Phosphate transport by locust rectum in vitro

1980 ◽  
Vol 58 (9) ◽  
pp. 1518-1523 ◽  
Author(s):  
E. W. Andrusiak ◽  
J. E. Phillips ◽  
J. Speight

The rectal cuticle is permeable to H2PO4−, but much less so to HPO42−. Everted rectal sacs of Schistocerca gregaria transport PO4 from lumen to hemocoel side against large concentration and electrical differences. This active process is not caused by solvent drag and it obeys Michaelis–Menten kinetics. Entry of 32PO4 into rectal tissue from the lumen is inhibited by arsenate. Much of the 32PO4 is converted to organic forms in the tissue but these do not enter the hemocoel compartment. Net rates of PO4 movement across the rectal wall in vitro are high enough to explain recovery of phosphate secreted in situ by Malpighian tubules of starved locusts. The location and possible mechanism of PO4 transport are discussed.

1964 ◽  
Vol 41 (1) ◽  
pp. 15-38
Author(s):  
I. WATER ◽  
J. E. PHILLIPS

1. The histology of the rectum of Schistocerca gregaria is described. 2. A method is described whereby net absorption of water from the rectum (in situ, isolated by ligation) can be measured. 3. Water is actively absorbed from the lumen of the rectum against an osmotic gradient and in the absence of a significant net flux of solute. 4. The maximum osmotic gradient developed is 2-3 times greater in locusts supplied with hypertonic saline than in locusts supplied with tap water. This indicates some ability to regulate water absorption in relation to requirement. 5. The ionic concentration of rectal fluid and the rate of salt absorption from the lumen have little effect on the maximum osmotic gradient developed across the rectal wall. 6. Possible mechanisms of active absorption of water are discussed.


1991 ◽  
Vol 159 (1) ◽  
pp. 433-447 ◽  
Author(s):  
ANDREW P. STAGG ◽  
JON F. HARRISON ◽  
JOHN E. PHILLIPS

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.


1997 ◽  
Vol 200 (11) ◽  
pp. 1627-1638 ◽  
Author(s):  
KA Collier ◽  
MJ O'Donnell

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.


1999 ◽  
Vol 202 (22) ◽  
pp. 3195-3203 ◽  
Author(s):  
G.M. Coast ◽  
J. Meredith ◽  
J.E. Phillips

The major stimulant of ileal fluid reabsorption in Locusta migratoria and Schistocerca gregaria corpora cardiaca, ion-transport peptide (ITP), had no stimulatory action on fluid secretion by isolated Malpighian tubules of S. gregaria, nor did it have a synergistic or antagonistic effect in combination with locustakinin (Lom-K) or Locusta-diuretic hormone (Locusta-DH). Stimulants of locust Malpighian tubules (Lom-K and Locusta-DH) had no action on either active transport of Cl(−) (measured as short-circuit current, I(sc)) or the rate of fluid reabsorption across S. gregaria ilea and recta in vitro. Thus, hormonal control of these major organs of the excretory system appears to be clearly separated. Lom-K and Locusta-DH acted synergistically to stimulate secretion by S. gregaria Malpighian tubules, and the diuretic response was more rapid than the response of the ileum and rectum to hindgut stimulants. Taken together, these data suggest that, in the initial phase of post-prandial diuresis, urine flow will exceed fluid uptake in the hindgut, thereby allowing excess water to be eliminated.


Author(s):  
J. P. Revel

Movement of individual cells or of cell sheets and complex patterns of folding play a prominent role in the early developmental stages of the embryo. Our understanding of these processes is based on three- dimensional reconstructions laboriously prepared from serial sections, and from autoradiographic and other studies. Many concepts have also evolved from extrapolation of investigations of cell movement carried out in vitro. The scanning electron microscope now allows us to examine some of these events in situ. It is possible to prepare dissections of embryos and even of tissues of adult animals which reveal existing relationships between various structures more readily than used to be possible vithout an SEM.


Author(s):  
D. Reis ◽  
B. Vian ◽  
J. C. Roland

Wall morphogenesis in higher plants is a problem still open to controversy. Until now the possibility of a transmembrane control and the involvement of microtubules were mostly envisaged. Self-assembly processes have been observed in the case of walls of Chlamydomonas and bacteria. Spontaneous gelling interactions between xanthan and galactomannan from Ceratonia have been analyzed very recently. The present work provides indications that some processes of spontaneous aggregation could occur in higher plants during the formation and expansion of cell wall.Observations were performed on hypocotyl of mung bean (Phaseolus aureus) for which growth characteristics and wall composition have been previously defined.In situ, the walls of actively growing cells (primary walls) show an ordered three-dimensional organization (fig. 1). The wall is typically polylamellate with multifibrillar layers alternately transverse and longitudinal. Between these layers intermediate strata exist in which the orientation of microfibrils progressively rotates. Thus a progressive change in the morphogenetic activity occurs.


Author(s):  
C. Jennermann ◽  
S. A. Kliewer ◽  
D. C. Morris

Peroxisome proliferator-activated receptor gamma (PPARg) is a member of the nuclear hormone receptor superfamily and has been shown in vitro to regulate genes involved in lipid metabolism and adipocyte differentiation. By Northern analysis, we and other researchers have shown that expression of this receptor predominates in adipose tissue in adult mice, and appears first in whole-embryo mRNA at 13.5 days postconception. In situ hybridization was used to find out in which developing tissues PPARg is specifically expressed.Digoxigenin-labeled riboprobes were generated using the Genius™ 4 RNA Labeling Kit from Boehringer Mannheim. Full length PPAR gamma, obtained by PCR from mouse liver cDNA, was inserted into pBluescript SK and used as template for the transcription reaction. Probes of average size 200 base pairs were made by partial alkaline hydrolysis of the full length transcripts. The in situ hybridization assays were performed as described previously with some modifications. Frozen sections (10 μm thick) of day 18 mouse embryos were cut, fixed with 4% paraformaldehyde and acetylated with 0.25% acetic anhydride in 1.0M triethanolamine buffer. The sections were incubated for 2 hours at room temperature in pre-hybridization buffer, and were then hybridized with a probe concentration of 200μg per ml at 70° C, overnight in a humidified chamber. Following stringent washes in SSC buffers, the immunological detection steps were performed at room temperature. The alkaline phosphatase labeled, anti-digoxigenin antibody and detection buffers were purchased from Boehringer Mannheim. The sections were treated with a blocking buffer for one hour and incubated with antibody solution at a 1:5000 dilution for 2 hours, both at room temperature. Colored precipitate was formed by exposure to the alkaline phosphatase substrate nitrobluetetrazoliumchloride/ bromo-chloroindlylphosphate.


2018 ◽  
Vol 12 (7-8) ◽  
pp. 38-45
Author(s):  
A. N. EFREMOV ◽  
N. V. PLIKINA ◽  
T. ABELI

Rare species are most vulnerable to man-made impacts, due to their biological characteristics or natural resource management. As a rule, the economic impact is associated with the destruction and damage of individual organisms, the destruction or alienation of habitats. Unfortunately, the conservation of habitat integrity is an important protection strategy, which is not always achievable in the implementation of industrial and infrastructural projects. The aim of the publication is to summarize the experience in the field of protection of rare species in the natural habitat (in situ), to evaluate and analyze the possibility of using existing methods in design and survey activities. In this regard, the main methodological approaches to the protection of rare species in the natural habitat (in situ) during the proposed economic activity were reflected. The algorithm suggested by the authors for implementing the in situ project should include a preparatory stage (initial data collection, preliminary risk assessments, technology development, obtaining permitting documentation), the main stage, the content of which is determined by the selected technology and a long monitoring stage, which makes it possible to assess the effectiveness of the taken measures. Among the main risks of in situ technology implementation, the following can be noted: the limited resources of the population that do not allow for the implementation of the procedure without prior reproduction of individuals in situ (in vitro); limited knowledge of the biology of the species; the possibility of invasion; the possibility of crossing for closely related species that сo-exist in the same habitat; social risks and consequences, target species or population may be important for the local population; financial risks during the recovery of the population. The available experience makes it possible to consider the approach to the conservation of rare species in situ as the best available technology that contributes to reducing negative environmental risks.


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