Age‑associated variation in the expression and function of TMEM16A calcium‑activated chloride channels in the cochlear stria vascularis of guinea pigs

Zhou,Ying; Song,Jia; Wang,Yan‑Ping; Zhang,Ai‑Mei; Tan,Chao‑Yang; Liu,Yan‑Hui; Zhang,Zhi‑Ping; Wang,Yang; Ma,Ke‑Tao; Li,Li; Si,Jun‑Qiang

doi:10.3892/mmr.2019.10423

Age‑associated variation in the expression and function of TMEM16A calcium‑activated chloride channels in the cochlear stria vascularis of guinea pigs

Authors:
- Ying Zhou
- Jia Song
- Yan‑Ping Wang
- Ai‑Mei Zhang
- Chao‑Yang Tan
- Yan‑Hui Liu
- Zhi‑Ping Zhang
- Yang Wang
- Ke‑Tao Ma
- Li Li
- Jun‑Qiang Si
View Affiliations

Affiliations: Department of Physiology, Medical College of Shihezi University, Shihezi, Xinjiang 832002, P.R. China
Published online on: June 25, 2019 https://doi.org/10.3892/mmr.2019.10423
Pages: 1593-1604
Copyright: © Zhou et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

The present study was designed to investigate the expression and function of transmembrane protein 16 (TMEM16A), a calcium‑activated chloride channel (CaCC), in the stria vascularis (SV) of the cochlea of guinea pigs at different ages, and to understand the role of CaCCs in the pathogenesis of presbycusis (age‑related hearing loss), the most common type of sensorineural hearing loss that occurs with natural aging. Guinea pigs were divided into the following groups: 2 weeks (young group), 3 months (youth group), 1 year (adult group), D‑galactose intervention (D‑gal group; aging model induced by subcutaneous injection of D‑galactose) and T16Ainh‑A01 (intraperitoneal injection of 50 µg/kg/day TMEM16A inhibitor T16Ainh‑A01 for 2 weeks). Differences in the hearing of guinea pigs between the various age groups were analyzed using auditory brainstem response (ABR), and immunofluorescence staining was performed to detect TMEM16A expression in the SV and determine the distribution. Reverse transcription‑quantitative PCR and western blot analyses were conducted to detect the mRNA and protein levels of TMEM16A in SV in the different age groups. Morris water maze behavior analysis demonstrated that spatial learning ability and memory were damaged in the D‑gal group. Superoxide dismutase activity and malondialdehyde content assays indicated that there was oxidative stress damage in the D‑gal group. The ABR thresholds gradually increased with age, and the increase in the T16Ainh‑A01 group was pronounced. Immunofluorescence analysis in the cochlear SV of guinea pigs in different groups revealed that expression of TMEM16A increased with increasing age (2 weeks to 1 year); fluorescence intensity was reduced in the D‑gal model of aging. As the guinea pigs continued to mature, the protein and mRNA contents of TMEM16A in the cochlea SV increased gradually, but were decreased in the D‑gal group. The findings indicated that CaCCs in the cochlear SV of guinea pigs were associated with the development of hearing in guinea pigs, and that downregulation of TMEM16A may be associated with age‑associated hearing loss.

View Figures

View References

1	Kong Weijia: Research progress of senile deafness. Chinese Medical Abstracts. 159–160. 2010.(In Chinese).
2	Viveki RG, Halappanavar AB, Joshi AV, Pujar K and Patil S: Sociodemographic and health profile of inmates of old age homes in and around Belgaum city, Karnataka. J Indian Med Assoc. 111:682–685. 2013.PubMed/NCBI
3	Vas V, Akeroyd MA and Hall DA: A Data-Driven Synthesis of Research Evidence for Domains of Hearing Loss, as Reported by Adults With Hearing Loss and Their Communication Partners. Trends Hear. 573–576. 2017.
4	Gates GA, Caspary DM, Clark W, Pillsbury HC, Brown SC and Dobie RA: Presbycusis. Otolaryngol Head Neck Surg. 100:1989. View Article : Google Scholar
5	Spicer SS and Schulte BA: Pathologic changes of presbycusis begin in secondary processes and spread to primary processes of strial marginal cells. Hear Res. 205:225–240. 2005. View Article : Google Scholar : PubMed/NCBI
6	Shi X: Pathophysiology of the cochlear intrastrial fluid-blood barrier (review). Hear Res. 338:52–63. 2016. View Article : Google Scholar : PubMed/NCBI
7	Kikuchi T, Kimura RS, Paul DL, Takasaka T and Adams JC: Gap junction systems in the mammalian cochlea. Brain Res Brain Res Rev. 32:163–166. 2000. View Article : Google Scholar : PubMed/NCBI
8	Forge A and Wright T: The molecular architecture of the inner ear. Br Med Bull. 63:5–24. 2002. View Article : Google Scholar : PubMed/NCBI
9	Forge A, Becker D, Casalotti S, Edwards J, Marziano N and Nickel R: Connexins and gap junctions in the inner ear. Audiol Neurootol. 7:141–145. 2002. View Article : Google Scholar : PubMed/NCBI
10	Spicer SS and Schulte BA: Novel structures in marginal and intermediate cells presumably relate to functions of apical versus basal strial strata. Hear Res. 200:87–101. 2005. View Article : Google Scholar : PubMed/NCBI
11	Shi X: Physiopathology of the cochlear microcirculation. Hear Res. 282:10–24. 2011. View Article : Google Scholar : PubMed/NCBI
12	Shi X, Han W, Yamamoto H, Tang W, Lin X, Xiu R, Trune DR and Nuttall AL: The cochlear pericytes. Microcirculation. 15:515–529. 2008. View Article : Google Scholar : PubMed/NCBI
13	Betsholtz C, Lindblom P and Gerhardt H: Role of pericytes in vascular morphogenesis. EXS. 115–125. 2005.PubMed/NCBI
14	Thomas WE: Brain macrophages: On the role of pencytes and perivascular cells. Brain Res Brain Res Rev. 31:42–57. 1999. View Article : Google Scholar : PubMed/NCBI
15	Kurz H, Fehr J, Nitschke R and Burkhardt H: Pericytes in the mature chorioallantoic membrane capillary plexus contain desmin and alpha-smooth muscle actin: Relevance for non-sprouting angiogenesis. Histochem Cell Biol. 130:1027–1040. 2008. View Article : Google Scholar : PubMed/NCBI
16	Peppiatt CM, Howarth C, Mobbs P and Attwell D: Bidirectional control of CNS capillary diameter by pericytes. Nature. 443:700–704. 2006. View Article : Google Scholar : PubMed/NCBI
17	Oishi K, Kamiyashiki T and Ito Y: Isometric contraction of microvascular pericytes from mouse brain parenchyma. Microvasc Res. 73:20–28. 2007. View Article : Google Scholar : PubMed/NCBI
18	Hall CN, Reynell C, Gesslein B, Hamilton NB, Mishra A, Sutherland BA, O'Farrell FM, Buchan AM, Lauritzen M and Attwell D: Capillary pericytes regulate cerebral blood flow in health and disease. Nature. 508:55–60. 2014. View Article : Google Scholar : PubMed/NCBI
19	Greenhalgh SN, Iredale JP and Henderson NC: Origins of fibrosis: Pericytes take centre stage. F1000Prime Rep. 5:372013. View Article : Google Scholar : PubMed/NCBI
20	Liu S, Agalliu D, Yu C and Fisher M: The role of pericytes in blood-brain barrier function and stroke. Curr Pharm Des. 18:3653–3662. 2012. View Article : Google Scholar : PubMed/NCBI
21	Greif DM and Eichmann A: Vascular biology: Brain vessels squeezed to death. Nature. 508:50–51. 2014. View Article : Google Scholar : PubMed/NCBI
22	Puro DG: Physiology and pathobiology of the pericyte-containing retinal microvasculature: New developments. Microcirculation. 14:1–10. 2007. View Article : Google Scholar : PubMed/NCBI
23	Rickheit G, Maier H, Strenzke N, Andreescu CE, De Zeeuw CI, Muenscher A, Zdebik AA and Jentsch TJ: Endocochlear potential depends on Cl- channels: Mechanism underlying deafness in Bartter syndrome IV. EMBO J. 27:2907–2917. 2008. View Article : Google Scholar : PubMed/NCBI
24	Wang Li and Zhang Hailing: Progress in research on function and molecular basis of Ca²⁺ activated Cl⁻ channel. Chin J Cell Biol. 477–484. 2012.(In Chinese).
25	Caputo A, Caci E, Ferrera L, Pedemonte N, Barsanti C, Sondo E, Pfeffer U, Ravazzolo R, Zegarra-Moran O and Galietta LJ: TMEM16A, a membrane protein associated with calcium-dependent chloride channel activity. Science. 322:590–594. 2008. View Article : Google Scholar : PubMed/NCBI
26	Schroeder BC, Cheng T, Jan YN and Jan LY: Expression cloning of TMEM16A as a calcium-activated chloride channel subunit. Cell. 134:1019–1029. 2008. View Article : Google Scholar : PubMed/NCBI
27	Hartzell HC, Yu K, Xiao Q, Chien LT and Qu Z: Anoctamin/TMEM16 family members are Ca2+-activated Cl- channels. J Physiol. 587:2127–2139. 2009. View Article : Google Scholar : PubMed/NCBI
28	Scudieri P, Sondo E, Ferrera L and Galietta LJ: The anoctamin family: TMEM16A and TMEM16B as calcium-activated chloride channels. Exp Physiol. 97:177–183. 2012. View Article : Google Scholar : PubMed/NCBI
29	Bernstein K, Vink JY, Fu XW, Wakita H, Danielsson J, Wapner R and Gallos G: Calcium-activated chloride channels anoctamin 1 and 2 promote murine uterine smooth muscle contractility. Am J Obstet Gynecol. 211:688.e1–e10. 2014. View Article : Google Scholar
30	Manoury B, Tamuleviciute A and Tammaro P: TMEM16A/Anoctamin 1 protein mediates calcium-activated chloride currents in pulmonary arterial smooth muscle cells. J Physiol. 588:2305–2314. 2010. View Article : Google Scholar : PubMed/NCBI
31	Song J, Wang Y, Liu YH, Ma KT, Li L and Jun-Qiang S: The expression of transmembrane protein 16A in guinea pig cochlea increased with years. J Xi'an Jiaotong Univ (Medical Sciences). 38:796–802. 2017.
32	Jin X, Shah S, Liu Y, Zhang H, Lees M, Fu Z, Lippiat JD, Beech DJ, Sivaprasadarao A, Baldwin SA, et al: Activation of the Cl- channel ANO1 by localized calcium signals in nociceptive sensory neurons requires coupling with the IP3 receptor. Sci Signal. 6:ra732013. View Article : Google Scholar : PubMed/NCBI
33	Henkel B, Drose DR, Ackels T, Oberland S, Spehr M and Neuhaus EM: Co-expression of anoctamins in cilia of olfactory sensory neurons. Chem Senses. 40:73–87. 2015. View Article : Google Scholar : PubMed/NCBI
34	Dauner K, Möbus C, Frings S and Möhrlen F: Targeted expression of anoctamin calcium-activated chloride channels in rod photoreceptor terminals of the rodent retina. Invest Ophthalmol Vis Sci. 54:3126–3136. 2013. View Article : Google Scholar : PubMed/NCBI
35	Wang HC, Lin CC, Cheung R, Zhang-Hooks Y, Agarwal A, Ellis-Davies G, Rock J and Bergles DE: Spontaneous activity of cochlear hair cells triggered by fluid secretion mechanism in adjacent support Cells. Cell. 163:1348–1359. 2015. View Article : Google Scholar : PubMed/NCBI
36	Yi E, Lee J and Lee CJ: Developmental role of Anoctamin-1/TMEM16A in Ca(2+)-dependent volume change in supporting cells of the mouse cochlea. Exp Neurobiol. 22:322–329. 2013. View Article : Google Scholar : PubMed/NCBI
37	Gao Y, Tao Y, Chu H, Chen J, Chen Q, Zhou L, Liu Y, Yu Y and Cui Y: Age-related expression of plasma membrane Ca(2+)-ATPase isoform 2 in the cochleas of C57BL/6J mice. Zhonghua Er Bi Yan Hou Tou Jing Wai Ke Za Zhi. 50:934–938. 2015.(In Chinese). PubMed/NCBI
38	Chen Qiu-jian, Yang Hai-di and Dai Tian-xing: Conservation of Hearing by Simultaneous Mutation of Na,K-ATPase and NKCC1. Journal of the Association for Research in Otolaryngology. 422–434. 2007.PubMed/NCBI
39	Li JL, Chu HQ, Zhou LQ, Xiong H, Wang Y, Chen QG, Chen J, Li ZY, Liu Y and Cui YH: Association of age-related hearing loss with ion transporter KCNQ1 and NKCC1 in cochlea of C57BL/6J mice. Zhonghua Er Bi Yan Hou Tou Jing Wai Ke Za Zhi. 46:139–143. 2011.(In Chinese). PubMed/NCBI
40	Liu YH, Zhang ZP, Wang Y, Song J, Ma KT, Si JQ and Li L: Electrophysiological properties of strial pericytes and the effect of aspirin on pericyte K+channels. Mol Med Rep. 17:2861–2868. 2018.PubMed/NCBI
41	Abulfadl YS, El-Maraghy NN, Ahmed AAE, Nofal S and Badary OA: Protective effects of thymoquinone on D-galactose and aluminum chloride induced neurotoxicity in rats: Biochemical, histological and behavioral changes. Neurol Res. 40:324–333. 2018. View Article : Google Scholar : PubMed/NCBI
42	Olszewski J: Guinea pig as often object to otoneurological experimental examinations. Otolaryngol Pol. 61:838–841. 2007.(In Polish). View Article : Google Scholar : PubMed/NCBI
43	Shirpoor A, Norouzi L, Khadem-Ansari MH, Ilkhanizadeh B and Karimipour M: The protective effect of vitamin E on morphological and biochemical alteration induced by pre and postnatal ethanol administration in the testis of male rat offspring: A three months follow-up study. J Reprod Infertil. 15:134–141. 2014.PubMed/NCBI
44	Wang Z, Lin Y, Chen W, Shang J and Wei T: Transplantation of bone marrow mesenchymal stem cell improves antioxidant capacity and immune activity of aging model rats. Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 33:151–154. 2017.(In Chinese). PubMed/NCBI
45	Cai R, Montgomery SC, Graves KA, Caspary DM and Cox BC: The FBN rat model of aging: Investigation of ABR waveforms and ribbon synapse changes. Neurobiol Aging. 62:53–63. 2018. View Article : Google Scholar : PubMed/NCBI
46	Slack JL, Bi W, Livak KJ, Beaubier N, Yu M, Clark M, Kim SH, Gallagher RE and Willman CL: Pre-clinical validation of a novel, highly sensitive assay to detect PML-RARalpha mRNA using real-time reverse-transcription polymerase chain reaction. J Mol Diagn. 3:141–149. 2001. View Article : Google Scholar : PubMed/NCBI
47	Zhang M, Gao CX, Wang YP, Ma KT, Li L, Yin JW, Dai ZG, Wang S and Si JQ: The association between the expression of PAR2 and TMEM16A and neuropathic pain. Mol Med Rep. 17:3744–3750. 2018.PubMed/NCBI
48	Manabe N, Kiso M, Shimabe M, Nakai-Sugimoto N and Miyamoto H: Abnormal accumulation of corpora lutea in ovaries of the senescence accelerated mouse prone (SAMP1). Int Cong Series. 1260:179–185. 2004. View Article : Google Scholar
49	Hongliang and Fangsheng: Three different methods for establishing mouse aging models. Chinese gerontology. 30:2607–2608. 2010.(In Chinese).
50	Zhou Yuel, Wang Yapin, Wang Jian wei, et al: Effects of sirtuin 1 in positive regulation of ginsenoside Rg1 on hematopoietic stem cell and progenitor cell senescence in vivo. Chinese Journal of Anatomy. 39:5–9. 2016.(In Chinese).
51	Ling Xin, Li Wen Li, Hai, Chun xu, et al: Oxidative damage which caused by γ-rays promotes agingγ. Carcinogenesis Distortion Mutation. 22:335–338. 2010.
52	Zhu YZ and Zhu HG: Establishment and measurement of D-galactose induced aging model. Fudan Univ J Med Sci. 34:617–619. 2007.
53	Wang Q, Zou L, Liu W, Hao W, Tashiro S, Onodera S and Ikejima T: Inhibiting NF-κB activation and ROS production are involved in the mechanism of silibinin's protection against D-galactose-induced senescence. Pharmacol Biochem Behav. 98:140–149. 2011. View Article : Google Scholar : PubMed/NCBI
54	Ni C, Zhang D, Beyer LA, Halsey KE, Fukui H, Raphael Y, Dolan DF and Hornyak TJ: Hearing dysfunction in heterozygous Mitf(Mi-wh)/+ mice, a model for Waardenburg syndrome type 2 and Tietz syndrome. Pigment Cell Melanoma Res. 26:78–87. 2013. View Article : Google Scholar : PubMed/NCBI
55	Tan WJT, Song L, Graham M, Schettino A, Navaratnam D, Yarbrough WG, Santos-Sacchi J and Ivanova AV: Novel role of the mitochondrial protein Fus1 in protection from premature hearing loss via regulation of oxidative stress and nutrient and energy sensing pathways in the inner ear. Antioxid Redox Signal. 27:489–509. 2017. View Article : Google Scholar : PubMed/NCBI
56	Nuttall HE, Moore DR, Barry JG, et al: The influence of cochlear spectral processing on the timing and amplitude of the speech-evoked auditory brain stem response[J]. Journal of Neurophysiology, 2015. 113(10): 3683–3691. 2015.
57	Muniak MA, Ayeni FE and Ryugo DK: Hidden hearing loss and endbulbs of Held: Evidence for central pathology before detection of ABR threshold increases. Hear Res. 364:104–117. 2018. View Article : Google Scholar : PubMed/NCBI
58	Konrad-Martin D, Dille MF, McMillan G, Griest S, McDermott D, Fausti SA and Austin DF: Age-related changes in the auditory brainstem response. J Am Acad Audiol. 23:18–35. 2012. View Article : Google Scholar : PubMed/NCBI
59	Alvarado JC, Fuentes-Santamaría V, Gabaldón-Ull MC and Juiz JM1: Age-Related Hearing Loss Is Accelerated by Repeated Short-Duration Loud Sound Stimulation Front Neurosci. 12:28–29. 2019.
60	Hibino H, Nin F, Tsuzuki C and Kurachi Y: How is the highly positive endocochlear potential formed? The specific architecture of the stria vascularis and the roles of the ion-transport apparatus. Pflugers Arch. 459:521–533. 2010. View Article : Google Scholar : PubMed/NCBI
61	Gratton MA and Schulte BA: Alterations in microvasculature are associated with atrophy of the stria vascularis in quiet-aged gerbils. Hear Res. 82:44–52. 1995. View Article : Google Scholar : PubMed/NCBI
62	Ocho S, Iwasaki S, Umemura K and Hoshino T: A new model for investigating hair cell degeneration in the guinea pig following damage of the stria vascularis using a photochemical reaction. Eur Arch Otorhinolaryngol. 257:182–187. 2000. View Article : Google Scholar : PubMed/NCBI
63	Wangemann P: Comparison of ion transport mechanisms between vestibular dark cells and strial marginal cells. Hear Res. 90:149–157. 1995. View Article : Google Scholar : PubMed/NCBI
64	Wangemann P: K+ cycling and the endocochlear potential. Hear Res. 165:1–9. 2002. View Article : Google Scholar : PubMed/NCBI
65	Takeuchi S and Ando M: Inwardly rectifying K+ currents in intermediate cells in the cochlea of gerbils: A possible contribution to the endocochlear potential. Neurosci Lett. 247:175–178. 1998. View Article : Google Scholar : PubMed/NCBI
66	Wu Jun and Han weiju: Structural changes in thestrial blood-labyrinth barrier of aged C57BL/6 mice. Cell Tissue Res. 685–696. 2015.
67	Bergers G and Song S: The role of pericytes in blood-vessel formation and maintenance. Neuro Oncol. 7:452–464. 2005. View Article : Google Scholar : PubMed/NCBI
68	Díaz-Flores L, Gutiérrez R, Varela H, Rancel N and Valladares F: Microvascular pericytes: A review of their morphological and functional characteristics. Histol Histopathol. 6:269–286. 1991.PubMed/NCBI
69	Gritli-Linde A, Vaziri Sani F, Rock JR, Hallberg K, Iribarne D, Harfe BD and Linde A: Expression patterns of the Tmem16 gene family during cephalic development in the mouse. Gene Expr Patterns. 9:178–191. 2009. View Article : Google Scholar : PubMed/NCBI
70	Jeon JH, Park JW, Lee JW, Jeong SW, Yeo SW and Kim IB: Expression and immunohistochemical localization of TMEM16A/anoctamin 1, a calcium-activated chloride channel in the mouse cochlea. Cell Tissue Res. 345:223–230. 2011. View Article : Google Scholar : PubMed/NCBI
71	Oh U and Jung J: Cellular functions of TMEM16/anoctamin. Pflugers Arch. 468:443–453. 2016. View Article : Google Scholar : PubMed/NCBI
72	Groebe K, Klemm-Manns M, Schwall GP, Hübenthal H, Unterluggauer H, Jansen-Dürr P, Tanguay RM, Morrow G and Schrattenholz A: Age-dependent prosttranslational modifications of voltage-dependent anion channel 1. Exp Gerontol. 45:632–637. 2010. View Article : Google Scholar : PubMed/NCBI
73	Chen J, Chu H, Xiong H, Chen Q, Zhou L, Bing D, Liu Y, Gao Y, Wang S, Huang X and Cui Y: Expression patterns of Ca(V)1.3 channels in the rat cochlea. Acta Biochim Biophys Sin (Shanghai). 44:513–518. 2012. View Article : Google Scholar : PubMed/NCBI
74	Chen J, Chu H, Xiong H, Yu Y, Huang X, Zhou L, Chen Q, Bing D, Liu Y, Wang S and Cui Y: Downregulation of Cav1.3 calcium channel expression in the cochlea is associated with age-related hearing loss in C57BL/6J mice. Neuroreport. 24:313–317. 2013. View Article : Google Scholar : PubMed/NCBI
75	Inui T, Mori Y, Watanabe M, Takamaki A, Yamaji J, Sohma Y, Yoshida R, Takenaka H and Kubota T: Physiological role of L-type Ca2+ channels in marginal cells in the stria vascularis of guinea pigs. J Physiol Sci. 57:287–298. 2007. View Article : Google Scholar : PubMed/NCBI
76	Crisan M, Yap S, Casteilla L, Chen CW, Corselli M, Park TS, Andriolo G, Sun B, Zheng B, Zhang L, et al: A perivascular origin for mesenchymal stem cells in multiple human organs. Cell Stem Cell. 3:301–313. 2008. View Article : Google Scholar : PubMed/NCBI
77	Ma KT, Li XZ, Li L, Zhang ZP, Zhao L, Zhu H and Si JQ: Comparison of electrophysiological properties of vascular smooth muscle cells in different arterioles in guinea pig. Sheng Li Xue Bao. 62:421–426. 2010.(In Chinese). PubMed/NCBI
78	Liu YH, Wang YP, Wang Y, Ma KT, Si JQ and Li L: Study on the electrophsiological properties in the stria vascularis pericytes in cochlear of guinea pig. Zhonghua Er Bi Yan Hou Tou Jing Wai Ke Za Zhi. 51:600–605. 2016.(In Chinese). PubMed/NCBI