Suzanne Dunel-Erb

The role of environmental sodium chloride relative to calcium in gill morphology of freshwater salmonid fish

SummaryUltrastructure, distribution and abundance of cell types were examined in the gills of two freshwater salmonid species, Salmo fario and Salmo gairdneri, in media of selected ion content. In plain hard water (PW) with high concentrations of Ca2+, Na+, and Cl-, gill chloride cells (CC) were confined to trailing edges and interlamellar regions of filaments whereas in mountain soft water (MW) with low concentrations of Ca2+, Na+, and Cl-, CC were more numerous on filaments and covered lamellae, particularly along trailing edges. CC also appeared on lamellae of PW trout acclimated to soft water in a pond. This proliferation was not alleviated when ambient Ca2+ levels were raised (MW + Ca2+) but regressed in elevated NaCl media (MW + NaCl). The regression process involved an initial covering of CC by pavement cells followed by cytolysis and then eventual disappearance of CC. In MW, mucous cells were distributed mainly on trailing edges and, to a lesser extent, leading edges of filaments; they were absent from lamellae regardless of external ion levels.The results of this study shed some light on the functional significance of CC in freshwater fish. It is suggested that proliferation of CC is an adaptive response to dilute freshwater (i.e. [NaCl]<0.1 mequiv·1-1).

Restoration of the jejunal mucosa in rats refed after prolonged fasting.

To investigate the importance of body fuel depletion on gut rehabilitation after food deprivation, we compared the kinetics of jejunal mucosa alteration and restoration in rats that were refed after reaching different stages in body fuel depletion. Rats (P2) were refed while still in the so-called phase II, where body protein utilization is minimized, whereas rats (P3) were refed when they had reached the stage of increasing protein utilization (phase III). There was a significant decrease in total mass of intestine (P2, -30%; P3, -40%) and jejunal mucosa (P2, -52%; P3, -60%), as well in the size of the crypts (P2, -15%; P3, -36%) and villi (P2, -37%; P3, -55%). Structural changes of the mucosa included disappearance of some villi and a reduction in the size and number of crypts. Despite the larger morphological alterations in P3, the restoration of mucosa was as fast and complete after only 3 days of refeeding for both P2 and P3 rats. The respective roles of the mitosis pressure and of the lamina propria dynamics were studied. The rapid reversibility of the gut mucosal alterations due to fasting might constitute an integrative process.

Gill epithelial cells kinetics in a freshwater teleost, Oncorhynchus mykiss during adaptation to ion-poor water and hormonal treatments.

The aim of this work was to determine the kinetics of the dramatic development of the gill chloride cells (CCs) during adaptation of the salmonid Oncorhynchus mykiss to an ion-poor environment.To monitor cell division, the incorporation in the mitotic cell DNA of bromo-deoxyuridine (BrdUrd) was visualized with a monoclonal antibody. The density of labelled nuclei was used as an index of cellular division (proliferation), concomitantly with morphometry of phenotypic changes monitored with SEM.In the filament epithelium, a phase of CC differentiation occurred within 12h after the transfer, followed by a delayed phase of cell proliferation (48h). In the lamellar epithelium, the present study demonstrates the absence of cell proliferation after ion-poor water transfer. The conclusion is that proliferation (mitosis) is important in the primary filament whereas differentiation and migration (from the filament) is the main mechanism for the appearance of CCs on the secondary lamellae.The present study suggests that cortisol promoted differentiation, but not division, of cells. CCs, presumably premature, were stained by anti-cortisol monoclonal antibody indicating the presence of cortisol. No mature CCs were stained.Growth hormone (oGH, ratGH) increased the rate of cell division both in lamellar and filament epithelium.

The vascular and epithelial serotonergic innervation of the actinopterygian gill filament with special reference to the trout, Salmo gairdneri

SummaryAntibodies against serotonin and 5-methoxytryptamine reveal indolaminergic neurons innervating the proximal part of the efferent arterial vasculature, the filament epithelia, the central venous sinus, and certain other serotonergic cells of the teleost gill filament. In the same area, acetylcholinesterase-positive and indoleaminergic neurons have already been described. We propose that these populations of neurons belong to a single neuronal type but express different agents. Our current results support this idea; in particular, they point to the presence of a single type of serotonin-containing nerve terminal, impinging on vascular smooth muscle. These results are in agreement with physiological data showing (i) the existence of non-cholinergic (atropine-resistant) vasoconstriction of the gill vasculature after nerve stimulation, and (ii) a potent vasoconstrictory action of infused serotonin. In addition, the above-mentioned serotonergic neurons have synaptic contacts with catecholaminergic nerve fibers, suggesting the existence of a modulatory relationship between the sympathetic and the cranial autonomic nerves supplying the teleost gill. Finally, these neurons show morphological relationships with a previously undescribed type of branchialserotonergic cell. The role of the parasympathetic nerve plexus of the teleost gill filament in the control of respiration and ionoregulation is discussed.

9 The Pseudobranch: Morphology and Function

Publisher Summary This chapter examines the morphology and function of the pseudobranch. Teleostean pseudobranch is a hemibranch and is maximally developed in highly evolved fish. In the teleosts, the pseudobranch is located in the cranial part of the subopercular cavity to which it is attached by one of its sides. Histological serial sections and cast preparations demonstrate that the pseudobranch has an arterio-arterial vasculature similar to that of the gill. The afferent pseudobranchial artery gives rise to successive arteries supplying filaments. Different types of pseudobranch organization have been observed according to the fresh- or saltwater origin of the fish, but it has been noticed that forms, sizes, and external structures vary widely within orders, families, and species without any obvious reason, whereas vascular organization and innervation keep a similar organization throughout. In some part of the pseudobranch with free lamellae, the epithelium consists of typical chloride cells in place of pseudobranchial cells. In the pseudobranch, chloride cells exhibit the characteristic differences of fresh- and saltwater species.

Neurons controlling the gill vasculature in five species of teleosts

SummaryA plexus of nerve fibers encompassing neuronal perikarya is present within the gill filament; it surrounds the proximal portion of the efferent filament artery and the efferent lamellar arterioles. This innervation resembles the pattern described for the area around the sphincter of the efferent filament artery: acetylcholinesterase-positive neurons and fibers, fast-fading yellow-fluorescent neurons and fibers, long-lasting green-fluorescent fibers. In addition, synaptic contacts between the different components suggest functional interrelationships. Nerves evidently control the efferent limb of the filament circulation including the sphincter of the efferent filament arteries, the proximal portion of the efferent filament arteries proper, and their corresponding efferent lamellar arterioles. However, the distal portion of this system is poorly innervated.

The sphincter of the efferent filament artery in teleost gills. I: Structure and parasympathetic innervation

Previous studies have shown the existence of a sphincter in the efferent filament artery of the teleost gill and its constrictory response to acetylcholine (ACH) and vagal stimulation. This study deals with the muscular organization of this sphincter and the distribution of its innervation as elucidated by degeneration methods and cytochemistry. The sphincter innervation is supplied by the protrematic vagus nerves. Nerve endings filled with cholinergic‐type vesicles are located in close association with the adventitial smooth muscle cells and display a strong acetylcholinesterase (ACHE) activity. Section of the protrematic vagus nerve induces a nearly complete degeneration of the sphincter innervation. ACHE‐positive nerve cell bodies are present both in the sphincter area and in the protrematic vagus nerve. These results suggest that innervation of the sphincter in the efferent filament artery is cholinergic through the activity of postganglionic axons of the parasympathetic system.

The sphincter of the efferent filament artery in teleost gills: II. Sympathetic innervation

In addition to the cholinergic innervation described in the sphincter of the efferent filament arteries (Bailly and Dunel‐Erb, ′86), an aminergic component has been demonstrated by specific techniques. The Falck fluorescence technique reveals a network of nerve fibers displaying a green fluorescence characteristic of catecholamines. At the ultrastructural level two types of fibers are present, one with clear vesicles and another with densecored vesicles. Axo‐axonal synaptic relationships exist between the two types. Results of 5‐ and 6‐OHDA (hydroxydopamine) treatments confirm the presence of an aminergic component.

Effects of fasting and refeeding on Jejunal morphology and cellular activity in rats in relation to depletion of body stores

Background: Intestinal mucosa atrophy following a period of starvation characterized by the mobilization of fat stores for energy expenditure (phase II) worsen after a long fast marked by an increase in protein catabolism (phase III). However, the morphology of the jejunum is completely restored after 3 days of refeeding. The aim of this study was to determine the mechanisms involved in the rapid jejunal restoration following the critical phase III. Methods: Jejunal structure was observed through conventional and environmental scanning electron microscopy, whilst cellular dynamics were studied using classical optic microscopy tools and immunohistochemistry. Results: Mucosal structural atrophy during fasting proved to worsen over the two phases. During phase II, apoptosis is still present at the tip of the villi, the number of mitosis in crypts showed a 30% decrease and a transient drop in cell migration is observed. During phase III, however, an 85% rise in mitosis was noticed along with an increase in cell migration and the disappearance of apoptotic cells at the villus tips. This increased cell renewal continues after food ingestion. Conclusions: Starved rats appeared to be in a phase of energy sparing in phase II, with depressed cellular events in the intestinal mucosa. In phase III, however, the preservation of functional cells and the early increase in crypt cell proliferation should prepare the mucosa to refeeding and could explain why jejunal repairs are complete after 3 days of refeeding following either phase II or phase III.

Observations of the intestinal mucosa using environmental scanning electron microscopy (ESEM); comparison with conventional scanning electron microscopy (CSEM)

In order to evaluate the potential use of environmental scanning electron microscopy (ESEM) in biology, structural changes of the jejunal villi of rats were studied after periods of fasting and refeeding, using a conventional scanning electron microscope (CSEM) and ESEM. While observation using the CSEM, involves chemical fixation, drying and coating, observation of fresh, unprepared materials can be directly realized with the ESEM. Environmental microscopy provides a relatively new technology for imaging hydrated materials without specimen preparation and conductive coating. Direct observation of biological samples in their native state is therefore possible with an ESEM. After fasting, the jejunal mucosa is dramatically reduced in size, splits and holes appearing at the tip of the villi. These changes were observed whatever the type of technique used. Artifacts due to the sample preparation for CSEM observation (drying, coating) can therefore be excluded. However, CSEM and ESEM must be used jointly. While, CSEM must be preferred for surface analysis involving high magnifications, ESEM observation, on the other hand, can prove valuable for determining the living aspect of the samples.