scholarly journals Active transport of brilliant blue FCF across the Drosophila midgut and Malpighian tubule epithelia

2019 ◽  
Author(s):  
Dawson B.H. Livingston ◽  
Hirva Patel ◽  
Andrew Donini ◽  
Heath A. MacMillan
Keyword(s):  
Traumatic Injury ◽  
Malpighian Tubule ◽  
Malpighian Tubules ◽  
Brilliant Blue ◽  
Barrier Dysfunction ◽  
Concentration Gradients ◽  
Barrier Disruption ◽  
Brilliant Blue Fcf ◽  
Summary Statement

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

Anti-diuretic activity of a CAPA neuropeptide can compromise Drosophila chill tolerance

2018 ◽  
Author(s):  
Heath A. MacMillan ◽  
Basma Nazal ◽  
Sahr Wali ◽  
Gil Y. Yerushalmi ◽  
Lidiya Misyura ◽  
...  
Keyword(s):  
Ion Transport ◽  
Cold Tolerance ◽  
Malpighian Tubule ◽  
Malpighian Tubules ◽  
Low Doses ◽  
Ion Balance ◽  
The Central Nervous System ◽  
Summary Statement ◽  
Cell Depolarization ◽  
High Doses

AbstractFor insects, chilling injuries that occur in the absence of freezing are often related to a systemic loss of ion and water balance that leads to extracellular hyperkalemia, cell depolarization, and the triggering of apoptotic signalling cascades. The ability of insect ionoregulatory organs (e.g. the Malpighian tubules) to maintain ion balance in the cold has been linked to improved chill tolerance, and many neuroendocrine factors are known to influence ion transport rates of these organs. Injection of micromolar doses of CAPA (an insect neuropeptide) have been previously demonstrated to improve Drosophila cold tolerance, but the mechanisms through which it impacts chill tolerance are unclear, and low doses of CAPA have been demonstrated to cause anti-diuresis in other insects, including dipterans. Here, we provide evidence that low (fM) and high (µM) doses of CAPA impair and improve chill tolerance, respectively, via two different effects on Malpighian tubule ion and water transport. While low doses of CAPA are anti-diuretic, reduce tubule K+ clearance rates and reduce chill tolerance, high doses facilitate K+ clearance from the haemolymph and increase chill tolerance. By quantifying CAPA peptide levels in the central nervous system, we estimated the maximum achievable hormonal titres of CAPA, and found evidence to suggest that CAPA may function as an anti-diuretic peptide in Drosophila. We provide the first evidence of a neuropeptide that can negatively affect cold tolerance in an insect, and the first evidence of CAPA as an anti-diuretic peptide in this ubiquitous insect model.Summary StatementMany insects ion balance in the cold. We show how one neuropeptide can slow ion transport and reduce the cold tolerance of a fly.

Acid-Base Variables in Malpighian Tubule Secretion and Response to Acidosis

1991 ◽  
Vol 159 (1) ◽  
pp. 433-447 ◽  
Author(s):  
ANDREW P. STAGG ◽  
JON F. HARRISON ◽  
JOHN E. PHILLIPS
Keyword(s):  
Schistocerca Gregaria ◽  
Malpighian Tubule ◽  
Malpighian Tubules ◽  
Acid Base ◽  
Acid Injection ◽  
Before And After ◽  
Acid Load ◽  
Acid Base Status ◽  
Tubule Fluid

1. Malpighian tubule fluid from Schistocerca gregaria adults, starved for 1 day, was collected in situ by cannulation of the gut, both before and after injecting 10 μmol of HCl or NaCl into the haemocoel. 2. Haemolymph pH at the neck remained depressed by 0.3 units for at least 6 h in HCl- as compared to NaCl-injected locusts. A lower haemolymph pH persisted near the acid injection site for several hours. 3. The pH of tubule fluid remained about 0.5 units more acid than haemolymph under all conditions. Thus, net tubular acid secretion was proportional to haemolymph acid-base status. 4. The greater acidity of tubular fluid after acid injection was associated with lower estimated bicarbonate concentrations and higher Pcoco2 without any change in total CO2 when compared to controls. 5. The combined contribution of bicarbonate, phosphate and urate to total buffering capacity of tubular fluid was estimated to be 75°, with bicarbonate responsible for 55° of the total. 6. The maximum rate of acid removal by all Malpighian tubules of starved locusts, including H+ trapped in ammonium ions, was calculated to be very small in relation to the acid load injected into the haemocoel Note: To whom reprint requests should be directed.

Analysis of epithelial transport by measurement of K+, Cl- and pH gradients in extracellular unstirred layers: ion secretion and reabsorption by Malpighian tubules of Rhodnius prolixus

1997 ◽  
Vol 200 (11) ◽  
pp. 1627-1638 ◽  
Author(s):  
KA Collier ◽  
MJ O'Donnell
Keyword(s):  
Epithelial Transport ◽  
Malpighian Tubule ◽  
Fluid Secretion ◽  
Rhodnius Prolixus ◽  
Malpighian Tubules ◽  
Unstirred Layer ◽  
Ph Gradients ◽  
In Situ Preparation

Summary The pH and concentrations of K+ and Cl- in the unstirred layer (USL) associated with the basolateral surfaces of the upper and lower Malpighian tubules of Rhodnius prolixus were measured using extracellular ion-selective microelectrodes. When stimulated with 5-hydroxytryptamine (5-HT) in vitro, the upper Malpighian tubule secretes Na+, K+, Cl- and water at high rates; the lower Malpighian tubule reabsorbs K+ and Cl- but not water. Concentrations of K+ and Cl- in the unstirred layer of the lower Malpighian tubule ([K+]USL, [Cl-]USL) were greater than those in the bathing saline, consistent with the accumulation of K+ and Cl- in the USL during 5-HT-stimulated KCl reabsorption. [K+]USL exceeded [K+]Bath by as much as 5.3-fold. Calculations of K+ flux based on measurements of [K+]USL at various distances from the tubule surface agreed well with flux calculated from the rate of fluid secretion and the change in K+ concentration of the secreted fluid during passage through the lower tubule. Concentrations of K+ in the unstirred layer of the upper Malpighian tubule were reduced relative to those in the bathing saline, consistent with depletion of K+ from the USL during 5-HT-stimulated secretion of K+ from bath to lumen. Changes in [K+]USL during 5-HT-stimulated K+ secretion from single upper Malpighian tubule cells could be resolved. Although differences between [K+]USL and [K+]Bath were apparent for upper and lower tubules in an in situ preparation, they were reduced relative to the differences measured using isolated tubules. We suggest that convective mixing of the fluids around the tubules by contractions of the midgut and hindgut reduces, but does not eliminate, differences between [K+]USL and [K+]Bath in situ. The USL was slightly acidic relative to the bath in 5-HT-stimulated upper and lower tubules; contributions to USL acidification are discussed. The results also show that the techniques described in this paper can resolve rapid and localized changes in ion transport across different regions of Malpighian tubules in response to stimulants or inhibitors of specific membrane transporters.

Fine Structure of the Malpighian Tubules of the Mosquito, Culex pipiens

1971 ◽  
Vol 29 ◽  
pp. 402-403
Author(s):  
Brendan Clifford
Keyword(s):  
Fine Structure ◽  
Culex Pipiens ◽  
Stellate Cell ◽  
Malpighian Tubule ◽  
Cell Types ◽  
Instar Larva ◽  
Malpighian Tubules ◽  
Calliphora Erythrocephala ◽  
Distinct Cell ◽  
Basal Plasma

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.

scholarly journals Pectin limits epithelial barrier disruption by Citrobacter rodentium through anti-microbial effects

2021 ◽  
Author(s):  
M. Beukema ◽  
K. Ishisono ◽  
J. de Waard ◽  
M. M. Faas ◽  
P. de Vos ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Epithelial Barrier ◽  
Citrobacter Rodentium ◽  
Barrier Dysfunction ◽  
Antimicrobial Effects ◽  
Barrier Disruption ◽  
Microbial Effects

Pectins inhibit the growth of C. rodentium in vitro, preventing attachment of C. rodentium to CMT93 epithelial cells. Through these antimicrobial effects, pectins protect the epithelium from C. rodentium-induced barrier dysfunction and damage.

Evaluating Differences in Transport Behavior of Sodium Chloride and Brilliant Blue FCF in Sand Columns

2015 ◽  
Vol 109 (3) ◽  
pp. 765-779 ◽  
Author(s):  
Jiazhong Qian ◽  
Yanan Wu ◽  
Yong Zhang ◽  
Yong Liu ◽  
Yuehan Lu ◽  
...  
Keyword(s):  
Sodium Chloride ◽  
Brilliant Blue ◽  
Transport Behavior ◽  
Brilliant Blue Fcf

Anti-diuresis in the blood-feeding insect Rhodnius prolixus Stål: the peptide CAP2b and cyclic GMP inhibit Malpighian tubule fluid secretion.

1997 ◽  
Vol 200 (17) ◽  
pp. 2363-2367 ◽  
Author(s):  
M C Quinlan ◽  
N J Tublitz ◽  
M J O'Donnell
Keyword(s):  
Cyclic Gmp ◽  
Physiological Role ◽  
Malpighian Tubule ◽  
Fluid Secretion ◽  
Blood Feeding ◽  
Rhodnius Prolixus ◽  
Malpighian Tubules ◽  
Tubule Fluid ◽  
Secretion Rates

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.

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