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neuroglobin mutation associated with hypoxia adaptation in Tibet chicken

Progress in Natural Science:Materials International 2007年第12期17卷 1419-1424页

作者： Zhang Lifan1, Liu Chong2, Lian Linsheng3, Zhao Chunjiang1, Bao Haigang1 and Wu Changxin1(1. College of Animal Science and Technology, China Agricultural University, Beijing 100094, China 2. College of Biological Science, China Agricultural University, Beijing 100094, China 3. College of Animal Science and Technology, Yunnan Agricultural University, Kunming 650201, China) College of Animal Science and Technology China Agricultural University Beijing 100094 China College of Biological Science China Agricultural University Beijing 100094 China College of Animal Science and Technology Yunnan Agricultural University Kunming 650201 China

Globin protein family plays an important role in storing and transporting oxygen. As a newly reported globin, the revealed function of neuroglobin includes binding and storing oxygen as well as facilitating the utilization of oxygen in neuronal cells. In the present study, mutations in exons of chicken Ngb gene were identified with the method of sequencing and created restriction site PCR in Tibet chicken and other four lowland chicken breeds. The mutations of Lys-2224(E4)-Asn and Ser-2279(E4)-Gly were identified in exon 4 of the gene. The Lys-2224(E4)-Asn mutation existed only in Tibet chicken and the mutation frequencies increased with increasing altitude. Analysis of the haplotype and diplotype of the two mutations in Tibet chicken populations of different altitudes showed that the frequencies of TG haplotype and TTGG diplotype also increased with increasing altitude, while the reverse tendency was found on GGAA diplotype. Under the hypoxic simulation incubation, the main haplotype was TG in living embryos and GA in dead embryos. The results showed that the Lys-2224(E4)-Asn mutation may be a specific mutation associated with hypoxia adaptation in Tibet chicken.

关键词： Tibet chicken neuroglobin hypoxia adaptation Ngb gene mutation.

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neuroglobin expression in rats after traumatic brain injury

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Neural Regeneration Research 2012年第25期7卷 1960-1966页

作者： Xin Lin Min Li Aijia Shang Yazhuo HU Xiao Yang Ling Ye Suyan Bian ZhongfengWang Dingbiao Zhou Department of Surgery Division of Nanlou General Hospital of Chinese PLA Beijing 100853 China Department of Traumatic-Aesthetic Surgery Huangsi Aesthetic Surgery Hospital Beijing 100120 China Department of Neurosurgery Division of Surgery General Hospital of Chinese PLA Beijing 100853 China Institute of Geriatrics General Hospital of Chinese PLA Beijing 100853 China Institute of Microbiology and Epidemiology Academy of Military Medical Sciences Beijing 100071 China Department of Cardiology Division of Nanlou General Hospital of Chinese PLA Beijing 100853 China Institute of Brain Science and State Key Laboratory of Medical Neurobiology Fudan University Shanghai 200032 China

In this study, we used a rat model of severe closed traumatic brain injury to explore the relationship between neuroglobin, brain injury and neuronal apoptosis. Real-time PCR showed that neuroglobin mRNA expression rapidly increased in the rat cerebral cortex, and peaked at 30 minutes and 48 hours following traumatic brain injury. Immunohistochemical staining demonstrated that neuroglobin expression increased and remained high 2 hours to 5 days following injury. The rate of increase in the apoptosis-related Bax/Bcl-2 ratio greatly decreased between 30 minutes and 1 hour as well as between 48 and 72 hours post injury. Expression of neuroglobin and the anti-apoptotic factor Bcl-2 greatly increased, while that of the proapoptotic factor decreased, in the cerebral cortex post severe closed traumatic brain injury. It suggests that neuroglobin might protect neurons from apoptosis after traumatic injury by regulating Bax/Bcl-2 pathway.

关键词： neuroglobin traumatic brain injury neuron apoptosis cerebral cortex Bax Bcl-2 neuralregeneration

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neuroglobin and neuroprotection:the role of natural and synthetic compounds in neuroglobin pharmacological induction

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Neural Regeneration Research 2021年第12期16卷 2353-2358页

作者： Lidia Ciccone Susanna Nencetti Simone Socci Elisabetta Orlandini Department of Pharmacy University of PisaPisaItaly Department of Earth Sciences University of PisaPisaItaly Research Center“E.Piaggio ”University of PisaPisaItaly

neuroglobin(Ngb)is a 17 kDa monomeric hexa-coordinated heme protein belonging to the globin *** is mainly expressed in neurons of the central and peripheral nervous system,although moderate levels of Ngb have been detected in non-nervous *** the past decade,Ngb has been studied for its neuroprotective role in a large number of neurological disorders such as Alzheimer’s disease,Huntington’s disease,brain ischemia and *** review discusses and summarizes the natural compounds and the small synthetic molecules capable of modulating Ngb expression that exhibits a protective role against various neurodegenerative diseases.

关键词： neuroglobin neuroglobin pharmacological induction neuroprotection neurodegenerative diseases natural compounds synthetic small molecules Alzheimer’s disease Huntington’s disease

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neuroglobin Facilitates Neuronal Oxygenation through Tropic Migration under Hypoxia or Anemia in Rat:How Does the Brain Breathe?

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Neuroscience Bulletin 2023年第10期39卷 1481-1496页

作者： Chun-Yang Li Hai-Feng Jiang Li Li Xiao-Jing Lai Qian-Rong Liu Shang-Bin Yu Cheng-La Yi Xiao-Qian Chen Department of Pathophysiology Tongji Medical CollegeKey Laboratory of Neurological DiseasesThe Ministry of Education(HUST)Huazhong University of Science and TechnologyWuhan 430030China School of Life Science and Technology Wuhan Polytechnic UniversityWuhan 430023China Department of Traumatic Surgery Tong-ji HospitalTong-ji Medical CollegeHuazhong University of Science and TechnologyWuhan 430030China

The discovery of neuroglobin(Ngb),a brain-or neuron-specific member of the hemoglobin family,has revolutionized our understanding of brain oxygen ***,how Ngb plays such a role remains far from ***,we report a novel mechanism by which Ngb might facilitate neuronal oxygenation upon hypoxia or *** found that Ngb was present in,co-localized to,and co-migrated with mitochondria in the cell body and neurites of *** induced a sudden and prominent migration of Ngb towards the cytoplasmic membrane(CM)or cell surface in living neurons,and this was accompanied by the *** vivo,hypotonic and anemic hypoxia induced a reversible Ngb migration toward the CM in cerebral cortical neurons in rat brains but did not alter the expression level of Ngb or its cytoplasm/mitochondria ***-down of Ngb by RNA interference significantly diminished respiratory succinate dehydrogenase(SDH)and ATPase activity in neuronal N2a ***-expression of Ngb enhanced SDH activity in N2a cells upon *** of Ngb at its oxygen-binding site(His64)significantly increased SDH activity and reduced ATPase activity in N2a *** together,Ngb was physically and functionally linked to *** response to an insufficient oxygen supply,Ngb migrated towards the source of oxygen to facilitate neuronal *** novel mechanism of neuronal respiration provides new insights into the understanding and treatment of neurological diseases such as stroke and Alzheimer’s disease and diseases that cause hypoxia in the brain such as anemia.

关键词： neuroglobin Hypoxia Ischemia Anemia Neuron Astrocyte

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Cerebroprotection with recombinant neuroglobin plasmid in a rat model of focal cerebral ischemia

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Neural Regeneration Research 2010年第1期5卷 52-57页

作者： Ji Zhu Wenyuan Tang Department of Neurosurgery First Affiliated Hospital of Chongqing Medical University Chongqing 400016 China

BACKGROUND： Adenovirus has been used to develop neuroglobin （Ngb） vectors. Although transfection efficiency is high, induced gene mutation, cytotoxicity, inflammation, and low exogenous gene content have limited its application. OBJECTIVE： To observe the effects of recombinant Ngb plasmid in a rat model of focal cerebral ischemia. DESIGN, TIME AND SETTING： Genetically engineered, randomized, controlled, animal experiment was performed at the Laboratory of Chongqing Medical University from May 2006 and January 2007. MATERIALS： 2, 3, 5-triphenyltetrazolium chloride was purchased from Shanghai Sangon Biological Engineering Technology and Services. Rabbit anti-rat Bcl-2 polyclonal antibody, rabbit anti-rat β-actin monoclonal antibody, and FITC-labeled goat anti-rabbit IgG were purchased from Sigma, USA. TUNEL apoptosis kit was purchased from Roche, Germany. METHODS： A total of 54 male, adult, Wistar rats were randomly assigned to 3 groups （n=18）： normal saline, plasmid control, and recombinant Ngb （pCDNA3.1 （＋）/Ngb）. Normal saline, plasmid pCDNA3.1 （＋）, and recombinant plasmid pCDNA3.1 （＋）/Ngb were separately injected into two sites in the rat cerebral cortex, and models of focal ischemia were established by occlusion of the right middle cerebral artery after 24 hours. MAIN OUTCOME MEASURES： Local ischemic damage was detected by 2, 3, 5- triphenyltetrazolium chloride staining, apoptosis in the penumbra was confirmed using the TUNEL method, and Bcl-2 protein expression in the penumbra was determined by indirect immunofluorescent staining and Western blot analysis. RESULTS： Compared with the normal saline and plasmid control groups, cerebral infarction size and the number of apoptotic cells in the pCDNA3.1 （＋）/Ngb group were significantly reduced （P 〈 0.01）. The percentage of Bcl-2-positive cells in the penumbra of the pCDNA3.1 （＋）/Ngb group was significantly increased （P 〈 0.01）. The relative expression level of Bcl-2 protein was increased by 40%-50%. CONCLUSION： Recombi

关键词： neuroglobin cerebral ischemia cell apoptosis ischemic penumbra Bcl-2

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Overexpression of the neuroglobin gene delivered by ultrasound-targeted microbubble destruction protects SH-SY5Y cells against cobalt chloride induced hypoxia

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Neural Regeneration Research 2011年第25期6卷 1947-1953页

作者： Qian Yang Dianwen Gao Qingzhu Nie Zhengang Cai Jian Du Lujuan Shan Yuejian Liu Department of Ophthalmology Shengjing Hospital Affiliated to China Medical University Shenyang 110004 Liaoning Province China Department of General Surgery First Hospital Affiliated to Dalian Medical University Dalian 116021 Liaoning Province China Experimental Research Center First Hospital Affiliated to Dalian Medical University Dalian 116021 Liaoning Province China

In this study, we examined the effects of neuroglobin gene （Ngb） transfection into SH-SY5Y cells, using ultrasound-targeted microbubble destruction （UTMD）, on cobalt chloride-induced hypoxia. With an ultrasound intensity of 0.8 W/cm2, a 60-second exposure duration, 50% duty cycle, and 20% microbubble concentration, pAcGFP1-C1-Ngb-transfected cells exhibited the highest cell viability and transfection efficiency. The efficiency of plasmid delivery was significantly higher with UTMD than transfection with plasmid alone, transfection with plasmid using microbubbles, or transfection of plasmid by ultrasound. In addition, during cobalt chloride-induced hypoxia, caspase-3 activity in pAcGFP1-C1-Ngb-transfected cells was significantly lower than in untransfected cells. Ngb protein and mRNA expression were significantly higher in cells transfected by UTMD than in cells transfected with the other methods. These results demonstrate that UTMD can very efficiently mediate exogenous gene delivery, and that Ngb overexpression protects cells against cobalt chloride-induced hypoxia.

关键词： ultrasound-targeted microbubble destruction neuroglobin gene therapy recombinant plasmid SH-SY5Y cells neural regeneration

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Determination of vinyl orientation of mouse neuroglobin by 2D nuclear Overhauser spectroscopy

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Chinese Chemical Letters 2007年第3期18卷 319-321页

作者： Su Yuan Zhou Xiao Yan Wei Department of Biomechanlcs Shanghai University of Sports Shanghai 200438 China

^1H NMR has been used to determine the 2-, 4-vinyl orientation of heme active site from oxidized mouse neuroglobin （mNgb）. The NOEs between 3-methyl and Hα, Hβ of 2-vinyl, together with the NOEs between 5-methyl and Hα, Hβ of 4-vinyl, allowed the unambiguous determination of trans and cis orientations for the 2- and 4-vinyl groups in the mNgb, respectively.

关键词： Heme Vinyl orientation neuroglobin

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Expression of neuroglobin in ocular hypertension induced acute hypoxic-ischemic retinal injury in rats

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International Journal of Ophthalmology(English edition) 2011年第4期4卷 393-395页

作者： Shao-Yang Shi Xue-Mei Feng You Li Xun Li and Xiao-Long Chen Department of Ophthalmology Shengjing Hospital of China Medical University Shenyang 11004 Liaoning Province China

AIM: To investigate the Expression of neuroglobin (Ngb) in the retina of rats with ocular hypertension induced acute retinal hypoxic-ischemic injury. METHODS: Seventy Wistar rats were divided into 7 groups randomly. The Experimental model was induced by elevation of intraocular pressure via anterior chamber canula insertion in the left eyes and the fellow eyes were preserved as normal controls. The retinal tissues were taken at 1, 5, 10, 15, 20, 30 and 60 minutes after hypoxic-ischemia injury. Protein was extracted, and then analyzed by Western-blot method. SPSS was used for statistical analysis. RESULTS: The time-depended expressions of Ngb were observed. The level of Ngb increased rapidly at 1 minute after ischemia and reached to the peak at 5 minutes, which had significant difference from that of control group (P 0.05). CONCLUSION: The expression of Ngb in retinal tissue increased rapidly after hypoxic-ischemic injury in rats, suggesting that Ngb may play an important role in the process of acute retinal hypoxic-ischemic injury.

关键词： neuroglobin hypoxia-ischemia retina

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neuroglobin co-localize and migrate with mitochondria in neural cells

Neuroglobin co-localize and migrate with mitochondria in neu...

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中国神经科学学会第十二届全国学术会议

作者： Han Xiao Xu Li Tao Yang Rui Zhao Albert Cheung Hoi Yu Lina Li Laboratory For Functional study of astrocytes Neuroscience Research Institute Key Laboratory for Neuroscience of the Ministry of Education

Human brain constitutes 2 % of the total body weight, but utilizes over 20 % of the body oxygen at rest. The discovery of neuroglobin（Ngb） in both neurons and astrocytes suggest that Ngb might be closely involved in brain oxygen homeostasis. In the present study, we have investigated the expression, distribution and mobility of Ngb compared to cytoglobin（Cygb）, a universal cellular respiratory globin, in primary astrocytes and neurons. Unlike Cygb, the expression levels of Ngb in astrocytes and neurons increased under both ischemia and rotenone treatment. A co-localization of either endogenous or recombinant Ngb with mitochondria was evidenced using immunostaining and western blot. Furthermore, the L136 P Ngb mutant enhances the colocalization of Ngb and mitochondria under both normoxic and ischemic conditions. A co-migration of L136 P Ngb with mitochondria in the neurite of neurons was also observed. More interestingly, the co-migration and co-localization of L136 P Ngb with mitochondrial was abolished when cultures were treated with cytoskeleton inhibitors such as cytochalasin D, colchicine and nocodazole. Overall, our data demonstrated that Ngb is a migratory protein in cytoplasm that co-localize with mitochondria under both physiological and pathological conditions. In addition, the co-migration of Ngb with mitochondria requires microfilament. Based on our investigation, we propose the hypothesis that Ngb might have distinctive roles during oxygen transportation in comparison to Cygb and serve as an ‘emergency and mobile’ intrinsic protective protein in nervous system.

关键词： neuroglobin mitochondria astrocytes neurons

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neuroglobin associated with protective roles in neurons and astrocytes

Neuroglobin associated with protective roles in neurons and ...

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中国神经科学学会第九届全国学术会议暨第五次会员代表大会

作者： Albert Cheung-Hoi YU Neuroscience Research Institute Department of NeurobiologySchool of Basic Medical Sciencesthe Key Laboratory for Neuroscience（Ministry of Education）the Key Laboratory for Neuroscience（Ministry of Public Health）Peking University

＜正＞Objective neuroglobin（Ngb） is a recently discovered globin which is expressed in the brain,retina and other nerve *** was identified as an oxygen transport protein in neurons and suggested to be able to protect brain against hypoxic/ischemic *** have previously reported that Ngb could also be expressed in primary cultures of cerebral cortical astrocytes in addition to neurons,and reducing Ngb protein level would enhance apoptosis in astrocytes under ischemic *** objective of our work is to further analyze the roles of Ngb in both astrocytes and *** were compared with those obtained from cytoglobin（Cygb）,another newly found globin *** Primary cultures of cerebral cortical astrocytes and neurons were prepared from newborn ICR *** chamber was used to induce in vitro *** coding region of mouse Ngb or Cygb gene were inserted into different vectors and transfected into primary cultures of astrocytes and neurons. Reverse transcription（RT）-PCR and Western blot analysis were used to detect Ngb/Cygb gene and protein levels. Different cytoskeleton inhibitors were used to treat primary cultures of astrocytes and ***（1） The subcellular distribution of over-expressed Ngb and Cygb in astrocytes was ***-EGFP in astrocytes showed dot-like distribution in the cytoplasm while Cygb-DsRed was homogeneous in the nuclei and cytoplasm.（2） Ngb, but not Cygb,was co-localized with mitochondria in primary cultures of astrocytes and neurons.（3） The expression of Ngb but not Cygb in mitochondria of astrocytes and neurons was increased under ischemic treatment.（4） Ngb-EGFP expression in mitochondria of astrocytes was decreased under microtubules inhibitors treatment which exerted no effect on Cygb-EGFP expression in *** This study indicated that Ngb may play a more important role than Cygb in oxygen transport functions under ischemia.

关键词： neuroglobin cytoglobin astrocytes neurons protection oxygen transport

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