The duct system of the lachrymal salt gland of the green sea turtle,Chelonia mydas

Summary The duct system of the lachrymal salt gland of the green sea turtle comprises central canals, secondary ducts and a sac-like main duct. Distally the central canals consist of large columnar cells with lateral membranes folded into plicae which interdigitate in adjacent cells to form complex intercellular spaces. More proximally the central canals, secondary ducts and main duct consist of epithelia which are stratified or pseudostratified. The cells of these epithelia are separated by wide and complex inter-cellular spaces: they are joined by frequent maculae adherentes junctions. Complex intracellular webs of tonofilaments are associated with these junctions. At the luminal border of the epithelia of the secondary and main ducts is a layer of mucocytes. The mucocytes increase in density towards the proximal extremity of the main duct and secrete a thick luminal layer of mucus. The duct system is very well vascularised. It is suggested that it is unlikely to be merely a passive conduit and that it may have a role in the modification of the fluid secreted by the gland.     

Secretory capacity of the lachrymal salt gland of hatchling sea turtles,Chelonia mydas

Summary The lachrymal salt glands ofChelonia mydas were functional when hatchlings emerged from the nest. Osmotic concentrations up to 720 mosmol kg^–1 were recorded in spontaneously produced tears (salt gland secretions). When injected with a Na⁺ load (1500–2700 mol (100 g)^–1) newly emerged hatchlings produced tears ranging in osmotic concentration from 1000–1900 mosmol kg^–1 with Na⁺ secretion rates from single glands of 200–475 mol (100 g·h)^–1. In these circumstances the rate of sodium excretion, via the salt glands, was equivalent to the sodium content of 0.2 to 0.5 ml of sea water per hour. Since the apparent drinking rate of hatchlings within the first two days of entering sea water was approximately 0.5 to 1.7 ml per day, the excretion of Na⁺ imbibed by drinking is well within the secretory capacity of the lachrymal salt glands.In feeding hatchlings extraordinarily high Na⁺ secretion rates were induced by Na⁺ loading. Hatchlings which were loaded with Na⁺ by injection (1500–5400 mol (100 g)^–1) produced tears having osmotic concentrations between 1500 and >2000 mosmol kg^–1. The Na⁺ secretion rates from single glands were 750–4185 mol (100 g·h)^–1 with extremely high short term rates of 10700 mol (100 g·h)^–1 (50 mol min^–1 for 28 g hatchlings).In terms of gland mass the highest long term secretion rate translates into 21 mmol of Na⁺ per gram of salt gland per hour and is the highest secretion rate yet recorded for a reptilian salt gland. This rate is almost three times the highest rate recorded for sea snakes (8 mmol g·h^–1) and is similar to rates commonly observed in avian salt glands (25 mmol g·h^–1).Secretion by the lachrymal salt glands was initiated by increased blood concentrations of Na⁺ or K⁺, K⁺ being as effective as Na⁺ but with the composition of the teras being virtually unchanged compared to tears from Na⁺ stimulated hatchlings. Preliminary experiments indicated that secretion was not initiated by increased Cl^– concentration in the blood or by increased volume or osmotic concentration of the blood.Abbreviation O.P. osmotic pressure     

Allometric study of the lachrymal and Harderian glands of the rat during postnatal life.

The authors performed an allometric study of the growth of the rat's lachrymal and Harderian glands, during postnatal life. From the analysis of the results, they could conclude: (1) the growth of these glands in relation to body weight, during postnatal life, could be considered similar, following the allometric law; (2) the differential growth of the glands occurred in two stages: from birth until the 15th day and from the 15th day until the final period of life studied; (3) the two stages of development were separated by a critical period, during which an abrupt modification of the allometric coefficient occurred; (4) during the first days of postnatal life, the development of the Harderian gland was characterized by a high rate of growth and, just after eyelid disjunction and during rest of postnatal life, by a rate of allometric of growth less than 1. It is interesting to observe that the lachrymal and Harderian glands' critical period of development on the 15th day of postnatal life coincides with the time at which the eyelids of the animal open.     

Signaling pathways involved in pilocarpine-induced mucin secretion in rat submandibular glands

We have studied the signaling pathways involved in pilocarpine-induced mucin release in rat submandibular slices. Pilocarpine produced a significant increment of PGE2 levels and a positive (r=0.8870) and significant (p=0.0077) correlation between PGE2 production and mucin released was determined. The participation of PGE2 was confirmed by the use of indomethacin (indo) and of acetyl salicylic acid (ASA), cyclooxygenase inhibitors, which inhibited pilocarpine-induced mucin release. The muscarinic receptors involved in the regulation of mucin release were identified as M1 and M4 by the use of the selective acetylcholine receptors (mAChR) antagonists, pirenzepine, AF-DX 116, 4-DAMP and tropicamide. The secretory process was dependent on both, intracellular and extracellular calcium pools since it was inhibited by thapsigargin and verapamil. Cyclic AMP, nitric oxide synthase and PKC also participated in pilocarpine-induced mucin release. It is concluded that pilocarpine, by activation the M1 and M4 mAChR subtypes induces an increase of intracellular Ca2+ concentration ([Ca2+]I) and elevates cAMP levels, which in turn stimulates COX, PKC and NOS and promotes mucin exocytosis. PGE2 released induces cAMP accumulation which, together with PKC are involved in the PGE2 increased Ca2+/cAMP-regulated exocytosis. Thus, cAMP accumulation induced by cholinergic stimulation is, in part, the result of PGE2 production.     

Ion secretion by salt glands of desert iguanas (Dipsosaurus dorsalis)

Unlike the NaCl-secreting salt glands of many birds and reptiles, the nasal salt glands of lizards can secrete potassium as well as sodium, with either chloride or bicarbonate as the accompanying anion. The factors responsible for initiating secretion by the gland and the rates of cation and anion secretion were studied in the desert iguana, Dipsosaurus dorsalis. Lizards were given combinations of ions for several days, and secreted salt was collected daily and analyzed for sodium, potassium, chloride, and bicarbonate. Maximum total cation secretion rate was 4.4+/-0.38 micromol/g/d. Cation secretion ranged from 24% to 100% potassium; even high NaCl loads did not abolish potassium secretion. Maximum bicarbonate secretion was about 0.5 micromol/g/d; chloride was the predominant anion. Secretion rate increased only in response to those treatments that included potassium and/or chloride; sodium ions and other osmotic loads (e.g., sucrose) did not increase secretion. This is in contrast to birds and some other reptiles with salt glands, which initiate NaCl secretion in response to any osmotic load. The specificity of the response of the salt gland of Dipsosaurus may be related to the ecological importance of dietary potassium and chloride for herbivorous desert lizards.     

Mitotic activity and nuclear/cytoplasmatic ratio of the lachrymal and Harderian glands of the rat during postnatal life.

The authors performed a study of the mitotic activity and the nuclear/cytoplasmic (N/C) ratio during postnatal life of the lachrymal and Harderian glands of the rat. Based on the results, they concluded: (1) during the first days of postnatal life the development of lachrymal and Harderian glands was characterized by an intense mitotic activity and a low N/C ratio; (2) the period prior to eyelid disjunction was characterized by a diminished mitotic activity and a progressive and slow increase of the N/C ratio; (3) after eyelid disjunction, mitotic activity was reduced and an abrupt increase of the N/C ratio occurred, more evident in the Harderian gland; (4) during the final period of postnatal life studied mitotic activity was absent and the N/C ratio presented a higher, more constant level, which was always higher for the Harderian gland.     

The effects of oxymetazoline on secretion of protein and some electrolytes by rat submandibular and parotid glands

Oxymetazoline is a potent secretagogue for the salivary glands of rats. In the parotid gland, it activates preferentially alpha-adrenoceptors. As for the submandibular glands, it activates alpha-adrenoceptors at relatively low doses but at higher doses it allows secretion of new types of proteins.     

Hormone-dependent dissociation of blood flow and secretion rate in the lingual salt glands of the estuarine crocodile, Crocodylus porosus

Salt and water balance in the estuarine crocodile, Crocodylus porosus, involves the coordinated action of both renal and extra-renal tissues. The highly vascularised, lingual salt glands of C. porosus excrete a concentrated sodium chloride solution. In the present study, we examined the in vivo actions of vasoactive intestinal peptide (VIP), B-type natriuretic peptide (BNP) and angiotensin II (ANG II) on the secretion rate and blood perfusion of the lingual salt glands. These peptides were selected for their vasoactive properties in addition to their reported actions on salt gland activity in birds and turtles and rectal gland activity in elasmobranchs. The femoral artery was cannulated in seven juvenile crocodiles for delivery of peptides and measurement of mean blood pressure and heart rate. In addition, secretion rate of, and blood flow to, the salt glands were recorded simultaneously using laser Doppler flowmetry. VIP stimulated salt secretion was coupled to an increase in blood flow and vascular conductance of the lingual salt glands. BNP was a potent stimulant of salt gland secretion, resulting in a maximal secretion rate of more than 15-fold higher than baseline; however, this was not coupled to an increase in perfusion rate, which remained unchanged. ANG II failed to stimulate salt gland secretion and there was a transient decrease in salt gland blood flow and vascular conductance. It is evident from this study that blood flow to, and secretion rate from, the lingual salt glands of C. porosus are regulated independently; indeed, it is apparent that maximal secretion from the salt glands may not require maximal blood flow.     

The ontogeny of pulmonary surfactant secretion in the embryonic green sea turtle (Chelonia mydas).

Pulmonary surfactant, consisting predominantly of phosphatidylcholine (PC), is secreted from Type II cells into the lungs of all air-breathing vertebrates, where it functions to reduce surface tension. In mammals, glucocorticoids and thyroid hormones contribute to the maturation of the surfactant system. It is possible that phylogeny, lung structure, and the environment may influence the development of the surfactant system. Here, we investigate the ontogeny of PC secretion from cocultured Type II cells and fibroblasts in the sea turtle, Chelonia mydas, following 58, 62, and 73 d of incubation and after hatching. The influence of glucocorticoids and thyroid hormones on PC secretion was also examined. Basal PC secretion was lowest at day 58 (3%) and reached a maximal secretion rate of 10% posthatch. Dexamethasone (Dex) alone stimulated PC secretion only at day 58. Triiodothyronine (T(3)) stimulated PC secretion in cells isolated from days 58 and 73 embryos and from hatchling turtles. A combination of Dex and T(3) stimulated PC secretion at all time points.     

Ionic currents and secretion in the exocrine glands

Exocrine glands extrude both proteins and salt. Fluid secretion is related to a modification of the membrane permeability of secreting cells. This permeability change may be measured as an increase of labelled ion fluxes or as a rise of membrane conductance. It involves Na+, K+, Cl- and Ca2+ ions. Intracellular Ca2+ acts as "second messenger" in the development of the electrical response. Recent recordings using the "patch-clamp" technique have revealed three types of ion channel activated by secretory agents. These channels are sensitive to internal Ca2+ ions. They are respectively selective to K+, Cl- and positively charged monovalent ions. Two models suggesting possible roles for these channels in the secretion process are presented. However, evaluation of such models is presently restricted by numerous uncertainties on the function of secreting cells in vivo. Information is notably lacking concerning the exact composition of the secreted fluid, and the exchanges between exocrine glands and blood circulation.     

The homology of cranial glands in turtles: with special reference to the nomenclature of salt glands

The cranial glands of ten species of turtles were studied by the use of histochemistry applied to serial sections of whole heads. The majority were stenohaline species, but one brackish water form, Malaclemys, was included. The results show that all species have two major orbital glands, an anterior Harderian gland, and a posterior lachrymal gland. The latter is seromucous in all species except Malaclemys terrapin in which the gland shows little evidence or organic secretion. External and medial nasal glands are found in all species studied, and also are seromucous glands. With these reslts, combined with a review of the literature the following conclusions are made. The Harderian gland is by definition the orbital gland opening through the medial surface of the nictitating membrane at or near the anterior canthus. It is of constant occurrence, and histological appearance, probably serving the same function. However, despite much recent study this function remains unknown. The lachrymal gland is defined as the orbital gland which opens through the lateral surface of the nictitating membrane, or medial surface of the lower eyelid, at or near the posterior canthus. It is of variable occurrence, absent in many reptiles, and has a histological structure which is also variable. In the stenohaline species it is apparently involved in organic secretion, while in the brackish water Malaclemys it may be involved in salt secretion, as it is in Cheloniidae. The nasal glands in turtles are probably homologous with the nasal salt glands of lizards and birds, but they do not appear to subserve the same function. In all species of turtles studied the nasal glands are seromucous. They are perhaps involved in the maintenance of the epithelium of the olfactory cavity.     

Control of salt gland activity in the hatchling green sea turtle, Chelonia mydas

 We studied the control of salt gland secretion in hatchling Chelonia mydas. The threshold salt load to activate salt secretion was between 400 μmol NaCl 100 g bodymass (BM)⁻¹ and 600 μmol NaCl 100 g BM⁻¹, which caused an increase in plasma sodium concentration of 13% to 19%. Following a salt load of 2700 μmol NaCl 100 g BM⁻¹, salt gland secretion commenced in 12 ± 1.3 min and reached maximal secretory concentration within 2–7 min. Maximal secretory rate of a single gland averaged 415 μmol Na 100 g BM⁻¹ h⁻¹. Plasma sodium concentration and total osmotic concentration after salt loading were significantly higher than pretreatment values within 2 min. Adrenalin (25 μg kg BM⁻¹) and the cholinergic agonist methacholine (1 mg kg BM⁻¹) inhibited salt gland activity. Atropine (10 mg kg BM⁻¹) reversed methacholine inhibition and stimulated salt gland secretion when administered with a subthreshold salt load. Arginine vasotocin produced a transient reduction in sodium secretion by the active gland, while atrial natriuretic factor, vasoactive intestinal peptide and neuropeptide Y had no measurable effect on any aspect of salt gland secretion. Our results demonstrated that secretion of the salt gland in C. mydas can be modified by neural and hormonal chemicals in vivo and that the cholinergic and adrenergic stimulation of an exocrine gland do not appear to have the typical, antagonist actions on the chelonian salt gland. Accepted: 28 September 1999