Studies on Locust Rectum: II. Identification of Specific Ion Transport Processes Regulated by Corpora Cardiaca and Cyclic-AMP

1980 ◽  
Vol 86 (1) ◽  
pp. 225-236
Author(s):  
J. H. SPRING ◽  
J. E. PHILLIPS

Please send reprint requests to J. E. Phillips. 1. The unidirectional fluxes of 36Cl− and 22Na+ across short-circuited locust recta bathed in a simple NaCl saline were followed with time. Unidirectional fluxes and net flux of 22Na+ to the haemocoel side all remained constant for at least 4 h and were unaffected by either corpora cardiaca homogenate (CC) or cAMP. 2. Both CC and cAMP stimulated influx and net flux of 36Cl− to the haemocoel side. Over the whole time course of the experiment, i.e. both before and after stimulation, net Cl− flux approximately equalled the shortcircuit current (ISC). 3. Neither CC nor cAMP caused substantial stimulation of ISC or transepithelial electropotential difference (PD) if all Cl− in the bathing saline was replaced by either sulphate or nitrate or acetate. 4. Acetate saline sustains ISC, PD and transepithalial resistance (R) at higher levels than does simple Cl-saline. 5. Experiments with Cl-free, SO4-salines suggest that alternate electrogenic transport processes can be slowly turned on when Cl− is absent, provided a complex saline which contains several organic constituents, or simple acetate saline, is present.

1997 ◽  
Vol 272 (2) ◽  
pp. R640-R647 ◽  
Author(s):  
O. A. Candia ◽  
T. Yorio

The amphibian skin represents an important organ for osmoregulation and, like the mammalian kidney, maintains acid-base balance by secreting protons or base. However, the lack of a reliable and accurate method to measure the contribution of unidirectional fluxes of HCO3- ions to this mechanism has been an obstacle for the determination of the role of bicarbonate in epithelial acid-base homeostasis. Recently, one of us developed a method that allows for the reliable determination of transepithelial fluxes of bicarbonate, and this method was applied to determine unidirectional fluxes of (14)CO2 and H(14)CO3 under a variety of conditions. We report that the combined CO2 and HCO3- mucosal-to-serosal flux under 5% CO2 was 40% larger than the opposing flux, giving a net flux in the mucosal-to-serosal direction. This net flux was inhibited by acetazolamide. In CO2-free conditions, there was no detectable net flux; however, acetazolamide and PGF(2alpha) attenuated the mucosal-to-serosal flux and established an apparent secretion of HCO3-. A model is presented that depicts twelve vectors or components to the CO2 plus HCO3- fluxes in the frog skin. This model can accurately reproduce the experimental values measured from unidirectional fluxes of CO2 and HCO3- under a variety of conditions and can explain the effects of PGF(2alpha) on unidirectional 14C-labeled fluxes as a consequence of inhibition of H+ secretion to the apical bath, similar to what was previously suggested by our laboratory using a different methodological approach. The present method, utilizing radiolabeled HCO3-, may be useful as a means to evaluate the mechanism of action of hormones and drugs that may regulate acid-base homeostasis by altering proton and bicarbonate transport processes.


1994 ◽  
Vol 188 (1) ◽  
pp. 159-174 ◽  
Author(s):  
S Riestenpatt ◽  
W Zeiske ◽  
H Onken

Split gill lamellae (epithelium plus cuticle) of hyperregulating Chinese crabs acclimated to fresh water were mounted in a modified Ussing chamber. Active and electrogenic absorption of sodium and chloride were measured as positive amiloride-sensitive and negative Cl--dependent short-circuit currents (INa, ICl), respectively. Both currents were characterized before and after treatment of the tissue with theophylline or dibutyryl cyclic AMP. Both drugs increased INa and ICl. A simple circuit analysis showed that INa stimulation reflected a marked increase in the transcellular Na+ conductance, whereas the respective electromotive force was unchanged. The Michaelis constant (KNa) for Na+ current saturation was decreased after INa stimulation, indicating an increased affinity of the transport mechanism for its substrate. Consequently, the affinity for the Na+ channel blocker amiloride decreased as expected for a competitive interaction between substrate and inhibitor. Analysis of the amiloride-induced current-noise revealed a marked increase in the number of apical Na+ channels after INa stimulation with theophylline, whereas there was little change in the single-channel current. Stimulation of Cl- absorption was accompanied by a substantial increase in both transcellular conductance and electromotive force, indicating an activation of the apical H+ pump that provides the driving force for active Cl- uptake via apical Cl-/HCO3- exchange and basolateral Cl- channels.


1991 ◽  
Vol 155 (1) ◽  
pp. 455-467
Author(s):  
R. BRENT THOMSON ◽  
N. AUDSLEY ◽  
JOHN E. PHILLIPS

The commonly used method of passing short-circuit current (Isc) across insect epithelia through Ag-AgCl electrodes, without the use of salt bridges, leads to significant OH− production at the cathode (lumen side) when high currents are applied. The alkalization of the lumen previously reported when cyclic AMP was added to short-circuited locust hindgut is a result of this phenomenon rather than cyclic-AMP-mediated stimulation of acid-base transport in the hindgut. When salt bridges are used to pass short-circuit current across locust hindgut, acid secretion (JH) into the lumen equals alkaline movement (JOH) to the haemocoel side, and JH is similar under both open- and short-circuit conditions. JH is similar (1.5 μequiv cm−2 h−1) in recta and ilea. Addition of cyclic AMP inhibits JH across the rectum by 42–66%, but has no effect on the ileum when salt bridges are used. Electrical parameters (Isc, Vt, Rt) reflecting hindgut Cl− transport (JCL) before and after stimulation with cyclic AMP are the same whether or not salt bridges are used. We found no evidence of any coupling between JCl and JH/JOH.


1982 ◽  
Vol 101 (1) ◽  
pp. 233-254
Author(s):  
D.F. HOULIHAN ◽  
D. SELL

The oxygen consumption of excised abdomens of cockroaches and locusts has been measured before and after the injection of fluids into the ligated recta. Fluid injection caused a transient stimulation of oxygen consumption of up to 30% of the resting rate. The extra amount of oxygen consumed is positively correlated with the osmolality of the fluid injected and the amount of fluid absorbed. Parallel experiments were carried out on the time course of fluid uptake; these experiments revealed a correlation first between a rapid increase in fluid absorption and stimulation of oxygen consumption, and secondly between the final resting rate of oxygen consumption and a slower absorption of fluid. Locusts take up fluid at double the rate of cockroaches and have double the stimulation in oxygen consumption following fluid injection. In locusts the increases in oxygen consumption can also be correlated with the net movement of Na+, K+and Cl− from the rectum. The stimulation of oxygen consumption during fluid uptake is discussed in relation to the local osmosis model for fluid uptake.


1993 ◽  
Vol 292 (1) ◽  
pp. 303-308 ◽  
Author(s):  
S Pavelka ◽  
J Hermanská ◽  
M Baudysová ◽  
J Houstĕk

Iodothyronine 5′-deiodinase (5′D) of mouse brown adipocytes differentiated in cell culture was characterized in detail with respect to the adrenergic control of its biosynthesis. The stimulation of 5′D required mRNA and protein synthesis and was dependent on the stage of differentiation of the cells. The maximum induction was observed around confluence (7-day-old cells), in pre- and post-confluent cells the 5′D activity was significantly less induced. The transient responsiveness of brown fat-cells to the stimulatory effect of adrenergic agents was reflected also in the time course of the induction of 5′D by different concentrations of agonists. The maximum response occurred regularly after an 8 h incubation and implicated a rather fast turnover of the induced enzyme. On the basis of the inhibitory effects of cycloheximide and actinomycin D, the half-life of the induced 5′D and its mRNA were estimated to be 1.5 and 3.3 h respectively. The noradrenaline-induced 5′D activity was shown to be that of the type II enzyme, insensitive to propylthiouracil (PTU). The estimated values of its apparent Km for thyroxine, Km for the co-substrate dithiothreitol, and Vmax. in the presence of 1 mM PTU were 2 nM, 2.6 mM, and 0.1 pmol of I-/h per mg of protein respectively. The 5′D activity was effectively induced by forskolin and dibutyryl cyclic AMP, as well as by isoprenaline, noradrenaline and CGP-12177, but not by phenylephrine, cirazoline or oxymetazoline. This indicates that, contrary to previous observations in vivo, stimulation of 5′D in cultured brown fat-cells involves elevated cyclic AMP levels and is mediated predominantly via beta-receptors, particularly via the so-called beta 3-adrenoceptors.


1971 ◽  
Vol 125 (1) ◽  
pp. 329-342 ◽  
Author(s):  
Radhey L. Singhal ◽  
M. R. Parulekar ◽  
R. Vijayvargiya ◽  
G. Alan Robison

1. The ability of exogenously administered cyclic AMP (adenosine 3′:5′-monophosphate) to exert andromimetic action on certain carbohydrate-metabolizing enzymes was investigated in the rat prostate gland and seminal vesicles. 2. Cyclic AMP, when injected concurrently with theophylline, produced marked increases in hexokinase, phosphofructokinase, glyceraldehyde phosphate dehydrogenase, pyruvate kinase, and two hexose monophosphate-shunt enzymes, as well as α-glycerophosphate dehydrogenase activity in accessory sexual tissues of castrated rats. The 6-N,2′-O-dibutyryl analogue of cyclic AMP caused increases of enzyme activity that were greater than those induced by the parent compound. 3. Time-course studies demonstrated that, whereas significant increases in the activities of most enzymes occurred within 4h after the injection of cyclic AMP, maximal increases were attained at 16–24h. 4. Increase in the activity of the various prostatic and vesicular enzymes was dependent on the dose of cyclic AMP; in most instances, 2.5mg of the cyclic nucleotide/rat was sufficient to elicit a statistically significant response. 5. Administration of cyclic AMP and theophylline also produced stimulation of enzyme activities in secondary sexual tissues of immature rats. 6. Cyclic AMP and theophylline did not affect significantly any of the enzymes studied in hepatic tissue. 7. Stimulation of various carbohydrate-metabolizing enzymes in the prostate gland and seminal vesicles by cyclic AMP was independent of adrenal function. 8. Concurrent treatment with actinomycin or cycloheximide prevented the cyclic AMP- and theophylline-induced increases in enzyme activities in both castrated and adrenalectomized–castrated animals. 9. Administration of a single dose of testosterone propionate (5.0mg/100g) to castrated rats caused a significant increase in cyclic AMP concentration in both accessory sexual tissues. 10. In addition, treatment with theophylline potentiated the effects of a submaximal dose of testosterone (1.0mg/100g) on all those prostatic and seminal-vesicular enzymes that are increased by exogenous cyclic AMP. 11. The evidence indicates that cyclic AMP may be involved in triggering the known metabolic actions of androgens on secondary sexual tissues of the rat.


1978 ◽  
Vol 234 (5) ◽  
pp. C139-C145 ◽  
Author(s):  
R. C. Deth

The ability of lanthanum (La3+) to block calcium efflux from smooth muscle cells of the rabbit aorta has been examined. La3+ promotes the very early phase of 45Ca efflux, which is extracellular in origin, and partially inhibits the latter, cellular portion. Stimulation of 45Ca efflux caused by the release of intracellular 45Ca with either 10(-4) M dinitrophenol or 10 mM caffeine was not reduced by pretreatment with 10 mM La for 40 min, whereas stimulation due to norepinephrine was abolished. It was concluded that during the use of the "La method" for measuring cellular 45Ca there is an underestimation due to unblocked 45Ca loss. This loss can be reduced by processing tissue at 2 degrees C, which inhibits transport processes. The time course of 45Ca uptake and the stimulation of uptake by high K+ are qualitatively but not quantitatively similar if tissues are washed at 37 and 2 degrees C. Tissues washed in La3+ at 2 degrees C for 60 min retain approximately double the cellular 45Ca of those washed at 37 degrees C. This methodology provides an improved correlation between estimates of cellular calcium derived from 45Ca uptake and 45Ca efflux experiments.


1991 ◽  
Vol 13 (4) ◽  
pp. 335-341 ◽  
Author(s):  
Gregory D. Potter ◽  
Joseph H. Sellin ◽  
Susan M. Burlingame

1981 ◽  
Vol 241 (6) ◽  
pp. E454-E459
Author(s):  
W. M. Burch ◽  
H. E. Lebovitz

We assessed whether hormones and metabolic factors known to stimulate anabolic processes in the embryonic chick pelvic cartilage would stimulate ornithine decarboxylase (ODC) activity. In vitro organ culture of these pelvic cartilages in time-course experiments with N(6)-monobutyryl cyclic AMP (BtcAMP), insulin, and 5% rat serum demonstrated maximal stimulation of ODC activity between 4 and 6 h with each factor. However, at 2 h insulin and serum significantly stimulated ODC activity (P less than 0.05) and BtcAMP did not. ODC was stimulated above control (100%) with the following factors: parathyroid hormone (PTH) (555 +/- 15%), BtcAMP (324 +/- 34%), 1-methyl-3-isobutylxanthine (MIX) (223 +/- 6%), prostaglandin E1 (PGE1) (227 +/- 15%), 3,3',5-triiodothyronine (T3) (184 +/- 22%), insulin (182 +/- 14%), multiplication-stimulating activity (MSA) (178 +/- 6%), 5% rat serum (253 +/- 57%). THe increase in ODC activity seen with BtcAMP and insulin was not due to a change in Km or a decreased rate of degradation of the enzyme. Actinomycin D (1 microgram/ml) inhibited stimulation of ODC activity by T3 and the cyclic AMP-mediated factors (PTH, BtcAMP, MIX, PGE1), but had only minimal effects on ODC stimulation by insulin, MSA, or serum. Amanitin inhibited both BtcAMP and T3 stimulation of ODC, but had no effect on insulin stimulation of ODC. Thus, hormones and metabolic factors known to stimulate anabolic processes in chick embryonic pelvic cartilage also increase ODC activity through at least two mechanisms: transcriptional (cyclic AMP-mediated and T3) and posttranscriptional (insulin, serum, MSA).


Sign in / Sign up

Export Citation Format

Share Document