1.Isolation and culture of porcine primary fetal progenitors and neurons from the developing dorsal telencephalon
Niroch Nawzad AUBID ; Yong LIU ; Juan Miguel Peralvo VIDAL ; Vanessa Jane HALL
Journal of Veterinary Science 2019;20(2):e3-
The development of long-term surviving fetal cell cultures from primary cell tissue from the developing brain is important for facilitating studies investigating neural development and for modelling neural disorders and brain congenital defects. The field faces current challenges in co-culturing both progenitors and neurons long-term. Here, we culture for the first time, porcine fetal cells from the dorsal telencephalon at embryonic day (E) 50 and E60 in conditions that promoted both the survival of progenitor cells and young neurons. We applied a novel protocol designed to collect, isolate and promote survival of both progenitors and young neurons. Herein, we used a combination of low amount of fetal bovine serum, together with pro-survival factors, including basic fibroblast growth factor and retinoic acid, together with arabinofuranosylcytosine and could maintain progenitors and facilitate in vitro differentiation into calbindin 1+ neurons and reelin+ interneurons for a period of 7 days. Further improvements to the protocol that might extend the survival of the fetal primary neural cells would be beneficial. The development of new porcine fetal culture methods is of value for the field, given the pig's neuroanatomical and developmental similarities to the human brain.
Brain
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Calbindins
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Cell Culture Techniques
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Congenital Abnormalities
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Cytarabine
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Fibroblast Growth Factor 2
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Humans
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In Vitro Techniques
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Interneurons
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Neurons
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Stem Cells
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Telencephalon
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Tretinoin
2.JNK/stress-activated protein kinase associated protein 1 is required for early development of telencephalic commissures in embryonic brains.
Ik Hyun CHO ; Kang Woo LEE ; Hye Yeong HA ; Pyung Lim HAN
Experimental & Molecular Medicine 2011;43(8):462-470
We previously reported that mice lacking JSAP1 (jsap1-/-) were lethal and the brain of jsap1-/- at E18.5 exhibited multiple types of developmental defects, which included impaired axon projection of the corpus callosum and anterior commissures. In the current study, we examined whether the early telencephalic commissures were formed abnormally from the beginning of initial development or whether they arose normally, but have been progressively lost their maintenance in the absence of JSAP1. The early corpus callosum in the brain of jsap1+/+ at E15.5-E16.5 was found to cross the midline with forming a distinct U-shaped tract, whereas the early axonal tract in jsap1-/- appeared to cross the midline in a diffuse manner, but the lately arriving axons did not cross the midline. In the brain of jsap1-/- at E17.5, the axon terminals of lately arriving collaterals remained within each hemisphere, forming an early Probst's bundle-like shape. The early anterior commissure in the brain of jsap1+/+ at E14.5-E15.5 crossed the midline, whereas the anterior commissure in jsap1-/- developed, but was deviated from their normal path before approaching the midline. The axon tracts of the corpus callosum and anterior commissure in the brain of jsap1-/- at E16.5-E17.5 expressed phosphorylated forms of FAK and JNK, however, their expression levels in the axonal tracts were reduced compared to the respective controls in jsap1+/+. Considering the known scaffolding function of JSAP1 for the FAK and JNK pathways, these results suggest that JSAP1 is required for the pathfinding of the developing telencephalic commissures in the early brains.
Adaptor Proteins, Signal Transducing/genetics/*metabolism
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Animals
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Brain/*embryology/*metabolism
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Female
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Focal Adhesion Kinase 1/genetics/metabolism
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Immunohistochemistry
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In Situ Nick-End Labeling
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JNK Mitogen-Activated Protein Kinases/genetics/metabolism
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Mice
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Mice, Knockout
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Nerve Tissue Proteins/genetics/*metabolism
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Pregnancy
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Telencephalon/*embryology/*metabolism
3.Effects of Chinese herbal compound on monoamine and neuronal amino acids in rat's telencephalon in the course of exhaustion and recovery process.
Hong-Zhen LIU ; Li ZENG ; Xi-Liang KONG ; Iei ZHU ; Yun-Chao MA
Chinese Journal of Applied Physiology 2011;27(4):439-443
OBJECTIVETo observe the effect of Chinese herbal compound on variance of neurotransmitters in rat telencephalon and to further discuss the mechanism underlying Chinese herbal compound in improving exercise capacity and promoting recovery from exercise-induced fatigue.
METHODS64 rats (8 week old) were randomly divided into medicine group (MG) and control group (CG). Chinese herbal compound was administered to rats of MG for 8 weeks. 8 weeks later, every group was divided into 4 subgroups and all were killed at different time point separately, and then neurotransmitter in rat brain was tested.
RESULTSThe exhaustion time of MG was significantly longer than that in CG (P < 0.01). In rest conditions, glutamic acid (GLU) of MG was significantly higher than that in CG (P < 0.01), while, there were no significant differences between MG and CG in other indexes. After fixed quantitative load exercise, the content of 5-hydroxytryptamineZZ(5-HT), 5-hydroindole acetic (5-HIAA), gamma-aminobutyric acid (GABA), Dopamine (DA) and 5-HT/5-HIAA were significantly lower than those in CG, while, GLU, GLU/GABA and DA/5-HT were significantly higher than those in CG. Compared with CG, exhaustion significantly (P < 0.05) decreased 5-HT, GABA and 5-HT/5-HIAA, and significantly (P<0.05) increased GLU, DA/5-HT and GLU/GABA level in MG. 12 h after exhaustion, in contrast to CG, level of 5-HT and 5-HT/5-HIAA in MG were significantly (P < 0.01) lower while GLU, DA, GABA and DA/5-HT were significantly (P < 0.01) higher.
CONCLUSIONDuring exhaustion exercise, Chinese herbal compound demonstrated strong inhibiting effect on synthesis of 5-HT, 5-HIAA, DA, GABA and promoting effect on GLU synthesis, this had been confirmed by the combined effect, including increase of excitatory transmitter and excitability of central nervous system and the prolongation of exhaustion time and promoting recovery from fatigue.
Amino Acids ; metabolism ; Animals ; Biogenic Monoamines ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Fatigue ; metabolism ; physiopathology ; Hydroxyindoleacetic Acid ; metabolism ; Male ; Neurons ; metabolism ; Physical Conditioning, Animal ; physiology ; Physical Exertion ; physiology ; Rats ; Rats, Wistar ; Serotonin ; metabolism ; Telencephalon ; drug effects ; metabolism
4.A Winged-Helix Transcription Factor Foxg1 Induces Expression of Mss4 Gene in Rat Hippocampal Progenitor Cells.
Ju Suk NAM ; Haijie YANG ; Nam Ho KIM ; Yuanjie SUN ; Byung Soo CHOI ; Sung Oh HUH
Experimental Neurobiology 2010;19(2):75-82
Foxg1 (previously named BF1) is a winged-helix transcription factor with restricted expression pattern in the telencephalic neuroepithelium of the neural tube and in the anterior half of the developing optic vesicle. Previous studies have shown that the targeted disruption of the Foxg1 gene leads to hypoplasia of the cerebral hemispheres with severe defect in the structures of the ventral telencephalon. To further investigate the molecular mechanisms by which Foxg1 plays essential roles during brain development, we have adopted a strategy to isolate genes whose expression changes immediately after introduction of Foxg1 in cultured neural precursor cell line, HiB5. Here, we report that seventeen genes were isolated by ordered differential displays that are up-regulated by over-expression of Foxg1, in cultured neuronal precursor cells. By nucleotide sequence comparison to known genes in the GeneBank database, we find that nine of these clones represent novel genes whose DNA sequences have not been reported. The results suggest that these genes are closely related to developmental regulation of Foxg1.
Animals
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Base Sequence
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Brain
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Cell Line
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Cerebrum
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Clone Cells
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Neural Tube
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Neurons
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Rats
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Stem Cells
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Telencephalon
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Transcription Factors
5.Intervention of selenium on neurogranin expression in filial cerebrum of mice with excess iodine.
Huai-Lan GUO ; Xue-Feng YANG ; Jian XU ; Jun WANG ; Dong YU ; Xiu-Fa SUN
Chinese Journal of Preventive Medicine 2007;41(1):21-24
OBJECTIVETo study the effects of excess iodine intake on neurogranin expression in cerebrum of filial mice and the intervention of selenium.
METHODSSixty BALB/c mice were divided randomly into four groups with different drinking water: control group (tap water, NC), excess iodine group (3000 microg/L I, EL +), supplementing selenium group (200 microg/L Se, Se +) and the excess iodine plus selenium (3000 microg/L + I 200 microg/L Se, EI + Se +) group. The mice were mated at the end of the fourth month. Serum T4 and T3 were determined on postnatal day 14 and 28. The expression level of neurogranin in filial cerebrum was measured by immunohistochemistry and Western blot.
RESULTSSerum T4 level in EI (68.78 +/- 11.10 nmol/ L) + was lower significantly than that in NC (100.85 +/- 11.47 nmol/ L) and EI + Se + (93.15 +/- 12.10 nmol/ L) on postnatal day 14. Western blot analysis showed that the relative level of neurogranin in EI + (0.621 +/- 0.041) was lower than that in NC (0.841 +/- 0.039) and EI + Se + (0.781 +/- 0.029) on postnatal day 14 (P < 0.05). No significant difference in serum T4 and neurogranin level between four groups on postnatal day 28.
CONCLUSIONExcess iodine intake might change the expression of neurogranin in filial cerebrum and the selenium supplementation might alleviate it.
Animals ; Female ; Iodine ; adverse effects ; Male ; Mice ; Mice, Inbred BALB C ; Neurogranin ; biosynthesis ; Selenium ; pharmacology ; Telencephalon ; metabolism ; Thyroxine ; blood ; Triiodothyronine ; blood
7.Morphological and histological studies on the telencephalon of the salamander Onychodactylus fischeri.
Huan-Huan WANG ; Li-Yan LI ; Li-Wen WANG ; Chuan-Cheng LIANG
Neuroscience Bulletin 2007;23(3):170-174
OBJECTIVETo probe into the morphological and histological characteristics of the telencephalon of Onychodactylus fischeri, and to enrich the comparable neurobiology.
METHODHE-staining method was used to describe the characters of the telencephalon of Onychodactylus fischeri.
RESULTSThe olfactory bulb of Onychodactylus fischeri locates in the rastral and lateral to the cerebral hemisphere, and six distinct layers can be identified from the lateral to the medial, quite similar to Batrachuperus tibetanus and Hynobius leechii. In the cerebrum, the primordial hippocampus developed better than the primordial piriform. The former belongs to archipallium and the latter is paleopallium. Ventral to the primordial hippocampus there is a septal area which cannot be divided into medial and lateral parts. In the ventrical wall, there is neither medial limiting sulcus nor lateral limiting sulcus to separate the primordial hippocampus and the septal area, or the primordial piriform and the corpus striatum. The corpus striatum of Onychodactylus fischeri is paleostriatum. There is choroids plexus anterior in the lateral ventricle. The cell group that located at two sides of the third ventricle is the amygdale. Besides, the shape and size of neurons within the telencephalon are poorly differentiated.
CONCLUSIONOnychodactylus fischeri is a relatively primitive type in the amphibian. The present data will help us to further understand the nerve system of tailed amphibian.
Animals ; Telencephalon ; cytology ; Urodela ; anatomy & histology
8.Spinal Cord Regeneration in Rat using Neural Stem Cell Differentiated from Human Telencephalon.
Sang Rim KIM ; Kwang Hoon CHUNG ; Kwang Bok LEE
The Journal of the Korean Orthopaedic Association 2007;42(3):324-331
PURPOSE: To evaluate the effect of neural stem cells differentiated from a human telencephalon on the neural regeneration in the severed spinal cord. MATERIALS AND METHODS: The 1st surgery involving the insertion of plastic membrane in the transected cord was performed to prevent spontaneous healing of adult female rats (n=20, 171-237 g) with a complete spinal cord transection. The media was inserted only after removing the previously inserted plastic membrane in the control group (n=6). In the experimental group (n=14), media and neural stem cell (1x) were transplanted after removing the membrane, and immunohistochemical staining was performed. The experimental group was perfused transcardially 5 weeks after the 2nd surgery, and the level of neural cell regeneration determined by immunohistochemical staining. In behavioral analysis, the Basso-Beatie-Bresnahan (BBB) scores of the control and experimental group were compared weekly from immediately after the injury until 5 weeks post-injury after the 2nd surgery. RESULTS: Immunohistochemical stain revealed no neural regeneration in the control group. On the other hand, the survival of transplanted human neural stem cells and remarkable neural regeneration (differentiate to neuron and astrocyte) were observed in the experimental group. In the BBB locomotor scale, the experimental group showed significant recovery in terms of control; and the score increased from postoperative 2 weeks to 3 weeks, and reached a plateau from 3 weeks to 5 weeks. CONCLUSION: The effect of neural stem cells differentiated from human telencephalon on cord regeneration does not produce functional recovery in the BBB locomotor scale, but there is slight recovery of the muscle function. The survival of transplanted human neural stem cells and the possibility of differentiation to neurons or astrocytes were observed.
Adult
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Animals
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Astrocytes
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Female
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Hand
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Humans*
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Membranes
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Neural Stem Cells*
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Neurons
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Plastics
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Rats*
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Regeneration
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Spinal Cord Injuries
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Spinal Cord Regeneration*
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Spinal Cord*
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Telencephalon*
9.Correlation of cell apoptosis with brain edema and elevated intracranial pressure in traumatic brain injury.
Xiao-feng YANG ; Wei-guo LIU ; Hong SHEN ; Jiang-biao GONG ; Jun YU ; Wei-wei HU ; Shi-ting LÜ ; Xiu-jue ZHENG ; Wei-ming FU
Chinese Journal of Traumatology 2005;8(2):96-100
OBJECTIVETo study the correlation between brain edema, elevated intracranial pressure (ICP) and cell apoptosis in traumatic brain injury (TBI).
METHODSIn this study, totally 42 rabbits in 7 groups were studied. Six of the animals were identified as a control group, and the remaining 36 animals were equally divided into 6 TBI groups. TBI models were produced by the modified method of Feeney. After the impact, ICP of each subject was recorded continuously by an ICP monitor until the animal was sacrificed at scheduled time. The apoptotic brain cells were detected by an terminal deoxynucleotide-transferase-mediated dUTP-digoxigenin nick end labeling (TUNEL) assay. Cerebral water content (CWC) was measured with a drying method and calculated according to the Elliott formula. Then, an analysis was conducted to determine the correlation between the count of apoptotic cells and the clinical pathological changes of the brain.
RESULTSApoptotic cell count began to increase 2 h after the impact, and reached its maximum about 3 days after the impact. The peak value of CWC and ICP appeared 1 day and 3 days after the impact, respectively. Apoptotic cell count had a positive correlation with CWC and ICP.
CONCLUSIONSIn TBI, occurrence of brain edema and ICP increase might lead to apoptosis of brain cells. Any therapy which can relieve brain edema and/or decrease ICP would be able to reduce neuron apoptosis, thereby to attenuate the secondary brain damage.
Animals ; Apoptosis ; Brain Edema ; etiology ; metabolism ; pathology ; Brain Injuries ; complications ; pathology ; physiopathology ; Cell Count ; Disease Models, Animal ; In Situ Nick-End Labeling ; Intracranial Hypertension ; etiology ; pathology ; physiopathology ; Male ; Necrosis ; genetics ; pathology ; Rabbits ; Reference Values ; Telencephalon ; metabolism ; Water ; metabolism
10.The Comparison of Regional Cerebral Glucose Metabolism Using Positron Emission Tomography during Sevoflurane and Propofol Anesthesia in Human.
Yong Bo JEONG ; In Cheol CHOI ; Ja Young JU ; Jae Seung KIM ; Seong Deok KIM
Korean Journal of Anesthesiology 2005;49(3):385-394
BACKGROUND: Recent increases in use of sevoflurane have made active researches on its effects in the cerebral metabolism. However, no specific data on brain glucose metabolism has been reported from human study. We compared the brain glucose metabolism during sevoflurane anesthesia with that of propofol anesthesia using positron emission tomography (PET) in the same human volunteers. METHODS: PET scan was performed two times at intervals of one week in each eight volunteers. One scan was performed in sevoflurane anesthesia, and the other was performed in propofol anesthesia. Each was titrated to the point of unconsciousness. The scan was obtained by the 18fluorodeoxyglucose technique. Relative cerebral glucose metabolic rate (rCMRg) was assessed with statistical parametric mapping. RESULTS: The regions of decreased rCMRg during sevoflurane aneshesia were the visual cortex, posterior parietal association area, primary somatosensory area, and premotor area. During propofol anesthesia the decreased regions were the visual inferotemporal area and prefrontal association area in addition to those area of sevoflurane anesthesia. The increased regions were the partial prefrontal association area, basal ganglia, cingulate, olfactory-limbic cortex, midbrain, and pons during sevoflurane anesthesia, and the primary motor area, insula, thalamus, medulla along with those area of sevoflurane during propofol anesthesia. CONCLUSION: Propofol suppressed the rCMRg of neocortex area more than sevoflurane, and sevoflurane suppressed the rCMRg of paleocortex, telencephalon more than propofol when the unconsciousness level was achieved by anesthesia. Sevoflurane produces different effects on relative brain glucose metabolism with propofol.
Anesthesia*
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Basal Ganglia
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Brain
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Electrons*
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Glucose*
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Healthy Volunteers
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Humans*
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Mesencephalon
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Metabolism*
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Neocortex
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Pons
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Positron-Emission Tomography*
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Propofol*
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Rabeprazole
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Telencephalon
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Thalamus
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Unconsciousness
;
Visual Cortex
;
Volunteers

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