Connexin43 is required for production of the aqueous humor in the murine eye

Calera, Mónica R.; Topley, Heather L.; Liao, Yongbo; Duling, Brian R.; Paul, David L.; Goodenough, Daniel A.

doi:10.1242/jcs.03202

Cited by 45 publications

(62 citation statements)

References 32 publications

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“…Cx43, Gas1, Otx1 and Bmp4 are expressed in the progenitors of the CB but not of the retina ( Fig. 3B-D; data not shown) (Calera et al, 2006;Martinez-Morales et al, 2001), and Otx1 and Bmp4 have been related to CB development (Larsen et al, 2009;Zhao et al, 2002). In Dicer1 loxP/loxP ;-Cre mutants, the expression domains of Cx43, Otx1 and Gas1 were markedly decreased compared with in control littermates ( Fig.…”

Section: Research Articlementioning

confidence: 87%

“…This coordinated development entails close contact of the two epithelial layers. Indeed, targeting the gap junction or cell-adhesion molecules Cx43 and nectin-1/nectin-3 (also known as Pvrl1/Pvrl3), localized at the junctions between the pigmented and non-pigmented layers, disrupts formation of the CB (Calera et al, 2006;Inagaki et al, 2005). Here, we show that exclusive ablation of Dicer1 from either the non-pigmented (Dicer1 loxP/loxP ;-Cre) or pigmented (Dicer1 loxP/loxP ;Tyrp2-Cre) layers seriously hampers the development of both epithelia, emphasizing the vital and reciprocal role of the two layers in the formation of each other (Fig.…”

Section: Cb Formation Requires Dicer1 Functionmentioning

confidence: 99%

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Roles for Dicer1 in the patterning and differentiation of the optic cup neuroepithelium

2011

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SUMMARYThe embryonic ocular neuroepithilium generates a myriad of cell types, including the neuroretina, the pigmented epithelium, the ciliary and iris epithelia, and the iris smooth muscles. As in other regions of the developing nervous system, the generation of these various cell types requires a coordinated sequence of patterning, specification and differentiation events. We investigated the roles of microRNAs (miRNAs) in the development of optic cup (OC)-derived structures. We inactivated Dicer1, a key mediator of miRNA biosynthesis, within the OC in overlapping yet distinct spatiotemporal patterns. Ablation of Dicer1 in the inner layer of the OC resulted in patterning alteration, particularly at the most distal margins. Following loss of Dicer1, this region generated a cryptic population of cells with a mixed phenotype of neuronal and ciliary body (CB) progenitors. Notably, inactivation of Dicer1 in the retinal progenitors further resulted in abrogated neurogenesis, with prolongation of ganglion cell birth and arrested differentiation of other neuronal subtypes, including amacrine and photoreceptor cells. These alterations were accompanied by changes in the expression of Notch and Hedgehog signaling components, indicating the sensitivity of the pathways to miRNA activity. Moreover, this study revealed the requirement of miRNAs for morphogenesis of the iris and for the regulation of CB cell type proliferation and differentiation. Together, analysis of the three genetic models revealed novel, stage-dependent roles for miRNAs in the development of the ocular sub-organs, which are all essential for normal vision.

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Section: Research Articlementioning

confidence: 87%

Section: Cb Formation Requires Dicer1 Functionmentioning

confidence: 99%

Roles for Dicer1 in the patterning and differentiation of the optic cup neuroepithelium

2011

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“…For a more detailed examination, we performed immunohistochemistry with Cx43, AQP1, and AQP4, which are expressed on the CE. 5,10 In the mouse eye, Cx43-positive staining was seen in the lens epithelium, RPE and CE, and it was especially evident in the gap junction between PE and NPE (Figs. 1A, 1E).…”

Section: The Peripheral Structure Of Optic Cup-like Aggregate From Momentioning

confidence: 99%

Induction of Functional 3D Ciliary Epithelium–Like Structure From Mouse Induced Pluripotent Stem Cells

Kinoshita

Suzuma

Kaneko

et al. 2016

Invest. Ophthalmol. Vis. Sci.

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Citation: Kinoshita H, Suzuma K, Kaneko J, Mandai M, Kitaoka T, Takahashi M. Induction of functional 3D ciliary epithelium-like structure from mouse induced pluripotent stem cells. Invest Ophthalmol Vis Sci. 2016;57:153-161. DOI:10.1167/ iovs.15-17610 PURPOSE. To generate ciliary epithelium (CE) from mouse induced pluripotent stem (iPS) cells.METHODS. Recently, a protocol for self-organizing optic cup morphogenesis in threedimensional culture was reported, and it was suggested that ocular tissue derived from neural ectoderm could be differentiated. We demonstrated that a CE-like double-layered structure could be induced in simple culture by using a modified Eiraku differentiation protocol.RESULTS. Differentiation of a CE-like double-layered structure could be promoted by glycogen synthase kinase 3b (GSK-3b) inhibitor. Connexin43 and aquaporin1 were expressed in both thin layers, and induced CE-like cells expressed ciliary marker genes, such as cyclinD2, zic1, tgfb2, aldh1a3, wfdc1, otx1, BMP4, and BMP7. Increases in cytoplasmic and nuclear bcatenin in aggregates of the CE-like double-layered structure were confirmed by Western blot analysis. In addition, tankyrase inhibitor prevented the induction of the CE-like double-layered structure by GSK-3b inhibitor. Dye movement from pigmented cells to nonpigmented cells in the mouse iPS cell-derived CE-like structure was observed in a fluid movement experiment, consistent with the physiological function of CE in vivo.CONCLUSIONS. We could differentiate CE from mouse iPS cells in the present study. In the future, we hope that this CE-like complex will become useful as a graft for transplantation therapy in pathologic ocular hypotension due to CE dysfunction, and as a screening tool for the development of drugs for diseases associated with CE function.Keywords: ciliary epithelium, iPS, GSK-3b, b-catenin T he ciliary body (CB) plays important roles in eye homeostasis. It secretes aqueous humor that maintains IOP, controls the lens shape through the ciliary muscle and ciliary zonules, and synthesizes glycoproteins of the vitreous body. The CB contains a double-layered epithelial structure comprising the inner nonpigmented epithelium (NPE), the outer pigmented epithelium (PE), and the underlying stroma. The NPE is continuous with the neural retina (NR) and the iris pigmented epithelium, while the PE lies between the retinal pigment epithelium (RPE) and the outer iris muscle. 1 The NPE and the PE, as well as the NR and RPE, are derived from neural ectoderm. During eye development, the ciliary epithelium (CE) folds to form the ciliary processes, while neural crest cellderived stroma cells underneath the CE also contribute to CB morphogenesis. 2 The PE rests on the connective tissue stroma, and the NPE is polarized with its basal lamina facing the posterior chamber of the eye. The PE and NPE interact via their apical membranes with gap junctions.Water can pass from PE to NPE via gap junctions following NaCl movement 3,4 and exit the NPE to the posterior chamber by using ba...

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“…In addition, we also examined Connexin43 and Collagen IX protein expression for CB secretion function in the control and Notch2 CKO mutant CBs. Connexin43 forms gap junctions between ICE and OCE and controls aqueous humor production (38), whereas Collagen IX is secreted by the CB and accumulates on the surface of the CB and the retina (39). Both the control and Notch2 CKO mutant CBs exhibit similar levels of Connexin43 (Conn43) expression in the OCE and secreted Collagen IX protein on the surface of the CB ( Fig.…”

Section: Notch2mentioning

confidence: 99%

Notch2 regulates BMP signaling and epithelial morphogenesis in the ciliary body of the mouse eye

Zhou

Tanzie

Yan

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

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The ciliary body (CB) of the mammalian eye is responsible for secreting aqueous humor to maintain intraocular pressure, which is elevated in the eyes of glaucoma patients. It contains a folded two-layered epithelial structure comprising the nonpigmented inner ciliary epithelium (ICE), the pigmented outer ciliary epithelium (OCE), and the underlying stroma. Although the CB has an important function in the eye, its morphogenesis remains poorly studied. In this study, we show that conditional inactivation of the Jagged 1 (Jag1)-Notch2 signaling pathway in the developing CB abolishes its morphogenesis. Notch2 is expressed in the OCE of the CB, whereas Jag1 is expressed in the ICE. Conditional inactivation of Jag1 in the ICE or Notch2 in the OCE disrupts CB morphogenesis, but neither affects the specification of the CB region. Notch2 signaling in the OCE is required for promoting cell proliferation and maintaining bone morphogenetic protein (BMP) signaling, both of which have been suggested to be important for CB morphogenesis. Although Notch and BMP signaling pathways are known to crosstalk via the interaction between their downstream transcriptional factors, this study suggests that Notch2 maintains BMP signaling in the OCE possibly by repressing expression of secreted BMP inhibitors. Based on our findings, we propose that Jag1-Notch2 signaling controls CB morphogenesis at least in part by regulating cell proliferation and BMP signaling.T he mammalian eye is composed of the anterior segment, the posterior retina and the vitreous. The anterior segment consists of cornea, lens, and ciliary body (CB), whereas the posterior retina contains six types of retinal neurons and Müller glial cells, which are organized into three distinct cell layers. The light passes through the cornea and is then focused by the lens and detected by photoreceptors in the retina. Both the aqueous humor anteriorly and the vitreous humor posteriorly function together to sustain intraocular pressure (IOP) in the eye and thereby maintain its shape. Pressure regulation is particularly important because abnormally high IOP is a major risk factor for glaucoma (1). In the eye, the CB is responsible for producing aqueous humor (2). Although high IOP is often attributed to the blockage of the drainage system for the vitreous, known as the trabecular meshwork, abnormal CB function might also contribute to high IOP formation because of excess aqueous humor production. Finally, contraction of the muscles in the CB controls lens accommodation for near versus far vision. Despite its important biological functions and potential medical significance, the formation and development of the CB remain poorly studied.The CB contains two layers of apically adhered epithelial sheets, the pigmented outer ciliary epithelium (OCE) and the nonpigmented inner ciliary epithelium (ICE), and the underlying stroma (2). It forms at the periphery of the developing optic cup and first segregates from the retina and then from the iris. One previous study suggested that blood vesse...

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Connexin43 is required for production of the aqueous humor in the murine eye

Cited by 45 publications

References 32 publications

Roles for Dicer1 in the patterning and differentiation of the optic cup neuroepithelium

Roles for Dicer1 in the patterning and differentiation of the optic cup neuroepithelium

Induction of Functional 3D Ciliary Epithelium–Like Structure From Mouse Induced Pluripotent Stem Cells

Notch2 regulates BMP signaling and epithelial morphogenesis in the ciliary body of the mouse eye

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