The initial stages in the action of an insecticidal delta-endotoxin of Bacillus thuringiensis var. israelensis on the epithelial cells of the malpighian tubules of the insect, Rhodnius prolixus

1988 ◽  
Vol 90 (1) ◽  
pp. 131-144
Author(s):  
S.H. Maddrell ◽  
N.J. Lane ◽  
J.B. Harrison ◽  
J.A. Overton ◽  
R.B. Moreton
Keyword(s):  
Bacillus Thuringiensis ◽  
Intercellular Junctions ◽  
Fluid Secretion ◽  
Malpighian Tubules ◽  
Ultrastructural Changes ◽  
Cytoplasmic Vacuolization ◽  
Large Numbers ◽  
Delta Endotoxin ◽  
High Concentrations ◽  
Very High

The effects of the 27 X 10(3) Mr insecticidal delta-endotoxin from Bacillus thuringiensis var. israelensis have been studied using, as a model system, isolated insect Malpighian tubules. At all concentrations of the toxin higher than 1 microgram ml-1 (4 X 10(−8) moll-1) applied to the outer surface of the tubules, fluid secretion failed within about 30 min. Except at very high concentrations, where failure always takes at least 30 s, there was an inverse relationship between the concentration of toxin and the time of failure of toxin-treated tubules. During exposure to toxin, the tubules were initially unaffected for a relatively long period and then rapid failure occurred. If the tubules were removed into toxin-free saline just before failure would have occurred, fluid secretion remained normal for at least 2 h, but on return to the origin toxin-containing saline failure was almost immediate. The toxin was found not to bind to the basement membrane. Ultrastructural changes became evident as tubule failure occurred. These initially involved modifications to the basal side of the cells, but later also to the luminal microvilli. Intercellular junctions became disassociated and cytoplasmic vacuolization occurred. The population of intramembranous particles in the basal membranes became reduced with time. Our findings suggest the following hypothesis for the initial stages in the interaction of the toxin with the tubules. Toxin molecules attach to the accessible cell membranes progressively and irreversibly. They do not readily associate by diffusing laterally in the membrane, so that toxic effects develop only when sufficiently large numbers of them attach close together. The molecules may then associate in some way as a complex, perhaps forming a pore in the membrane. Relatively few such pores lead rapidly to cell failure and death.

The Retention of Amino Acids in the Haemolymph During Diuresis in Rhodnius

1980 ◽  
Vol 87 (1) ◽  
pp. 315-330
Author(s):  
S.H.P. MADDRELL ◽  
B.O.C. GARDINER
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Fluid Secretion ◽  
Malpighian Tubules ◽  
High Rate ◽  
Excretory System ◽  
Low Permeability ◽  
High Concentrations ◽  
Active Reabsorption ◽  
Very High

The haemolymph of Rhodnius is rich in amino acids. During the rapid diuresis after a blood meal, no more than trace amounts of amino acids are lost in the urine. There is no significant reabsorption of amino acids in the excretory system. That they escape elimination can instead be attributed to a combination of the low permeability of the Malpighian tubules to amino acids, the very high rate of fluid secretion by the tubules, and the dilution of the haemolymph by an expansion in its volume after feeding. Amino acid losses are low in spite of the fact that the tubules actively accumulate high concentrations of amino acids in their cells and passive losses from these stores augment to some extent the flux of amino acids into the lumen. At times other than during diuresis, fluid secretion by the Malpighian tubules is slow. Calculations show that haemolymph solutes can then passively reach the higher concentrations in the lumen that are required for the operation of the excretory system (which relies on unselective passive entry and active reabsorption of useful substances). An advantage of the extraordinarily high rate of fluid secretion during diuresis is that fluid excretion can be rapidly completed. There is then little time for significant amounts of haemolymph solute to be lost passively.

scholarly journals A Preliminary Report on the Use of Bench-Top X-Ray Micro-Computerised Tomography to Study the Malpighian Tubules of the Overwintering Seven Spotted LadybirdCoccinella septempunctataL. (Coleoptera: Coccinellidae)

2012 ◽  
Vol 2012 ◽  
pp. 1-6 ◽  
Author(s):  
Duncan Bell ◽  
Lewis Woolnough ◽  
David Mortimore ◽  
Nick Corps ◽  
Diana M. Hudson ◽  
...  
Keyword(s):  
Coccinella Septempunctata ◽  
Ct Scanning ◽  
Small Sample ◽  
Malpighian Tubules ◽  
Micro Ct ◽  
Computerised Tomography ◽  
X Ray ◽  
High Concentrations ◽  
Very High ◽  
Metallic Pollutants

The application of micro-CT scanning techniques on a small sample of “Seven-spot ladybirds”Coccinella septempunctata,collected in December 2009, identified an accumulation of material with a very high, relative X-ray attenuation value in the malpighian tubules of most but not all of the individuals sampled. The passage of metals such as cadmium in soil through a food chain to finally accumulate in high concentrations in ladybirds and lacewings has been previously reported. The identification of the dense material found in our sample of ladybirds, its origin, and the process by which it accumulates in, and is processed by, the malpighian tubules is the challenge ahead. The authors speculate that a straightforward means of monitoring levels of metallic pollutants in the environment might emerge.

scholarly journals Action of activated 27,000 Mr toxin from Bacillus thuringiensis var. israelensis on Malpighian tubules of the insect, Rhodnius prolixus

1989 ◽  
Vol 94 (3) ◽  
pp. 601-608
Author(s):  
S.H. Maddrell ◽  
J.A. Overton ◽  
D.J. Ellar ◽  
B.H. Knowles
Keyword(s):  
Bacillus Thuringiensis ◽  
Cell Membrane ◽  
Basal Cell ◽  
Time Course ◽  
Fluid Secretion ◽  
Rhodnius Prolixus ◽  
Malpighian Tubules ◽  
Bathing Solution ◽  
Concentrated Solutions ◽  
Intracellular Milieu

The action of activated 27,000 Mr toxin from Bacillus thuringiensis var. israelensis (Bti toxin) on Malpighian tubules of Rhodnius prolixus has been investigated. Its binding to the tubules is slowed by low temperature but is not prevented even at 0 degree C. The binding is less effective at pH 10 than at pH7. Pretreatment of the tubules with 0.1 mmol l-1 ouabain or bumetanide or 1 mumol l-1 5-hydroxytryptamine did not affect the toxicity of the toxin. The toxin causes very large changes in the trans-epithelial potential difference; it changes from 40 mV, lumen negative, often to more than 100 mV, lumen positive. This reflects an initial collapse of the potential of the basal cell membrane, followed by a large positive-going potential change at the luminal cell membrane. Just prior to the effects of the toxin on rapid fluid secretion, the basal cell membrane becomes permeable to sucrose molecules. Raffinose at 170 mmol l-1 in the bathing solution does not protect the tubules from Bti toxin action but dextran, Mr5000, at 60 mmol l-1 significantly delayed failure of fluid secretion and, even more, the onset of staining of the tubule cells with Trypan Blue. Exposing tubules to saline that is calcium-free and/or magnesium-free, or has a composition adjusted to be similar to that of the intracellular milieu, does not affect the time course of failure of fluid secretion induced by the toxin. There is no evidence that effective aggregates of Bti toxin molecules are formed in concentrated solutions.(ABSTRACT TRUNCATED AT 250 WORDS)

scholarly journals ION AND FLUID SECRETION BY DIFFERENT SEGMENTS OF THE MALPIGHIAN TUBULES OF THE BLACK FIELD CRICKET TELEOGRYLLUS OCEANICUS

1993 ◽  
Vol 177 (1) ◽  
pp. 1-22
Author(s):  
A. T. Marshall ◽  
P. Cooper ◽  
G. D. Rippon ◽  
A. E. Patak
Keyword(s):  
Cyclic Amp ◽  
Fluid Secretion ◽  
Distal Segment ◽  
Malpighian Tubules ◽  
Secretion Rate ◽  
Osmotic Concentration ◽  
Corpora Cardiaca ◽  
Diuretic Hormone ◽  
High Concentrations ◽  
Tubule Fluid

Cricket Malpighian tubules have two morphologically distinct segments, a thin distal segment, which occupies approximately 10 % of the total tubule length, and a main segment. The two segments differ in secretion rates and response to corpora cardiaca extract. The secreted fluids differ in osmotic concentration and elemental composition. The distal segment secretes fluid at a rate (per mm length) which is approximately twice that of the main segment under control conditions. After stimulation by corpora cardiaca extract (Cc) the rate from the main segment approximately doubles whilst the distal segment rate remains unchanged. Fluid from the main segment and the whole tubule is slightly hypo-osmotic to the medium (5–11 mosmol kg-1) under control conditions, whereas that from the distal segment is slightly hyperosmotic (12 mosmol kg-1). On stimulation with Cc, the whole tubule fluid becomes slightly hyperosmotic (12 mosmol kg-1), that from the main segment remains slightly hypo-osmotic (3 mosmol kg-1) but fluid from the distal segment becomes very hyperosmotic (55 mosmol kg-1). Differences between the tubule fluid and the medium osmolality are indicated in parentheses. Fluid from the main segment has high concentrations of K (166 mmol l-1), Cl (111 mmol l-1), Na (41 mmol l-1) and P (83 mmol l-1), whereas that from the distal segment has high concentrations of K (101 mmol l-1) and Cl (137 mmol l-1). On stimulation with Cc, the elemental concentrations in fluids from the main segments and whole tubules do not change significantly but the K and Cl concentrations in distal segment fluid increase (182 and 188 mmol l-1 respectively). The Mg present in whole tubule fluid is derived largely from the distal segment. The ionic composition accounts for the observed osmotic concentrations in fluid from whole tubules, main segments and stimulated distal segments, but not for the concentrations in fluid from unstimulated distal segments. The fluid from unstimulated distal segments contains an unidentified organic solute accounting for approximately 90 mosmol kg-1 of the osmotic concentration. The distal segment contributes 22 % and 11 % of the fluid volume, 26 % Cl, 14 % K and 12 % Cl, 11 % K in control and Cc-stimulated tubules respectively. Considerably higher values are observed in individual tubules. The distal segment makes a significant contribution to the total ion output of the tubule. The cyclic AMP content of tubule segments treated with corpora cardiaca extract was found to increase in both main and distal segments. When expressed in terms of protein content there was no difference between segments. However, in terms of total cell volume, the cells of the distal segment had a tenfold greater cyclic AMP content than those of the main segment. This is consistent with a 10- to 20-fold higher secretion rate of K by the distal segment. It is suggested that the distal segment, whilst having a higher length-specific fluid secretion rate than the main segment, is, nevertheless, concerned primarily with ion and solute secretion since it is unresponsive to diuretic hormone. The prime role of the main segment, which does respond to diuretic hormone, is fluid secretion. There appear to be major differences in hydraulic conductivity between the two segments.

Freezing survival by isolated Malpighian tubules of the New Zealand alpine weta Hemideina maori.

1998 ◽  
Vol 201 (2) ◽  
pp. 227-236 ◽  
Author(s):  
D S Neufeld ◽  
L P Leader
Keyword(s):  
New Zealand ◽  
Membrane Potential ◽  
Basolateral Membrane ◽  
Fluid Secretion ◽  
Malpighian Tubules ◽  
High Concentration ◽  
Freeze Thaw ◽  
Thaw Cycle ◽  
Freeze Thaw Cycle ◽  
High Concentrations

The ability of isolated Malpighian tubules from a freeze-tolerant insect, the New Zealand alpine weta (Hemideina maori), to withstand freezing was assessed by measuring post-freeze membrane potentials and rates of fluid secretion. The hemolymph of cold-acclimated Hemideina maori was found to contain relatively high concentrations of the cryoprotectants trehalose (>300 mmol l-1) and proline (41 mmol l-1). Survival of isolated Malpighian tubules was correspondingly high when a high concentration of trehalose was present in the bathing saline. Tubules allowed to recover for 20 min from a 1 h freeze to -5 degrees C in saline containing 400 mmol l-1 trehalose had a basolateral membrane potential of -53 mV compared with a potential of -63 mV in tubules not exposed to a freeze/thaw cycle. Fluid secretion in tubules that had experienced a freeze/thaw cycle in saline containing 400 mmol l-1 trehalose was 9.9+/-2.6 nl h-1 compared with 18.7+/-5.0 nl h-1 (means +/- s.e.m., N=18) in tubules that had not been frozen. Tubules frozen in saline containing a lower concentration of trehalose (200 mmol l-1) or in glucose (400 mmol l-1) showed a similar ability to survive freezing to -5 degrees C. In contrast, freezing for 1 h at -5 degrees C in saline containing 400 mmol l-1 sucrose produced a 57 % decrease in membrane potential and an 88 % decrease in secretion rate. Tubules held in saline lacking high concentrations of sugars showed no survival after freezing to -5 degrees C for 1 h. When frozen to -15 degrees C, tubules appeared to survive best in saline with the highest trehalose concentration (400 mmol l-1). Freezing damage was not simply the result of exposure to cold, since tubules chilled (unfrozen) to -5 degrees C for 1 h were not compromised even when the bathing saline lacked a high sugar concentration. Exposure of tubules to a combination of low temperature and high osmolality mimicked damage caused by actual freezing: the membrane potential showed a 60 % recovery when the test was performed in saline containing trehalose, but showed no recovery in saline containing sucrose.

scholarly journals When two cells are better than one: specialized stellate cells provide a privileged route for uniquely rapid water flux in Drosophila renal tubule

2019 ◽  
Author(s):  
Pablo Cabrero ◽  
Selim Terhzaz ◽  
Anthony J. Dornan ◽  
Saurav Ghimire ◽  
Heather L. Holmes ◽  
...  
Keyword(s):  
Water Permeability ◽  
Plasma Membranes ◽  
Water Flux ◽  
Malpighian Tubule ◽  
Stellate Cells ◽  
Fluid Secretion ◽  
Malpighian Tubules ◽  
Principal Cells ◽  
Very High

AbstractInsects are highly successful, in part through an excellent ability to osmoregulate. The renal (Malpighian) tubules can secrete fluid faster on a per-cell basis than any other epithelium, but the route for these remarkable water fluxes has not been established. In Drosophila melanogaster, we show that 4 members of the Major Intrinsic Protein family are expressed at very high level in the fly renal tissue; the aquaporins Drip and Prip, and the aquaglyceroporins Eglp2 and Eglp4. As predicted from their structure and by their transport function by expressing these proteins in Xenopus oocytes, Drip, Prip and Eglp2 show significant and specific water permeability, whereas Eglp2 and Eglp4 show very high permeability to glycerol and urea. Knockdowns of any of these genes impacts tubule performance resulting in impaired hormone-induced fluid secretion. The Drosophila tubule has two main secretory cell types: active cation-transporting principal cells with the aquaglyceroporins localize to opposite plasma membranes and small stellate cells, the site of the chloride shunt conductance, with these aquaporins localising to opposite plasma membranes. This suggests a model in which cations are pumped by the principal cells, causing chloride to follow through the stellate cells in order to balance the charge. As a consequence, osmotically obliged water follows through the stellate cells. Consistent with this model, fluorescently labelled dextran, an in vivo marker of membrane water permeability, is trapped in the basal infoldings of the stellate cells after kinin diuretic peptide stimulation, confirming that these cells provide the major route for transepithelial water flux. The spatial segregation of these components of epithelial water transport may help to explain the unique success of the higher insects.Significance statementThe tiny insect renal (Malpighian) tubule can transport fluid at unparalleled speed, suggesting unique specialisations. Here we show that strategic allocation of Major Intrinsic Proteins (MIPs) to specific cells within the polarized tubule allow the separation of metabolically intense active cation transport from chloride and water conductance. This body plan is general to at least many higher insects, providing a clue to the unique success of the class Insecta.

scholarly journals Specialized stellate cells offer a privileged route for rapid water flux in Drosophila renal tubule

2020 ◽  
Vol 117 (3) ◽  
pp. 1779-1787 ◽  
Author(s):  
Pablo Cabrero ◽  
Selim Terhzaz ◽  
Anthony J. Dornan ◽  
Saurav Ghimire ◽  
Heather L. Holmes ◽  
...  
Keyword(s):  
Water Permeability ◽  
Plasma Membranes ◽  
Water Flux ◽  
Stellate Cells ◽  
Fluid Secretion ◽  
Cell Types ◽  
Renal Tissue ◽  
Malpighian Tubules ◽  
Very High

Insects are highly successful, in part through an excellent ability to osmoregulate. The renal (Malpighian) tubules can secrete fluid faster on a per-cell basis than any other epithelium, but the route for these remarkable water fluxes has not been established. In Drosophila melanogaster, we show that 4 genes of the major intrinsic protein family are expressed at a very high level in the fly renal tissue: the aquaporins (AQPs) Drip and Prip and the aquaglyceroporins Eglp2 and Eglp4. As predicted from their structure, and by their transport function by expressing these proteins in Xenopus oocytes, Drip, Prip, and Eglp2 show significant and specific water permeability, whereas Eglp2 and Eglp4 show very high permeability to glycerol and urea. Knockdowns of any of these genes result in impaired hormone-induced fluid secretion. The Drosophila tubule has 2 main secretory cell types: active cation-transporting principal cells, wherein the aquaglyceroporins localize to opposite plasma membranes, and small stellate cells, the site of the chloride shunt conductance, with these AQPs localizing to opposite plasma membranes. This suggests a model in which osmotically obliged water flows through the stellate cells. Consistent with this model, fluorescently labeled dextran, an in vivo marker of membrane water permeability, is trapped in the basal infoldings of the stellate cells after kinin diuretic peptide stimulation, confirming that these cells provide the major route for transepithelial water flux. The spatial segregation of these components of epithelial water transport may help to explain the unique success of the higher insects in regulating their internal environments.

Stereological Analyses of Hepatocyte Ultrastructure Monitor Arsenic Liver Burdens

1980 ◽  
Vol 38 ◽  
pp. 442-443
Author(s):  
E. M. B. Sorensen ◽  
R. R. Mitchell ◽  
L. L. Graham
Keyword(s):  
Drainage System ◽  
Water Levels ◽  
Ultrastructural Changes ◽  
Hepatocyte Ultrastructure ◽  
Lepomis Cyanellus ◽  
Large Numbers ◽  
Activation Data ◽  
Water Analyses ◽  
Municipal Lake ◽  
Green Sunfish

Endemic freshwater teleosts were collected from a portion of the Navosota River drainage system which had been inadvertently contaminated with arsenic wastes from a firm manufacturing arsenical pesticides and herbicides. At the time of collection these fish were exposed to a concentration of 13.6 ppm arsenic in the water; levels ranged from 1.0 to 20.0 ppm during the four-month period prior. Scale annuli counts and prior water analyses indicated that these fish had been exposed for a lifetime. Neutron activation data showed that Lepomis cyanellus (green sunfish) had accumulated from 6.1 to 64.2 ppm arsenic in the liver, which is the major detoxification organ in arsenic poisoning. Examination of livers for ultrastructural changes revealed the presence of electron dense bodies and large numbers of autophagic vacuoles (AV) and necrotic bodies (NB) (1), as previously observed in this same species following laboratory exposures to sodium arsenate (2). In addition, abnormal lysosomes (AL), necrotic areas (NA), proliferated rough endoplasmic reticulum (RER), and fibrous bodies (FB) were observed. In order to assess whether the extent of these cellular changes was related to the concentration of arsenic in the liver, stereological measurements of the volume and surface densities of changes were compared with levels of arsenic in the livers of fish from both Municipal Lake and an area known to contain no detectable level of arsenic.

The metachor as a characteristic of the association of electrolytes in aqueous solutions

1984 ◽  
Vol 49 (5) ◽  
pp. 1061-1078 ◽  
Author(s):  
Jiří Čeleda ◽  
Stanislav Škramovský
Keyword(s):  
Aqueous Solutions ◽  
Apparent Volume ◽  
Solutions Of Electrolytes ◽  
Molal Volumes ◽  
The Difference ◽  
High Concentrations ◽  
Experimental Values ◽  
Suitable Characteristic ◽  
Association Of Ions ◽  
Very High

Based on the earlier paper introducing a concept of the apparent parachor of a solute in the solution, we have eliminated in the present work algebraically the effect which is introduced into this quantity by the additivity of the apparent molal volumes. The difference remaining from the apparent parachor after substracting the contribution corresponding to the apparent volume ( for which the present authors suggest the name metachor) was evaluated from the experimental values of the surface tension of aqueous solutions for a set of 1,1-, 1,2- and 2,1-valent electrolytes. This difference showed to be independent of concentration up to the very high values of the order of units mol dm-3 but it was directly proportional to the number of the free charges (with a proportionality factor 5 ± 1 cm3 mol-1 identical for all studied electrolytes). The metachor can be, for this reason, a suitable characteristic for detection of the association of ions and formation of complexes in the solutions of electrolytes, up to high concentrations where other methods are failing.

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