1.Changes of Na(+) channels in rat hippocampal CA1 neurons in early development after birth.
Jia-Qi QIAO ; Ai-Li LIU ; Tiao-Tiao LIU ; Xin TIAN
Acta Physiologica Sinica 2013;65(2):201-209
The purpose of this research is to investigate the critical period of voltage-gated Na(+) channel development in hippocampal CA1 neurons. Changes of Na(+) currents in acutely isolated hippocampal CA1 neurons of rats at different ages (0-4 weeks after birth) were recorded using the whole-cell patch-clamp technique. The results indicated that the maximum current density of Na(+) channels was increasing with age, and the amplitudes in 1, 2, 3 and 4 weeks respectively grew by (42.76 ± 4.91)%, (146.80 ± 7.63)%, (208.79 ± 5.28)% and (253.72 ± 5.74)% (n = 10, P < 0.05) compared with that in 0 week. The current density in CA1 neurons of 1-2 weeks after birth increased more significantly than those of other groups. The activation curve of Na(+) channel shifted to the left. The half-activation voltages (mV) in 0-2 weeks were -39.06 ± 0.65, -43.41 ± 0.52, -48.29 ± 0.45 (n = 10, P < 0.05), respectively, showing significant age-dependent decrease, and there were no significant changes in other groups. The slope factors of activation curve for each group did not change significantly. There were no regular changes in inactivation curve and no significant changes in half-inactivation voltage. The slope factors of inactivation curve in 1-2 weeks were: 5.77 ± 0.56, 4.42 ± 0.43 (n = 10, P < 0.05). The inactivation rate of the second week after birth was faster than that of the first week, and there were no significant changes during 0-1 week and 2-4 weeks. The recovery from inactivation curve of Na(+) channel shifted to the left. The recovery time declined in 1-3 weeks. Changes of action potential properties were consistent with Na(+) current. These results suggest that the period of 1-2 weeks after birth may be the critical development period of voltage-gated Na(+) channel in hippocampal CA1 neurons. During this time, the distribution of Na(+) channel increases significantly; the activation curve of Na(+) channel shifts to the left; inactivation rate increases as well as recovery time shortens.
Action Potentials
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Animals
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CA1 Region, Hippocampal
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cytology
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Neurons
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physiology
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Patch-Clamp Techniques
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Rats
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Sodium Channels
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physiology
2.Dopamine-dependent long-term depression in hippocampus of rat induced by exposure to spatial novelty..
Na LIU ; Bin XUE ; Hua XING ; Lin XU ; Shan-Xiang JIANG
Acta Physiologica Sinica 2009;61(6):511-516
To study the role of long-term depression (LTD) in the mechanisms of learning and memory in hippocampus of rat, recordings were taken from freely moving animals that had undergone chronic implantation of a recording electrode in the hippocampus CA1 region and a bipolar stimulating electrode in the ipsilateral Schaffer collateral-commissural pathway. The recording electrode was inserted 3.8 mm posterior to bregma and 2.8 mm right of the midline, and the stimulating electrode was inserted 4.8 mm posterior to bregma and 3.8 mm right of the midline via holes drilled through the skull. The entire assembly was connected with a rubber socket on the animal's head and then stabilized with dental cement. The correct placement of the electrodes into the hippocampal CA1 area was confirmed via electrophysiological criteria and postmortem histological analysis. After 2 weeks of surgery recovery, the rats were placed in the familiar recording chamber for 3 days. The field excitatory postsynaptic potentials (fEPSPs) were evoked by stimulating with a square wave constant current pulse of 0.2 ms duration, at a frequency of 0.033 Hz and at a stimulation intensity adjusted to given an fEPSP amplitude of 50% of the maximum, and the baseline of fEPSPs were recorded for 3 days in the familiar recording environment at the same time each day. A novelty environment that was made of clear Perspex (40 cm x 40 cm x 40 cm) was prepared and we examined whether exposure to a novelty spatial environment facilitated the expression of activity-dependent persistent decrease in synaptic transmission (namely LTD) at CA1 synapses in the rat hippocampus. The results showed that brief exposure to a novelty environment for 10 min facilitated the expression of LTD in the hippocampal CA1 area under no other exogenous high- or low-frequency stimulation protocol. This facilitatory effect was dependent on the activation of D1/D5 receptors: the D1/D5 receptors antagonist SCH23390 prevented the decrease of synaptic transmission in the hippocampus during the novelty exploration. These data provided important evidence that LTD may underlie certain forms of learning and memory and that dopamine participates in the synaptic plasticity in the process of hippocampal spatial information storage.
Animals
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CA1 Region, Hippocampal
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physiology
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Dopamine
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physiology
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Electrodes
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Excitatory Postsynaptic Potentials
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Exploratory Behavior
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Neuronal Plasticity
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Rats
3.Inhibitory effects of epileptic spikes on theta rhythm in rat pilocarpine model of temporal lobe epilepsy.
Man-Ling GE ; Bao-Qiang GUO ; Xue CHEN ; Ying SUN ; Sheng-Hua CHEN ; Ying ZHENG ; Hui-Juan ZHANG ; Wei SUN
Acta Physiologica Sinica 2014;66(2):118-128
Epilepsy clinically has an inhibitory impact on cognitive function, but whether it is associated with epileptogenesis is unclear. Since the epileptic spike characterizes temporal lobe epilepsy (TLE), the present study was aimed to analyze the transient effects of sporadic spikes (SSs) on theta rhythm during epileptogenesis. The local field potentials (LFPs) were recorded in CA1 area in four rats with the pilocarpine injections during exploration, and theta phase stability and power were globally estimated around SSs, also during prolonged period without SS (both as experiments) as well as pre-injections (control). Finally, the LFPs were simulated by changing the average excitatory and inhibitory synaptic gain values (including slow and fast inhibition loops) with the help of simplified dynamical model of CA1 networks, and then theta phase stability was evaluated in several cases. It was found that the SSs could have negative impacts on theta rhythm both transiently and persistently, which may be dependent on the temporal courses leading to epilepsy, being acuter in early stage than later stage, but even in latent stage, theta power was strong. The simulations partly demonstrated that the synaptic imbalance concomitant with the occurrence of SSs might be related to the dynamics of theta phase stability. The results indicate that the SSs might have persistent negative impacts on the cognition rhythm, and the effects might alter during epileptogenesis, leading to the cognitive dysfunction.
Animals
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CA1 Region, Hippocampal
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physiopathology
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Epilepsy, Temporal Lobe
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chemically induced
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physiopathology
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Pilocarpine
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Rats
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Theta Rhythm
4.A design of raster plot for illustrating dynamic neuronal activity during deep brain stimulation.
Lu HUANG ; Zhaoxiang WANG ; Zhouyan FENG
Journal of Biomedical Engineering 2019;36(2):177-182
Deep brain stimulation (DBS), which usually utilizes high frequency stimulation (HFS) of electrical pulses, is effective for treating many brain disorders in clinic. Studying the dynamic response of downstream neurons to HFS and its time relationship with stimulus pulses can reveal important mechanisms of DBS and advance the development of new stimulation modes (e.g., closed-loop DBS). To exhibit the dynamic neuronal firing and its relationship with stimuli, we designed a two-dimensional raster plot to visualize neuronal activity during HFS (especially in the initial stage of HFS). Additionally, the influence of plot resolution on the visualization effect was investigated. The method was then validated by investigating the neuronal responses to the axonal HFS in the hippocampal CA1 region of rats. Results show that the new design of raster plot is able to illustrate the dynamics of indexes (such as phase-locked relationship and latency) of single unit activity (i.e., spikes) during periodic pulse stimulations. Furthermore, the plots can intuitively show changes of neuronal firing from the baseline before stimulation to the onset dynamics during stimulation, as well as other information including the silent period of spikes immediately following the end of HFS. In addition, by adjusting resolution, the raster plot can be adapted to a large range of firing rates for clear illustration of neuronal activity. The new raster plot can illustrate more information with a clearer image than a regular raster plot, and thereby provides a useful tool for studying neuronal behaviors during high-frequency stimulations in brain.
Action Potentials
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Animals
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Axons
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physiology
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CA1 Region, Hippocampal
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physiology
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Deep Brain Stimulation
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Neurons
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physiology
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Rats
5.Effects of electroacupuncture with different courses on the synaptic structure and synaptic function-related proteins in mice with radiation-induced brain injury.
Xin WU ; Yan-Hui LI ; Wen-Jing ZHANG ; Song-Jiang ZHANG ; Jian-Feng GAO
Acta Physiologica Sinica 2021;73(6):909-916
The aim of the present study was to investigate the effects of different courses of electroacupuncture on synaptic structure and synaptic function-related proteins expression in the hippocampal CA1 region of radiation-induced brain injury mice. Sixty C57BL/6J male mice were randomly divided into control group, radiation-induced brain injury model group, 1-week electroacupuncture group (EA1), 2-week electroacupuncture group (EA2), 3-week electroacupuncture group (EA3), and electroacupuncture-control (EA-Ctrl) group. The mice in model group were exposed to X-ray irradiation (8 Gy, 10 min) to establish radiation-induced brain injury model. The mice in EA groups were acupunctured at electroacupuncture points (Baihui, Fengfu and bilateral Shenshu) for 1 week, 2 weeks and 3 weeks respectively after radiation. Immunohistochemistry was used to observe synaptic structure in hippocampal CA1 region. The expressions of brain-derived neurotrophic factor (BDNF), synapsin-1 and postsynaptic density 95 (PSD95) in the hippocampal CA1 region of each group were detected by RT-PCR and Western blotting. The results showed that the nuclear gap in model and EA-Ctrl groups was significantly decreased compared to control group, however nucleus to cytoplasm ratio was significantly increased. The synaptic cleft, postsynaptic density (PSD) thickness, the mitochondrial surface density, volume density and specific surface area were significantly reduced. Compared with model group, the nucleus to cytoplasm ratio of EA2 group was significantly decreased, the PSD thickness and mitochondrial volume density were significantly increased; the nuclear gap of EA3 group was significantly increased, nucleus to cytoplasm ratio was significantly decreased, synaptic cleft and PSD thickness were significantly increased, and the mitochondrial surface density and specific surface area were all increased significantly. In addition, compared with the control group, the gene and protein expressions of BDNF, synapsin-1 and PSD95 in the hippocampal CA1 region of the model group and EA-Ctrl group were significantly decreased. However, compared with the model group, the gene expression of synapsin-1 in EA groups was significantly up-regulated, the gene expression of BDNF in EA1 and EA2 groups was significantly up-regulated, and the gene expression of PSD95 in EA2 group was significantly up-regulated. Moreover, the protein expressions of BDNF, synapsin-1 and PSD95 of EA groups were significantly up-regulated compared with the model group. These results indicate that the synaptic structure and the expression of synaptic function-related proteins in hippocampal CA1 region were injured by radiation exposure, whereas electroacupuncture intervention can significantly improve the synaptic structure and function damage caused by radiation.
Animals
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Brain Injuries
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CA1 Region, Hippocampal
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Electroacupuncture
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Hippocampus
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Male
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Mice
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Mice, Inbred C57BL
6.Division of CA1, CA3 and DG regions of the hippocampus of Wistar rat.
Xiao-Cai SUN ; Li LI ; Min ZHANG ; Wen-Bin LI ; Qing-Jun LI ; Li ZHAO
Chinese Journal of Applied Physiology 2012;28(2):189-192
OBJECTIVEDivision of the CA1, CA3 and dentate gyrus (DG) regions of the hippocampus of Wistar rat under the stereomicroscope.
METHODSTwenty-four Wistar rats were randomly assigned to three groups. (1) The brain was sectioned coronally (n = 6). The sections were stained with thionin and the morphology of cells in each region of the hippocampus was observed under microscopy. (2) The hippocampus was dissected out and observed on the whole. Then, the CA1, CA3 and DG regions of the hippocampus were divided. Every region divided was sectioned, and the morphology of cells was observed. (3) Rats with brain ischemia or not were also decapitated and the HSP 70 expressions were observed in CA1, CA3 + DG regions by Western blot and immunohistochemical staining (n = 12).
RESULTS(1) The CA1, CA3 and DG regions of the hippocampus could be clearly observed in coronal section of the brain stained by thionin. (2) Under the stereomicroscope, the CA1 and DG regions of the hippocampus could be separated along the hippocampal fissure between them in ventral surface of the hippocampus. The CA3 and DG regions of the hippocampus could be separated along a fissure between them. The appearance of cells in the sections of the divided CA1, CA3 and DG specimens is consistent with that in the brain coronal sections, respectively. (3) The results of Western blot indicated that the HSP 70 expression of the brain ischemia group was up-regulated significantly in CA3 + DG regions compared with the sham group. However, HSP 70 expression has no significant changes in CA1 region. The above results were consistent with those of the immunohistochemical staining.
CONCLUSIONThe CA1, CA3 and DG regions of the hippocampus of Wistar rat could be divided under stereomicroscope, and the divided each region was sensible for detection of protein using Western blot.
Animals ; CA1 Region, Hippocampal ; anatomy & histology ; metabolism ; CA3 Region, Hippocampal ; anatomy & histology ; metabolism ; Dentate Gyrus ; anatomy & histology ; metabolism ; Male ; Rats ; Rats, Wistar
7.Regulation of astroglia on synaptic plasticity in the CA1 region of rat hippocampus.
Laixun, TAN ; Shenggang, SUN ; Shenhan, DUAN ; Xilin, WANG
Journal of Huazhong University of Science and Technology (Medical Sciences) 2005;25(5):484-7
The regulation of astroglia on synaptic plasticity in the CA1 region of rat hippocampus was examined. Rats were divided into three groups: the newly born (< 24 h), the juvenile (28-30 days) and the adult groups (90 - 100 days), with each group having 20 animals. The CA1 region of rat hippocampus was immunohistochemically and electron-microscopically examined, respectively, for the growth of astroglia and the ultrastructure of synapses. The high performance liquid chromatography was employed to determine the cholesterol content of rat hippocampus. In the newly-born rats, a large number of neurons were noted in the hippocampal CA1 region of the newly-born rats, and few astroglia and no synaptic structure were observed. In the juvenile group, a few astroglias and some immature synapses were found, which were less than those in adult rats (P < 0.01). The cholesterol content was 2.92 +/- 0.03 mg/g, 11.20 +/- 3.41 mg/g and 12.91 +/- 1.25 mg/g for newly born, the juvenile and the adult groups, respectively, with the differences among them being statistically significant (P < 0.01). Our study suggests that the astrocytes may play an important role in the synaptic formation and functional maturity of hippocampal neurons, which may be related to the secretion of cholesterol from astrocytes.
Age Factors
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Animals, Newborn
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Astrocytes/cytology
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Astrocytes/metabolism
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Astrocytes/*physiology
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CA1 Region, Hippocampal/*physiology
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CA1 Region, Hippocampal/*ultrastructure
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Cell Communication/physiology
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Cholesterol/metabolism
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Neuronal Plasticity/*physiology
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Random Allocation
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Rats, Wistar
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Synapses/*physiology
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Synapses/ultrastructure
8.Expressions of the pERK1/2 and the cFos Proteins at an Early Stage of Transient Global Ischemia-reperfusion Injury in the Hippocampus of Rats.
Chang Seok KO ; Young Hyun YUN ; Hun Cheol AHN ; Jeong Woo CHOI ; Dae Han WI ; Su Jin YU ; Jae Hwang PARK
Journal of the Korean Society of Emergency Medicine 2004;15(6):475-486
PURPOSE: This study was to evaluate temporal changes in the expressions of the phosphorylated extracellular-regulated kinase1/2 (pERK1/2), the phosphorylated MAPK/ERK kinase1/2 (pMEK1/2) and the cFos proteins in the hippocampus of rats following transient global ischemia. METHODS: Transient global ischemia was induced in the forebrains of Sprague-Dawley rats by using a 4-vessel occlusion for 20 min under anesthetic condition. Hematoxyline-eosin staining showed typical microscopic findings that represented neuronal cell death in hippocampal CA1 regions 5 days after transient global ischemia. Four-vessel occlusion-reperfusion produced ischemic injury in major forebrain structures, such as the striatum, the cortex and the hippocampus, in the finding of triphenyltetrazolium chloride (TTC) staining. RESULTS: A high density of pERK1/2 immunoreactivity existed in the pyramidal-cell layers of the CA2-3 regions and in the granular-cell layers of the dentate gyrus 5 min after ischemia. Following ischemia, expression of the pMEK1/2 protein showed temporal changes similar to that of the pERK1/2 protein. A significant expression of the cFos protein was noted in the pyramidal-cell layers of the CA2-3 regions and in the granular-cell layers of the dentate gyrus 2 hours after global ischemia. CONCLUSION: Intracellular signaling cascades of the ERK or the cFos protein take part in early cellular events in the hippocampus of rats in response to ischemic insult.
Animals
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CA1 Region, Hippocampal
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Cell Death
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Dentate Gyrus
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Hippocampus*
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Ischemia
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Neurons
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Prosencephalon
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Rats*
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Rats, Sprague-Dawley
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Reperfusion Injury*
9.Age-dependent differences in myelin basic protein expression in the hippocampus of young, adult and aged gerbils.
Ji Hyeon AHN ; Tae Kyeong LEE ; Joon Ha PARK ; Jeong Hwi CHO ; In Hye KIM ; Jae Chul LEE ; Seongkweon HONG ; Yong Hwan JEON ; Il Jun KANG ; Young Joo LEE ; Moo Ho WON ; Choong Hyun LEE
Laboratory Animal Research 2017;33(3):237-243
Myelin degeneration is one of the characteristics of aging and degenerative diseases. This study investigated age-related alterations in expression of myelin basic protein (MBP) in the hippocampal subregions (dentate gyrus, CA2/3 and CA1 areas) of gerbils of various ages; young (1 month), adult (6 months) and aged (24 months), using western blot and immunohistochemistry. Western blot results showed tendencies of age-related reductions of MBP levels. MBP immunoreactivity was significantly decreased with age in synaptic sites of trisynaptic loops, perforant paths, mossy fibers, and Schaffer collaterals. In particular, MBP immunoreactive fibers in the dentate molecular cell layer (perforant path) was significantly reduced in adult and aged subjects. In addition, MBP immunoreactive mossy fibers in the dentate polymorphic layer and in the CA3 striatum radiatum was significantly decreased in the aged group. Furthermore, we observed similar age-related alterations in the CA1 stratum radiatum (Schaffer collaterals). However, the density of MBP immunoreactive fibers in the dentate granular cell layer and CA stratum pyramidale was decreased with aging. These findings indicate that expression of MBP is age-dependent and tissue specific according to hippocampal layers.
Adult*
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Aging
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Blotting, Western
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CA1 Region, Hippocampal
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Gerbillinae*
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Hippocampus*
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Humans
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Immunohistochemistry
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Myelin Basic Protein*
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Myelin Sheath*
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Perforant Pathway
10.Characterization of nestin expression in astrocytes in the rat hippocampal CA1 region following transient forebrain ischemia.
Jeong Min CHO ; Yoo Jin SHIN ; Jang Mi PARK ; Jin KIM ; Mun Yong LEE
Anatomy & Cell Biology 2013;46(2):131-140
Recent studies have suggested that nestin facilitates cellular structural remodeling in vasculature-associated cells in response to ischemic injury. The current study was designed to investigate the potential role of post-ischemic nestin expression in parenchymal astrocytes. With this aim, we characterized ischemia-induced nestin expression in the CA1 hippocampal region, an area that undergoes a delayed neuronal death, followed by a lack of neuronal generation after transient forebrain ischemia. Virtually all of the nestin-positive cells in the ischemic CA1 hippocampus were reactive astrocytes. However, induction of nestin expression did not correlate simply with astrogliosis, but rather showed characteristic time- and strata-dependent expression patterns. Nestin induction in astrocytes of the pyramidal cell layer was rapid and transient, while a long-lasting induction of nestin was observed in astrocytes located in the CA1 dendritic subfields, such as the stratum oriens and radiatum, until at least day 28 after ischemia. There was no detectable expression in the stratum lacunosum moleculare despite the evident astroglial reaction. Almost all of the nestin-positive cells also expressed a transcription factor for neural/glial progenitors, i.e., Sox-2 or Sox-9, and some cells were also positive for Ki-67. However, all of the nestin-positive astrocytes expressed the calcium-binding protein S100beta, which is known to be expressed in a distinct, post-mitotic astrocyte population. Thus, our data indicate that in the ischemic CA1 hippocampus, nestin expression was induced in astroglia that were becoming reactive, but not in a progenitor/stem cell population, suggesting that nestin may allow for the structural remodeling of these cells in response to ischemic injury.
Animals
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Astrocytes
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CA1 Region, Hippocampal
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Hippocampus
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Intermediate Filament Proteins
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Ischemia
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Nerve Tissue Proteins
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Neurons
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Prosencephalon
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Pyramidal Cells
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Rats
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Transcription Factors