Immunohistochemical and ultrastructural investigation of the human vestibular dark cell area: Roles of subepithelial capillaries and T lymphocyte-melanophage interaction in an immune surveillance system

Masuda, Masataka; Yamazaki, Kazuto; Kanzaki, Jin; Hosoda, Yasuhiro

doi:10.1002/(sici)1097-0185(199710)249:2<153::aid-ar1>3.0.co;2-z

Cited by 13 publications

(9 citation statements)

References 29 publications

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“…An intimate relation between the VDCs and the microvascular network in the rat was verified by TEM observations, which correlates with a previous study in man [14].…”

Section: Discussionsupporting

confidence: 90%

Vestibular tributaries to the vein of the vestibular aqueduct

Hansen

Qvortrup

Friis

2010

Acta Oto-Laryngologica

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A microvascular network was observed in close relation to the VDCs in the utricle and the crista ampullaris of the lateral semicircular canal in the vestibular apparatus. One major vein emanated from these networks, which emptied into the vein of the vestibular aqueduct. Veins draining the saccule and the common crus of the superior and posterior semicircular canals were likewise observed to merge with the vein of the vestibular aqueduct.

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“…An intimate relation between the VDCs and the microvascular network in the rat was verified by TEM observations, which correlates with a previous study in man [14].…”

Section: Discussionsupporting

confidence: 90%

Vestibular tributaries to the vein of the vestibular aqueduct

Hansen

Qvortrup

Friis

2010

Acta Oto-Laryngologica

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“…A comparative study by Middleton et al (Middleton et al, 2005) found that vWFis the gold standard for EC identification in human tissue. vWF expression in vestibular capillary ECs, previously reported by Masuda et al (Masuda et al, 1997), was pre-validated in our immunofluorescence labeling. In our study, we validated the EC cell line from vestibular tissue based on expression of vWF and Glut1, the latter a second well-known EC marker protein, as well as on the basis of features such as a flat and generally cuboidal morphology, as shown in Figure 5.…”

Section: Discussionsupporting

confidence: 72%

“…Despite the variety of protein expression, Von Willebrand factor is considered the gold standard for EC identification (Middleton et al, 2005). Expression of vWF in endothelial cells in human vestibular capillaries was previously reported by Masuda M et al (Masuda et al, 1997). In the present study, we identified ECs from the vestibular system in confocal microscope images on the basis of vWF expression.…”

Section: Resultssupporting

confidence: 61%

Culture media-based selection of endothelial cells, pericytes, and perivascular-resident macrophage-like melanocytes from the young mouse vestibular system

et al. 2017

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The vestibular blood-labyrinth barrier (BLB) is comprised of perivascular-resident macrophage-like melanocytes (PVM/Ms) and pericytes (PCs), in addition to endothelial cells (ECs) and basement membrane (BM), and bears strong resemblance to the cochlear BLB in the stria vascularis. Over the past few decades, in vitro cell-based models have been widely used in blood-brain barrier (BBB) and blood-retina barrier (BRB) research, and have proved to be powerful tools for studying cell-cell interactions in their respective organs. Study of both the vestibular and strial BLB has been limited by the unavailability of primary culture cells from these barriers. To better understand how barrier component cells interact in the vestibular system to control BLB function, we developed a novel culture medium–based method for obtaining EC, PC, and PVM/M primary cells from tiny explants of the semicircular canal, sacculus, utriculus, and ampullae tissue of young mouse ears at post-natal age 8 - 12 d. Each phenotype is grown in a specific culture medium which selectively supports the phenotype in a mixed population of vestibular cell types. The unwanted phenotypes do not survive passaging. The protocol does not require additional equipment or special enzyme treatment. The harvesting process takes less than 2 h. Primary cell types are generated within 7 - 10 d. The primary culture ECs, PCs, and PVM/M shave consistent phenotypes more than 90% pure after two passages (~ 3 weeks). The highly purified primary cell lines can be used for studying cell-cell interactions, barrier permeability, and angiogenesis.

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“…It is currently speculated that the dark cell area may contribute to the maintenance of homeostasis of both the endolymph and perilymph or that it may be a significant functional site for maintenance of the microenvironment of the vestibular organs of the mammalian inner ear because epithelial dark cells contain various enzymes, such as Na + ‐K + ‐ATPase (Wangemann et al, 1996), Ca 2+ ‐ATPase (Wangemann et al, 1996), and carbonic anhydrase (Watanabe and Ogawa, 1984). In humans, this dark cell area consists of epithelial dark cells, a large number of subepithelial melanocytes, a relatively small number of other stromal cells, and melanophages, usually detected next to the subepithelial blood vessels (Masuda et al, 1994, 1995, 1997).…”

Section: Discussionmentioning

confidence: 99%

Connexin 26 distribution in gap junctions between melanocytes in the human vestibular dark cell area

et al. 2001

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We have previously demonstrated the presence of gap junctions between melanocytes in the human vestibular organ and have speculated that melanocytes function in maintaining the homeostasis of the microenvironment of the inner ear. The purpose of the present study was to characterize the expression and ultrastructural localization of connexin (Cx) protein in melanocytes of the human vestibular organs. Surgical material was obtained from patients operated on for vestibular schwannoma and was processed for light microscopy, confocal laser scanning microscopy, conventional TEM, and immuno TEM. The specimens were labeled with anti‐Cx26, Cx32, and Cx43 antibodies and examined by light microscopy. Specimens were also labeled with anti‐Cx26 antibody and examined by laser microscopy and immuno‐TEM methods. The specimens examined in this study were mainly dark cell areas from the human vestibular organ, whose epithelial and subepithelial layers are rich in melanocytes. Light‐microscopic immunohistochemical studies showed positive labeling for Cx26 protein between subepithelial melanocytes, and Cx32 was also detected. Use of anti‐Cx26 antibody and confocal laser scanning microscopy revealed high levels of Cx26 around the subepithelial melanocytes. Post‐embedding immuno‐gold transmission electron microscopy showed significant aggregation of gold particles (33.97 ± 8.01% of total gold particles) around the gap junctions of the subepithelial melanocytes. The results of this study indicated that melanocytes are connected through gap junctions that mainly contain Cx26. This suggested that the melanocytes in the human vestibular organ may play a role in transporting material between the endolymph and perilymph. Anat Rec 262:137–146, 2001. © 2001 Wiley‐Liss, Inc.

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Immunohistochemical and ultrastructural investigation of the human vestibular dark cell area: Roles of subepithelial capillaries and T lymphocyte-melanophage interaction in an immune surveillance system

Cited by 13 publications

References 29 publications

Vestibular tributaries to the vein of the vestibular aqueduct

Vestibular tributaries to the vein of the vestibular aqueduct

Culture media-based selection of endothelial cells, pericytes, and perivascular-resident macrophage-like melanocytes from the young mouse vestibular system

Connexin 26 distribution in gap junctions between melanocytes in the human vestibular dark cell area

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