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The HNK-1 epitope in the inner connective tissue layer of the human ciliary body in exfoliation syndrome and various types of glaucoma.

Possible changes in the expression of the HNK-1 carbohydrate epitope in the inner connective tissue layer of the human ciliary body, located between the ciliary epithelium and muscle, was studied using 2 formalin-fixed, paraffin-embedded eyes with exfoliation syndrome, 33 eyes with different types of glaucoma, and 21 morphologically normal control eyes. A strong immunoreaction delineating cell process was observed in this layer with monoclonal antibodies HNK-1 and VC1.1 recognizing the HNK-1 epitope in control specimens, whereas partly granular immunoreaction was present in eyes with exfoliation syndrome. Exfoliation material was also immunoreactive. In all types of advanced glaucoma, the immunoreaction was mostly granular in nature and greatly diminished. No difference in HNK-1 immunoreactivity between control and glaucoma eyes was seen in the retina and ciliary epithelium. Elevated intraocular pressure, either directly or by decreasing blood flow to the ciliary body, may cause degenerative or metabolic changes in the inner connective tissue layer cells that bear or secrete molecules sharing the HNK-1 epitope. The partly granular immunoreactivity in eyes with exfoliation syndrome only indicates changes in this epitope even without an increase in intraocular pressure.

Adult↗

Effect of mitomycin C on ciliary body and intraocular pressure with various application depths: an experimental study.

BACKGROUND: To evaluate the effects of mitomycin C (MMC) on intraocular pressure (IOP) and ciliary body via transmission electron microscopy when applied under conjunctiva or different depths of sclera, without performing any filtering surgery. METHODS: Thirty-six eyes of 36 New Zealand albino rabbits were used in this study. MMC was prepared in a concentration of 0.4 mg/mL and 0.05 cc (20 microg) was soaked in preprepared sterile surgical sponges. Six groups each consisting of six eyes were formed and IOP was measured preoperatively. Group 1 was the control group: the superior conjunctiva was opened and only irrigated with balanced salt solution (BSS). In group 2, MMC soaked sponges were applied under the conjunctiva. In groups 3 and 4, a scleral flap of approximately 1/3 scleral thickness was prepared and in groups 5 and 6, and a scleral flap of approximately 2/3 scleral thickness was prepared, all with a standard size of 4 x 4 mm. MMC soaked sponges were applied under these areas for 5 min in eyes in groups 3 and 5 followed by an irrigation of the relevant areas with 10 cc BSS, whereas only irrigation with BSS was done in groups 4 and 6 as control groups. No filtering procedure was performed in any of the eyes. Eyes were enucleated on the 30th day following measurement of IOP and the ciliary body regions were evaluated using transmission electron microscopy. Kruskal-Wallis test was used for the statistical assessment of IOP between groups. RESULTS: The deep scleral flap group (group 5) showed statistically significantly more IOP reduction than both the superficial scleral flap group (group 3; P = 0.004) and the subconjunctival group (group 2; P = 0.002) on postoperative day 30. Electron microscopic evaluation of the surgical groups revealed a wide range of different histopathological effects due to different MMC application methods. The histopathological changes were more evident in the group 5, where MMC was applied under deep scleral flap. CONCLUSIONS: Subscleral application of MMC seems to provide greater IOP decrease than subconjunctival application, possibly caused by a more significant ciliary body toxicity. This may be the beginning of a non-penetrating, easy to perform and safe method to decrease IOP in glaucoma patients, which the authors call 'toxic ciliary ablation surgery'. However, the long-term results and complications must be assessed with further studies.

Animals↗

Prejunctional modulation of norepinephrine release in the human iris-ciliary body.

PURPOSE: To characterize the prejunctional mechanisms that control the impulse-evoked release of norepinephrine in the isolated, superfused human iris-ciliary body. METHODS: Human iris-ciliary body tissue segments were preincubated with 3H-norepinephrine, superfused and electrically-stimulated in vitro to evoke the discharge of 3H-norepinephrine. The effects of prejunctional modulators on evoked 3H-norepinephrine overflow were evaluated. RESULTS: Stimulation-evoked (but not spontaneous) 3H-norepinephrine release was inhibited by alpha 2-adrenergic, muscarinic, dopaminergic, neuropeptide Y, and prostaglandin agonists and was enhanced by angiotensin II. Agonist-induced effects on 3H-norepinephrine overflow were blocked by selective antagonists, where available. Yohimbine and atropine alone enhanced 3H-norepinephrine output, suggesting that prejunctional alpha 2-adrenergic and muscarinic receptors undergo tonic activation by endogenously released neurotransmitters. CONCLUSIONS: Human ocular sympathetic nerves express inhibitory alpha 2-adrenergic, muscarinic, dopaminergic, prostaglandin, and neuropeptide Y receptors and facilitatory angiotensin II receptors that control the impulse-evoked release of 3H-norepinephrine. These receptors may be useful targets for pharmacologic manipulation of the adrenergic system in vivo.

Adolescent↗

Glycoconjugates of the rat ciliary body epithelium: a lectin histochemical and protein blotting study.

The present study sought to identify and partially characterize the glycoconjugates specific to the double-layered ciliary body epithelium of the rat eye by lectin histochemistry and lectin blottings. Hydrated paraffin sections of Carnoy-fixed Sprague-Dawley rat eyes were stained with a panel of 21 different biotinylated lectins, followed by streptavidin-peroxidase and the glucose oxidase-diaminobenzidine-nickel staining procedure. The results of lectin histochemistry revealed that the inner epithelial layer was rich in GlcNAc(beta1,4)GlcNAc, alpha-Gal, Gal(beta1,3)GalNAc, GalNAc(alpha1,3)GalNAc/Gal, GalNAc(alpha1,6)Gal, Fuc(alpha1,2)Gal(beta1,4)GlcNAc and Gal(beta1,4)GlcNAc(beta1,2)Man(alpha1,6) sugar residues as shown by its positive reactivities with S-WGA, PWA, DSA, GS-I-B4, PNA, DBA, SBA, WFA, UEA-I, LTA and PHA-E. The reactivities of GS-I-B4, PNA, DBA and SBA were restricted to the inner layer at the tips of the ciliary processes. On the other hand, the outer epithelial layer was stained evenly by DSA and Jacalin, and partly by MAA, showing that this epithelial layer was rich in GlcNAc(beta1,4)GlcNAc, Gal(beta1,3)GalNAc and NeuAc(alpha2,3)Gal disaccharides. These lectin binding patterns of the ciliary body epithelium suggest a topographical and functional difference in this double cell-layered epithelium. Their possible roles in the secretion of aqueous humour and production of ciliary zonule are discussed. Some identified lectin markers specific to these two cell layers may be useful for further experimental studies. Glycoproteins extracted from the dissected ciliary body were separated by SDS-PAGE electrophoresis and analyzed by protein blottings with 8 different lectins. The results showed that at least 10 major membrane-bound glycoproteins, with molecular weights ranging from 30 to 150 kD, rich in beta-GlcNAc, beta-Gal, alpha/beta-GalNAc and NeuAc(alpha2,6)Gal residues, were present in the microsomal fraction.

Acetylgalactosamine↗

Influences on the density of beta-adrenergic receptors in the cornea and iris--ciliary body of the rabbit.

By measurement of the specific binding of 3H-dihydroalprenolol, the densities of beta-adrenergic receptors on membranes prepared from homogenized corneas and iris--ciliary bodies of rabbits were studied. Sympathetic denervation, as a result of subconjunctival treatment with 6-hydroxydopamine, causes an increase in the density of beta-adrenergic receptors in membranes prepared from the ipsilateral iris--ciliary body but not the cornea. Topical treatment with epinephrine for 5 days causes a decrease in the density of beta-adrenergic receptors in membranes prepared from cornea and iris-ciliary body, whereas similar treatment with timolol causes an increase in the density of beta-adrenergic receptors. In the cornea, the decrease in receptor density that occurs following in vivo treatment with epinephrine is associated with a decreased ability to synthesize cyclic AMP, whereas the increase in receptor density that occurs following in vivo treatment with timolol is not associated with an altered ability to synthesize cyclic AMP. Our results indicate that the density of beta-adrenergic receptors in the anterior segment of the eye is inversely related to the level of adrenergic stimulation to the tissue but that the ability of a tissue to synthesize cyclic AMP does not necessarily parallel the change in receptor density.

Animals↗

Adreno-cholinergic modulation of junctional communications between the pigmented and nonpigmented layers of the ciliary body epithelium.

PURPOSE: Cell-to-cell communications between the epithelial layers of the ciliary body may be critical for aqueous humor production. The aim of this study was to identify pharmacologic agents that affect this path. METHODS: Whole New Zealand rabbit ciliary bodies were mounted in Ussing-type chambers with Ca2+(-)free and Ca2+(-)rich Tyrode's in the nonpigmented (NPE; aqueous) and pigmented (PE; serosa) epithelial side hemichambers, respectively. The NPE of the PE were then permeabilized, either selectively to monovalent ions with amphotericin B or nonselectively to small solutes with digitonin. Resultant active transport activities were tracked as short circuit currents (ISCS). RESULTS: Permeabilization of the NPE with either 10 microM amphotericin B or 10 micro M digitonin led to an aqueous-to-serosa-positive ISC. This ISC was inhibited by serosal-side ouabain and heptanol, indicating movement of Na+ from permeabilized NPE to the PE by the interlayer junctional path, followed by PE-to-serosa active Na+ transport. Permeabilization of the PE with amphotericin B elicited an ISC in the opposite direction, This ISC was abolished by aqueous-side ouabain and by heptanol, consistent with sequential PE to NPE Na+ translocation, followed by active, NPE-to-aqueous transport. Acetylcholine, epinephrine, norepinephrine, and the alpha 1-adrenergic agonist phenylephrine, but not brominidine, an alpha 2-adrenergic agonist, each caused an approximately 50% reduction of these currents. The inhibitions were fully dependent on serosal-side Ca2+ and were blocked by one calmodulin inhibitor, trifluoperazine, but not by another, calmidazolium. CONCLUSIONS: The above observations provide evidence that cholinergic or alpha 1-adrenergic activation of the PE causes Ca2+(-)dependent inhibition of the NPE-PE junctional path. A triflouperazine-sensitive entity, which may be distinct from calmodulin, is involved in the inhibition.

Adrenergic Agonists↗

Expression of vascular endothelial growth factor a, matrix metalloproteinase 9 and extravascular matrix patterns in iris and ciliary body melanomas.

PURPOSE: It was the aim of this study to assess the expression of vascular endothelial growth factor (VEGF)-A, matrix metalloproteinase (MMP)-9 and extravascular matrix patterns (EMPs) in iris and ciliary body melanomas and their correlations with histopathologic parameters. METHODS: The study was conducted on 3 iris and 15 ciliary body melanomas. All tumors were subjected to immunohistochemical techniques for VEGF-A and MMP-9 expressions, the presence of EMPs was assessed, and routine paraffin sections were stained with hematoxylin-eosin. Cell type, tumor localization, degree of pigmentation, necrosis, mitotic index, lymphocytic infiltration and sclera invasion were analyzed using light microscopy. RESULTS: The mean patient age at the time of treatment was 43 years (range 19-69, median 39.5); 10 (55.6%) patients were males and 8 (44.4%) females. Histopathological cell types were spindle cells in 55.6%, mixed cells in 16.7%, and epithelioid cell types in 27.8% of tumors. Positive reaction for VEGF-A and MMP-9 was present in 66.7 and 72.3% of the tumors, respectively. Microvascular loops and/or networks were seen in 33.4% of the tumors, with the remaining 66.7% of tumors displaying one or more of the other patterns. Metastatic disease developed in only 1 patient during follow-up. Tumor cell type, tumor size, mitotic rate, degree of pigmentation and EMPs were not correlated with metastasis. CONCLUSIONS: This study suggests that VEGF-A and MMP-9 were positive in the majority of iris and ciliary body melanomas. No correlation was found between VEGF-A and MMP-9 immunoreactivity and EMPs and occurrence of metastases in cases of anterior uveal melanoma.

Adult↗

Identification of a novel element in the human eye: the inner connective tissue layer of the ciliary body characterized with antibodies to the HNK-1 epitope.

PURPOSE: To characterize the nature and the developmental distribution of the HNK-1 epitope in the inner connective tissue layer of the human ciliary body, located between the ciliary epithelium and muscle with two monoclonal antibodies to the HNK-1 epitope common to many cell adhesion molecules. METHODS: Nine fetal (gestational age 13-40 wk) and 32 postnatal human eyes (age 3 mo to 78 yr) were studied by immunohistochemistry with monoclonal antibodies HNK-1 and VC1.1 to the HNK-1 epitope. Antibodies to cytoskeletal elements were used to characterize the cells in this region. RESULTS: The HNK-1-immunopositive cells appeared underneath the pigment epithelium of the pars plicata by the 20th gestational week, spread into the pars plana after the 28th week, and reached the ora serrata during the first year of life. The immunoreaction was constantly present in all adult eyes examined; they were sharply demarcated from the iris, ciliary muscle, and choroid. The HNK-1-positive subepithelial layer was not labeled with monoclonal antibodies V9 or Vim 3B4 to vimentin, monoclonal antibodies CAM 5.2 and CY-90 to cytokeratin 8 and 18, or monoclonal antibodies DE-U-10 and D33 to desmin in adult eyes, but was uniformly positive for vimentin in fetal eyes. The HNK-1 epitope was distributed along cell membranes or adjacent extracellular matrix of stromal cells. CONCLUSION: The HNK-1-positive stromal region is a constant and conspicuous element of the human eye that may have a role in structurally stabilizing the ciliary body, perhaps in relation to accommodation or aqueous secretion.

Adolescent↗

Na+ and Cl- fluxes, and effects of pharmacological agents on the short-circuit current of the isolated rabbit iris-ciliary body.

Unidirectional Na+ and Cl- fluxes were measured across the isolated rabbit iris-ciliary body under short-circuited conditions. Na+ fluxes were in the range of 9-13 mueq/hr X cm2, and Cl- fluxes varied between 7-12 mueq/hr X cm2. A statistically significant net Na+ or Cl- flux could not be found. Ouabain, 5X10(-5) M, did not change the Na+ or Cl- flux in either direction despite a marked effect on the short-circuit current (SCC). There was a disagreement between the electrical conductance calculated from unidirectional fluxes and electrical measurements, suggesting the presence of electrically silent exchange mechanisms. Theophylline and isobutyl methylxanthine stimulated the SCC, whereas epinephrine, trifluormethazolamide and diisothiocyanostilbene disulfonic acid inhibited the SCC. Furosemide had a minor inhibitory effect, and the Ca2+ ionophore A23187 was without effect. Amphotericin B produced a substantial stimulation of the SCC from the aqueous side but an inhibition of the SCC from the blood side. This dual effect is consistent with the presence of Na+ -K+ pumps in the basolateral membranes of both the pigmented and non-pigmented cell layers of the ciliary body.

1-Methyl-3-isobutylxanthine↗

Ultrabiomicroscopic study of the effects of brimonidine, apraclonidine, latanoprost and ibopamine on the chamber angle and ciliary body.

The authors used the Zeiss Humphrey (Mod840, 50 MHz) ultrabiomicroscope to evaluate the changes that Brimonidine, Apraclonidine, Latanoprost and Ibopamine cause in the anterior chamber and on the ciliary body of healthy subjects. The eyes of 60 volunteers, separated into 4 groups according to drug instilled, were studied and the parameters analyzed were: anterior chamber depth (ACD), pupillary diameter (PD), angle opening at 500 microns from the scleral spur (AOD500), trabecular iris angle (TIA) and iris thicknesses (ID1 and ID3). The study showed the miotic effect of Brimonidine (S) that was accompanied by an angle opening (S). Apraclonidine and Latanoprost caused no statistically significant changes in the angle or in the ciliary body. Ibopamine caused mydriasis (S). The UBM, therefore, showed itself to be useful also in the study of the mechanism of action of drugs on the angular structures and on the ciliary body.

Adrenergic alpha-Agonists↗

Comparison of interactions of R-(+)- and S-(-)-isomers of beta-adrenergic partial agonists, befunolol and carteolol, with high affinity site of beta-adrenoceptors in isolated rabbit ciliary body and guinea-pig taenia caeci.

The stereoselectivities of beta-adrenergic partial agonists for the high affinity binding site of beta-adrenoceptors in the rabbit ciliary body and the guinea-pig taenia caeci were studied. The pA2 values of the S-(-)-isomers of befunolol and carteolol against S-(-)-isoprenaline, which were calculated from the shift of each concentration - response curve in increasing cyclic AMP levels, were significantly larger than those of the R-(+)-isomers in the guinea-pig taenia caeci, while the pA2 values of the S-(-)-isomers were not significantly larger than those of the R-(+)-isomers in the rabbit ciliary body. The pK1 values determined from the binding experiments were in good agreement with the pA2 values from the increases in cyclic AMP levels. These results suggest that the high affinity binding site of beta-adrenoceptors in the guinea-pig taenia caeci may be able to discriminate stereoselectively between the R-(+)- and S-(-)-isomers, while in the rabbit ciliary body there is no stereo-selectivity between the two enantiomers.

Adrenergic beta-Antagonists↗

Plateau iris syndrome associated with multiple ciliary body cysts. Report of three cases.

OBJECTIVE: To describe plateau iris syndrome associated with multiple neuroepithelial cysts of the pars plicata. METHODS: Case reports of 3 patients with plateau iris syndrome who were found to have multiple bilateral ciliary body cysts on ultrasound biomicroscopic examination. RESULTS: Ultrasound biomicroscopy revealed classic features of plateau iris syndrome in each patient but also showed multiple neuroepithelial cysts of the ciliary body in each eye. CONCLUSION: Plateau iris syndrome may be associated with multiple ciliary body cysts.

Aged↗

Autoradiographic localization of carbonic anhydrase in the rabbit ciliary body.

The high binding affinity of acetazolamide for carbonic anhydrase (K1 congruent to 10(-8) M) was employed to demonstrate the distribution of the enzyme in the rabbit ciliary body by incubating the tissue with 3H-acetazolamide (1.5 Ci/mmol). Specificity of binding was ascertained by displacing 3H-acetazolamide with a high concentration of unlabeled ethoxzolamide (K1 congruent to 10(-9) M). Wedges of the globe anterior to the ora serrata were incubated in bicarbonate buffered physiological saline, 95% O2/5% CO2, at 0 degrees C for 2 hr with either 3H-acetazolamide (0.2 microM), 3H-acetazolamide and unlabeled ethoxzolamide (100 microM), or physiological saline alone. They were then washed for 2 hr in fresh physiological saline and processed for autoradiography. The autoradiographs showed the label localized in both pigmented and nonpigmented layers of ciliary body epithelium in the pars plicata and in the iridial processes. The epithelia of both crests and troughs showed localization of label. In contrast, no concentration of label was found in the stroma of the ciliary body, including vascular endothelium, and in the epithelia of the pars plana. In sections that were incubated with 3H-acetazolamide in the presence of an excess of unlabeled ethoxzolamide, no localization of label occurred. These findings suggest that the epithelia of the pars plicata, but not those of the pars plana, contain carbonic anhydrase. This is consistent with hypotheses restricting aqueous humor formation to the pars plicata.

Acetazolamide↗

Drug responses of adenylate cyclase in iris-ciliary body determined by adenine labelling.

The intracellular adenine nucleotide pool of rabbit iris-ciliary body was labelled by uptake of 3H-adenine in vitro. A variety of agents were tested for their ability to stimulate or inhibit the incorporation of radioactivity into cyclic AMP formed from ATP labelled with 3H-adenine. Isoproterenol, vasoactive intestinal peptide, forskolin, and prostaglandin E2 stimulated incorporation of label 3-10-fold in 15-20 min compared with paired tissues not treated with hormone, whereas histamine, serotonin, substance P, and bradykinin were inactive. Clonidine, alpha-methylnorepinephrine, and dopamine decreased the rate of incorporation of label into the cyclic-AMP pool in tissues that showed high spontaneous basal rates. In low-basal tissues these drugs were inactive by themselves but clonidine and alpha-methylnorepinephrine blocked the stimulation effected by isoproterenol. The findings indicate that several receptor-coupled adenylate cyclase systems are present in ICB and that dual adrenergic control of adenylate cyclase through positive and negative coupling of adrenergic receptors probably occurs. The negatively coupled adrenergic receptors appear to be similar to the alpha 2-subclass of adrenergic receptor described in other tissues. These observations suggest a role for the large number of alpha 2-adrenergic-binding sites found in albino rabbit iris-ciliary body by ligand binding assays.

Adenine Nucleotides↗

Prejunctional alpha 2-adrenoceptors and adenylyl cyclase regulation in the rabbit iris-ciliary body.

Agents that elevate intracellular cyclic AMP (cAMP) have been found to enhance the synaptic discharge of norepinephrine (NE) from sympathetic nerve terminals in the rabbit iris-ciliary body and other peripheral tissues. We explored the hypothesis that prejunctional alpha 2-adrenergic receptors that mediate feedback inhibition of NE release may be coupled to adenylyl cyclase inhibition. To indirectly monitor cAMP changes in sympathetic axon terminals, we analyzed the cAMP-mediated activation of tyrosine hydroxylase, a sympathetic marker protein that undergoes acute phosphorylation and activation by cAMP-dependent protein kinase A. Tyrosine hydroxylase activity was assayed in situ by incubation of rabbit iris-ciliary body tissue segments in buffered Krebs-Ringer solution containing the substrate tyrosine (100 microM) and the DOPA decarboxylase inhibitor brocresine (30 microM). Intraneuronal DOPA accumulation was quantified by HPLC with electrochemical detection. Tyrosine hydroxylase activity was increased approximately 2 fold by incubation with forskolin (10 microM) plus IBMX (0.5 mM) or with 8-Bromo-cAMP (3 mM). Simultaneous addition of the alpha 2-adrenergic agonist clonidine (1 microM) attenuated the response to forskolin/IBMX, but had no effect on the response to 8-Br-cAMP. Clonidine-mediated inhibition of the forskolin/IBMX response was abolished by treatment of tissues with N-ethylmaleimide (NEM), an alkylating agent that inactivates pertussis toxin-sensitive G proteins (Gi) that couple receptors to adenylyl cyclase inhibition. These findings suggest that prejunctional alpha 2-adrenoceptors in the rabbit iris-ciliary body are negatively coupled to adenylyl cyclase. This mechanism may contribute to autofeedback regulation of NE biosynthesis and release.

Adenylyl Cyclases↗

Adenocarcinoma of the ciliary body pigment epithelium in a child.

Tumors arising from the pigment epithelium of the ciliary body are rare and most commonly occur in white adults. We describe a tumor that was clinically indistinguishable from a melanoma or medulloepithelioma occurring in a 7-year-old Japanese boy. Three months after initial evaluation, the eye was enucleated because of intractable intraocular pressure elevation and pain. On pathologic examination the tumor was found to fill approximately one third of the anterior chamber. Light microscopic and ultrastructural findings were consistent with an adenocarcinoma of the ciliary body pigment epithelium. To our knowledge, this is the youngest patient described with this tumor.

Adenocarcinoma↗

Reattachment of the detached ciliary body with suturing for treatment of contusional ocular hypotension.

Ocular hypotension resulting from a detached ciliary body after ocular contusion usually is best treated surgically. We report a simple, safe, and relatively effective procedure used to suture the ciliary body onto the inner surface of the sclera in eight such cases. Intraocular pressure (IOP) rose from unmeasurable levels to 10 to 12 mm Hg in five of these cases. Although in the three other cases IOP did not increase immediately after the operation, in one case it rose to 10 to 12 mm Hg 1 month postoperatively, and in the other two cases it rose to a normal level after additional laser therapy.

Adolescent↗

[Ciliary body changes associate with aldose reductase in galactosemic rats (2)].

Morphological changes of the ciliary body epithelium in rats fed a 50% galactose diet for 18 months with or without aldose reductase inhibitor (ARI: FR74366, Fujisawa) were investigated on coronal sections by both the light microscopic observation and the measurement of the area of non-pigmented epithelium (non-PE) as the ratio of non-PE/PE. The measurement was performed with color image analyzer. Structural change (hypertrophy) was observed only in non-pigmented epithelial cells of galactosemic rats. In contrast, no hypertrophic changes were seen in galactosemic rats treated with FR74366. These results strongly suggest that aldose reductase is associated with hypertrophic change of non-pigmented epithelial cells of the ciliary body as one of the diabetic complications.

Aldehyde Reductase↗