Comparison of intraocular pressure lowering by topical and systemic carbonic anhydrase inhibitors in the rabbit.

We compared the effect on intraocular pressure (IOP) of maximal doses of a topical carbonic anhydrase inhibitor (CAI) at acidic and alkaline pH where it is maximally effective with full systemic CA inhibition in ocular normotensive New Zealand Albino rabbits. Tonometric IOP levels were measured hourly during 3 hour control period. Topical MK-417 (pKa 5.8, 8.3), a close congener of MK-507 (Dorzolamide) was given as a 1.4% solution at pH 4.5 (n=6) and pH 9.2 (n=6). MK-417 was instilled to the left eye with the right eye used as an untreated control. One hour later methazolamide was given intravenously at 10 mg/kg, a dose known to give full inhibition of the enzyme. Control IOP (mm Hg) was 19.12+/-0.50. One hour following MK-417, the left eye IOP was 13.40 +/-0.70 (pH 4.5) and 13.25+/-0.70 (pH 9.2). The right eye pressure was unchanged. Methazolamide injection at this time gave no further drop in the left eye IOP at either pH. IOP in the right eye fell to 14.00+/-0.70 so that 2 hours after methazolamide injection, the 2 eyes had the same pressure. In conclusion, topical CAI in sufficient dose and correct pH yields IOP lowering equivalent to a maximally effective dose of systemic CAI in rabbits.     

Postjunctional adrenergic receptors in the rabbit eye: effects on uveal flow and intraocular pressure in isolated arterially perfused eyes

In uveal vessels of isolated arterially perfused rabbit eyes there is direct evidence for the presence of post-junctional alpha 1-receptors and indirect evidence for alpha 2-receptors. Since vasoconstriction by epinephrine could not be blocked by an alpha 1-antagonist, only 30% by an alpha 2-antagonist and almost fully by the combination or by phentolamine, we suggest the presence of an intermediate alpha-type receptor which is neither alpha 1 nor alpha 2 but has characteristics of either. All alpha-types produce vasoconstriction and a fall in IOP. Of the beta-types only beta 1-receptors can be demonstrated. They mediate vasodilatation and a rise in IOP. Neither beta 2- nor non-selective beta-agonists or -antagonists (e.g. timolol) affect IOP or uveal flow.     

Phorbol ester: effect on intraocular pressure, adenylate cyclase, and protein kinase in the rabbit eye

Protein kinase C was identified as a major protein kinase enzyme activity in rabbit ciliary processes. Phorbol myristate acetate (4 beta-PMA) in the presence of Ca2+ activated protein kinase C but did not directly affect the cyclic AMP-dependent protein kinase enzyme isolated from ciliary processes. To elucidate possible roles of protein kinase C, PMA was injected intravitreally into rabbit eyes. Fifty pmoles of PMA produced approximately a 40% decrease of the intraocular pressure relative to the control eye lasting for more than 72 hr. A reduction of intraocular pressure was still elicited by this dose of PMA in animals pretreated with systemic indomethacin given to suppress a possible inflammatory response. The biologically inactive analogue, 4 alpha-phorbol didecanoate (100 pmoles/eye) had no significant effect on intraocular pressure. In vivo and in vitro treatment with PMA had no significant effect on adenylate cyclase in ciliary process membranes assayed in vitro. However, protein kinase C isolated from rat brain, when added together with cofactors to membranes in vitro, augmented adenylate cyclase activation by isoproterenol, vasoactive intestinal peptide and aluminum fluoride. A slight increase in the basal activity and in the forskolin response was not statistically significant. The effect of protein kinase C to increase responsiveness of ciliary process adenylate cyclase was totally dependent on the presence of Ca2+ and was augmented by addition of PMA. These findings indicate modulation of adenylate cyclase activity by protein kinase C acting at the level of the G-proteins and suggest a possible role for this enzyme in water and electrolyte transport in the ciliary processes.     

Comparison of effects of topical ibopamine and epinephrine on the circadian rhythm of intraocular pressure of the rabbit eye as measured by telemetry.

The purpose of this study was to measure the effect of topical ibopamine and epinephrine on intraocular pressure in undisturbed rabbits. Six pigmented rabbits were outfitted with a telemetric transducer system connected via catheter to the vitreous cavity of one eye for continuous monitoring of intraocular pressure. After entrainment to a 12 hour light, 12 hour dark cycle and after stabilization of the circadian rhythm of intraocular pressure, the animals were tested with 2% ibopamine and 2% epinephrine. Each drug was instilled during the light phase and during the dark phase of the circadian cycle. When administered during the light phase, both drugs caused a transient pressure rise followed by prolonged hypotension. When administered during the dark phase, neither drug caused a pressure rise but both drugs caused prolonged hypotension. It was concluded that ibopamine and epinephrine cause identical intraocular pressure changes in the normal rabbit eye. The effect was dependent on the timing of administration during the circadian cycle. Since ibopamine is a pro-drug of deoxyepinephrine (N-methyl dopamine, epinine), its effects are assumed to be due to this metabolite, a metabolite that is structurally similar to epinephrine.     

Effects of prostaglandin E2 on intraocular pressure, anterior chamber depth and blood flow volume of the iris and the ciliary body in rabbit eyes]

The effects of prostaglandin E2 (PGE2) on intraocular pressure (IOP), anterior chamber depth, and blood flow volume of the iris and the ciliary body of albino rabbits were investigated. Swelling and marked capillary dilatation of the ciliary body were observed after topical application of PGE2. IOP increased immediately by 200 micrograms, 400 micrograms, and 1,000 micrograms of PGE2, reached a peak within 30 minutes; then decreased gradually. The anterior chamber depth decreased only at 5 and 10 minutes after 1,000 micrograms of PGE2. The blood flow volume of the iris and the ciliary body immediately increased, and kept it up for 3 hours. Changes of IOP may be associated with an increase of blood flow attendant upon capillary dilatation of the ciliary body, and an increase of aqueous outflow due to breakdown of the blood-aqueous barrier. In addition to morphological and functional changes of the ciliary body, further factors which promote vitreous volume expansion may play an essential part in the development of malignant glaucoma.     

Continual monitoring of intraocular pressure: effect of central venous pressure, respiration, and eye movements on continual recordings of intraocular pressure in the rabbit, dog, and man.

A new method has been devised for continual monitoring of intraocular pressure by radiotelemetry. The use of this instrument for monitoring intraocular pressure by a variety of ophthalmic conditions is described.     

The ocular penetration of topical forskolin and its effects on intraocular pressure, aqueous flow rate and cyclic AMP level in the rabbit eye

The ocular penetration of 14C-forskolin in suspension was studied using albino rabbits. The effects of topical forskolin suspension on cyclic AMP (cAMP) synthesis, aqueous flow and intraocular pressure (IOP) were also studied. It was shown that only 0.03% of the instilled forskolin penetrated the ocular tissue. The calculated kep value for forskolin was 0.2 X 10(-4) cm/hr. The peak concentrations were calculated to be 4 X 10(-7) mole/liter, 4.6 X 10(-7) and 2.7 X 10(-7) mole/1,000 g tissue in aqueous, iris and ciliary body, respectively, after instillation of 1% forskolin suspension. Topical 1% forskolin suspension caused cAMP increase in the aqueous humor 30 minutes after instillation, but cAMP returned to baseline level 60 minutes after instillation. The cAMP level in the ciliary body was not increased by forskolin. Aqueous flow did not change, and the IOP was slightly decreased 45 and 60 minutes after instillation of forskolin suspension. The in vivo least-effective concentration of forskolin in the ciliary epithelium was considered to be about 2.7 X 10(-7) mole/1,000 g tissue. The weak IOP lowering effect of topical forskolin suspension was considered to be due to its poor ocular penetration. However, slight modification of molecular structure might increase ocular penetration. Present results suggest only a slight increase in penetrative potential would be needed to make forskolin effective in antiglaucoma therapy.     

Beta adrenergic effects on ciliary epithelial permeability, aqueous humor formation and pseudofacility in the normal and sympathectomized rabbit eye

Relatively selective beta 1- and beta 2-adrenergic agonists (tazolol and terbutaline) and antagonists (metoprolol and butoxamine) were topically applied to albino rabbit eyes and to isolated ciliary epithelium. The beta 2-agonist, terbutaline, was the most effective agent in reducing intraocular pressure although all compounds induced changes in aqueous humor formation rate and pseudofacility. The effects on the isolated ciliary epithelium were determined on tissues derived from both normal and superior cervical ganglionectomized eyes. The beta 1- and beta 2-agonists as well as the beta 1-antagonist induced large permeability increases while the beta 2-antagonist induced a small permeability increase in normal tissues. In ganglionectomized tissues the beta 1-antagonist effect was decreased and the beta 2-antagonist effect enhanced while the agonist effects were almost unchanged. The results show that both beta 1- and beta 2-receptors exist in the ciliary epithelium and that stimulation of either receptor leads to an increase in fluid permeability and alterations in aqueous dynamics.     

Comparison of the effects of adrenergic agonists and alpha-, beta 1-, beta 2-antagonists on the intraocular pressure and adenylate cyclase activity in the ciliary processes of the rabbit

The hypotensive effect of different adrenergic agonists and antagonists were screened both in normo- and hypertensive rabbit eyes. The drugs were applicated topically and intraocular pressure (IOP) was monitored constantly with a manometric method. Subsequently the inhibitory effect of the antagonists on isoproterenol-stimulated adenylate cyclase activity in the ciliary processes was analyzed in vitro. In normotensive eyes agonist isoproterenol (beta) and antagonists labetalol (alpha beta), metoprolol (beta 1) and acebutolol (beta 1) decreased significantly IOP. In hypertensive eyes only isoproterenol and labetalol decrease IOP markedly. Timolol (beta) did not decrease IOP, although it inhibited adenylate cyclase activity as actively as labetalol. The other antagonists showed no inhibitory effect on in this respect. The results indicated that alpha-activity (labetalol, alpha beta) seems to potentiate the beta-activity (timolol, beta) in decreasing IOP. Whether this alpha-effect is a presynaptic effect (adrenergic denervation destroys it) is not yet clear. Similarly the beta 1-effect (metoprolol), without affecting adenylate cyclase activity, decreased IOP as well. This might be due to pre-synaptic release of endogenous transmitter(s) which in turn stimulate post-synaptic adenylate cyclase and induce the decreased IOP. Besides this, the possible role of blood vessels and CNS must also be kept in mind.     

Effect of ionic changes, amino acids, and metabolic inhibitors on hydraulic conductivity of the isolated rabbit iris-ciliary body

The hydraulic conductivity (Lp) of the rabbit ciliary epithelium has been measured under various conditions. Of the ionic changes in the bathing medium imposed upon the tissue, replacement of sodium and either depletion of bicarbonate or deletion of potassium all caused an increase in Lp. Chloride replacement had no effect on Lp. The changes in Lp occur more slowly than the changes in electrical parameters caused by the same perturbations and indicate that ions and fluid move across this tissue by different routes. Of the 10 amino acids tested, only three increased Lp; 2 basic and 1 neutral amino acid. No correlation existed between posterior chamber: plasma ratios, or the nature of the amino acid (i.e., basic, neutral, or acidic) and the effect on Lp. Of the metabolic inhibitors, only iodoacetic acid induced an increase in Lp. The transport inhibitor, ouabain, at concentrations up to 10(-4) M for 4 hours was without effect. The effects of these agents on electrophysiological parameters, therefore, must be confined to net ionic pathways. Inulin and sucrose permeabilities were determined under either a zero or 10 mmHg hydrostatic pressure gradient using radioactive tracers. Application of a hydrostatic pressure gradient increased sucrose permeability by ten-fold but only increased inulin permeability by 20%. This data, together with control tissues run in parallel with many of the experimental tissues, indicates that the tissue is not highly leaky and does not change characteristics with time.(ABSTRACT TRUNCATED AT 250 WORDS)     

The influence of 12(R)-hydroxyeicosatetraenoic acid on ciliary epithelial sodium, potassium-adenosine triphosphatase activity and intraocular pressure in the rabbit.

Ciliary epithelium was isolated from the rabbit eye and used as a source of plasma membrane material for sodium, potassium-adenosine triphosphatase (Na,K-ATPase) measurements. In the presence of 12(R)-hydroxyeicosatetraenoic acid [12(R)HETE], Na,K-ATPase (ouabain-sensitive ATPase) activity was reduced from 22.5 to 16.3 microM phosphate released/mg protein/hr. Ouabain-insensitive ATPase activity was not altered by 12(R)HETE. No changes in ciliary epithelium ATPase activity were observed in the presence of 12(S)HETE. In parallel studies with conscious rabbits, 12(R)HETE applied topically to the eye was found to lower intraocular pressure (IOP). It is possible that the IOP-lowering effect of 12(R)HETE may be, in part, associated with its ability to suppress the Na,K-ATPase activity of the ciliary epithelium.     

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.     

Comparison of the effects of 0.5% timolol maleate, 2% carteolol hydrochloride, and 0.3% metipranolol on intraocular pressure and perimetry findings and evaluation of their ocular and systemic effects

PURPOSE: To compare the effects of 0.5% timolol maleate, 2% carteolol, and 0.3% metipranolol on intraocular pressure (IOP) in 45 patients with primary open-angle glaucoma (POAG) and ocular hypertension. A secondary goal of this study was to evaluate the ocular and systemic side effects of these medications. METHODS: Measurements of IOP were taken at baseline (pretreatment) and 2, 6, and 12 hours after instillation on treatment days 15, 30, 60, and 90. Mean sensitivity (MS) and mean defect (MD) values of perimetry before and after treatment and the effects of the three beta blockers on serum lipid profiles were determined. Ocular and systemic side effects were recorded. RESULTS: The most prominent IOP lowering effect was noted with metipranolol at 2 and 6 hours on day 15, and with timolol maleate at 12 hours on day 15 and at all hours of the subsequent days on which measurements were taken. Timolol maleate produced a significant decrease in IOP at 12 hours on day 15 compared with carteolol. There was not a statistically significant difference between the MS and MD values on perimetry before and after treatment for any treatment. There was a statistically significant decrease in levels of total cholesterol and high-density lipoprotein (HDL) cholesterol and a significant increase in triglyceride levels; these changes were observed for all treatments. CONCLUSION: The effects of the three medications were not statistically different from each other in terms of IOP reduction and visual field changes. Careful monitoring of blood lipid levels is necessary with long-term treatment with beta blockers, because these agents reduced serum levels of HDL and total cholesterol while increasing triglycerides. Such changes in lipid levels could lead to increased incidence of complications, particularly in patients with atherosclerosis or coronary heart disease.