Search

Human tear fluid has plasminogen activator activity. The type of plasminogen activator activity in unstimulated and stimulated tears was determined, using antibodies that specifically neutralize tissue plasminogen activator or urokinase. All plasminogen activator activity was tissue plasminogen activator-related in both types of tears. Correlations between activities of beta-hexosaminidase, lysozyme and lactate dehydrogenase with tissue plasminogen activator activity indicate that the contribution to plasminogen activator activity of conjunctival and corneal epithelium is more important in unstimulated tears than stimulated tears. In stimulated tears the tissue plasminogen activator activity originates mainly from the lacrimal gland. It is suggested that a constant concentration of plasminogen activator is released from the lacrimal gland and that this concentration is independent of the secretion rate of tear fluid and that the release from the conjunctiva is due to desquamation of cells.

A variety of enzymes were identified in rat tears and lacrimal gland fluid. The use of a tapered glass cannula in the opening of the excretory duct was found to be an useful method to collect samples of rat lacrimal gland fluid, i.e., the fluid directly originated from the main excretory duct of the lacrimal gland, uncontaminated by secretions from the Harderian gland and from desquamating conjunctival and corneal epithelial cells. Based upon comparison of the enzyme pattern in tears, lacrimal gland fluid and the lacrimal gland tissue, we concluded that the lacrimal gland is the primary tissue source for peroxidase (POD), amylase (AMY) and total protein in rat tears, while plasminogen activator and lactatedehydrogenase (LDH) may be present in tears primarily as secretion products from other ocular tissue sources. beta-hexosaminidase (beta-hex) is released from the lacrimal gland and from other ocular tissue sources as well.

The excretory duct of the lacrimal gland of rabbits and guinea pigs was cannulated in situ for collection of pure lacrimal gland fluid, not contaminated by secretions from the Harderian gland or contributions of desquamating cells of the conjunctival and corneal epithelium. Tears as well as lacrimal gland fluid of both species showed a species-specific and molecular weight-dependent pattern after sodium dodecylsulphate-polyacrylamide (SDS-PAA) gradient slab gel electrophoresis. The most striking difference in both species was a protein corresponding to serum albumin present in tears and almost lacking in lacrimal gland fluid. Likewise, a variety of enzymes, total protein and PGE2 were measured in tears and lacrimal gland fluid. For rabbit tears the lacrimal gland is the primary tissue source of lysozyme (LZM), beta-hexosaminidase (beta-hex), angiotensin-converting enzyme (ACE), plasminogen activator (PA) and total protein, while lactate dehydrogenase (LDH) and the greater part of prostaglandin E2 (PGE2) are present in rabbit tears mainly as products from other ocular tissue sources. In guinea pig tears peroxidase (POD), ACE, PA and less PGE2 are exceted by the lacrimal gland, amylase (AMY), LDH and a substantial amount of PGE2 are added to the guinea pig tears by other ocular tissue sources. Beta-hex and total protein are released from the lacrimal gland and from other ocular tissue sources as well.

The role of calcium in stimulus-secretion coupling in rat lacrimocytes was investigated. Monolayers of lacrimocytes released peroxidase into the incubation medium upon stimulation with calcium ionophore A 23187, L-norepinephrine, L-phenylephrine and tyramine. Our lacrimocyte preparation can be considered free of endogenous catecholamines, because lacrimocytes derived both from normal and from reserpinized rats responded to tyramine with respect to peroxidase secretion. Phenoxybenzamine, cocaine, verapamil and sodiumorthovanadate failed to inhibit the L-phenylephrine-evoked peroxidase secretion. The stimulatory effects of A 23187, L-norepinephrine and L-phenylephrine required extracellular Ca2+ and could be blocked by phentolamine. The action of tyramine was insensitive to all these measures. It is suggested that the stimulus-secretion of peroxidase from lacrimocytes may consist of a Ca2+-dependent phase for stimulants as A 23187, L-norepinephrine and L-phenylephrine and a Ca2+-independent phase for the non-specific tyramine action.

The identification of alpha-adrenergic receptors and subdivision into alpha 1- or alpha 2-subtypes were studied by measuring the specific binding of the radioligands [3H]-prazosin as well as [3H]-clonidine to membranes prepared from homogenized rat lacrimal glands. The absence of high-affinity binding for [3H]-prazosin as well as for [3H]-clonidine indicates that rat lacrimal glands do not possess a substantial amount of alpha 1- and alpha 2-adrenoceptors. The binding data correspond with the characterization by pharmacological means. Monolayers of lacrimocytes were incubated with various selective alpha 1-, alpha 2-, beta-adrenergic agonists and antagonists or other substances, and peroxidase discharge was measured over a period of 1 hr. Among various substances only L-norepinephrine, L-phenylephrine, tyramine and ionophore A 23187 were stimulants of peroxidase secretion, whereas the adrenergic-stimulated secretory response was only suppressed by phentolamine.

Free and lysosomal activities of 10 acid hydrolases have been determined in human lacrimal gland tissue. The enzyme activities appeared mainly in the lysosomal fraction, and the proportions correspond very well with those found in the tear fluid. Acetylsalicylic acid at blood levelsof about 1 mM produced a significant lowering of the beta-hexosaminidase concentration in tears. It is suggested that acetylsalicylic acid may have a stabilizing effect on lacrimal lysosomes, resulting in a diminished release of lysosomal enzyme from the lacrimal gland.