Knocking out p75 neurotrophin receptor suppresses mineralization in the femurs of mice

Yingying Wang; Qing Chu; Kun Yang; Xiujie Wen; Gang Li; Junyu Liu; Junyu Liu; Luchuan Liu

doi:10.16016/j.1000-5404.201702160

您当前的位置：

首页 >

文章列表页 >

Knocking out p75 neurotrophin receptor suppresses mineralization in the femurs of mice

Basic Medicine | 更新时间：2026-01-09

- Knocking out p75 neurotrophin receptor suppresses mineralization in the femurs of mice
- Journal of Third Military Medical University Vol. 39, Issue 12, Pages: 1245-1250(2017)
- 作者机构：
  
  1.400042 重庆，第三军医大学大坪医院野战外科研究所口腔科
  2.710032 西安，第四军医大学口腔医院牙周病黏膜病科
  3.400037 重庆，第三军医大学新桥医院口腔科
  4.400016 重庆，重庆医科大学附属第一医院口腔科
- 作者简介：
- 基金信息：
  
  the General Program of National Natural Science Foundation of China(81470032);the State Key Laboratory Program of Military Stomatology(2014KB14)
- DOI：10.16016/j.1000-5404.201702160
  CLC： R322.71;R336;R394.1
- Received：28 February 2017，
  
  Revised：2017-03-16，
  
  Published：30 June 2017
- 稿件说明：
移动端阅览
Yingying Wang, Qing Chu, Kun Yang, et al. Knocking out p75 neurotrophin receptor suppresses mineralization in the femurs of mice[J]. Journal of Third Military Medical University, 2017, 39(12): 1245-1250.
DOI：

Yingying Wang, Qing Chu, Kun Yang, et al. Knocking out p75 neurotrophin receptor suppresses mineralization in the femurs of mice[J]. Journal of Third Military Medical University, 2017, 39(12): 1245-1250. DOI： 10.16016/j.1000-5404.201702160.

摘要

目的

对比野生型和p75 neurotrophin receptor(p75NTR)敲除小鼠股骨骨质的改变，探讨p75NTR敲除对小鼠股骨矿化发育的影响。

方法

取同窝p75NTR

+/+

(野生型)和p75NTR

-/-

(敲除型)雄性小鼠，出生后8周龄时行股骨Micro-CT扫描，检测股骨松质骨的骨量(BV)、骨体积分数(BV/TV)、骨小梁数(Tb.N)、骨小梁厚度(Tb.Th)、骨表面积/骨量(BS/BV)、骨小梁分离度(Tb.Sp)和皮质骨的骨量(Ct.BV)、骨皮质厚度(Ct.TbTh)、骨皮质的骨表面积/骨量(Ct.BS/BV)；出生后40 d分别取同窝两种基因型雄性小鼠行钙黄绿素荧光双标实验，以检测股骨的矿化沉积速率；提取4周龄大小鼠股骨总RNA及蛋白质，RT-PCR和Western blot检测ALP、Runx2的表达。

结果

Micro-CT结果显示：p75NTR

-/-

小鼠股骨的BV、BV/TV、Tb.N、Tb.Th、Ct.BV、Ct.TbTh明显低于同窝p75NTR

+/+

小鼠(

0.05)，同时，p75NTR

-/-

小鼠的BS/BV、Tb.Sp、Ct.BS/BV明显高于p75NTR

+/+

小鼠(

0.01)；钙黄绿素荧光双标检测结果显示：p75NTR

-/-

小鼠股骨的矿化速率明显低于同窝p75NTR

+/+

小鼠(

0.01)；RT-PCR及Western blot结果显示：p75NTR

-/-

小鼠的ALP、Runx2表达均明显低于p75NTR

+/+

小鼠(

0.01)。

结论

p75NTR敲除对小鼠股骨的矿化发育有一定的抑制作用。

Abstract

Objective

To compare the quantitative parameters of the femoral bone between wild-type mice and p75 neurotrophin receptor knockou

t (p75NTR

-/-

) mice and explore the impact of p75NTR knockout on mineralization and development of the femoral bone in mice.

Methods

The left femurs of p75NTR

-/-

and wild-type (p75NTR

+/+

) male mice (8 weeks old) were scanned with micro-CT for measurement of the cancellous bone volume (BV)

bone volume fraction (BV/TV)

number of bone trabecula (Tb.N)

trabecular thickness (Tb.Th)

bone surface area/bone volume ratio (BS/BV)

trabecular separation (Tb.Sp)

cortical bone volume (Ct.BV)

cortical bone thickness (Ct.Tb.Th)

and cortical bone surface area/ bone volume ratio (Ct.BS/BV). The mineral apposition rate (MAR) of the femurs of the p75NTR

-/-

mice and their wild-type littermates was analyzed on day 40 after birth using calcein double fluorescent labeling. At the 4th week

the femurs were taken to detect the expression of Runx2 and ALP using real-time PCR and Western blotting.

Results

Micro-CT imaging data showed significantly lower BV

BV/TV

Tb.N

Tb.Th

Ct.BV and Ct.TbTh (

0.05) and significantly higher BS/BV

Tb.Sp and Ct.BS/BV (

0.01) in p75NTR

-/-

mice than in p75NTR

+/+

mice. Calcein double fluorescent labeling showed that the MAR of the femur was significantly lower in in p75NTR

-/-

mice (

0.01). PCR and Western blotting results showed significantly decreased expressions of Runx2 and ALP in the femurs of p75NTR

-/-

mice as compared with those in p75NTR

+/+

mice (

0.01).

Conclusion

Knocking out p75NTR suppresses mineralization in the femoral bone of mice.

关键词

Keywords

references

Rohner D, Bucher P, Hammer B. Prefabricated fibular flaps for reconstruction of defects of the maxillofacial skeleton: planning, technique, and long-term experience[J]. Int J Oral Maxillofac Implants, 2013, 28(5): e221-e229. DOI: 10.11607/jomi.te01.

Imai S, Tokunaga Y, Maeda T, et al . Calcitonin gene-related peptide, substance P, and tyrosine hydroxylase-immunoreactive innervation of rat bone marrows: an immunohistochemical and ultrastructural investigation on possible efferent and afferent mechanisms[J ] . J Orthop Res, 1997, 15(1): 133-140. DOI: 10.1002/jor.1100150120.

Akiyama Y, Mikami Y, Watanabe E, et al . The P75 neurotrophin receptor regulates proliferation of the human MG63 osteoblast cell line[J ] . Differentia tion, 2014, 87(3/4): 111-118. DOI: 10.1016/j.diff.2014.01.002.

Kuci Z, Seiberth J, Latifi-Pupovci H, et al . Clonal analysis of multipotent stromal cells derived from CD271 + bone marrow mononuclear cells: functional heterogeneity and different mechanisms of allosuppression[J ] . Haematologica, 2013, 98(10): 1609-1616. DOI: 10.3324/haematol.2013.092700..

Moscatelli I, Pierantozzi E, Camaioni A, et al . p75 neurotrophin receptor is involved in proliferation of undifferentiated mouse embryonic stem cells[J ] . Exp Cell Res, 2009, 315(18): 3220-3232. DOI: 10.1016/j.yexcr.2009.08.014.

Tomellini E, Lagadec C, Polakowska R, et al . Role of p75 neurotrophin receptor in stem cell biology: more than just a marker[J ] . Cell Mol Life Sci, 2014, 71(13): 2467-2481. DOI: 10.1007/s00018-014-1564-9.

Cattoretti G, Schiró R, Orazi A, et al . Bone marrow stroma in humans: anti-nerve growth factor receptor antibodies selectively stain reticular cells in vivo and in vitro [J ] .Blood, 1993, 81(7): 1726-1738..

Lee K F, Li E, Huber L J, et al . Targeted mutation of the gene encoding the low affinity NGF receptor p75 leads to deficits in the peripheral sensory nervous system[J ] . Cell, 1992, 69(5): 737-749.

Quirici N, Soligo D, Bossolasco P, et al . Isolation of bone marrow mesenchymal stem cells by anti-nerve growth factor receptor antibodies[J ] . Exp Hematol, 2002, 30(7): 783-791.

鞠迎新, 温秀杰, 杨琨, 等. p75神经营养受体在胎鼠颌突外胚间充质干细胞体外矿化过程中表达变化的研究[J].中华口腔医学杂志, 2016, 51(7): 426-431. DOI: 10.3760/cma.j.issn.1002-0098.2016.07.010.

Ju Y X, Wen X J, Yang K, et al . Expression of p75 neurotrophin receptor during the mineralization of ectomesenchymal stem cells in vitro [J ] . Chin J Stomatol, 2016, 51(7): 426-431. DOI: 10.3760/cma.j.issn.1002-0098.2016.07.010..

Wen X, Liu L, Deng M, et al . Characterization of p75(+) ectomesenchymal stem cells from rat embryonic facial process tissue[J ] . Biochem Biophys Res Commun, 2012, 427(1): 5-10.DOI: 10.1016/j.bbrc.2012.08.109.

Alexander D, Schfer F, Munz A, et al . LNGFR induction during osteogenesis of human jaw periosteum-derived cells[J ] . Cell Physiol Biochem, 2009, 24(3/4): 283-290. DOI: 10.1159/000233253.

Austin C P, Battey J F, Bradley A, et al . The knockout mouse project[J ] . Nat Genet, 2004, 36(9): 921-924. DOI: 10.1038/ng0904-921.

Gregory S G, Sekhon M, Schein J, et al .A physical map of the mouse genome[J ] . Nature, 2002, 418(6899): 743-750.

Mikami Y, Ishii Y, Watanabe N, et al . CD271/p75(NTR) inhibits the differentiation of mesenchymal stem cells into osteogenic, adipogenic, chondrogenic, and myogenic lineages[J ] . Stem Cells Dev, 2011, 20(5): 901-913. DOI: 10.1089/scd.2010.0299.

Mikami Y, Suzuki S, Ishii Y, et al . The p75 neurotrophin receptor regulates MC3T3-E1 osteoblastic differentiation[J ] . Differentiation, 2012, 84(5): 392-399. DOI: 10.1016/j.diff.2012.07.001.

Smith T M. Experimental determination of the periodicity of incremental features in enamel[J]. J Anat, 2006, 208(1): 99-113. DOI: 10.1111/j.1469-7580.2006.00499.x.

Yang K, Wang Y, Ju Y, et al . p75 neurotrophin receptor regulates differential mineralizat ion of rat ectomesenchymal stem cells[J ] . Cell Prolif, 2017, 50(1): e12290. DOI: 10.1111/cpr.12290.

Komori T. Regulation of osteoblast differentiation by transcription factors[J]. J Cell Biochem, 2006, 99(5): 1233-1239. DOI: 10.1002/jcb.20958.

Metsis M. Genes for neurotrophic factors and their receptors: structure and regulation[J]. Cell Mol Life Sci, 2001, 58(8): 1014-1020. DOI: 10.1007/PL00000916.

Xing Y, Nie X, Chen G, et al . Comparison of P75 NTR-positive and -negative etcomesenchymal stem cell odontogenic differentiation through epithelial-mesenchymal interaction[J ] . Cell Prolif, 2016, 49(2): 185-194. DOI: 10.1111/cpr.12248.

Views

下载量

CSCD

Alert me when the article has been cited

提交

Tools

Publicity Resources

Naringin improves lipopolysaccharide-induced depression-like behaviors in mice

Effects of Necdin on odontogenic differentiation and mineralization of rat dental germ ectodermal mesenchymal stem cells

The role of cathepsin K in cardiac dysfunction in mice with alcoholic cardiomyopathy and its damage mechanism

Chaihu Oral Liquid improves hippocampal neurogenesis and short-term memory defects in adult BTBR autism mice

Alpha-ketoglutarate alleviates cholestatic liver injury in mice

Related Author

Chuan WU

Anqi KONG

Ruxin XIE

Jianghui LI

Yun CAI

Xiaotang FAN

张艺馨

袁洪燕

Related Institution

Department of Military Cognitive Psychology, Faculty of Medical Psychology, Army Medical University (Third Military Medical University)

State Key Laboratory of Trauma, Burns and Combined Injury, Department of Rehabilitation Medicine, Laboratory of Trauma, Center of Bone Metabolism and Repair, Army Medical University (Third Military Medical University)

Department of Rehabilitation, Institute of Surgery Research, Third Affiliated Hospital, Army Medical University (Third Military Medical University)

Rehabilitation Center, University-town Affiliated Hospital of Chongqing Medical University

Department of Clinical Hematology, Faculty of Pharmacy and Laboratory Medicine, Army Medical University (Third Military Medical University)

⁰