Histological study of the thyroid tissue in carp fish (Cyprinus Carpio) in Summer and Winter

This study is designed to investigate the effect of temperature on thyroid gland of 24 adult healthy female carp fish (12/ season). Neutral buffered formalin solution10% was used for fixation and routine histological techniques. Hemotoxline and Eosine and PAS stains were, also used. The results revealed that the thyroid tissue in carp fish was closely associated with the renal and sub pharyngeal tissue. Spleen and liver did not show any thyroid follicles. The thyroid tissue of carp fish consisted of disseminate thyroid follicles not surrounded by apparent capsule. Different sizes and shapes of thyroid follicles were observed. The follicular cells form tight epithelium. Para follicular cells were distributed among the follicles. The seasonal thyroid follicles were often filled with colloid summer and winter. In conclusion the study was classified glands units of the thyroid tissue into two states: The first stat of the low activity in winter its cells were epithelial flat to squamous and follicles almost filled with homogeneous, while the state of activity in the Summer, follicular homogenous colloid cells were cuboidal follicular and sessile to high contain homogeneous diluted colloid.

Lectin-like Ox-LDL Receptor Is Expressed in Human INT-407 Intestinal Cells: Involvement in the Transcytosis of Pancreatic Bile Salt–dependent Lipase

We have recently shown that the pancreatic bile salt–dependent lipase (BSDL) can be taken up by intestinal cells and transported to the blood circulation. This mechanism likely involves (specific) receptor(s) able to bind BSDL and located at the apical intestinal cell membrane. In this study, using Int407 human intestinal cells cultured to form a tight epithelium, we attempted to characterize (the) BSDL receptor(s). We found that an apical 50-kDa protein was able to bind BSDL. Further, we have demonstrated that Int407 cells expressed the lectin-like oxidized-LDL receptor (LOX-1), the upregulation of which by oxidized-LDL potentiates the transcytosis of BSDL, whereas carrageenan and to a lesser extent polyinosinic acid and fucoidan decrease the enzyme transcytosis. The mAb JTX92, which blocks the LOX-1 receptor function, also impaired the BSDL transcytosis. To confirm these results, the cDNA encoding the human intestinal receptor LOX-1 has been cloned, inserted into vectors, and transfected into Int407 cells. Overexpression of LOX-1 by these cells leads to a substantial increase in the BSDL transcytosis. Globally, these data support the view that LOX-1 could be an intestinal receptor for BSDL, which is implicated in the transcytosis of this enzyme throughout Int407 cells.

Cultured branchial epithelia from freshwater fish gills

We have developed a method for the primary culture of gill epithelial cells from freshwater rainbow trout on permeable supports, polyethylene terephthalate membranes ('filter inserts'). Primary cultures of gill cells (6-9 days in Leibowitz L-15 culture medium plus foetal bovine serum and glutamine) are trypsinized and the cells seeded onto the inserts. After 6 days of growth with L-15 medium on both surfaces (approximately isotonic to trout plasma), the cells form a tight epithelium as judged from a progressive rise in transepithelial resistance which reaches a stable plateau for a further 6 days, as long as L-15 exposure is continued on both surfaces. The cultured epithelium (approximately 8 µm thick) typically consists of 2-4 overlapping cell layers organized as in the lamellae in vivo, with large intercellular spaces, multiple desmosomes and putative tight junctions. The cells appear to be exclusively pavement-type cells with an apical surface glycocalyx, an abundance of rough endoplasmic reticulum, no selective DASPEI staining and relatively few mitochondria. Transepithelial resistance (approximately 3.5 k cm2), permeability to a paracellular marker (polyethylene glycol-4000; 0.17x10(-6) cm s-1) and unidirectional flux of Na+ and Cl- (approximately 300 nmol cm-2 h-1) all appear realistic because they compare well with in vivo values; net fluxes of Na+ and Cl- are zero. The preparation acidifies the apical medium, which accumulates a greater concentration of ammonia. Upon exposure to apical freshwater, resistance increases six- to elevenfold and a basolateral-negative transepithelial potential (TEP) develops as in vivo. These responses occur even when mannitol is used to prevent changes in apical osmotic pressure. Net Na+ and Cl- loss rates are low over the first 12 h (-125 nmol cm-2 h-1) but increase substantially by 48 h. The elevated resistance and negative TEP gradually attenuate but remain significantly higher than pre-exposure values after 48 h of apical freshwater exposure. The preparation may provide a valuable new tool for characterizing some of the mechanisms of active and passive ion transport in the pavement cells of the freshwater gill.

What Is New About the Structure of the Epithelial Na+ Channel ?

The epithelial Na+ channel (ENaC) in the apical membrane of tight epithelium represents the first member of a new family of ion channels. The channel is formed by the association of three homologous subunits, a-, b-, and g-ENaC, that functionally complement to give full activity to the channel complex.

Volume-activated chloride permeability can mediate cell volume regulation in a mathematical model of a tight epithelium.

Cell volume regulation during anisotonic challenge is investigated in a mathematical model of a tight epithelium. The epithelium is represented as compliant cellular and paracellular compartments bounded by mucosal and serosal bathing media. Model variables include the concentrations of Na, K, and Cl, hydrostatic pressure, and electrical potential, and the mass conservation equations have been formulated for both steady-state and time-dependent problems. Ionic conductance is represented by the Goldman constant field equation (Civan, M.M., and R.J. Bookman. 1982. Journal of Membrane Biology. 65:63-80). A basolateral cotransporter of Na, K, and Cl with 1:1:2 stoichiometry (Geck, P., and E. Heinz. 1980. Annals of the New York Academy of Sciences. 341:57-62.) and volume-activated basolateral ion permeabilities are incorporated in the model. MacRobbie and Ussing (1961. Acta Physiologica Scandinavica. 53:348-365.) reported that the cells of frog skin exhibit osmotic swelling followed by a volume regulatory decrease (VRD) when the serosal bath is diluted to half the initial osmolality. Similar regulation is achieved in the model epithelium when both a basolateral cotransporter and a volume-activated Cl permeation path are included. The observed transepithelial potential changes could only be simulated by allowing volume activation of the basolateral K permeation path. The fractional VRD, or shrinkage as percent of initial swelling, is examined as a function of the hypotonic challenge. The fractional VRD increases with increasing osmotic challenge, but eventually declines under the most severe circumstances. This analysis demonstrates that the VRD response depends on the presence of adequate intracellular chloride stores and the volume sensitivity of the chloride channel.

Ion transport and electrophysiology in rabbit cecum

We investigated the basic bioelectrical properties and the transport of Na+, Rb+, and Cl- in the isolated cecal epithelium of the rabbit. Intracellular microelectrode recordings provided first estimates of the individual membrane potentials and resistances. The cecal epithelium was found to be a moderately tight epithelium with a high transepithelial potential and a relative high paracellular resistance. It has, however, a low transepithelial resistance caused by a high rate of transcellular ion transport and a high transcellular conductance. Under short-circuit conditions Na+ and Cl- are absorbed, whereas Rb+ is secreted. Na+ absorption seems to be mediated via an electrogenic mechanism with low amiloride sensitivity and is partially linked to Cl- transport. Cl- absorption is not influenced by secretagogues, and Rb+ secretion occurs via a transcellular route. A basolateral K+ conductance was also found. The results show rabbit cecum as an epithelium with a high electrolyte transport capacity, distinctly different from the neighboring proximal colon, and suited to absorb the large quantities of electrolytes that are present in the luminal liquid.

Kinetics of Extracellular Solute Movement in the Isolated Midgut of Tobacco Hornworm (Manduca Sexta)

Measurements were made of the uptake of 14C-labelled sucrose in the short-circuited isolated midgut of fifth instar larvae of the tobacco hornworm. The longterm volume of distribution after bilateral exposure indicates that the extracellular space is about 0.26 ml g− wet tissue, much less than has been previously suggested. The results of unilateral exposure yield a value of about 0.04 ml g−1 for luminal goblet cell cavities and 0.22 ml g−1 for interstitial space. The kinetics of uptake suggest that the interstitial space is best modelled as a distributed volume rather than as a single compartment. This mathematical result fits the physical picture of an interstitial fluid in long, narrow channels which communicate freely with the haemolymph but are nearly impermeable to sucrose at the luminal end; that is, they are sealed by septate junctions. The electrical conductance of the epithelium is high, but the transepithelial permeability to sucrose is very much lower than to K+. Thus, the tissue is a physically tight epithelium.

Cadmium inhibition of Ca2+ uptake in rainbow trout gills

The effects of cadmium (Cd2+) on calcium (Ca2+) transport in the gills of rainbow trout (Salmo gairdneri) were studied. The gill epithelium of freshwater fish represents a model for a Ca2+-transporting tight epithelium. Unidirectional Ca2+ fluxes in the gills were estimated in an isolated saline-perfused head preparation. Ca2+ influx was not affected when up to 10 microM Cd were added to the ventilatory water at the start of flux determinations (in vitro exposure). However, after 16 h in vivo preexposure of the fish to 0.1 microM Cd in the water, a 79% inhibition of Ca2+ influx was observed. Ca2+ efflux was not affected when up to 10 microM Cd were added to the ventilatory water during the flux determination. Ca2+ efflux in fish preexposed to 0.1 microM Cd for 16 h was also not affected; a preexposure to 1 microM Cd, however, resulted in a 173% increase in Ca2+ efflux rates. Tracer retention in the gill tissue indicated that both Ca2+ and Cd2+ enter the gill epithelium via a lanthanum (La3+)-inhibitable pathway. It is concluded that Cd2+ readily enters the branchial epithelial cells, similarly as Ca2+ does via La3+-sensitive apical Ca2+ channels. The inhibitory action of Cd2+ on transepithelial Ca2+ influx seems to result from an inhibition of the basolateral Ca2+ transport, occurring after a critical intracellular Cd2+ concentration has been reached.