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Osteoblasts are inherently programmed to repel sensory innervation

Bone Research 2020年第3期8卷 343-355页

作者： Luis Leitao Estrela Neto Francisco Conceicao Ana Monteiro Marina Couto Cecilia JAlves Daniela MSousa Meriem Lamghari Instituto de Investigacao e Inovacao em Saude(i3S) Universidade do Porto4200-135 PortoPortugal Instituto de Engenharia Biomedica(INEB) Universidade do Porto4200-135 PortoPortugal Instituto de Ciencias Biomedicas Abel Salazar(ICBAS) Universidade do Porto4050-313 PortoPortugal

Tissue innervation is a complex process controlled by the expression profile of signaling molecules secreted by tissue-resident cells that dictate the growth and guidance of axons.Sensory innervation is part of the neuronal network of the bone tissue with a defined spatiotemporal occurrence during bone development.Yet,the current understanding of the mechanisms regulating the map of sensory innervation in the bone tissue is still limited.Here,we demonstrated that differentiation of human mesenchymal stem cells to osteoblasts leads to a marked impairment of their ability to promote axonal growth,evidenced under sensory neurons and osteoblastic-lineage cells crosstalk.The mechanisms by which osteoblast lineage cells provide this nonpermissive environment for axons include paracrine-induced repulsion and loss of neurotrophic factors expression.We identified a drastic reduction of NGF and BDNF production and stimulation of Sema3A,Wnt4;and Shh expression culminating at late stage of OB differentiation.We noted a correlation between Shh expression profile,OB differentiation stages,and OB-mediated axonal repulsion.Blockade of Shh activity and signaling reversed the repulsive action of osteoblasts on sensory axons.Finally,to strengthen our model,we localized the expression of Shh by osteoblasts in bone tissue.Overall,our findings provide evidence that the signaling profile associated with osteoblast phenotype differentiating program can regulate the patterning of sensory innervation,and highlight osteoblast-derived Shh as an essential player in this cue-induced regulation.

关键词： stimulation innervation reversed

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A paracrine role for white thermogenic adipocytes in innervation: an evidence-based hypothesis

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Neural Regeneration Research 2019年第2期14卷 217-221页

作者： Aejin Lee Ouliana Ziouzenkova Department of Human Sciences The Ohio State University

White adipose tissue(WAT) stores energy and also plays an important endocrine role in producing adipokines for communication with the peripheral and central nervous system. WAT consists of the major lipogenic unilocular adipocytes and the minor populations of beige and brite multilocular adipocytes. These multilocular adipocytes express thermogenic genes and have phenotypic similarity with thermogenic brown adipose tissue. According to a current paradigm, multilocular adipocytes have a thermogenic function in WAT. In this mini review, we discuss data revealing heterogeneity among multilocular cell subsets in WAT and their functions beyond thermogenesis. We propose a hypothetical neuroendocrine role for multilocular adipocytes subsets in the formation of adaptive sensory-sympathetic circuits between the central nervous system and adipose tissue, which activate lipolysis and thermogenesis in WAT in high energy demand situations.

关键词： obesity thermogenesis innervation vitamin A aldehyde dehydrogenase paracrine efferent afferent brown adipose tissue

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innervation of the ovary in amphioxus: ultrastructural and immunohistochemical study

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Chinese Science Bulletin 1998年第11期43卷 925-928页

作者： WelschU Third Institute of Oceanography SOA Xiamen China Department of Anatomy University of München Germany The Fourth Military Medical University Xi’an China

The distribution of unmyelinated nerve fiber in the ovary of amphioxus was found with transmission electron microscopic technique for the first time. The fiber is located under the ovary coat, and in close contact with it. There are two types of synaptic vesicles in the terminals of nerve fiber: one is dense-cored vesicle, the other is clear vesicle. In addition, the nerve terminals contact with follicle cells of ovary can be seen. Using immunohistochemical method, it is further demonstrated that the unmyelinated nerve fiber may be a noradrenergic nerve fiber which is located on the ovary coat and follicle cell.

关键词： amphioxus ovary innervation ultrastructure immunohistochemistry.

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Differential innervation of Secretory Coils and Ducts in Human Eccrine Sweat Glands

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Chinese Medical Journal 2018年第16期131卷 1964-1968页

作者： Zhan Ouyang Hai-Hong Li Ming-Jun Zhang Si-Tian Xie Liu-Hang-Hang Cheng Department of Burn and Plastic Surgery The Second Affiliated Hospital Shantou University Medical College Shantou Guangdong 515041 China

Background： Previous studies demonstrate that eccrine sweat glands are innervated by both cholinergic and adrenergic nerves. However, it is still unknown whether the secretory coils and ducts ofeccrine sweat glands are equally innervated by the sympathetic nerve fibers. To welt understand the mechanisms on sweat secretion and reabsorption, the differential innervation of secretory coils and ducts in human eccrine sweat glands was investigated in the study. Methods： From June 2016 to June 2017, six human skins were fixed, paraffin-embedded, and cut into 5 μm-thick sections, followed by costaining for nerve fiber markers protein gene product 9.5 （PGP 9.5）, tyrosine hydroxylase （TH） and vasoactive intestinal peptide （VIP）, and eccrine sweat gland markers K7, S100P, and K14 by combining standard immunofluorescence with tyramide signal amplification （IF-TSA）. Stained sections were observed under the microscope, photographed, and analyzed. Results： The fluorescent signals of PGP 9.5, TH, and VIP were easily visualized, by 1F-TSA, as circular patterns surrounding eccrine sweat glands, but only PGP 9.5 could be observed by standard IF. The IF-TSA method is more sensitivity than standard IF in detecting antigens expressed at low levels. PGP 9.5, TH, and VIP appeared primarily surrounding the secretory coils and sparsely surrounding the sweat ducts. Conclusion： Sweat secretion is mainly controlled by autonomic nerves whereas sweat reabsorption is less affected by nerve activity.

关键词： Duct Eccrine Sweat Glands innervation Nerve Fibers Secretory Coil Tyramide Signal Amplification

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Adenomyosis uterine innervation in mice correlates to nerve growth factor expression,inflammation,and vascularization

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Neural Regeneration Research 2010年第1期5卷 15-20页

作者： Yan Li Shaofen Zhang Xian Xia Shien Zou Department of Gynaecology Obstetrics and Gynaecology H~spital of Fudan University Shanghai 200011 China

BACKGROUND： Studies have shown that abnormal innervation is an important factor impacting occurrence and development of pathological pain in endometriosis. OBJECTIVE： To observe uterine innervation of adenomyosis mice and to analyze the cause of innervation changes due to nerve growth factor （NGF） expression, inflammation, and vascularization. DESIGN, TIME AND SETTING： This randomized, controlled, animal experiment was performed at the Research Institute of Obstetrics and Gynecology Hospital, and Central Laboratory of Zhongshan Hospital, Fudan University from March to December 2008. MATERIALS： Tamoxifen was provided by Fudan Forward, China. Rabbit anti-mouse NGF was purchased from Santa Cruz Corporation, USA; rabbit anti-protein gene product 9.5 （PGP9.5） and rabbit anti-substance P （SP） were purchased from Chemicon, USA. METHODS： A total of 40 newborn ICR mice were randomly assigned to adenomyosis model and control groups, with 20 animals in each group. Mice in the adenomyosis model group were orally administrated 2.7 μmol/kg tamoxifen on days 2-5 after birth, while the controls were not treated. MAIN OUTCOME MEASURES： Both uteri from all mice were harvested at days 135-145 after birth Expressions of polyclonal PGP9.5 and SP were immunohistochemically detected to demonstrate pan- and sensory nerve fibers. Microvessel density was quantified in the endometrium and myometrium using immunochemical staining for polyclonal rabbit anti-CD31, which stained vessels. Gene expression for NGF, high-affinity tyrosine kinase receptor （trkA）, p75 neuretrophin receptor （p75NTR）, bradykinin receptor-1 （BKR-1）, and 2 （BKR-2）, as well as substance P receptor （neurokininl receptor, NK1-R）, were detected by reverse transcription-polymerase chain reaction. NGF-13 protein expression was detected by Western blot analysis. RESULTS： More nerve fibers were stained with PGP9.5 in the endometrium and myometrium, and with SP in the endometrium, in adenomyosis mice compared with controls （P 〈 0.01 and P 〈 0.05）. Microvessel density in the myom

关键词： adenomyosis innervation nerve growth factor inflammation vascularization

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innervation frequency response of sounding muscles in Mingming cicada

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Chinese Science Bulletin 1995年第1期40卷 71-75页

作者：蒋锦昌杨新宇唐欢徐慕玲陈浩 Institute of Biophysics Chinese Academy of Sciences Beijing 100101 China

In cicadas the songs are produced by vibrations of the sounding membrane driven by sounding muscles. The innervations of sounding movements are synchro-controlling type for some species and asynchro-controlling type for other species. The contracting movements of sounding muscles on both sides are synchronous for some species and alternate for other species, and the contracting frequencies are from several decade to several hundred and generally below 500 Hz. On the basis of studies on the song property and innervation of sound production in the Mingnfing cicada, the innervation frequency-response of movements in sounding muscles is studied in this note. The results provide the important basis for expounding the sounding mechanism of vari-toned songs in cicadas.

关键词： Mingming cicada (Oncotympana maculaticollis Motsch) sounding muscle sounding nerve innervation frequency responese.

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Changes in corneal innervation and pain responses in fungal keratitis

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International Journal of Ophthalmology(English edition) 2020年第1期13卷 1-6页

作者： Chang-Jie Ren Yi-Fan Zhou Yuan Wu Xu-Dong Peng Cui Li Qian Wang Guo-Qiang Zhu Jia You Jie Zhang Gui-Qiu Zhao Jing Lin Department of Ophthalmology the Affiliated Hospital of Qingdao University

AIM: To characterize changes in the cornea nerve and pain responses in fungal keratitis(FK).METHODS: A retrospective analysis of in vivo confocal microscopy images of 11 FK corneas was performed, and the results were compared with those for 11 normal corneas. Subbasal corneal nerves were analyzed for total nerve number, main nerve trunk number, branching patterns and tortuosity. C57 BL/6 mice were infected with Aspergillus fumigatus. Disease severity was determined through clinical scoring and slit lamp photography. Corneas were harvested at 1, 3, 5, and 7 d post infection(p.i.) and assessed for β III tubulin. Corneal mechanical sensitivity thresholds were detected by von Frey test. β-endorphin(β-EP) and μ receptor protein expression was detected through Western blotting.RESULTS: Total nerve number, main nerve trunk number, and nerve branching were significantly lower in FK patients than in controls, but tortuosity was not significantly different. In infected mice, subbasal nerve density decreased from 1 d p.i., reaching a minimum at 5 d p.i. Clinical scores rose at 1 d p.i., peaked at 3 d p.i., and decreased at 5 d p.i. Mechanical sensitivity thresholds showed the same trends. β-EP and μ receptor protein expression increased after infection.CONCLUSION: Corneal nerve density is lower in FK patients and Aspergillus fumigatus-infected mice than in controls. Pain sensitivity decreases with postinfection corneal ulcer aggravation. β-EP and μ receptor proteins are both upregulated in infected mouse corneas.

关键词： keratitis pain fungal innervation subbasal nerve mice

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Transferrin receptor 1 plays an important role in muscle development and denervation-induced muscular atrophy

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Neural Regeneration Research 2021年第7期16卷 1308-1316页

作者： Ying Li Juan-Xian Cheng Hai-Hong Yang Li-Ping Chen Feng-Jiao Liu Yan Wu Ming Fan Hai-Tao Wu Department of Neurobiology Beijing Institute of Basic Medical SciencesBeijingChina Department of Anesthesiology the General Hospital of Western Theater CommandChengduSichuan ProvinceChina Chinese Institute for Brain Research(CIBR) BeijingChina Key Laboratory of Neuroregeneration Coinnovation Center of NeuroregenerationNantong UniversityNantongJiangsu ProvinceChina

Previous studies demonstrate an accumulation of transferrin and transferrin receptor 1(TfR1) in regenerating peripheral nerves.However, the expression and function of transferrin and TfR1 in the denervated skeletal muscle remain poorly understood.In this study, a mouse model of denervation was produced by complete tear of the left brachial plexus nerve.RNA-sequencing revealed that transferrin expression in the denervated skeletal muscle was upregulated, while TfR1 expression was downregulated.We also investigated the function of TfR1 during development and in adult skeletal muscles in mice with inducible deletion or loss of TfR1.The ablation of TfR1 in skeletal muscle in early development caused severe muscular atrophy and early death.In comparison, deletion of TfR1 in adult skeletal muscles did not affect survival or glucose metabolism, but caused skeletal muscle atrophy and motor functional impairment, similar to the muscular atrophy phenotype observed after denervation.These findings suggest that TfR1 plays an important role in muscle development and denervation-induced muscular atrophy.This study was approved by the Institutional Animal Care and Use Committee of Beijing Institute of Basic Medical Sciences, China(approval No.SYXK 2017-C023) on June 1, 2018.

关键词： brachial plexus nerve innervation iron motor dysfunction muscle atrophy signal skeletal muscle transferrin

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Dopaminergic effects on in vitro osteogenesis

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Bone Research 2015年第3期3卷 130-139页

作者： Dong Joon Lee Henry C Tseng Sing Wai Wong Zhengyan Wang Meng Deng Ching-Chang Ko NC Oral Health Institute School of Dentistry University of North Carolina Duke Eye Center and Department of Ophthalmology Duke University Medical Center Oral Biology Program School of Dentistry University of North Carolina Department of Pediatric Dentistry School of Dentistry University of North Carolina Department of Orthodontics School of Dentistry University of North Carolina

Multiple growth factors（e.g., BMP2, TGF-b1, FGF2） and isolated genes have been shown to improve osteoblastic proliferation and mineralization, advancing bone tissue engineering. Among these factors, both polydopamine（PDA） and dopamine（DA） monomer have recently been reported to increase osteoblast proliferation and mineralization in vitro. Although a well-characterized neurotransmitter, DA＇s role in the bone is unknown. We hypothesize that DA can directly act on osteoblasts, and examined whether osteoblasts express DA receptors that respond to exogenous DA. m RNAs and protein cell lysates were obtained from MC3T3-E1 cells during osteogenic differentiation phase. Reverse transcription polymerase chain reaction and western blot analysis were used to examine the expression of DA receptors, D1–D5. Dose-response effect and time course of DA treatment on cell proliferation, mineralization, and osteogenic differentiation were investigated at pre-determined days. Real-time PCR was performed to investigate whether DA affects osteogenic gene expression（ALP, BSP, OC, OSX, RUNX2, and Collagen1a2） with or without receptor antagonists（SCH233390 and GR103691）. Two-way ANOVA was used for statistical analysis. All five DA receptors（D1, D2, D3, D4, and D5） m RNAs and proteins were expressed in MC3T3-E1 cells. DA treatment increased cell proliferation for up to 7 days（P, 0.05）. Osteogenic mineralization was significantly greater in the DA-treated group than control group（P, 0.05）. Finally, expression of all the osteogenic genes was inhibited by DA receptor antagonists for D1, D3, and D5. Our findings suggest that MC3T3-E1 osteoblasts express functional DA receptors that enhance proliferation and mineralization. PDA is not biologically inert and has important implications in orthopedic applications. Furthermore, osteoblast differentiation might be regulated by the nervous system, presumably during bone development, remodeling, or repair.

关键词： DIFFERENTIATION RECEPTORS CYTOTOXICITY innervation EXPRESSION DELETION TISSUE

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Mechanical tension promotes skin nerve regeneration by upregulating nerve growth factor expression

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Neural Regeneration Research 2013年第17期8卷 1576-1581页

作者： Hu Xiao Dechang Wang Ran Huo Yibing Wang Yongqiang Feng Qiang Li Department of Burn and Plastic Surgery Shandong Provincial Hospital Affiliated to Shandong University

This study aimed to explore the role of mechanical tension in hypertrophic scars and the change in nerve density using hematoxylin-eosin staining and S100 immunohistochemistry, and to observe the expression of nerve growth factor by western blot analysis. The results demonstrated that mechanical tension contributed to the formation of a hyperplastic scar in the back skin of rats, in conjunction with increases in both nerve density and nerve growth factor expression in the scar tissue. These experimental findings indicate that the cutaneous nervous system plays a role in hypertrophic scar formation caused by mechanical tension.

关键词： neural regeneration innervation nerve growth factor hypertrophic scar keloid mechanical tension wound healing surgery grants-supported paper neuroregeneration

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