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Liu, Wei
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Publications (10 of 38) Show all publications
Liu, W., Loewenheim, H., Santi, P. A., Glueckert, R., Schrott-Fischer, A. & Rask-Andersen, H. (2018). Expression of trans-membrane serine protease 3 (TMPRSS3) in the human organ of Corti. Cell and Tissue Research, 372(3), 445-456
Open this publication in new window or tab >>Expression of trans-membrane serine protease 3 (TMPRSS3) in the human organ of Corti
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2018 (English)In: Cell and Tissue Research, ISSN 0302-766X, E-ISSN 1432-0878, Vol. 372, no 3, p. 445-456Article in journal (Refereed) Published
Abstract [en]

TMPRSS3 (Trans-membrane Serine Protease 3) is a type II trans-membrane serine protease that has proteolytic activity essential for hearing. Mutations in the gene cause non-syndromic autosomal recessive deafness (DFNB8/10) in humans. Knowledge about its cellular distribution in the human inner ear may increase our understanding of its physiological role and involvement in deafness, ultimately leading to therapeutic interventions. In this study, we used super-resolution structured illumination microscopy for the first time together with transmission electron microscopy to localize the TMPRSS3 protein in the human organ of Corti. Archival human cochleae were dissected out during petroclival meningioma surgery. Microscopy with Zeiss LSM710 microscope achieved a lateral resolution of approximately 80 nm. TMPRSS3 was found to be associated with actin in both inner and outer hair cells. TMPRSS3 was located in cell surface-associated cytoskeletal bodies (surfoskelosomes) in inner and outer pillar cells and Deiters cells and in subcuticular organelles in outer hair cells. Our results suggest that TMPRSS3 proteolysis is linked to hair cell sterociliary mechanics and to the actin/microtubule networks that support cell motility and integrity.

Keywords
Cochlea, Trans-membrane Serine Protease 3 (TMPRSS3), Immunohistochemistry, Super-resolution structured illumination microscopy (SR-SIM), Human
National Category
Cell Biology
Identifiers
urn:nbn:se:uu:diva-356851 (URN)10.1007/s00441-018-2793-2 (DOI)000432109000001 ()29460002 (PubMedID)
Projects
OTOSTEM
Available from: 2018-08-16 Created: 2018-08-16 Last updated: 2018-08-16Bibliographically approved
Liu, W., Molnar, M., Garnham, C., Benav, H. & Rask-Andersen, H. (2018). Macrophages in the Human Cochlea: Saviors or Predators-A Study Using Super-Resolution Immunohistochemistry. Frontiers in Immunology, 9, Article ID 223.
Open this publication in new window or tab >>Macrophages in the Human Cochlea: Saviors or Predators-A Study Using Super-Resolution Immunohistochemistry
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2018 (English)In: Frontiers in Immunology, ISSN 1664-3224, E-ISSN 1664-3224, Vol. 9, article id 223Article in journal (Refereed) Published
Abstract [en]

The human inner ear, which is segregated by a blood/labyrinth barrier, contains resident macrophages [CD163, ionized calcium-binding adaptor molecule 1 (IBA1)-, and CD68-positive cells] within the connective tissue, neurons, and supporting cells. In the lateral wall of the cochlea, these cells frequently lie close to blood vessels as perivascular macrophages. Macrophages are also shown to be recruited from blood-borne monocytes to damaged and dying hair cells induced by noise, ototoxic drugs, aging, and diphtheria toxin-induced hair cell degeneration. Precise monitoring may be crucial to avoid self-targeting. Macrophage biology has recently shown that populations of resident tissue macrophages may be fundamentally different from circulating macrophages. We removed uniquely preserved human cochleae during surgery for treating petroclival meningioma compressing the brain stem, after ethical consent. Molecular and cellular characterization using immunofluorescence with antibodies against IBA1, TUJ1, CX3CL1, and type IV collagen, and super-resolution structured illumination microscopy (SR-SIM) were made together with transmission electron microscopy. The super-resolution microscopy disclosed remarkable phenotypic variants of IBA1 cells closely associated with the spiral ganglion cells. Monitoring cells adhered to neurons with "synapse-like" specializations and protrusions. Active macrophages migrated occasionally nearby damaged hair cells. Results suggest that the human auditory nerve is under the surveillance and possible neurotrophic stimulation of a well-developed resident macrophage system. It may be alleviated by the non-myelinated nerve soma partly explaining why, in contrary to most mammals, the human's auditory nerve is conserved following deafferentiation. It makes cochlear implantation possible, for the advantage of the profoundly deaf. The IBA1 cells may serve additional purposes such as immune modulation, waste disposal, and nerve regeneration. Their role in future stem cell-based therapy needs further exploration.

Place, publisher, year, edition, pages
FRONTIERS MEDIA SA, 2018
Keywords
human, cochlea, macrophages, ionized calcium-binding adaptor molecule 1, structured illumination microscopy, immunohistochemistry
National Category
Neurosciences
Identifiers
urn:nbn:se:uu:diva-348109 (URN)10.3389/fimmu.2018.00223 (DOI)000424911600001 ()29487598 (PubMedID)
Funder
Science for Life Laboratory - a national resource center for high-throughput molecular bioscienceEU, FP7, Seventh Framework Programme, 603029
Available from: 2018-04-11 Created: 2018-04-11 Last updated: 2018-04-11Bibliographically approved
Agrawal, S., Schart-Moren, N., Liu, W., Ladak, H. M., Rask-Andersen, H. & Li, H. (2018). The secondary spiral lamina and its relevance in cochlear implant surgery. Upsala Journal of Medical Sciences, 123(1), 9-18
Open this publication in new window or tab >>The secondary spiral lamina and its relevance in cochlear implant surgery
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2018 (English)In: Upsala Journal of Medical Sciences, ISSN 0300-9734, E-ISSN 2000-1967, Vol. 123, no 1, p. 9-18Article in journal (Refereed) Published
Abstract [en]

Objective: We used synchrotron radiation phase contrast imaging (SR-PCI) to study the 3D microanatomy of the basilar membrane (BM) and its attachment to the spiral ligament (SL) (with a conceivable secondary spiral lamina [SSL] or secondary spiral plate) at the round window membrane (RWM) in the human cochlea. The conception of this complex anatomy may be essential for accomplishing structural preservation at cochlear implant surgery.

Material and methods: Sixteen freshly fixed human temporal bones were used to reproduce the BM, SL, primary and secondary osseous spiral laminae (OSL), and RWM using volume-rendering software. Confocal microscopy immunohistochemistry (IHC) was performed to analyze the molecular constituents.

Results: SR-PCI reproduced the soft tissues including the RWM, Reissner's membrane (RM), and the BM attachment to the lateral wall (LW) in three dimensions. A variable SR-PCI contrast enhancement was recognized in the caudal part of the SL facing the scala tympani (ST). It seemed to represent a SSL allied to the basilar crest (BC). The SSL extended along the postero-superior margin of the round window (RW) and immunohistochemically expressed type II collagen.

Conclusions: Unlike in several mammalian species, the human SSL is restricted to the most basal portion of the cochlea around the RW. It anchors the BM and may influence its hydro-mechanical properties. It could also help to shield the BM from the RW. The microanatomy should be considered at cochlear implant surgery.

Place, publisher, year, edition, pages
TAYLOR & FRANCIS LTD, 2018
Keywords
Basilar membrane, cochlea, human, secondary spiral lamina, synchrotron-phase contrast imaging
National Category
Otorhinolaryngology
Identifiers
urn:nbn:se:uu:diva-354541 (URN)10.1080/03009734.2018.1443983 (DOI)000428060300002 ()29537931 (PubMedID)
Funder
Swedish Research Council, 2017-03801
Available from: 2018-06-20 Created: 2018-06-20 Last updated: 2018-06-20Bibliographically approved
Liu, W., Li, H., Edin, F., Brännström, J., Glueckert, R., Schrott-Fischer, A., . . . Rask-Andersen, H. (2017). Molecular composition and distribution of gap junctions in the sensory epithelium of the human cochlea a super-resolution structured illumination microscopy (SR-SIM) study. Upsala Journal of Medical Sciences, 122(3), 160-170
Open this publication in new window or tab >>Molecular composition and distribution of gap junctions in the sensory epithelium of the human cochlea a super-resolution structured illumination microscopy (SR-SIM) study
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2017 (English)In: Upsala Journal of Medical Sciences, ISSN 0300-9734, E-ISSN 2000-1967, Vol. 122, no 3, p. 160-170Article in journal (Refereed) Published
Abstract [en]

Background: Mutations in the GJB2 gene, which encodes the Connexin26 (Cx26) protein, are the most common cause of childhood hearing loss in American and European populations. The cochlea contains a gap junction (GJ) network in the sensory epithelium and two connective tissue networks in the lateral wall and spiral limbus. The syncytia contain the GJ proteins beta 2 (GJB2/Cx26) and beta 6 (GJB6/Cx30). Our knowledge of their expression in humans is insufficient due to the limited availability of tissue. Here, we sought to establish the molecular arrangement of GJs in the epithelial network of the human cochlea using surgically obtained samples. Methods: We analyzed Cx26 and Cx30 expression in GJ networks in well-preserved adult human auditory sensory epithelium using confocal, electron, and super -resolution structured illumination microscopy (SR-SIM). Results: Cx30 plaques (<5 mu m) dominated, while Cx26 plaques were subtle and appeared as 'mini junctions' (2-300 nm). 3-D volume rendering of Z-stacks and orthogonal projections from single optical sections suggested that the GJs are homomeric/homotypic and consist of assemblies of identical GJs composed of either Cx26 or Cx30. Occasionally, the two protein types were co-expressed, suggesting functional cooperation. Conclusions: Establishing the molecular composition and distribution of the GJ networks in the human cochlea may increase our understanding of the pathophysiology of Cx-related hearing loss. This information may also assist in developing future strategies to treat genetic hearing loss.

Keywords
Cochlea, confocal microscopy, connexin 26/30, human, SR-SIM
National Category
Otorhinolaryngology
Identifiers
urn:nbn:se:uu:diva-340976 (URN)10.1080/03009734.2017.1322645 (DOI)000414107800002 ()28513246 (PubMedID)
Available from: 2018-02-12 Created: 2018-02-12 Last updated: 2018-02-12Bibliographically approved
Senn, P., Roccio, M., Hahnewald, S., Frick, C., Kwiatkowska, M., Ishikawa, M., . . . Loewenheim, H. (2017). NANOCI-Nanotechnology Based Cochlear Implant With Gapless Interface to Auditory Neurons. Paper presented at 14th International Conference on Cochlear Implants and other Implantable Auditory Technologies, MAY 11-14, 2016, Toronto, CANADA. Otology and Neurotology, 38(8), E224-E231
Open this publication in new window or tab >>NANOCI-Nanotechnology Based Cochlear Implant With Gapless Interface to Auditory Neurons
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2017 (English)In: Otology and Neurotology, ISSN 1531-7129, E-ISSN 1537-4505, Vol. 38, no 8, p. E224-E231Article in journal (Refereed) Published
Abstract [en]

Cochlear implants (CI) restore functional hearing in the majority of deaf patients. Despite the tremendous success of these devices, some limitations remain. The bottleneck for optimal electrical stimulation with CI is caused by the anatomical gap between the electrode array and the auditory neurons in the inner ear. As a consequence, current devices are limited through 1) low frequency resolution, hence suboptimal sound quality and 2), large stimulation currents, hence high energy consumption (responsible for significant battery costs and for impeding the development of fully implantable systems). A recently completed, multinational and interdisciplinary project called NANOCI aimed at overcoming current limitations by creating a gapless interface between auditory nerve fibers and the cochlear implant electrode array. This ambitious goal was achieved in vivo by neurotrophin-induced attraction of neurites through an intra-cochlear gel-nanomatrix onto a modified nanoCI electrode array located in the scala tympani of deafened guinea pigs. Functionally, the gapless interface led to lower stimulation thresholds and a larger dynamic range in vivo, and to reduced stimulation energy requirement (up to fivefold) in an in vitro model using auditory neurons cultured on multi-electrode arrays. In conclusion, the NANOCI project yielded proof of concept that a gapless interface between auditory neurons and cochlear implant electrode arrays is feasible. These findings may be of relevance for the development of future CI systems with better sound quality and performance and lower energy consumption. The present overview/review paper summarizes the NANOCI project history and highlights achievements of the individual work packages.

Keywords
Auditory nerve regeneration, BDNF, Cochlear implant, Gapless interface, Guinea pig, Hearing loss, Hydrogel, Multi-electrode array, Neuron-electrode interface
National Category
Otorhinolaryngology
Identifiers
urn:nbn:se:uu:diva-335874 (URN)10.1097/MAO.0000000000001439 (DOI)000411032100003 ()28806330 (PubMedID)
Conference
14th International Conference on Cochlear Implants and other Implantable Auditory Technologies, MAY 11-14, 2016, Toronto, CANADA
Funder
EU, European Research Council, 281056
Available from: 2017-12-14 Created: 2017-12-14 Last updated: 2017-12-14Bibliographically approved
Liu, W., Schrott-Fischer, A., Glueckert, R., Benav, H. & Rask-Andersen, H. (2017). The Human "Cochlear Battery" - Claudin-11 Barrier and Ion Transport Proteins in the Lateral Wall of the Cochlea. Frontiers in Molecular Neuroscience, 10, Article ID 239.
Open this publication in new window or tab >>The Human "Cochlear Battery" - Claudin-11 Barrier and Ion Transport Proteins in the Lateral Wall of the Cochlea
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2017 (English)In: Frontiers in Molecular Neuroscience, ISSN 1662-5099, Vol. 10, article id 239Article in journal (Refereed) Published
Abstract [en]

Background: The cochlea produces an electric field potential essential for hair cell transduction and hearing. This biological "battery" is situated in the lateral wall of the cochlea and contains molecular machinery that secretes and recycles K+ ions. Its functioning depends on junctional proteins that restrict the para-cellular escape of ions. The tight junction protein Claudin-11 has been found to be one of the major constituents of this barrier that maintains ion gradients (Gow et al., 2004; Kitajiri et al., 2004a). We are the first to elucidate the human Claudin-11 framework and the associated ion transport machinery using super-resolution fluorescence illumination microscopy (SR-SIM). Methods: Archival cochleae obtained during meningioma surgery were used for SR-SIM together with transmission electron microscopy after ethical consent. Results: Claudin-11-expressing cells formed parallel tight junction lamellae that insulated the epithelial syncytium of the stria vascularis and extended to the suprastrial region. Intercellular gap junctions were found between the barrier cells and fibrocytes. Conclusion: Transmission electron microscopy, confocal microscopy and SR-SIM revealed exclusive cell specialization in the various subdomains of the lateral wall of the human cochlea. The Claudin-11-expressing cells exhibited both conductor and isolator characteristics, and these micro-porous separators may selectively mediate the movement of charged units to the intrastrial space in a manner that is analogous to a conventional electrochemical "battery." The function and relevance of this battery for the development of inner ear disease are discussed.

Keywords
human, cochlea, stria vascularis, spiral ligament, Claudin-11, structured illumination microscopy
National Category
Neurosciences Otorhinolaryngology
Identifiers
urn:nbn:se:uu:diva-332662 (URN)10.3389/fnmol.2017.00239 (DOI)000407621000001 ()28848383 (PubMedID)
Available from: 2017-11-09 Created: 2017-11-09 Last updated: 2018-01-13Bibliographically approved
Hellberg, V., Gahm, C., Ehrsson, H., Liu, W., Rask-Andersen, H. & Laurell, G. (2016). In Response to Immunohistochemical Localization of OCT2 in the Cochlea of Various Species [Letter to the editor]. The Laryngoscope, 126(6), E232-E232
Open this publication in new window or tab >>In Response to Immunohistochemical Localization of OCT2 in the Cochlea of Various Species
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2016 (English)In: The Laryngoscope, ISSN 0023-852X, E-ISSN 1531-4995, Vol. 126, no 6, p. E232-E232Article in journal, Letter (Refereed) Published
National Category
Otorhinolaryngology
Identifiers
urn:nbn:se:uu:diva-301136 (URN)10.1002/lary.25879 (DOI)000379980200009 ()26864569 (PubMedID)
Available from: 2016-08-22 Created: 2016-08-18 Last updated: 2017-11-28Bibliographically approved
Cai, Y., Edin, F., Jin, Z., Alexsson, A., Gudjonsson, O., Liu, W., . . . Li, H. (2016). Strategy towards independent electrical stimulation from cochlear implants: Guided auditory neuron growth on topographically modified nanocrystalline diamond. Acta Biomaterialia, 31, 211-220
Open this publication in new window or tab >>Strategy towards independent electrical stimulation from cochlear implants: Guided auditory neuron growth on topographically modified nanocrystalline diamond
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2016 (English)In: Acta Biomaterialia, ISSN 1742-7061, E-ISSN 1878-7568, Vol. 31, p. 211-220Article in journal (Refereed) Published
Abstract [en]

Cochlear implants (CI) have been used for several decades to treat patients with profound hearing loss. Nevertheless, results vary between individuals, and fine hearing is generally poor due to the lack of discrete neural stimulation from the individual receptor hair cells. A major problem is the deliverance of independent stimulation signals to individual auditory neurons. Fine hearing requires significantly more stimulation contacts with intimate neuron/electrode interphases from ordered axonal re-growth, something current CI technology cannot provide.

Here, we demonstrate the potential application of micro-textured nanocrystalline diamond (NCD) surfaces on CI electrode arrays. Such textured NCD surfaces consist of micrometer-sized nail-head-shaped pillars (size 5 5 lm2) made with sequences of micro/nano-fabrication processes, including sputtering, photolithography and plasma etching.

The results show that human and murine inner-ear ganglion neurites and, potentially, neural progenitor cells can attach to patterned NCD surfaces without an extracellular matrix coating. Microscopic methods revealed adhesion and neural growth, specifically along the nail-head-shaped NCD pillars in an ordered manner, rather than in non-textured areas. This pattern was established when the inter-NCD pillar distance varied between 4 and 9 lm.

The findings demonstrate that regenerating auditory neurons show a strong affinity to the NCD pillars, and the technique could be used for neural guidance and the creation of new neural networks. Together with the NCD’s unique anti-bacterial and electrical properties, patterned NCD surfaces could provide designed neural/electrode interfaces to create independent electrical stimulation signals in CI electrode arrays for the neural population.

National Category
Medical Materials Engineering and Technology
Identifiers
urn:nbn:se:uu:diva-266956 (URN)10.1016/j.actbio.2015.11.021 (DOI)000370086100019 ()26593784 (PubMedID)
Funder
EU, FP7, Seventh Framework Programme, 603029
Available from: 2015-11-14 Created: 2015-11-14 Last updated: 2017-12-01Bibliographically approved
Liu, W., Edin, F., Blom, H., Magnusson, P., Schrott-Fischer, A., Glueckert, R., . . . Rask-Andersen, H. (2016). Super-resolution structured illumination fluorescence microscopy of the lateral wall of the cochlea: the Connexin26/30 proteins are separately expressed in man. Cell and Tissue Research, 365(1), 13-27
Open this publication in new window or tab >>Super-resolution structured illumination fluorescence microscopy of the lateral wall of the cochlea: the Connexin26/30 proteins are separately expressed in man
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2016 (English)In: Cell and Tissue Research, ISSN 0302-766X, E-ISSN 1432-0878, Vol. 365, no 1, p. 13-27Article in journal (Refereed) Published
Abstract [en]

Globally 360 million people have disabling hearing loss and, of these, 32 million are children. Human hearing relies on 15,000 hair cells that transduce mechanical vibrations to electrical signals in the auditory nerve. The process is powered by the endo-cochlear potential, which is produced by a vascularized epithelium that actively transports ions in conjunction with a gap junction (GJ) system. This "battery" is located "off-site" in the lateral wall of the cochlea. The GJ syncytium contains the GJ protein genes beta 2 (GJB2/connexin26 (Cx26)) and 6 (GJB6/connexin30 (Cx30)), which are commonly involved in hereditary deafness. Because the molecular arrangement of these proteins is obscure, we analyze GJ protein expression (Cx26/30) in human cochleae by using super-resolution structured illumination microscopy. At this resolution, the Cx26 and Cx30 proteins were visible as separate plaques, rather than being co-localized in heterotypic channels, as previously suggested. The Cx26 and Cx30 proteins thus seem not to be co-expressed but to form closely associated assemblies of GJ plaques. These results could assist in the development of strategies to treat genetic hearing loss in the future.

Keywords
Human cochlea, Connexin (as elsewhere) 26/30, Structured illumination microscopy
National Category
Cell and Molecular Biology
Identifiers
urn:nbn:se:uu:diva-299834 (URN)10.1007/s00441-016-2359-0 (DOI)000378877600003 ()26941236 (PubMedID)
Available from: 2016-07-29 Created: 2016-07-28 Last updated: 2018-05-18Bibliographically approved
Hayashi, H., Edin, F., Li, H., Liu, W. & Rask-Andersen, H. (2016). The effect of pulsed electric fields on the electrotactic migration of human neural progenitor cells through the involvement of intracellular calcium signaling. Brain Research, 1652, 195-203
Open this publication in new window or tab >>The effect of pulsed electric fields on the electrotactic migration of human neural progenitor cells through the involvement of intracellular calcium signaling
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2016 (English)In: Brain Research, ISSN 0006-8993, E-ISSN 1872-6240, Vol. 1652, p. 195-203Article in journal (Refereed) Published
Abstract [en]

Endogenous electric fields (EFs) are required for the physiological control of the central nervous system development. Application of the direct current EFs to neural stem cells has been studied for the possibility of stem cell transplantation as one of the therapies for brain injury. EFs generated within the nervous system are often associated with action potentials and synaptic activity, apparently resulting in a pulsed current in nature. The aim of this study is to investigate the effect of pulsed EF, which can reduce the cytotoxicity, on the migration of human neural progenitor cells (hNPCs). We applied the mono-directional pulsed EF with a strength of 250mV/mm to hNPCs for 6h. The migration distance of the hNPCs exposed to pulsed EF was significantly greater compared with the control not exposed to the EF. Pulsed EFs, however, had less of an effect on the migration of the differentiated hNPCs. There was no significant change in the survival of hNPCs after exposure to the pulsed EF. To investigate the role of Ca(2+) signaling in electrotactic migration of hNPCs, pharmacological inhibition of Ca(2+) channels in the EF-exposed cells revealed that the electrotactic migration of hNPCs exposed to Ca(2+) channel blockers was significantly lower compared to the control group. The findings suggest that the pulsed EF induced migration of hNPCs is partly influenced by intracellular Ca(2+) signaling.

Keywords
Pulsed electric field, Human neural progenitor cell, Electrotactic migration, Intracellular calcium signaling, Time-lapse video microscopy
National Category
Neurology
Identifiers
urn:nbn:se:uu:diva-310911 (URN)10.1016/j.brainres.2016.09.043 (DOI)000388059700023 ()27746154 (PubMedID)
Funder
EU, European Research Council, 281056 603029
Available from: 2016-12-20 Created: 2016-12-20 Last updated: 2017-11-29Bibliographically approved
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