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At least 19 recordsLinked to original sources

Muscarinic cholinergic inhibition of adenylate cyclase in the rabbit iris-ciliary body and ciliary epithelium.

The effects of cholinergic agents on hormone-stimulated cyclic AMP (cAMP) accumulation were investigated in iris-ciliary body segments, excised ciliary processes, and isolated ciliary epithelium from albino rabbit eyes. In all three tissue preparations, the cholinergic agonist carbamylcholine markedly inhibited the stimulation of cAMP biosynthesis by vasoactive intestinal peptide VIP--a potent activator of nonpigmented ciliary epithelial adenylate cyclase. Carbamylcholine also attenuated cAMP increases mediated by isoproterenol, prostaglandin E2, and forskolin. The effects of carbamylcholine on VIP-induced cAMP synthesis were concentration dependent (EC50 = 23 nM), mimicked by selective muscarinic cholinergic agonists (oxotremorine, pilocarpine), and antagonized by atropine. Carbamylcholine- and clonidine-mediated inhibition of VIP-stimulated cAMP accumulation in ciliary processes were nonadditive, indicating that inhibitory muscarinic and alpha 2-adrenergic receptors coexist on VIP-responsive target cells. These findings suggest that the cholinergic system may have a direct role in modulation of ciliary epithelial adenylate cyclase and aqueous humor secretion.

Adenosine Triphosphate↗

Synthesis and composition of glycosaminoglycans by explant cultures of human ciliary body and ciliary processes in serum-containing and serum-free defined media.

Freshly isolated ciliary body explants and tips of ciliary processes derived from human eyes were cultured in serum-containing or serum-free defined medium. These cultures synthesized tissue-bound and medium-released ("excreted") glycosaminoglycans (GAGs), as evidenced by the incorporation of 14C-glucosamine and enzymatic characterization of these labelled GAGs (hyaluronic acid, heparan sulphate, chondroitin sulphate, dermatan sulphate). The GAG synthesis and excretion rate was enhanced by serum. It is suggested that ciliary epithelium performs this function of GAG synthesis also under in vivo conditions.

Adolescent↗

Effect of heptanol on the short circuit currents of cornea and ciliary body demonstrates rate limiting role of heterocellular gap junctions in active ciliary body transport.

Rabbit ciliary body and cornea were mounted in Ussing-type chambers in Tyrode's under voltage clamp and the effects of heptanol, a gap junction inhibitor, on the short circuit current generated by each of the respective epithelia were determined. Studies were carried out either in control conditions or following amphotericin B permeabilization of either the basolateral membrane of the nonpigmented epithelium of the ciliary body or the apical membrane of the corneal epithelium, respectively. Previous studies have shown that, following these permeabilizations, short circuit currents are established, reflecting aqueous (or tear)-to-serosa Na+ fluxes, and that Na+ translocation through gap junctions connecting the individual layers of these tissues constitutes the major rate limiting step. Heptanol inhibited most of the short circuit current of the amphotericin B-modified ciliary body and cornea and of the unmodified ciliary body epithelium (control). In all these cases, the apparent IC50 was about 0.8 M. In the unmodified corneal epithelium, where ion translocation across the apical membrane constitutes the main rate limiting step for active secretion, 0.4 or 0.8 mM heptanol induced short circuit current increases; partial inhibition was observed only at high concentrations known to cause maximal inhibition of junctional permeability. Heptanol also enhanced the volume regulatory decrease of cultured human NPE cells, a process dependent on cell swelling-induced stimulation of Cl- and K+ permeabilities. Combined with our previous results demonstrating the lack of heptanol effects on other epithelial functions, these data suggest that the effect of heptanol on the active ciliary body transepithelial transport is primarily due to inhibition of the nonpigmented-pigmented junctional path and that this path is a potential site of rate limitation for the secretory process.

Alcohols↗

Growth factors induce neurogenesis in the ciliary body.

The ciliary body of the eye is a nonneural tissue that is derived from the anterior rim of the optic cup, an extension of the neural tube. This tissue normally does not contain neurons and functions to produce components of the aqueous humor. We found that intraocular injections of insulin, EGF, or FGF2 stimulate NPE cells to proliferate and differentiate into neurons. These growth factors had region-specific effects along the radial axis of the ciliary body, with insulin and EGF stimulating proliferation of NPE cells close to the retina, while FGF2 stimulated the proliferation of NPE cells further toward the lens. Similar region-specific effects were observed for accumulations of neurons in the NPE in response to injections of different growth factors. The neurons derived from NPE cells express neurofilament, beta3 tubulin, RA4, calretinin, Islet1, or Hu, and a few produced long axonal projections, several millimeters in length that extend across the ciliary body. Our results suggest that the ciliary body has the capacity to generate retinal neurons, but normally neurogenesis is actively inhibited.

Animals↗

BMP signaling is required for development of the ciliary body.

The ciliary body in the eye secretes aqueous humor and glycoproteins of the vitreous body and maintains the intraocular pressure. The ciliary muscle controls the shape of the lens through the ciliary zonules to focus the image onto the retina. During embryonic development, the ciliary epithelium is derived from the optic vesicle, but the molecular signals that control morphogenesis of the ciliary body are unknown. We report that lens-specific expression of a transgenic protein, Noggin, can block BMP signaling in the mouse eye and result in failure in formation of the ciliary processes. Co-expression of transgenic BMP7 restores normal development of the ciliary epithelium. Ectopic expression of Noggin also promotes differentiation of retinal ganglion cells. These results indicate that BMP signaling is required for development of the ciliary body and may also play a role in regulation of neuronal differentiation in the developing eye.

Animals↗

Expression of Na,K-ATPase alpha subunit isoforms in the human ciliary body and cultured ciliary epithelial cells.

We have analyzed the expression of Na,K-ATPase alpha subunit isoforms in the transporting ciliary processes of the human eye and in cultured cells derived from non-pigmented (NPE) and pigmented (PE) ciliary epithelium. Northern hybridization analysis shows that the mRNAs encoding all the three distinct forms of Na,K-ATPase alpha subunit [alpha 1, alpha 2, and alpha 3] are expressed in the human ciliary processes in vivo. Immunohistochemical analysis using antibodies specific for each of the three alpha subunit isoforms confirms that these polypeptides are present in the microsomal fraction from the human ciliary processes. The monoclonal antibody McB2, which is specific to the Na,K-ATPase alpha 2 subunit isoform, has been found to decorate specifically the basolateral membrane domains of NPE cells but not of the PE cells, suggesting its expression in vivo only in the ocular NPE ciliary epithelium. However, cultured cells derived from the NPE and PE layers exhibit a different pattern of expression of mRNA and protein for the Na,K-ATPase alpha subunit isoforms when compared to the tissue. Both the NPE and PE cells express alpha 1 and alpha 3 mRNA and polypeptide, whereas alpha 2 mRNA and polypeptide are undetectable in these cells. The established cell lines derived from the NPE layer express comparable levels of the alpha 1 and alpha 3 isoforms of Na,K-ATPase as detected in the primary culture. However, the established NPE cell lines are also distinguishable from the normal PE cells when analyzed by Western blot analysis with A x 2 antibodies. The results presented here clearly show that the NPE and PE cells in the ciliary body have a distinct expression of Na,K-ATPase alpha subunit isoforms as compared to cultured cells.

Blotting, Western↗

Expression of a chemokine by ciliary body epithelium in horses with naturally occurring recurrent uveitis and in cultured ciliary body epithelial cells.

OBJECTIVE: To determine whether a chemokine (RANTES)-like protein expressed by ciliary epithelium plays a role in uveitis. SAMPLE POPULATION: 3 clinically normal horses intradermal, 5 eyes from 5 horses with recurrent uveitis, and 10 normal eyes from 5 age- and sex-matched horses. PROCEDURE: Cross-reactivity and sensitivity of recombinant human (rh)-regulated upon activation, normal T-cell expressed and secreted (RANTES) protein were evaluated in horses by use of intradermal hypersensitivity reactions and a chemotaxis assay. Aqueous humor and ciliary body of eyes from clinically normal horses and horses with uveitis were examined for RANTES expression by use of an ELISA and reverse transcription-polymerase chain reaction (RT-PCR). Expression of RANTES mRNA and protein content of primary cultures of equine ciliary pigmented epithelial cells (RT-PCR) and culture supernatant (ELISA) were measured 6 or 24 hours, respectively, after cultures were stimulated with interleukin-1beta and tumor necrosis factor-alpha. RESULTS: Strong reactions to intradermal hypersensitivity testing and significant chemotaxis of equine leukocytes to rh-RANTES wereas observed. Aqueous humor of eyes from horses with uveitis contained increased concentrations of rh-RANTES-like protein (mean +/- SD, 45.9+/-31.7 pg/ml), compared with aqueous humor from clinically normal horses (0 pg/ml). Ciliary body from horses with uveitis expressed RANTES mRNA, whereas ciliary body from clinically normal horses had low mRNA expression. Stimulated ciliary pigmented epithelial cells expressed increased amounts of rh-RANTES-like protein (506.1+/-298.3 pg/ml) and mRNA, compared with unstimulated samples. CONCLUSIONS AND CLINICAL RELEVANCE: Ciliary epithelium may play a role in recruitment and activation of leukocytes through expression of RANTES.

Animals↗

Immunoregulatory properties of bone marrow-derived cells in the iris and ciliary body.

Iris and ciliary body of mouse eyes have been examined for the presence of bone marrow-derived cells possessing the capability of functioning as antigen-presenting cells (APC). We have determined that iris and ciliary body contain significant numbers of cells bearing T200, indicating their bone marrow origin. Most of these express the F4/80 marker typically found on mature macrophages. However, approximately one-third of the cells express Ia and a similar number express Mac-1 markers. Virtually none of the cells express Thy-1 or surface immunoglobulin. Whole preparations of excised iris/ciliary body, or single cell suspensions prepared from these tissues were then assayed for their capacity to induce proliferation among allogeneic lymphocytes. It was discovered that iris/ciliary body tissues or cells did not function as alloantigen-presenting cells, although tissue and cells derived from the corneal limbus were allostimulatory. In addition, iris/ciliary body tissues and cells displayed the ability to suppress mixed lymphocyte reactions to which they had been added as regulatory cells. We conclude that normal iris and ciliary body contain bone marrow-derived cells that fail to function as alloantigen-presenting cells. However, cells were present that have the capacity to inhibit alloimmune lymphocyte proliferation. The strategic location of inhibitory cells in the tissues that line the anterior chamber of the eye raises the possibility that these cells may play a role in the phenomenon of immunological privilege that is characteristic of this site.

Animals↗

Expression of protein gene product 9.5 in lepromatous eyes showing ciliary body nerve damage and a "dying back" phenomenon in the posterior ciliary nerves.

BACKGROUND/AIM: Peripheral nerve destruction is the hallmark of leprosy. Ocular complications form a substantial part of the clinical manifestations but histopathology of nerve destruction within ocular structures has not been shown satisfactorily. The role of protein gene product (PGP) 9.5 in identifying nerve destruction in the ciliary body and posterior ciliary nerves of lepromatous eyes is shown. METHODS: Serial sections from two lepromatous eyes and two non-lepromatous eyes were stained with PGP 9.5. Histopathological comparison was done on the expression of the PGP 9.5 stain in nerves within the ciliary body, posterior ciliary nerves adjacent to the optic nerve, and nerves tracking through the sclera. RESULTS: In non-lepromatous eyes, PGP 9.5 was expressed in nerves within the ciliary body, the nerves within the sclera, and posterior ciliary nerves adjacent to the optic nerve. In lepromatous eyes no PGP 9.5 was expressed, signifying nerve destruction. CONCLUSIONS: Nerve destruction in lepromatous eyes has been confirmed histopathologically by the absence of or patchy staining with PGP 9.5. Nerve destruction in the ciliary body can extend to the posterior ciliary nerves by an ascending axonopathy. This "dying back" phenomenon is akin to the "glove and stocking" anaesthesia found in lepromatous leprosy.

Adult↗