Expression characteristics of metabotropic glutamate receptor 4 in cochlear hair cells during early postnatal stages in mice

CHEN Zhiji; LIU Xuelai; ZHOU Xiaoqing; SHE Yuting; FENG Menglong; KUANG Shaojing; FANG Hongyan; YUAN Wei

doi:10.16016/j.2097-0927.202502004

您当前的位置：

首页 >

文章列表页 >

Expression characteristics of metabotropic glutamate receptor 4 in cochlear hair cells during early postnatal stages in mice

Basic Medicine | 更新时间：2025-06-11

- Expression characteristics of metabotropic glutamate receptor 4 in cochlear hair cells during early postnatal stages in mice
- Journal of Army Medical University Vol. 47, Issue 11, Pages: 1227-1234(2025)
- 作者机构：
  
  重庆市人民医院（重庆大学附属人民医院）耳鼻咽喉头颈外科，重庆
- 作者简介：
- 基金信息：
  
  the Open Research Project of National Clinical Research Center for Otorhinolaryngology(2024KF008);the Postdoctoral Project of Chongqing Natural Science Foundation(CSTB2023NSCQ-BHX0196);the General Program of Chongqing Science-Technology and Health Commission Joint Medical Research Project(2024MSXM101);the Key Special Program for Technological Innovation and Application Development of Chongqing(CSTB2023TIAD-KPX0059)
- DOI：10.16016/j.2097-0927.202502004
  CLC： R322.922;R329.28;R339.16
- Received：05 February 2025，
  
  Revised：19 March 2025，
  
  Published：15 June 2025
- 稿件说明：
移动端阅览
陈知己，刘雪莱，周小清，等.出生后早期代谢型谷氨酸受体4在小鼠耳蜗毛细胞中的表达特征［J］.陆军军医大学学报，2025，47（11）：1227-1234.DOI：10.16016/j.2097-0927.202502004.

CHEN Z J，LIU X L，ZHOU X Q，et al.Expression characteristics of metabotropic glutamate receptor 4 in cochlear hair cells during early postnatal stages in mice［J］.J Army Med Univ，2025，47（11）：1227-1234.DOI：10.16016/j.2097-0927.202502004.
陈知己，刘雪莱，周小清，等.出生后早期代谢型谷氨酸受体4在小鼠耳蜗毛细胞中的表达特征［J］.陆军军医大学学报，2025，47（11）：1227-1234.DOI：10.16016/j.2097-0927.202502004. DOI：

CHEN Z J，LIU X L，ZHOU X Q，et al.Expression characteristics of metabotropic glutamate receptor 4 in cochlear hair cells during early postnatal stages in mice［J］.J Army Med Univ，2025，47（11）：1227-1234.DOI：10.16016/j.2097-0927.202502004. DOI：

摘要

目的

探究代谢型谷氨酸受体4（metabotropic glutamate receptor 4，mGluR4）在小鼠耳蜗基底膜的表达定位，及其从出生后至毛细胞突触成熟的表达变化。

方法

10只健康的3月龄C57BJ/6L野生型小鼠（5只雄鼠，5只雌鼠，体质量25~35 g）配对繁殖，分别选取出生后P1~P30不同日龄的小鼠，采用分频听觉脑干反应（auditory brainstem response， ABR）检测P12、P14、P16、P21、P30日龄小鼠在45.2、32.0、22.6、16.0、11.3、8.0、5.6和4.0 kHz的听反应阈，用免疫荧光标记联合激光共聚焦显微镜扫描耳蜗基底膜铺片。观察mGluR4与突触相关蛋白Ctbp2和Snap25的表达定位关系，以及统计随突触成熟发生的荧光斑数量变化。

结果

在小鼠ABR检测中发现，P12日龄小鼠在所有频率最大刺激声强度下均未能诱发出分化良好的波形，70%的P14日龄小鼠能够被至少1个频率的刺激声诱发出分化良好的Ⅰ波。在P14、P16、P21、P30日龄小鼠ABR测试中，50%的小鼠无法被45.2 kHz刺激声诱发出ABR波形，87.5%的小鼠无法被32.0 kHz刺激声诱发出ABR波形，其他频率的听反应阈随小

鼠日龄增加逐步降低。在基底膜铺片免疫荧光染色检测中发现，mGluR4表达于出生后的小鼠耳蜗基底膜，其荧光强度在内毛细胞（inner hair cell， IHC）强于外毛细胞（outer hair cell，OHC）。在P11小鼠的IHC中即能够检测到清晰明显的mGluR4荧光斑点，其存在于细胞表达Snap25的突触囊泡胞吐活性区，但与突触蛋白Ctbp2无明显共标。在突触区mGluR4的荧光斑点信号统计中，P30小鼠荧光斑点数量多于P14小鼠（

0.05）。

结论

在小鼠出生后早期，mGluR4在IHC突触区表达逐渐增加。随着突触功能成熟其可能参与囊泡胞吐调控，并可能通过自身药理特性抑制谷氨酸递质释放，影响突触可塑性。

Abstract

Objective‍　‍To investigate the spatiotemporal expression pattern of metabotropic glutamate receptor 4 (mGluR4) in the mouse cochlear basilar membrane and its dynamic changes during postnatal development until hair cell synaptic maturation. Methods‍　‍Ten healthy 3-month-old C57BL/6L wild-type mice (5 males

5 females

25～35 g) were bred to obtain offspring. Cochlear samples from postnatal days 1~30 (P1~P30) were analyzed. Frequency-specific auditory brainstem response (ABR) thresholds were measured at 45.2

32.0

22.6

16.0

11.3

8.0

5.6

and 4.0 kHz in P12

P14

P16

P21

and P30 mice. Immunofluorescence assay combined with confocal laser scanning microscopy was used to scan cochlear whole-mount preparations. Spatiotemporal co-localization profiling of mGluR4 and synaptic markers Ctbp2 and Snap25 was observed

and quantitative comparisons of fluorescent puncta density were performed. Results‍　‍ABR testing revealed undifferentiated waveforms in all P12 mice

while 70% of P14 mice exhibited well-defined Wave I. In P14

P16

P21 and P30

mice undergoing ABR testing

50% failed to exhibit ABR waveforms when stimulated by 45.2 kHz tones

and 87.5% showed no response to 32.0 kHz stimuli. Auditory response thresholds for other frequencies progressively decreased with increasing postnatal age. Immunolocalization demonstrated stronger mGluR4 expression in inner hair cells (IHCs) versus outer hair cells (OHCs). Distinct mGluR4 puncta were observed at IHC presynaptic active zones (co-localized with Snap25) as early as P11

but showed minimal overlap with CtBP2-labeled ribbons. Quantit

ative analysis revealed significantly higher mGluR4 puncta density at P30 compared to P14 synapses (

0.05). Conclusion‍　‍Postnatal upregulation of mGluR4 at IHC synaptic regions correlates with functional maturation. These findings suggest mGluR4 may modulate vesicular exocytosis regulation and exert pharmacological inhibition on glutamate release

potentially influencing synaptic plasticity during auditory pathway refinement.

关键词

Keywords

references

VOORN R A ， VOGL C . Molecular assembly and structural plasticity of sensory ribbon synapses—a presynaptic perspective ［J］. Int J Mol Sci ， 2020 ， 21 （ 22 ）： 8758 . DOI： 10.3390/ijms21228758 http://dx.doi.org/10.3390/ijms21228758 .

COATE T M ， KATIE SCOTT M ， GURJAR M . Current concepts in cochlear ribbon synapse formation ［J］. Synapse ， 2019 ， 73 （ 5 ）： e22087 . DOI： 10.1002/syn.22087 http://dx.doi.org/10.1002/syn.22087 .

SEBE J Y ， CHO S ， SHEETS L ， et al . Ca 2+ -permeable AMPARs mediate glutamatergic transmission and excitotoxic damage at the hair cell ribbon synapse ［J］. J Neurosci ， 2017 ， 37 （ 25 ）： 6162 - 6175 . DOI： 10.1523/JNEUROSCI.3644-16.2017 http://dx.doi.org/10.1523/JNEUROSCI.3644-16.2017 .

LU Y . Metabotropic glutamate receptors in auditory processing ［J］. Neuroscience ， 2014 ， 274 ： 429 - 445 . DOI： 10.1016/j.neuroscience.2014.05.057 http://dx.doi.org/10.1016/j.neuroscience.2014.05.057 .

DOMIN H ， BURNAT G . mGlu4R， mGlu7R， and mGlu8R allosteric modulation for treating acute and chronic neurodegenerative disorders ［J］. Pharmacol Rep ， 2024 ， 76 （ 6 ）： 1219 - 1241 . DOI： 10.1007/s43440-024-00657-7 http://dx.doi.org/10.1007/s43440-024-00657-7 .

MIELECKI D ， SALIŃSKA E . Group Ⅲ metabotropic glutamate receptors： guardians against excitotoxicity in ischemic brain injury， with implications for neonatal contexts ［J］. Pharmacol Rep ， 2024 ， 76 （ 6 ）： 1199 - 1218 . DOI： 10.1007/s43440-024-00651-z http://dx.doi.org/10.1007/s43440-024-00651-z .

MASILAMONI G J ， SMITH Y . Metabotropic glutamate receptors： targets for neuroprotective therapies in Parkinson disease ［J］. Curr Opin Pharmacol ， 2018 ， 38 ： 72 - 80 . DOI： 10.1016/j.coph.2018.03.004 http://dx.doi.org/10.1016/j.coph.2018.03.004 .

曹德茂，申宝玺，武永康，等 . 谷氨酸受体以及兴奋性毒性研究进展［J］. 中华神经创伤外科电子杂志， 2017 ， 3 （ 2 ）： 109 - 113 . DOI： 10.3877/cma.j.issn.2095-9141.2017.02.012 http://dx.doi.org/10.3877/cma.j.issn.2095-9141.2017.02.012 .

CAO D M ， SHEN B X ， WU Y K ， et al . Research progress of glutamate receptors and excitability toxicity ［J］. Chin J Neurotraumatic Surg Electron Ed ， 2017 ， 3 （ 2 ）： 109 - 113 . DOI： 10.3877/cma.j.issn.2095-9141.2017.02.012 http://dx.doi.org/10.3877/cma.j.issn.2095-9141.2017.02.012 .

张策，乔健天，刘荣建 . 突触前代谢型谷氨酸受体调节神经递质的释放［J］. 生理科学进展， 2002 ， 33 （ 4 ）： 293 - 298 .

ZHANG C ， QIAO J T ， LIU R J . Suppression of neurotransmitter release by presynaptic metabotropic glutamate receptors ［J］. Prog Physiol Sci ， 2002 ， 33 （ 4 ）： 293 - 298 .

UPRETI C ， ZHANG X L ， ALFORD S ， et al . Role of presynaptic metabotropic glutamate receptors in the induction of long-term synaptic plasticity of vesicular release ［J］. Neuropharmacology ， 2013 ， 66 ： 31 - 39 . DOI： 10.1016/j.neuropharm.2012.05.004 http://dx.doi.org/10.1016/j.neuropharm.2012.05.004 .

VAN HOOK M J ， BABAI N ， ZURAWSKI Z ， et al . A presynaptic group Ⅲ mGluR recruits Gβγ/SNARE interactions to inhibit synaptic transmission by cone photoreceptors in the vertebrate retina ［J］. J Neurosci ， 2017 ， 37 （ 17 ）： 4618 - 4634 . DOI： 10.1523/JNEUROSCI. 2948-16.2017 http://dx.doi.org/10.1523/JNEUROSCI.2948-16.2017 .

ZHANG Z C ， LIU Y F ， LUAN Y ， et al . Activation of type 4 metabotropic glutamate receptor regulates proliferation and neuronal differentiation in a cultured rat retinal progenitor cell through the suppression of the cAMP/PTEN/AKT pathway ［J］. Front Mol Neurosci ， 2020 ， 13 ： 141 . DOI： 10.3389/fnmol.2020.00141 http://dx.doi.org/10.3389/fnmol.2020.00141 .

李春晖，杜波，丁大连，等 . 小鼠耳蜗的形态学发育过程［J］. 解剖科学进展， 2008 ， 14 （ 2 ）： 148 - 150 . DOI： 10.16695/j.cnki.1006-2947.2008.02.026 http://dx.doi.org/10.16695/j.cnki.1006-2947.2008.02.026 .

LI C H ， DU B ， DING D L ， et al . Development of corti organ in Kunming mouse ［J］. Prog Anat Sci ， 2008 ， 14 （ 2 ）： 148 - 150 . DOI： 10.16695/j.cnki.1006-2947.2008.02.026 http://dx.doi.org/10.16695/j.cnki.1006-2947.2008.02.026 .

PANGRSIC T ， VOGL C . Balancing presynaptic release and endocytic membrane retrieval at hair cell ribbon synapses ［J］. FEBS Lett ， 2018 ， 592 （ 21 ）： 3633 - 3650 . DOI： 10.1002/1873-3468.13258 http://dx.doi.org/10.1002/1873-3468.13258 .

ZURAWSKI Z ， THOMPSON GRAY A D ， BRADY L J ， et al . Disabling the Gβγ-SNARE interaction disrupts GPCR-mediated presynaptic inhibition， leading to physiological and behavioral phenotypes ［J］. Sci Signal ， 2019 ， 12 （ 569 ）： eaat8595 . DOI： 10.1126/scisignal.aat8595 http://dx.doi.org/10.1126/scisignal.aat8595 .

MIELECKI D ， BRATEK-GEREJ E ， SALIŃSKA E . Metabotropic glutamate receptors-guardians and gatekeepers in neonatal hypoxic-ischemic brain injury ［J］. Pharmacol Rep ， 2024 ， 76 （ 6 ）： 1272 - 1285 . DOI： 10.1007/s43440-024-00653-x http://dx.doi.org/10.1007/s43440-024-00653-x .

FREITAS G A ， NISWENDER C M . GRM7 gene mutations and consequences for neurodevelopment ［J］. Pharmacol Biochem Behav ， 2023 ， 225 ： 173546 . DOI： 10.1016/j.pbb.2023.173546 http://dx.doi.org/10.1016/j.pbb.2023.173546 .

HAIDER H F ， FLOOK M ， APARICIO M ， et al . Biomarkers of presbycusis and tinnitus in a Portuguese older population ［J］. Front Aging Neurosci ， 2017 ， 9 ： 346 . DOI： 10.3389/fnagi.2017.00346 http://dx.doi.org/10.3389/fnagi.2017.00346 .

YU P P ， JIAO J ， CHEN G S ， et al . Effect of GRM7 polymorphisms on the development of noise-induced hearing loss in Chinese Han workers： a nested case-control study ［J］. BMC Med Genet ， 2018 ， 19 （ 1 ）： 4 . DOI： 10.1186/s12881-017-0515-3 http://dx.doi.org/10.1186/s12881-017-0515-3 .

WELLS H R R ， NEWMAN T A ， WILLIAMS F M K . Genetics of age-related hearing loss ［J］. J Neurosci Res ， 2020 ， 98 （ 9 ）： 1698 - 1704 . DOI： 10.1002/jnr.24549 http://dx.doi.org/10.1002/jnr.24549 .

SUN S H ， BABOLA T ， PREGERNIG G ， et al . Hair cell mechanotransduction regulates spontaneous activity and spiral ganglion subtype specification in the auditory system ［J］. Cell ， 2018 ， 174 （ 5 ）： 1247 - 1263.e15 . DOI： 10.1016/j.cell.2018.07.008 http://dx.doi.org/10.1016/j.cell.2018.07.008 .

ZHANG D ， REN M H ， BI Z H ， et al . Lypd1-DTR/+： a new mouse model for specifically damaging the type ic spiral ganglion neurons of the cochlea ［J］. Neurosci Bull ， 2023 ， 39 （ 9 ）： 1459 - 1462 . DOI： 10.1007/s12264-023-01071-4 http://dx.doi.org/10.1007/s12264-023-01071-4 .

EYOB W ， GEORGE A K ， HOMME R P ， et al . Regulation of the parental gene GRM4 by circGrm4 RNA transcript and glutamate-mediated neurovascular toxicity in eyes ［J］. Mol Cell Biochem ， 2021 ， 476 （ 2 ）： 663 - 673 . DOI： 10.1007/s11010-020-03934-0 http://dx.doi.org/10.1007/s11010-020-03934-0 .

原晶晶，闫占峰，周沫，等 . GluR4在单侧聋大鼠耳蜗核不同发育时间的表达［J］. 听力学及言语疾病杂志， 2023 ， 31 （ 06 ）： 546 - 550 . DOI： 10.3969/j.issn.1006-7299.2023.06.014 http://dx.doi.org/10.3969/j.issn.1006-7299.2023.06.014 .

YUAN J J ， YAN Z F ， ZHOU M ， et al . Expression of GluR4 in cochlear nucleus of unilateral deafness rats at different developmental stages ［J］. J Audiol Speech Pathol ， 2023 ， 31 （ 06 ）： 546 - 550 . DOI： 10.3969/j.issn.1006-7299.2023.06.014 http://dx.doi.org/10.3969/j.issn.1006-7299.2023.06.014 .

KLOTZ-WEIGAND L ， ENZ R . Metabotropic glutamate receptors at ribbon synapses in the retina and cochlea ［J］. Cells ， 2022 ， 11 （ 7 ）： 1097 . DOI： 10.3390/cells11071097 http://dx.doi.org/10.3390/cells11071097 .

MA X Y ， GUO J M ， FU Y Y ， et al . G protein-coupled receptors in cochlea： Potential therapeutic targets for hearing loss ［J］. Front Mol Neurosci ， 2022 ， 15 ： 1028125 . DOI： 10.3389/fnmol.2022.1028125 http://dx.doi.org/10.3389/fnmol.2022.1028125 .

王露露，谢飞，赵清远，等 . 人Waardenburg综合征点突变SOX10 p.R106W小鼠模型的构建及听力表型初步研究［J］. 陆军军医大学学报， 2023 ， 45 （ 24 ）： 2512 - 2520 . DOI： 10.16016/j.2097-0927.202309001 http://dx.doi.org/10.16016/j.2097-0927.202309001 .

WANG L L ， XIE F ， ZHAO Q Y ， et al . Construction and otological phenotypic characterization of a mouse model harboring orthologous human Waardenburg syndrome mutation SOX10 p.R106W ［J］. J Army Med Univ ， 2023 ， 45 （ 24 ）： 2512 - 2520 . DOI： 10.16016/j.2097-0927.202309001 http://dx.doi.org/10.16016/j.2097-0927.202309001 .

Views

下载量

CSCD

Alert me when the article has been cited

提交

Tools

Publicity Resources

Stereological study on changes of spinal synapses in hippocampal dentate gyrus from chronic unpredictable stress rats

Screening and identification of synaptic regulatory genes in the inner ear of mice with presbycusis

The study of the protection of telmisartan on the cognitive function in diabetic rats

Effects of shear stress on morphology of different microvascular endothelial cells

Expression of synaptophysin in rat retina following acute high intraocular pressure

Related Author

Tang Yong

Zhu Yanqing

Qi Yingqiang

Jiang Lin

Huang Chunxia

Gao Yuan

Tang Jing

Liang Xin

Related Institution

重庆医科大学：基础医学院组织学与胚胎学教研室、干细胞与组织工程研究室，附属第一医院老年科，基础医学院生理学教研室

Department of Otolaryngology, Head and Neck Surgery, First Affiliated Hospital, Army Medical University (Third Military Medical University)

Endocrinology group , Department of Geriatrics , the First Affiliated Hospital , Chongqing Medical University

Department of Otorhinolaryngology, Southwest Hospital, Third Military Medical University

Department of Anatomy and Neurobiology, Xiangya School of Medicine, Central South University

⁰