be a more potent endogenous analgesic at the spinal cord level.

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