The permeability properties of septate junctions in Malpighian tubules of Rhodnius

1987 ◽  
Vol 88 (2) ◽  
pp. 251-265 ◽  
Author(s):  
H.B. Skaer ◽  
S.H. Maddrell ◽  
J.B. Harrison

This paper describes the structural characteristics and permeability properties of the smooth septate junctions between the upper Malpighian tubule cells of a blood-sucking bug, Rhodnius prolixus. The permeability of the paracellular route was tested only for solutes that could be demonstrated not to cross the epithelium via the cellular route. The intercellular clefts were readily permeated by sucrose, inulin and polyethylene glycol (PEG), showing a higher permeability to molecules of smaller radius (PEG versus sucrose). Negatively charged molecules permeated the clefts more readily than positively charged ones. The effects of pH, urea and luminal flow rate on permeability were studied. The results are discussed in relation to the physiological tightness of the Malpighian tubules to certain solutes and to its function as an excretory epithelium.

2002 ◽  
Vol 205 (11) ◽  
pp. 1645-1655 ◽  
Author(s):  
Juan P. Ianowski ◽  
Robert J. Christensen ◽  
Michael J. O'Donnell

SUMMARYIntracellular ion activities (aion) and basolateral membrane potential (Vbl) were measured in Malpighian tubule cells of Rhodnius prolixus using double-barrelled ion-selective microelectrodes. In saline containing 103mmoll-1Na+, 6mmoll-1 K+ and 93mmoll-1Cl-, intracellular ion activities in unstimulated upper Malpighian tubules were 21, 86 and 32mmoll-1, respectively. In serotonin-stimulated tubules, aCl was unchanged, whereas aNa increased to 33mmoll-1 and aK declined to 71mmoll-1. Vbl was -59mV and -63mV for unstimulated and stimulated tubules, respectively. Calculated electrochemical potentials(Δμ/F) favour passive movement of Na+ into the cell and passive movement of Cl- out of the cell in both unstimulated and serotonin-stimulated tubules. Passive movement of K+ out of the cell is favoured in unstimulated tubules. In stimulated tubules, Δμ/F for K+ is close to 0 mV.The thermodynamic feasibilities of Na+-K+-2Cl-, Na+-Cl-and K+-Cl- cotransporters were evaluated by calculating the net electrochemical potential (Δμnet/F) for each transporter. Our results show that a Na+-K+-2Cl- or a Na+-Cl- cotransporter but not a K+-Cl- cotransporter would permit the movement of ions into the cell in stimulated tubules. The effects of Ba2+ and ouabain on Vbl and rates of fluid and ion secretion show that net entry of K+ through ion channels or the Na+/K+-ATPase can be ruled out in stimulated tubules. Maintenance of intracellular Cl- activity was dependent upon the presence of both Na+ and K+ in the bathing saline. Bumetanide reduced the fluxes of both Na+ and K+. Taken together, the results support the involvement of a basolateral Na+-K+-2Cl- cotransporter in serotonin-stimulated fluid secretion by Rhodnius prolixus Malpighian tubules.


1988 ◽  
Vol 137 (1) ◽  
pp. 265-276 ◽  
Author(s):  
S. H. Maddrell ◽  
J. A. Overton

Ouabain, at all concentrations higher than 2 × 10(−7) mol l-1, stimulates the rate at which the Malpighian tubules of the insect, Rhodnius, transport sodium ions and fluid into the lumen. An effect on paracellular movement of sodium ions is unlikely because ouabain makes the electrical potential of the lumen more positive, which would slow diffusion of sodium into the lumen. Radioactive ouabain binds to the haemolymph-facing sides of the tubule cells but not to the luminal face. This binding is reduced in the presence of elevated levels of potassium or of non-radioactive ouabain. Bound ouabain is only slowly released on washing in ouabain-free saline. The evidence suggests that there is a Na+/K+-ATPase on the outer (serosal) membranes of the tubules. Such a pump would transport sodium in a direction opposed to the flow of ions and water involved in fluid transport; poisoning it with ouabain would remove this brake, and fluid flow and sodium transport would increase, as observed.


Author(s):  
J.S. Ryerse

Relatively little is known about the structure or function of the basal lamina in insect cells. I report here the ultrastructural organization of the basal lamina in Malpighian tubule yellow region primary cells of larval stage skipper butterflies (Calpodes ethlius). The Malpighian tubules selectively filter the blood and produce a primary urine which flows down the tubule lumen into the alimentary canal for excretion. The basal lamina provides the only barrier between the Malpighian tubule cells and the blood which freely circulates within the open body cavity and it may therefore regulate which molecules and ions gain access to the tubule cells. As a first step in these studies on the structure, composition and function of the basal lamina in insect cells the ultrastructure of the basal lamina in Calpodes Malpighian tubules after conventional TEM tissue processing and poststaining has been compared with that observed following en bloc staining with a variety of electron dense stains and probes with specificity for basal lamina components.


1998 ◽  
Vol 201 (24) ◽  
pp. 3411-3418
Author(s):  
J. A. Riegel ◽  
S. H. P. Maddrell ◽  
R. W. Farndale ◽  
F. M. Caldwell

External application of the 3',5'-cyclic monophosphates of inosine,cytidine, uridine and thymidine stimulated the fluid secretion rate (FSR)of Malpighian tubules isolated from Drosophila melanogaster. The evidence suggested that the cyclic nucleotides acted intracellularly in some capacity. Receptors of the 'purinergic' type appeared not to be major contributors to fluid secretion; of three purinergic agonists tried,adenosine, adenosine 5'-monophosphate (AMP) and adenosine 5'-triphosphate(ATP), only adenosine had an effect, but this was not observed consistently. None of the purinergic agonists interfered with the stimulation of the FSR by adenosine 3',5'-cyclic monophosphate (cAMP). The maximum stimulation of the fluid-secretion rate by any cyclic nucleotide was approximately double the unstimulated (control) rate. Tubules stimulated to less than maximal FSR by one cyclic nucleotide could be stimulated maximally by an appropriate concentration of another cyclic nucleotide. Malpighian tubules bathed in solutions that contained either[3H]cAMP or [3H]cGMP accumulated radioactivity to a level many times that in the medium. Accumulation of radioactivity by tubules bathed in 430 nmol l-1 [3H]cAMP was suppressed by 1 mmol l-1 non-radioactive cyclic nucleotides in the order cAMP>>cGMP>cIMP>cCMP; neither cTMP nor cUMP suppressed the accumulation of [3H]cAMP. Approximately 35 % of the[3H]cAMP and 80 % of the [3H]cGMP that entered the Malpighian tubule cells was metabolised to compounds that were not identified. It was concluded that cyclic nucleotides enter the Malpighian tubule cells by at least one transport mechanism which is particularly sensitive to purine-based nucleotides.


Author(s):  
Brendan Clifford

An ultrastructural investigation of the Malpighian tubules of the fourth instar larva of Culex pipiens was undertaken as part of a continuing study of the fine structure of transport epithelia.Each of the five Malpighian tubules was found to be morphologically identical and regionally undifferentiated. Two distinct cell types, the primary and stellate, were found intermingled along the length of each tubule. The ultrastructure of the stellate cell was previously described in the Malpighian tubule of the blowfly, Calliphora erythrocephala by Berridge and Oschman.The basal plasma membrane of the primary cell is extremely irregular, giving rise to a complex interconnecting network of basal channels. The compartments of cytoplasm entrapped within this system of basal infoldings contain mitochondria, free ribosomes, and small amounts of rough endoplasmic reticulum. The mitochondria are distinctive in that the cristae run parallel to the long axis of the organelle.


1979 ◽  
Vol 37 (1) ◽  
pp. 373-389
Author(s):  
H.B. Skaer ◽  
J.B. Harrison ◽  
W.M. Lee

Smooth septate junctions in the midgut of Musca domestica and in Malpighian tubules of both Musca and Rhodnius prolixus are described. Details of the structures revealed after standard fixation, fixation in the presence of the stain, lanthanum hydroxide, and after freeze-fracture are discussed in the light of models previously put forward to explain the interrelations of the images obtained by these different methods. The organization of the junction between cells of the midgut varies in the apical-to-basal axis. At the apical border the septa (or ridges in freeze-fracture replicas) are packed tightly and follow an undulating but strictly parallel course. This packing loosens towards the middle of the junction until, at its basal extremity, the septa (ridges in replicas) are widely separated and follow independent meandering courses. That these features are found both in lanthanum-infiltrated specimens and freeze-fracture replicas allows a correlation to be made between the septa and the freeze-fracture ridges. The functional significance of these smooth septate junctions is discussed.


1997 ◽  
Vol 200 (17) ◽  
pp. 2363-2367 ◽  
Author(s):  
M C Quinlan ◽  
N J Tublitz ◽  
M J O'Donnell

Rhodnius prolixus eliminates NaCl-rich urine at high rates following its infrequent but massive blood meals. This diuresis involves stimulation of Malpighian tubule fluid secretion by diuretic hormones released in response to distention of the abdomen during feeding. The precipitous decline in urine flow that occurs several hours after feeding has been thought until now to result from a decline in diuretic hormone release. We suggest here that insect cardioacceleratory peptide 2b (CAP2b) and cyclic GMP are part of a novel mechanism of anti-diuresis. Secretion rates of 5-hydroxytryptamine-stimulated Malpighian tubules are reduced by low doses of CAP2b or cyclic GMP. Maximal secretion rates are restored by exposing tubules to 1 mmol l-1 cyclic AMP. Levels of cyclic GMP in isolated tubules increase in response to CAP2b, consistent with a role for cyclic GMP as an intracellular second messenger. Levels of cyclic GMP in tubules also increase as urine output rates decline in vivo, suggesting a physiological role for this nucleotide in the termination of diuresis.


PROTOPLASMA ◽  
2017 ◽  
Vol 254 (6) ◽  
pp. 2189-2199 ◽  
Author(s):  
Saška Lipovšek ◽  
Franc Janžekovič ◽  
Tone Novak

2019 ◽  
Author(s):  
Dawson B.H. Livingston ◽  
Hirva Patel ◽  
Andrew Donini ◽  
Heath A. MacMillan

AbstractUnder conditions of stress, many animals suffer from epithelial barrier disruption that can cause molecules to leak down their concentration gradients, potentially causing a loss of organismal homeostasis, further injury or death. Drosophila is a common insect model, used to study barrier disruption related to aging, traumatic injury, or environmental stress. Net leak of a non-toxic dye (Brilliant blue FCF) from the gut lumen to the hemolymph is often used to identify barrier failure under these conditions, but Drosophila are capable of actively transporting structurally-similar compounds. Here, we examined whether cold stress (like other stresses) causes Brilliant blue FCF (BB-FCF) to appear in the hemolymph of flies fed the dye, and if so whether Drosophila are capable of clearing this dye from their body following chilling. Using in situ midgut leak and transport assays as well as Ramsay assays of Malpighian tubule transport, we tested whether these ionoregulatory epithelia can actively transport BB-FCF. In doing so, we found that the Drosophila midgut and Malpighian tubules can mobilize BB-FCF via an active transcellular pathway, suggesting that elevated concentrations of the dye in the hemolymph may occur from increased paracellular permeability, reduced transcellular clearance, or both.Summary StatementDrosophila are able to actively secrete Brilliant blue FCF, a commonly used marker of barrier dysfunction


1957 ◽  
Vol 34 (2) ◽  
pp. 276-289
Author(s):  
B. A. KILBY ◽  
ELISABETH NEVILLE

1. Homogenates of fat-body of Schistocerca gregaria Forsk. were shown to catalyse transamination reactions between α-ketoglutarate and numerous α-amino acids. The aspartate/glutamate and alanine/glutamate transaminases were the most active. They were present in both the ‘soluble’ and the mitochondrial fractions of fat-body cells and also in Malpighian tubules and mid-gut wall. The other transaminases in the fat-body were confined to the mitochondrial fraction. 2. Fat-body, Malpighian tubule and mid-gut wall homogenates were able to convert glutamic acid into glutamine, a compound which could also act as an amino-group donor in some transamination reactions. 3. A glutamate-cytochrome c reductase system which involved diphosphopyridine nucleotide was present in fat-body. 4. Fat-body contained an active arginase, but urease could not be detected. A D-amino-acid oxidase was present, together with a less active L-amino-acid oxidase. 5. In general, it appears that amino-acid metabolism in the locust resembles that in higher animals.


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