Aqueous Humor Formation Research Articles

Aqueous humor dynamics

Intraocular pressure is mainly controlled by the rate of aqueous humor production and the resistance to its outflow. Aqueous humor formation is the result of fast unidirectional secretion and slow contradirectional reabsorption. The most striking evidence of endogenous regulation of the ciliary epithelial secretion is provided by the observation of the circadian rhythm. Aqueous humor formation is mainly controlled by the interaction of inhibiting alpha(2)-adrenoreceptors and stimulating beta-adrenoceptors. The role of the central nervous system in the regulation of this process remains unclear and the precise mechanism of outflow regulation is not fully understood. The aqueous humor passes into the anterior chamber and leaves the eye by two routes: the direct outflow pathway through the trabecular meshwork or the indirect outflow pathway through the ciliary muscle. Further biochemical, biological and pharmacological investigations are necessary to determine the fundamental process of aqueous humor formation, outflow and regulation.

An evaluation of the fixed-combination of latanoprost and timolol for use in open-angle glaucoma and ocular hypertension

Glaucoma is one of the leading causes of irreversible blindness worldwide. Although there is no cure for this chronic disease, medical treatment is aimed at reducing levels of intraocular pressure (IOP) using ocular hypotensive agents. Very often, patients require more than one IOP-reducing drug, resulting in complex medication regimens that may be difficult to maintain and that can lead to non-compliance. A fixed-combination (FC) ophthalmic solution consisting of the prostaglandin, latanoprost (0.005%), and the β-blocker, timolol (0.5%), is now available. The primary mechanism of action of latanoprost is to increase uveoscleral outflow whereas timolol lowers IOP levels by decreasing the formation of aqueous humor in the ciliary epithelium. Due to the unique mechanism of action of latanoprost, once-daily dosing of one drop of FC latanoprost/timolol results in additional IOP reduction compared with either drug administered separately. FC latanoprost/timolol is well-tolerated and has a safety profile similar to that of its individual components. This combination drug provides a safe, effective and convenient alternative for the treatment of patients with elevated IOP levels uncontrolled with monotherapy.

Aqueous humor dynamics in monkeys after topical 8-iso PGE(2).

To determine in normotensive cynomolgus monkeys, the effects of topical 8-iso prostaglandin (PG)E(2) on intraocular pressure (IOP), aqueous humor formation (AHF), uveoscleral outflow (Fu), and total and trabecular outflow facility. IOP was measured by Goldmann applanation tonometry under ketamine anesthesia after single or twice-daily topical treatments with 8-iso PGE(2). With animals under pentobarbital anesthesia, AHF and flow to blood (equated to trabecular outflow) were determined by anterior chamber perfusion with radioactively labeled albumin solution. Fu and trabecular outflow facility were calculated from these measurements. Total outflow facility was measured by two-level, constant-pressure perfusion. IOP was not significantly changed after single or multiple 10- micro g doses of 8-iso PGE(2). The 25- micro g dose significantly decreased IOP by 2 to 3 mm Hg compared to the contralateral vehicle-treated control 4 to 6 hours after a single dose and by 3 to 5 mm Hg within 1.5 hours after twice-daily treatments for 4 to 5 days. Total outflow facility corrected for control eye washout was increased by an apparent 37% (P < 0.02, n = 7) from 2 to 3.5 hours after the ninth dose, largely due to outlier values obtained in one monkey. Isotope studies performed after twice-daily treatments totaling 9 to 29 doses showed no change in AHF, trabecular outflow facility, or total outflow facility. Relative to AHF, trabecular outflow was significantly decreased, and the calculated Fu was significantly increased when all data were analyzed. The present findings are consistent with lowering of IOP by 8-iso PGE(2), primarily by increasing Fu. A direct effect on the trabecular meshwork was not indicated by these in vivo studies.

Electron microprobe analysis of ouabain-exposed ciliary epithelium: PE-NPE cell couplets form the functional units

Aqueous humor is secreted by the bilayered ciliary epithelium. Solutes and water enter the pigmented ciliary epithelial (PE) cell layer, cross gap junctions into the nonpigmented ciliary epithelial (NPE) cell layer, and are released into the aqueous humor. Electrical measurements suggest that heptanol reduces transepithelial ion movement by interrupting PE-NPE communication and that gap junctions may be a regulatory site of aqueous humor formation. Several lines of evidence also suggest that net ciliary epithelial transport is strongly region dependent. Divided rabbit iris-ciliary bodies were incubated in chambers under control and experimental conditions, quick-frozen, cryosectioned, and freeze-dried. Elemental intracellular contents of NPE and PE cells were determined by electron probe X-ray microanalysis. With or without heptanol, ouabain produced concentration- and time-dependent changes more markedly in anterior than in posterior epithelium. Without heptanol, there were considerable cell-to-cell variations in Na gain and K loss. However, contiguous NPE and PE cells displayed similar changes, even when nearby cell pairs were little changed by ouabain in aqueous, stromal, or both reservoirs. In contrast, with heptanol present, ouabain added to aqueous or both reservoirs produced much larger changes in NPE than in PE cells. The results indicate that 1) heptanol indeed interrupts PE-NPE junctions, providing an opportunity for electron microprobe analysis of the sidedness of modification of ciliary epithelial secretion; 2) Na and K undergo faster turnover in anterior than in posterior epithelium; and 3) PE-NPE gap junctions differ from PE-PE and NPE-NPE junctions in permitting ionic equilibration between adjoining ouabain-stressed cells.

Chirurgie du glaucome

Surgical treatment is mainly used for primary glaucomas, but some surgical procedures can also be applied to secondary glaucomas. Many surgical and physical techniques are aimed at reducing the intraocular pressure. These techniques can be divided into those that improve aqueous outflow by shunting aqueous humor out of the eye into the subconjonctival space (filtering procedures), and those that decrease the rate of aqueous humor formation by partial destruction of the ciliary epithelium (cyclodestructive procedures).

An evaluation of the fixed-combination of latanoprost and timolol for use in open-angle glaucoma and ocular hypertension

Glaucoma is one of the leading causes of irreversible blindness worldwide. Although there is no cure for this chronic disease, medical treatment is aimed at reducing levels of intraocular pressure (IOP) using ocular hypotensive agents. Very often, patients require more than one IOP-reducing drug, resulting in complex medication regimens that may be difficult to maintain and that can lead to non-compliance. A fixed-combination (FC) ophthalmic solution consisting of the prostaglandin, latanoprost (0.005%), and the β-blocker, timolol (0.5%), is now available. The primary mechanism of action of latanoprost is to increase uveoscleral outflow whereas timolol lowers IOP levels by decreasing the formation of aqueous humor in the ciliary epithelium. Due to the unique mechanism of action of latanoprost, once-daily dosing of one drop of FC latanoprost/timolol results in additional IOP reduction compared with either drug administered separately. FC latanoprost/timolol is well-tolerated and has a safety profile similar to that of its individual components. This combination drug provides a safe, effective and convenient alternative for the treatment of patients with elevated IOP levels uncontrolled with monotherapy.

Studies on the mechanism of action of timolol and on the effects of suppression and redirection of aqueous flow on outflow facility

Long-term use of drugs that suppress aqueous humor formation, such as timolol and dorzolamide, or that redirect aqueous humor outflow from the trabecular meshwork, such as prostaglandin F 2α analogues, could cause underperfusion of the trabecular meshwork and a secondary decrease in outflow facility. We investigated the mechanism of suppression of aqueous humor formation by timolol in monkey eyes by measuring aqueous humor ascorbate levels. We also determined whether suppression of aqueous humor formation with and without redirection of aqueous humor away from the trabecular meshwork could lead to a subsequent reduction in outflow facility, and whether this reduction was correlated with increased fibronectin levels in anterior chamber aqueous humor. In cynomolgus monkeys, unilateral dose/aqueous humor formation response curves were generated for timolol, dorzolamide, and a combination of timolol+dorzolamide. Aqueous humor formation and/or outflow facility were measured in both eyes after approximately four days, four weeks and seven weeks of twice daily treatment with 3·5 μg timolol+1·0 mg dorzolamide to one eye and 30% DMSO to the other. In some monkeys, 5 μg prostaglandin F 2α-isopropyl ester (PG) was added to timolol+dorzolamide for 4-week treatments. Intraocular pressure and corneal endothelial transfer coefficients ( k a) were also measured at four weeks. Aqueous humor fibronectin levels were determined in four monkeys after approximately 9·5 weeks of timolol+dorzolamide treatment. Aqueous humor formation, intraocular pressure, and aqueous humor ascorbate levels were also determined in rhesus monkeys at baseline and after a single unilateral topical administration of 25 μg timolol. Compared to baseline for the same eye, aqueous humor formation was significantly decreased in treated eyes at all doses of timolol and at 1·8 and 4 mg dorzolamide. Compared to the opposite control eye, aqueous humor formation was lower in treated eyes after 3·5 and 5 μg timolol and after all doses of dorzolamide. Aqueous humor formation after treatment with 3·5 μg timolol+1·0 mg dorzolamide was decreased in treated vs. control eyes, after four days and was suppressed in both treated and control eyes after four weeks of treatment, but not when PG was added. There was no difference in k a values with or without the addition of PG. Intraocular pressure was significantly lower in both treated and control eyes vs. baseline after ∼6·5 weeks treatment with timolol+dorzolamide when taken 2 hr after the last dose and after ∼3·5 weeks treatment with timolol+dorzolamide+PG when measured 6 hr after the last dose. Outflow facility after treatment with timolol+dorzolamide was unchanged after four days, tended to be lower in the treated vs. control eyes after four and seven weeks, and was significantly lower in treated vs. control eyes after four weeks treatment with timolol+dorzolamide+PG (0·352±0·052 vs. 0·515±0·096 μl min −1 mmHg −1, p≤0·02). Both treated vs. control eye aqueous humor fibronectin levels were below the level of detection for our assay (0·01 μg ml −1). The 25 μg timolol dose decreased ipsilateral, but not contralateral intraocular pressure (12·6±1·7 vs. 15·2±0·9; p<0·05) and aqueous humor formation (1·40±0·08 vs. 2·03±0·09 μg ml −1, p≤0·01). There was no difference in anterior chamber ascorbate levels in treated vs. control eyes or compared to their respective baselines. Our findings indicate that timolol affects neither ciliary epithelial transport of ascorbate nor aqueous fibronectin levels. Our data also indicate that decreasing aqueous humor formation over a period of time can lead to reduction in outflow facility, particularly when combined with therapy that redirects aqueous from the trabecular meshwork. Future intraocular pressure-lowering therapies for glaucoma may better be directed at enhancing flow through the trabecular pathway as opposed to decreasing aqueous humor formation or rerouting aqueous humor away from the trabecular meshwork.

The fall and rise of active chloride transport: implications for regulation of intraocular pressure.

Early study of transepithelial salt transfer focused on Cl(-) and not Na(+), partly because Cl(-) was readily measureable. The advent of flame photometry and tracer techniques brought Na(+) to the fore, especially since short-circuited frog skin (Rana temporaria) produces baseline net movement of Na(+) and not of Cl(-). Zadunaisky was among the first to describe what is currently termed secondary active Cl(-) transport, helping stimulate interest in Cl(-) handling by other tissues, notably the thick ascending limb of the loop of Henle important in renal counter-current multiplication. More recently, molecules responsible for electroneutral and electrogenic Cl(-) transfer have been cloned, and specific diseases resulting from their faulty expression have been identified. The clinical importance of transepithelial Cl(-) transfer is illustrated by studies of aqueous humor formation by the eye's bilayered ciliary epithelium. NaCl is taken up from the stroma by the pigmented ciliary epithelial (PE) layer, diffuses through gap junctions into the nonpigmented ciliary epithelial (NPE) layer, and is released into the aqueous humor largely through Na(+) pumps and Cl(-) channels. ATP released by NPE cells can be ecto-enzymatically metabolized to adenosine. Adenosine can mediate paracrine/autocrine stimulation of Cl(-) channels and aqueous humor secretion by occupying A(3) adenosine receptors (ARs). A(3)AR agonists indeed elevate, and A(3)AR antagonists lower, intraocular pressure (IOP) in wild-type mice. A(3)AR knockout mice have low IOP and their responses to A(3)AR agonists and antagonists are blunted; this suggests that reducing Cl(-)-channel activity with A(3)AR antagonists may provide a novel approach for treating glaucoma.

Purinergic signaling in the rabbit ciliary body epithelium

In the past forty years, a wealth of information has accumulated that points to the presence of adenosine and adenine nucleotides in the anterior segment of the eye and a number of hypotheses have been introduced to describe the possible role of these agents in the regulation of aqueous humor flow. However, in the absence of a generally accepted model for the cellular and molecular mechanisms of aqueous humor formation by the ciliary body epithelium, efforts to identify the signal transduction pathway(s) responsible for regulation of the ion and water transport have not been successful. This article briefly reviews the evidence for (i). the presence in aqueous humor of adenine nucleotides, cyclic adenosine monophosphate and adenosine, their metabolic product, (ii). the possible role of these agents in the regulation of aqueous humor dynamics, and (iii). the expression of ecto-nucleotidases, receptors, and second messengers that may mediate such regulation. Finally, a model for the regulation of aqueous humor formation by adenosine and ATP is proposed.

Regulation of Na,K-ATPase expression by endothelin-1 in transformed human ciliary non-pigmented epithelial (HNPE) cells.

Endothelin-1 (ET-1) (1-100 nM) decreases the activity of Na,K-ATPase, a key enzyme responsible for aqueous humor formation, in transformed human non-pigmented ciliary epithelial (HNPE) cells. The present study sought to determine if ET-1 alters the expression of the catalytically active alpha subunit of Na,K-ATPase in HNPE cells and identify mechanisms underlying these effects. We report that acute (15 and 30 min) treatment with ET-1 results in an increase in mRNA expression of the alpha 1 subunit of Na,K-ATPase. Similar to ET-1's effects, ouabain (100 microM), a selective inhibitor of Na,K-ATPase, and monensin (10 microM), a sodium ionophore, also increased Na,K-ATPase expression in HNPE cells. The increase in Na,K-ATPase expression by short-term treatment with ouabain and monensin was dependent on their ability to elevate intracellular sodium concentrations. However, acute ET-1 treatment mediated increase in Na,K-ATPase expression was independent of changes in intracellular sodium. A prolonged (24 hr) ET-1 treatment results in an increase in both mRNA and protein levels of the alpha 1 subunit of Na,K-ATPase. These observations suggest that ET-1 could play a homeostatic role in modulating aqueous humor formation by having differential effects on the activity and expression of Na,K-ATPase by the ciliary epithelium in the eye.

Real-time direct measurement of nitric oxide in bovine perfused eye trabecular meshwork using a clark-type electrode.

NO was detected in bovine trabecular meshwork (TM). Bovine eyes were perfused (posterior ciliary artery). In some eyes (operated eyes) a NO electrode was inserted adjacent to the TM (scleral flap). Vascular perfusion/intraocular pressures (VPP/IOP) were recorded. In operated eyes, epinephrine (1 nM-100 microM) increased NO (maximally 979.9 +/- 117.6 nM, mean +/- SDM). Timolol (1 mM) shifted the epinephrine-NO concentration-response curve rightward (2.94 log units) without significantly changing the maximal response (903.0 +/- 67.7 nM, mean +/- SDM). The non-selective NO synthase (NOS) inhibitor L-NMMA (100 microM) virtually abolished the NO response to epinephrine. L-NMMA alone (1 microM-100 microM) significantly reduced tonic NO generation (maximally 109.5 +/- 24.9 nM, mean +/- SDM), whereas timolol alone (1 microM-1 mM) had no effect. In unoperated eyes, epinephrine (1 nM-100 microM) reduced IOP (maximally 2.56 +/- 0.64 mmHg, mean +/- SDM). Epinephrine (100 microM) mildly increased VPP (4.6 +/- 1.3 mmHg, mean +/- SDM). Baseline aqueous humor formation rate (11.5 +/- 3.2 microl/min, mean +/- SDM) was unaffected. Effluent perfusate (effusate) total NO(2)(-) was determined by enzymatically reducing all NO(3)(-) to NO(2)(-), then assessing resultant NO(2)(-) (Griess assay). Epinephrine (1 nM-1 microM) increased effusate NO(2)(-) (maximally 15.8 +/- 4.9 microM, mean +/- SDM). Timolol (1 mM) reduced, and L-NMMA (100 microM) virtually abolished effusate NO(2)(-) response to epinephrine. L-NMMA alone (1 microM-100 microM) reduced tonic effusate NO(2)(-) (maximally from 5.8 +/- 1.6 microM to 1.1 +/- 0.9 microM, mean +/- SDM), whereas timolol alone (1 microM-1 mM) had no effect. NO is generated tonically in bovine TM and increases in response to epinephrine.

Aqueous humor dynamics and trabecular meshwork and anterior ciliary muscle morphologic changes with age in rhesus monkeys.

To determine in rhesus monkeys the age-dependence of uveoscleral outflow (Fu) and morphology of the trabecular meshwork (TM) and anterior ciliary muscle (CM). Intraocular pressure (IOP) was measured by Goldmann applanation tonometry in monkeys under ketamine anesthesia. After anterior chamber cannulation under pentobarbital anesthesia, aqueous humor formation (AHF), anterior chamber volume, trabecular outflow, and Fu were determined isotopically. The CM and TM were examined by light and electron microscopy. IOP increased significantly with age in monkeys aged 3 to 29 years. AHF and anterior chamber volume were unchanged. Fu was decreased, and trabecular outflow increased in monkeys aged 25 to 29 years compared with the remaining monkeys. Morphologically, there was a significant increase in the thickness of the elastic fibers of the trabeculum ciliare covering the anterior tips of the CM, and an increase in extracellular material between the muscle tips. The number of TM cells decreased with age, whereas the amount of fibrillar material and sheath-derived plaques increased. This increase was less pronounced in the middle filtering portion of the cribriform region than in the anterior and posterior portions. The decline in Fu in very old rhesus monkeys with normal IOP parallels that seen in normotensive aging humans. This may be correlated with thickening of the elastic fiber sheath in the CM tips in addition to other morphologic changes. The TM findings are analogous to those in the aging human eye and are consistent with the age-related decrease in outflow facility reported in both humans and monkeys.

Effects of ion transport and channel-blocking drugs on aqueous humor formation in isolated bovine eye.

To investigate the role of active chloride secretion by the ciliary epithelium in the formation of aqueous humor (AH), by using the in vitro perfused eye. Bovine eyes collected from an abattoir were cannulated through the ophthalmic artery and perfused with oxygenated Krebs' solution at 37 degrees C. Aqueous humor formation (AHF) was measured by the fluorescein-dilution technique. Drugs were added to the perfusate and/or to the anterior chamber. NaK-adenosine triphosphatase (ATPase) inhibitor, ouabain (1.0 mM), produced a significant reduction in AHF by 46% and 42% when added to the stromal or aqueous side, respectively. When added to both sides (1.0 mM), it produced a reduction of 61%. Bumetanide (0.1 mM), a specific inhibitor of Na-K-2Cl cotransport, and furosemide (0.1 mM), a nonspecific anion transport inhibitor, produced 35% and 45% reductions when applied to the stromal side. DIDS (0.001-0.1 mM), which is believed to inhibit the Cl-HCO(3) exchanger, Na-HCO(3) cotransporter, and chloride channel, produced a dose-dependent reduction when added to the stromal side. The inhibition was 55% by the highest concentration used. 5-Nitro-2-(3-phenylpropylamino)-benzoic acid (NPPB; 0.1 mM), a chloride channel blocker in the nonpigmented cells, produced a 25% reduction when applied to the aqueous side. Acetazolamide (0.1 mM), a carbonic anhydrase inhibitor, applied to the stromal side, produced 31% reduction. At least 60% of the AH is formed by active secretion in bovine eyes. Transport of anions through the ciliary epithelium (CE), particularly the chloride ion, plays a crucial role in AHF.

Effects of multiple dosing of epinephrine on aqueous humor dynamics in human eyes.

Numerous studies have provided conflicting evidence to explain the ocular hypotensive mechanism of action of epinephrine. Although epinephrine has been shown consistently to increase outflow facility, its effects on aqueous flow and uveoscleral outflow are not as clear. The purpose of this study was to clarify the effects of multiple doses of topical epinephrine on aqueous humor dynamics in human eyes. This was done by evaluating the four main parameters that determine steady state intraocular pressure. These parameters were assessed at baseline and after a week of twice-daily treatment of epinephrine hydrochloride 2% to one eye. Twenty-six human volunteers were enrolled in the study. Intraocular pressure was measured by pneumatonometry, aqueous flow and trabecular outflow facility by fluorophotometry, episcleral venous pressure by venomanometry and uveoscleral outflow by mathematical calculation. In epinephrine-treated eyes compared to baseline, intraocular pressure and aqueous flow were reduced from 21.2 +/- 0.3 to 17.1 +/- 0.2 mmHg (19%, p = .01) and 3.3 +/- 0.2 to 2.9 +/- 0.2 microl/min (12%, p = .03), respectively. Trabecular outflow facility obtained by fluorophotometry was increased from 0.18 +/- 0.02 to 0.26 +/- 0.03 microl/min/mmHg (44%, p = .02). Topical epinephrine did not significantly affect uveoscleral outflow or episcleral venous pressure. In conclusion, multiple topical doses of epinephrine lowered intraocular pressure in human volunteers by reducing aqueous humor formation and increasing trabecular outflow facility. The increase in uveoscleral outflow suggested by other studies was not observed.

PGE(2), Ca(2+), and cAMP mediate ATP activation of Cl(-) channels in pigmented ciliary epithelial cells.

Purines regulate intraocular pressure. Adenosine activates Cl(-) channels of nonpigmented ciliary epithelial cells facing the aqueous humor, enhancing secretion. Tamoxifen and ATP synergistically activate Cl(-) channels of pigmented ciliary epithelial (PE) cells facing the stroma, potentially reducing net secretion. The actions of nucleotides alone on Cl(-) channel activity of bovine PE cells were studied by electronic cell sorting, patch clamping, and luciferin/luciferase ATP assay. Cl(-) channels were activated by ATP > UTP, ADP, and UDP, but not by 2-methylthio-ATP, all at 100 microM. UTP triggered ATP release. The second messengers Ca(2+), prostaglandin (PG)E(2), and cAMP activated Cl(-) channels without enhancing effects of 100 microM ATP. Buffering intracellular Ca(2+) activity with 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'- tetraacetic acid or blocking PGE(2) formation with indomethacin inhibited ATP-triggered channel activation. The Rp stereoisomer of 8-bromoadenosine 3',5'-cyclic monophosphothioate inhibited protein kinase A activity but mimicked 8-bromoadenosine 3',5'-cyclic monophosphate. We conclude that nucleotides can act at >1 P2Y receptor to trigger a sequential cascade involving Ca(2+), PGE(2), and cAMP. cAMP acts directly on Cl(-) channels of PE cells, increasing stromal release and potentially reducing net aqueous humor formation and intraocular pressure.

Timolol may inhibit aqueous humor secretion by cAMP-independent action on ciliary epithelial cells.

The beta-adrenergic antagonist timolol reduces ciliary epithelial secretion in glaucomatous patients. Whether inhibition is mediated by reducing cAMP is unknown. Elemental composition of rabbit ciliary epithelium was studied by electron probe X-ray microanalysis. Volume of cultured bovine pigmented ciliary epithelial (PE) cells was measured by electronic cell sizing; Ca(2+) activity and pH were monitored with fura 2 and 2',7'-bis(2-carboxyethyl)-5(6)-carboxyfluorescein, respectively. Timolol (10 microM) produced similar K and Cl losses from ciliary epithelia in HCO/CO(2) solution but had no effect in HCO/CO(2)-free solution or in HCO/CO(2) solution containing the carbonic anhydrase inhibitor acetazolamide. Inhibition of Na(+)/H(+) exchange by dimethylamiloride in HCO/CO(2) solution reduced Cl and K comparably to timolol. cAMP did not reverse timolol's effects. Timolol (100 nM, 10 microM) and levobunolol (10 microM) produced cAMP-independent inhibition of the regulatory volume increase (RVI) in PE cells and increased intracellular Ca(2+) and pH. Increasing Ca(2+) with ionomycin also blocked the RVI. The results document a previously unrecognized cAMP-independent transport effect of timolol. Inhibition of Cl(-)/HCO exchange may mediate timolol's inhibition of aqueous humor formation.

Intraocular pressure and vascular effects of sodium azide in bovine perfused eye.

The effects of the nitrovasodilator, sodium azide, on intraocular pressure (IOP) and ciliary vascular tone were compared. IOP was measured in the bovine isolated eye that was perfused via the ciliary artery. Separately, vasodilator effects were assessed after raising the vascular tone using noradrenaline (10 microM). Aqueous humor formation (AHF) rate was estimated by a fluorescein dilution method. Cyclic GMP in the ciliary processes was measured by radioimmunoassay. When compared with controls, sodium azide (10 nmole bolus dose) was found to lower IOP (2.2 +/- 0.3 mm Hg; P < 0.01) via a reduction in AHF (12.19 +/- 0.26 microl/min to 6.36 +/- 0.53 microl/min; P < 0.001). Azide (1 micromole) also reduced ciliary vascular resistance (81.0 +/- 5.5%; P < 0.01). However, the drug was 20x more potent as an ocular hypotensive than as a vasodilator (ED50 0.28 nmole on IOP, 5.55 nmole on vascular effect). Azide (10 nmole) also increased levels of ciliary cyclic GMP (127 +/- 17 fmol/mg protein to 233 +/- 27 fmol/mg protein; P < 0.01). The IOP-lowering effect of azide does not appear to depend on its ability to activate guanylyl cyclase (GC) in vascular smooth muscle, but rather is likely a consequence of direct activation of ciliary epithelial GC.

Update on topical carbonic anhydrase inhibitors.

Topical carbonic anhydrase inhibitors are a novel addition to the armamentarium of medical glaucoma treatment; dorzolamide has been available since 1995 and brinzolamide since 1998. They lower intraocular pressure by inhibiting carbonic anhydrase, a key enzyme for aqueous humor formation. Intraocular pressure-lowering activity of the substances appears to be the same and is similar to that of most other agents, but it does not reach the activity of the unselective beta-blocker timolol or the prostaglandin latanoprost. On concomitant treatment, additivity is reached with all other topical agents. A possible improvement of blood flow may offer an additional benefit, but its significance for the long-term outcome for human glaucoma remains to be shown. Side effects are mostly local. A more physiologic pH of brinzolamide appears to be advantageous.

Effects of serotonergic compounds on aqueous humor dynamics in monkeys

Purpose. The effects of several serotonergic agonists on aqueous humor formation (AHF), total outflow facility (OF) and intraocular pressure (IOP) were investigated in living cynomolgus monkeys. Methods. We determined the effect of a single topical unilateral 300 µg or 3 mg dose of the 5-HT agonists serotonin, 5-carboxamidotryptamine (5-CT), sumatripan, gepirone, and 8-hydroxy-2(di-n-propylaminotetralin) (8-OH-DPAT) and a 450 µg dose of flesinoxan on IOP (Goldmann applanation tonometry), AHF (scanning ocular fluorophotometry) and total OF (8-OH-DPAT only, topically and intracamerally). Results. Serotonin, 5-CT, sumatripan or gepirone had no significant effect on IOP or AHF. 8-OH-DPAT caused an AHF increase of ~70% over 6 hr in both ipsilateral drug- and contralateral vehicle-treated eyes, but no significant change in IOP compared with baseline measured on a separate occasion in the same animals. 8-OH-DPAT did not increase protein levels or rate of entry of systemically administered fluorescein in the anterior chamber aqueous humor compared to historic controls, and no difference was seen between ipsilateral and contralateral eyes. Flesinoxan had no effect on IOP and produced an insignificant 25% increase in flow in treated eyes compared to baseline. Conclusion. The results for 8-OH-DPAT and possibly flexinoxan indicate the presence of a secretion-stimulating 5-HT1A receptor in monkey ciliary epithelium that has little effect on IOP. OF was unchanged following 8-OH-DPAT administered topically or following intracameral exchange.

Effects of AGN 192024, a new ocular hypotensive agent, on aqueous dynamics

PURPOSE: To determine the mechanism of intraocular pressure lowering for the Ocular Hypotensive Lipid, AGN 192024 (Allergan, Inc, Irvine, California). METHODS: Twenty-five normal human volunteers between the ages of 21 and 48 took part in a randomized, double-masked, placebo-controlled, paired-comparison study in which intraocular pressure, aqueous humor flow, and tonographic resistance to outflow were studied. Measurements of aqueous flow were made during the day and at night while subjects slept. Intraocular pressure was measured with the Goldmann tonometer, and resistance to outflow was measured by electronic recording Schiötz tonography. RESULTS: Intraocular pressure was decreased by 20% on day 3 in AGN 192024-treated eyes in comparison with placebo-treated eyes in normal subjects ( P < .001). Aqueous humor flow was stimulated 13% during the day ( P = .007) and 14% at night ( P = .014) by the drug. Tonographic resistance to outflow was decreased 26% by AGN 192024 ( P < .001), and apparent resistance to outflow (the ratio of intraocular pressure to aqueous flow) was decreased 31% ( P < .001). Assuming that AGN 192024 does not cause prolonged lowering of episcleral venous pressure, the results show that pressure-insensitive outflow is enhanced by 50%, whereas tonographic facility of outflow (reciprocal of resistance) was enhanced 35%. CONCLUSIONS: AGN 192024 is an ocular hypotensive agent that works by enhancing both pressure-sensitive and pressure-insensitive aqueous humor outflow without diminishing aqueous humor formation.