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Gene and protein expression profiles of olfactory ensheathing cells from olfactory bulb versus olfactory mucosa

Neural Regeneration Research 2022年第2期17卷 440-449页

作者： Yuan-Xiang Lan Ping Yang Zhong Zeng Neeraj Yadav Li-Jian Zhang Li-Bin Wang He-Chun Xia School of Clinical Medicine Ningxia Medical UniversityYinchuanNingxia Hui Autonomous RegionChina Ningxia Human Stem Cell Institute General Hospital of Ningxia Medical UniversityYinchuanNingxia Hui Autonomous RegionChina Department of Neurosurgery General Hospital of Ningxia Medical UniversityYinchuanNingxia Hui Autonomous RegionChina Clinical Laboratory Center General Hospital of Ningxia Medical UniversityYinchuanNingxia Hui Autonomous RegionChina School of International Education Ningxia Medical UniversityYinchuanNingxia Hui Autonomous RegionChina Biochip Research Center General Hospital of Ningxia Medical UniversityYinchuanNingxia Hui Autonomous RegionChina

olfactory ensheathing cells(OECs) from the olfactory bulb(OB) and the olfactory mucosa(OM) have the capacity to repair nerve injury. However, the difference in the therapeutic effect between OB-derived OECs and OM-derived OECs remains unclear. In this study, we extracted OECs from OB and OM and compared the gene and protein expression profiles of the cells using transcriptomics and non-quantitative proteomics techniques. The results revealed that both OB-derived OECs and OM-derived OECs highly expressed genes and proteins that regulate cell growth, proliferation, apoptosis and vascular endothelial cell regeneration. The differentially expressed genes and proteins of OB-derived OECs play a key role in regulation of nerve regeneration and axon regeneration and extension, transmission of nerve impulses and response to axon injury. The differentially expressed genes and proteins of OM-derived OECs mainly participate in the positive regulation of inflammatory response, defense response, cytokine binding, cell migration and wound healing. These findings suggest that differentially expressed genes and proteins may explain why OB-derived OECs and OM-derived OECs exhibit different therapeutic roles. This study was approved by the Animal Ethics Committee of the General Hospital of Ningxia Medical University(approval No. 2017-073) on February 13, 2017.

关键词： biological process cellular component gene Gene Ontology Kyoto Encyclopedia of Genes and Genomes molecular function olfactory bulb olfactory ensheathing cells olfactory mucosa protein

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Efficient RT-QuIC seeding activity for α-synuclein in olfactory mucosa samples of patients with Parkinson’s disease and multiple system atrophy

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Translational Neurodegeneration 2019年第1期8卷 290-303页

作者： Chiara Maria Giulia De Luca Antonio Emanuele Elia Sara Maria Portaleone Federico Angelo Cazzaniga Martina Rossi Edoardo Bistaffa Elena De Cecco Joanna Narkiewicz Giulia Salzano Olga Carletta Luigi Romito Grazia Devigili Paola Soliveri Pietro Tiraboschi Giuseppe Legname Fabrizio Tagliavini Roberto Eleopra Giorgio Giaccone Fabio Moda Fondazione IRCCS Istituto Neurologico Carlo Besta Unit of Neurology 5 and NeuropathologyMilanItaly Fondazione IRCCS Istituto Neurologico Carlo Besta Unit of Neurology I-Parkinson and Movement Disorders UnitMilanItaly Department of Health Sciences Universitàdegli Studi di MilanoOtolaryngology UnitSan Paolo HospitalMilanItaly Department of Neuroscience Scuola Internazionale Superiore di Studi Avanzati(SISSA)Laboratory of Prion BiologyTriesteItaly Fondazione IRCCS Istituto Neurologico Carlo Besta Scientific DirectorateMilanItaly

Background:Parkinson’s disease(PD)is a neurodegenerative disorder whose diagnosis is often challenging because symptoms may overlap with neurodegenerative parkinsonisms.PD is characterized by intraneuronal accumulation of abnormalα-synuclein in brainstem while neurodegenerative parkinsonisms might be associated with accumulation of eitherα-synuclein,as in the case of Multiple System Atrophy(MSA)or tau,as in the case of Corticobasal Degeneration(CBD)and Progressive Supranuclear Palsy(PSP),in other disease-specific brain regions.Definite diagnosis of all these diseases can be formulated only neuropathologically by detection and localization ofα-synuclein or tau aggregates in the brain.Compelling evidence suggests that trace-amount of these proteins can appear in peripheral tissues,including receptor neurons of the olfactory mucosa(OM).Methods:We have set and standardized the experimental conditions to extend the ultrasensitive Real Time Quaking Induced Conversion(RT-QuIC)assay for OM analysis.In particular,by using human recombinantα-synuclein as substrate of reaction,we have assessed the ability of OM collected from patients with clinical diagnoses of PD and MSA to induceα-synuclein aggregation,and compared their seeding ability to that of OM samples collected from patients with clinical diagnoses of CBD and PSP.Results:Our results showed that a significant percentage of MSA and PD samples inducedα-synuclein aggregation with high efficiency,but also few samples of patients with the clinical diagnosis of CBD and PSP caused the same effect.Notably,the final RT-QuIC aggregates obtained from MSA and PD samples owned peculiar biochemical and morphological features potentially enabling their discrimination.Conclusions:Our study provide the proof-of-concept that olfactory mucosa samples collected from patients with PD and MSA possess important seeding activities forα-synuclein.Additional studies are required for(i)estimating sensitivity and specificity of the technique and for(ii)evaluating its application for the diagno

关键词： RT-QuIC olfactory mucosa Parkinson’s disease Neurodegenerative parkinsonisms α-Synuclein

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Isolation,culture,and purification of olfactory mucosa-derived olfactory ensheathing cells using modified differential attachment with low concentration serum

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Neural Regeneration Research 2008年第8期3卷 817-821页

作者： Huaqing Yang Qiang Li Kunzheng Wang Bin Wang Hui Qiang Wei Wang Jianxiang Yao Department of Orthopedics Second Hospital of Xi＇an Jiaotong University Xi＇an 710004 Shaanxi Province China Department of Orthopedics Second Hospital of Tsinghua University Beijing 100049 China

BACKGROUND：Studies have demonstrated that olfactory mucosa can promote the regeneration and formation of axonal medullary sheath of injured neurons. To date, olfactory ensheathing cells （OECs） utilized in basic and clinical research arise primarily from the olfactory bulb mucosa. However, little is known regarding culture, purification, and biological properties of OECs . OBJECTIVE： To isolate and culture OECs utilized modified, differential attachment in combination with neurotrophic factor 3 （NT3） and low concentration serum to explore an optimal in vitro culture method for OECs.DESIGN, TIME AND SETTING： Single-sample observation was performed at the Medical Experimental Center of Stomatology College, Xi＇an Jiaotong University between March 2006 and December 2007. MATERIALS： Twelve samples from aborted embryos, 4-6 months, were used to isolate OECs; rabbit-anti-human p75NTR and glial fibrillary acidic protein （GFAP） antibody were provided by Sigma, USA. METHODS： The differential time was six hours. This was repeated twice, based on Nash＇s differential attachment. Attached OECs were cultured in DMEM-F12 culture medium containing 10% fetal bovine serum （FBS） or 2.5% FBS and NT3. MAIN OUTCOME MEASURES： OEC morphology was observed, and p75NTR and GFAP immunocyto-chemistry was used for identification and purity detection. RESULTS： Some cells attached after three days in culture. Several cells possessed short neurites with good refractivity. Some shuttle-shaped fibroblasts could be seen. On day six, more cells attached, exhibiting a three-dimensional appearance. Many cells appeared dipolar or tripolar, with slender neurites, and fibroblasts were clustered. On day nine, the number of dipolar or tripolar cell bodies with slender neurites was increased, and fibroblasts were clustered. On day 15, fibroblasts occupied the majority of the bottom of the culture bottle, with several OECs surviving at the upper layer. OECs were positive for P75NTR and GFAP expression, as identified by an immunocytologically stained brown ce

关键词： cell culture olfactory ensheathing cells olfactory mucosa purification

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Effect of Mancozeb Inhalation on the olfactory mucosa in Rats

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International Journal of Otolaryngology and Head & Neck Surgery 2022年第5期11卷 227-233页

作者： Berzan Haznedar Engin Deveci Ertuğrul Gök Department of Otolaryngology Gazi Yaş argil Training and Research Hospital Diyarbak&#305 r Republic of Turkey Department of Histology and Embryology Medical School Dicle University Diyarbak&#305 r Republic of Turkey Department of Forensic Medicine Medical School Dicle University Diyarbak&#305 r Republic of Turkey

Background: Mancozeb, (ethylene-bis-dithiocarbamate), is an important fungicide useful against a wide range of fungus affecting ornamental plants, crops, and fruits. We aimed to evaluate the changes in the olfactory mucosa due to mancozeb toxicity and, if cytokine is active, IL-6 immunoactivity. Material and Method: In experimental group, the mancozeb (500 mg/kg) was administered with inhalation to 10 male Wistar Albino rats for five days a week. The control group (n = 10) received distilled water with spray at the same time period. The experiment was terminated after three weeks. Samples were placed in 10% formaldehyde for fixation and placed in paraffin, sections of 5 μm were prepared from paraffin blocks and stained with Hematoxylin-Eosin. Interleukin-6 (IL-6) primary antibody were used for immunohistochemical analysis. Result: In the mancozeb group, olfactory epithelial cell degeneration and apoptosis, inflammation, dilatation and congestion in the vessels were observed. IL-6 expression was increased in vascular endothelium and inflammatory cells. Conclusions: Mancozeb was thought that the increase in IL-6 expression due to the increase in cell degeneration signal was thought to affect the development of cell apoptosis and angiogenesis, and that the use of mancozeb might adversely affect the olfactory mechanism.

关键词： Mancozeb olfactory mucosa IL-6 Rat

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Nogo-A expression in injured spinal cord following human olfactory mucosa-derived olfactory ensheathing cells transplantation

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Neural Regeneration Research 2011年第16期6卷 1217-1221页

作者： Bin Wang Qiang Li Xijing He Weixiong Wang Department of Emergency Surgery the Second Affiliated Hospital of Guangzhou Medical University Guangzhou 510260 Guangdong Province China Department of Orthopedics Qingdao Municipal Hospital Qingdao 266011 Shandong Province China 3The Second Department of Orthopedics the Second Affiliated Hospital of Xi'an Jiaotong University Xi'an 710004 Shaanxi Province China

Transplantation of olfactory bulb-derived olfactory ensheathing cells （OECs） promotes motor functional recovery in rats with acute spinal cord injury, possibly by Nogo-A expression changes at the injury site. The present study transplanted OECs derived from the olfactory mucosa （OM） of rats OM-derived OEC （OM-OEC） transplantation significantly reduced the increase of Nogo-A protein and mRNA expression caused by spinal cord injury, supporting the hypothesis that OM-OECs improve spinal cord regeneration by reducing Nogo-A expression.

关键词： Spinal cord injury olfactory ensheathing cells olfactory mucosa cell transplantation neural regeneration

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olfactory swab sampling optimization for α-synuclein aggregate detection in patients with Parkinson’s disease

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Translational Neurodegeneration 2022年第1期11卷 390-400页

作者： Matilde Bongianni Mauro Catalan Daniela Perra Elena Fontana Francesco Janes Claudio Bertolotti Luca Sacchetto Stefano Capaldi Matteo Tagliapietra Paola Polverino Valentina Tommasini Giulia Bellavita Elham Ataie Kachoie Roberto Baruca Andrea Bernardini Mariarosaria Valente Michele Fiorini Erika Bronzato Stefano Tamburin Laura Bertolasi Lorenzo Brozzetti Maria Paola Cecchini Gianluigi Gigli Salvatore Monaco Paolo Manganotti Gianluigi Zanusso Department of Neurosciences Biomedicineand Movement SciencesPoliclinico G.B.RossiUniversity of Verona37134 VeronaItaly Neurology Unit Department of MedicineSurgery and Health SciencesOspedale CattinaraUniversity of Trieste34128 TriesteItaly Neurology Unit University of Udine Academic Hospital33100 UdineItaly Department of Surgical Sciences DentistryGynecology and PediatricsUniversity of Verona37134 VeronaItaly Biocrystallography Laboratory Department of BiotechnologyUniversity of Verona37134 VeronaItaly Otolaryngology Unit Department of MedicineSurgery and Health SciencesOspedale CattinaraUniversity of Trieste34128 TriesteItaly.

Background: In patients with Parkinson’s disease (PD), real-time quaking-induced conversion (RT-QuIC) detection of pathological α-synuclein (α-syn) in olfactory mucosa (OM) is not as accurate as in other α-synucleinopathies. It is unknown whether these variable results might be related to a different distribution of pathological α-syn in OM. Thus, we investigated whether nasal swab (NS) performed in areas with a different coverage by olfactory neuroepithelium, such as agger nasi (AN) and middle turbinate (MT), might affect the detection of pathological α-syn. Methods: NS was performed in 66 patients with PD and 29 non-PD between September 2018 and April 2021. In 43 patients, cerebrospinal fluid (CSF) was also obtained and all samples were analyzed by RT-QuIC for α-syn. Results: In the first round, 72 OM samples were collected by NS, from AN (NSAN) or from MT (NSMT), and 35 resulted positive for α-syn RT-QuIC, including 27/32 (84%) from AN, 5/11 (45%) from MT, and 3/29 (10%) belonging to the non-PD patients. Furthermore, 23 additional PD patients underwent NS at both AN and MT, and RT-QuIC revealed α-syn positive in 18/23 (78%) NSAN samples and in 10/23 (44%) NSMT samples. Immunocytochemistry of NS prepara-tions showed a higher representation of olfactory neural cells in NSAN compared to NSMT. We also observed α-syn and phospho-α-syn deposits in NS from PD patients but not in controls. Finally, RT-QuIC was positive in 22/24 CSF samples from PD patients (92%) and in 1/19 non-PD. Conclusion: In PD patients, RT-QuIC sensitivity is significantly increased (from 45% to 84%) when NS is performed at AN, indicating that α-syn aggregates are preferentially detected in olfactory areas with higher concentration of olfactory neurons. Although RT-QuIC analysis of CSF showed a higher diagnostic accuracy compared to NS, due to the non-invasiveness, NS might be considered as an ancillary procedure for PD diagnosis.

关键词： Parkinson disease Alpha-synuclein Real-time quaking-induced conversion assay olfactory mucosa Cerebrospinal fluid

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Transplantation of olfactory mucosa as a Scaffold for Axonal Regeneration Following Spinal Cord Contusion in Rats

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Neuroscience & Medicine 2013年第2期4卷 112-116页

作者： Koichi Iwatsuki Toshiki Yoshimine Yoshiyuki Sankai Masao Umegaki Yu-Ichiro Ohnishi Masahiro Ishihara Takashi Moriwaki Noriko Oda Department of Neurosurgery Medical SchoolOsaka UniversityOsakaJapan Faculty of Engineering‚Information and Systems University of TsukubaTsukubaJapan

Object: The inability of the spinal cord to regenerate after SCI is due to the extremely limited regenerative capacity of most central nervous system (CNS) axons, along with the hostile environment of the adult CNS, which does not support axonal growth. It seems that for successful axonal regeneration to take place, a supportive local environment is required after the injury. We have previously reported that transplantation of the olfactory mucosa is effective in restoring functional recovery in rats following spinal cord transaction. In this study, we examined histological features of olfactory mucosa grafts in rats subjected to a spinal cord contusion protocol. Respiratory mucosa was utilized as a control, as we have previously found that respiratory mucosa does not support neuronal generation. Methods: The rats spinal cords were crash-injured by dropping a 10-g metal rod from a height of 7.5 cm, and a couple of weeks later, the injury sites were exposed, and both olfactory and respiratory mucosae were inserted into the posterior sulcuses of the spinal cord. The each number of olfactory and mucosa transplanted rats were five. The Basso, Beattie, and Bresnahan (BBB) score was observed. Immunohistochemical study for neurofilament was performed. Results: olfactory mucosa transplanted rats following spinal cord injury can support at least partial hind limb motor recovery compared with respiratory mucosa transplanted rats and we identified numerous axons surrounding the transplanted olfactory mucosa cells, and penetrating the olfactory mucosa at the transplant site. Conclusion: olfactory mucosa might be a suitable scaffold for axonal regeneration.

关键词： olfactory mucosa Spinal Cord Injury Transplantation Scaffold

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