The tubulo-glomerular feedback mechanism — a determinant for the autoregulation of the glomerular filtration rate in superficial and juxtamedullary nephrons

Summary The intrinsic myogenic hypothesis and the tubuloglomerular feedback mechanism (TGF) give the presently most cherished explanation to the autoregulation of renal blood flow and glomerular filtration rate. A series of experiments was performed on young, normohydrated rats in order to evaluate the importance of TGF as an autoregulatory factor of the single nephron glomerular filtration rate (SNGFR) in superficial and juxtamedullary nephron populations. Micropuncture techniques were applied to tubular structures of the renal surface and on the papilla for the measurement of hydrostatic pressures and SNGFR. The SNGFR was also measured with a modified Hanssen technique. A TV-technique was used to record the urine free flow rate in the loop of Henle.The net driving forces for glomerular filtration at the afferent end of the glomerular capillaries were estimated to be 19 and 47 mm Hg for superficial and juxtamedullary nephrons respectively, when the urine flow at the macula densa was zero. The SNGFR of the two nephron populations amounted to 29.6 and 84.1 nl·min^–1·g^–1 K.W., as measured with the micropuncture technique. With a modified Hanssen technique the corresponding values were 25.8 and 27.7 nl·min^–1. g^–1 K.W. (kidney weight).The SNGFR was found to be well autoregulated when the urine flow at the macula densa was intact, but not when the urine flow was interrupted.The flow rate in the loop of Henle was in free flow conditions 7.3 nl·min^–1·g^–1 K.W. which shall be compared with 19.2 nl·min^–1·g^–1 K.W. when the urine flow to the macula densa was zero.We conclude that SNGFR is mainly autoregulated by the TGF-mechanism in young, normohydrated rats at lower arterial pressures. In normal conditions TGF is highly activated for juxtamedullary nephrons, but not for the superficial ones. The high urine flow rate in the loop of Henle at reduced flow rates at the macula densa may invalidate the use of loop blockade in studies of water and solute transfer across the loop walls.     

Factors controlling the release of renin

Summary Blood was collected by micropuncture from the efferent arteriole and superficial venules of the cat's kidney. Blood samples were also collected from the femoral artery and the renal vein. The blood renin concentrations (ng Al ml^–1 2 h^–1) in a basal state were 37.2±3.5 (S.E.M.) (n=60) (artery), 32.5±5.2 [7] (efferent arteriole), 53.5±4 (116) (superficial venule), 54.2±5.4 (43) (renal vein). The corresponding values after haemorrhage were 389±98 (21), 345±115(6), 963±208 (37), 907±290 (17), ng Al ml^–1 2 h^–1. The efferent arteriolar renin did not differ from that in the artery but the concentrations in superficial venular blood and renal vein were higher than arterial concentration. Thus renin entered the circulation between the efferent arteriole and the superficial venule. The blood renin concentration of different superficial venules at a steady arterial renin concentration varied markedly. Into certain venules there appeared to be little or no renin secretion, into others a marked renin secretion, suggesting a heterogeneity of renin secretion by the different nephrons.When the flow of tubular fluid to the macula densa of a group of nephrons was blocked, the renin concentration fell and was significantly less than the renin concentration in venules draining non blocked nephrons and less than in the renal vein.These results suggest that the juxtaglomerular apparatus of the nephrons do not release renin in a synchronous fashion. The release appears to be episodic and is inhibited when flow to the macula densa is ceased. This implies that a high sodium concentration at the macula densa stimulates renin release.Supported by the National Heart Foundation of Australia     

Micropuncture studies on the filtration rate of single superficial and juxtamedullary glomeruli in the rat kidney

Summary Single nephron filtration rates of superficial and juxtamedullary nephrons were determined in high and low sodium rats. Single nephron GFR was calculated from TF/P inulin and tubular flow rate in superficial nephrons and single juxtamedullary GFR from corresponding data in long loops of Henle. In low sodium rats superficial nephron GFR was 23.5±6.4 (SD)×10^–6 ml/min×g KW, juxtamedullary nephron GFR was 58.2±13.6 and total kidney GFR (C _In) was 0.94±0.16 ml/min×g KW. Using these single nephron values, total kidney GFR and a total number of 30,000 glomeruli per kidney, the number of superficial and juxtamedullary glomeruli was calculated to be 23,267 and 6,733, respectively. During high sodium diet superficial nephron GFR increased to 38.1±11.3 and single juxtamedullary GFR decreased to 16.5±6.6, total kidney GFR increasing to 1.01±0.24. Calculation again revealed the same distribution of the two nephron types. End-proximal TF/P inulin in superficial nephrons was 2.36±0.36 in low sodium and 2.31±0.28 in high sodium rats. Loops of Henle TF/P inulin and intratubular flow rate were inversely related: the highest TF/P inulin values and lowest intratubular flow rates were found in the descending limb. These data quantify the distribution of superficial and juxtamedullary nephrons on a functional basis and suggest a mechanism by which the kidney adjusts sodium excretion by altering the contribution of each nephron type to total kidney GFR.Supported by the Deutsche Forschungsgemeinschaft and the U.S. Department of the Army, through its European Research Office.     

Phosphate reabsorption in juxtamedullary nephron terminal segments

Summary Adult Munich Wistar rats undergoing mild salt diuresis (NaCl 20 g·l^–1, 0.1 ml·min^–1) were injected with tracer doses of³H-Inulin and³²P-sodium phosphate in thin descending and ascending limbs of Henle's loop, collecting ducts accessible at the surface of the papilla and early distal superficial tubules. Kidneys were prepared for simultaneous papillary microinjection and urinary flow collection. Expressed in percent of the amounts injected, unidirectional phosphate reabsorption fluxes were 5±1% and 3±1% for injections into early distal superficial tubules and collecting ducts, respectively. By contrast, the flux was 21.7±3% for injections into either the descending or ascending thin limbs of juxtamedullary nephrons. We conclude from these results that in the rat, a significant amount of phosphate is reabsorbed by the juxtamedullary distal tubules and/or the subsequent arcades connecting the juxtamedullary distal tubules to the collecting ducts.     

Valine oxidation in the rat medullary thick ascending limb

In the kidney, a branched-chain amino acid transferase (BCAAT) activity has been localized mainly in the medullary thick ascending limb (MTAL) of the rat nephron. BCAAT is the first enzyme involved in the metabolic pathway of the three branched-chain amino acids (BCAA): leucine, isoleucine and valine. The present work has been designed to study valine catabolism. Valine and leucine oxidation in MTAL were compared by measuring the rate of ¹⁴CO₂ produced when these substrates were incubated as sole substrates at a final concentration of 1 mM. Since glucose is also metabolized in MTAL, valine and leucine oxidation were quantified also in the presence of glucose (1 mM). The results show that: (1) valine oxidation was greater than that of leucine (63.0±4.7 vs 39.7±5.2 pmol · h^–1 · (g^–1 protein, respectively; P<0.001). As previously shown, leucine oxidation was found to be increased in the presence of glucose whereas glucose oxidation decreased. In contrast, the presence of glucose strongly diminished valine oxidation (19.2±1.9 vs 63.1±4.7 pmol · h^–1 · (g^–1 protein; P<0.001) whereas glucose oxidation was increased in the presence of valine (268.2±14.9 vs 229.6±16.2 pmol · h^–1 · g^–1 protein; P<0.05). We conclude that in rat MTAL, under near physiological conditions (in the presence of glucose, as in vivo), leucine is a preferential respiratory fuel as compared to valine. However, valine supports energetic salt transport and facilitates glucose oxidation.     

Role of the macula densa in the control of renal renin gene expression in two-kidney/one-clip rats

This study was designed to examine whether macula densa function is involved in the changes of renal renin gene expression upon acute hypoperfusion of one kidney. To block macula densa function, rats with free access to salt and water were subcutaneously infused with furosemide (12 mg/day) for 6 days. Then, 4 days after the start of the infusion, the left renal arteries were clipped with 0.2-mm silver clips and renin mRNA levels in ipsilateral and contralateral kidneys, as well as plasma renin activities (PRA), were determined 48 h after clipping. In non-clipped animals furosemide increased PRA from 10 to 47 ng angiotensin I · h^–1 · ml^–1 and raised renin mRNA levels in both kidneys 2.5-fold. In vehicle-infused animals, clipping of the left renal artery increased PRA to 37 ng angiotensin I · h^–1 · ml^–1 and led to a 5-fold rise of renin mRNA levels in the ipsilateral kidneys and to a suppression to 20% of the control values in the contralateral kidneys. PRA values in clipped and furosemide-infused animals were 45 ng angiotensin I · h^–1 · ml^–1. In these animals renin mRNA levels increased in the ipsilateral kidneys to similar absolute values as in vehicle-infused rats, whilst contralateral renin mRNA levels fell to about 25% of the respective controls. These findings indicate that the stimulations of renin gene expression by inhibition of macula densa salt transport and by renal artery clipping are not additive, suggesting that the macula densa mechanism may participate in the stimulation of renin gene expression upon hypoperfusion. The macula densa mechanism, however, appears to be not essentially involved in the suppression of renin gene expression in the contralaterals to stenosed kidneys.     

Radioactive microsphere distribution and single glomerular blood flow in the normal rabbit kidney

Summary Intracortical distribution of blood flow was studied in the rabbit kidney with 15 m labelled microspheres (M) injected into the left ventricule. M injection did not alter renal function. Thanks to arterial filling of left kidney with silicone rubber, efferent vascular patterns of the glomeruli could be precisely identified. Glomeruli of different populations were sampled by microdissection and their radioactivity measured. Assuming that intracortical distribution of M reflected distribution of flow to the glomeruli, individual glomerular blood flows (GBF) were determined. In hydropenic rabbits, GBF was higher in deep glomeruli providing vasa recta (G4) (193±14 nl·min^–1) than in most other superficial (G1 and G2) and deep glomeruli (G3) (190±27, 113±10 and 127±9 nl·min^–1 respectively; . The exact significance of GBF found in superficial glomeruli with straight ascending efferent arterioles supplying aglomerular suscapsular cortex (G1) was questioned because of the possible axial streaming of spheres. Afferent medulary blood flow was calculated to represent 9.0±0.9% of total renal blood flow.Attaché de Recherches I.N.S.E.R.M.     

Tubuloglomerular feedback and autoregulation of glomerular filtration rate in Wistar-Kyoto spontaneously hypertensive rats

Experiments were done in Wistar-Kyoto spontaneously hypertensive rats (SHR) to examine the efficiency of autoregulatory adjustments of kidney and nephron filtration rate (GFR) to acute changes in blood pressure (BP) over a broad blood pressure range. When BP of the SHR was reduced from 158±7 to 118 ±3 mm Hg by aortic clamping, kidney-GFR remained unchanged from 1.19±0.11 to 1.17±013 ml·min^–1·g^–1 kidney weight (KW), respectively. Single nephron GFR (SNGFR) measured at early distal tubule sites was similarly unchanged with the same BP change, 27.9±1.5 vs. 24.9±2.1 nl·min^–1·g^–1 KW (P>0.10). Proximal and distal estimates of SNGFR were significantly different from each other at high BP (7 nl·min^–1·g^–1,P<0.025), but were not different at low BP (2.0 nl·ml^–1·g^–1,P>0.10). Studies assessing tubuloglomerular feedback activity were done with orthograde perfusion of the loop of Henle using recollections of early proximal flow rate (EPFR) as an index of change of glomerular filtration rate. A change in perfusion rate from 0 to 45 nl·min^–1 induced a reduction in early proximal flow rate of 40.5 ±4.5%. Juxtaglomerular renin activity of superficial nephrons was 36.2±4.3 in the SHR, a value insignificantly different from 23.7±4.4 ng Angiotensin II amide·0.1 ml^–1·h^–1. 5 glomeruli^–1 in normal controls (P>0.05). The SHR appears to behave as a normal animal with respect to tubologlomerular feedback and autoregulatory renal vascular adjustments. Like normal rat models, the SHR demonstrated dependence on maintenance of distal filtrate delivery to achieve single nephron GFR autoregulation.Financial support for these studies and for Dr. Ploth were made available by funds from the Deutsche Forschungsgemeinschaft     

Macula densa cells sense luminal NaCl concentration via furosemide sensitive Na+2Cl−K+ cotransport

The macula densa cells of the juxtaglomerular apparatus probably serve as the sensor cells for the signal which leads to the appropriate tubuloglomerular feedback response. The present study reports basolateral membrane voltage (PD_bl) measurements in macula densa cells. We isolated and perfused in vitro thick ascending limb segments with the glomerulus, and therefore the macula densa cells, and the early distal tubule still attached. Macula densa cells were impaled with microelectrodes under visual control. PD_bl was recorded in order to examine how these cells sense changes in luminal NaCl concentrations. The addition of furosemide, a specific inhibitor of the Na⁺2Cl^–K⁺ cotransporter in the thick ascending limb, to the lumen of the perfused thick ascending limb hyperpolarized PD_bl from –55±5 mV to –79±4 mV (n=7). Reduction of NaCl in the lumen perfusate from 150 mmol/l to 30 mmol/l also hyperpolarized PD_bl from –48±3 mV to –66±5 mV (n=4). A Cl^– concentration step in the bath from 150 mmol/l to 30 mmol/l resulted in a 24±4 mV (n=4) depolarization of PD_bl. This depolarization of PD_bl was absent when furosemide was present during the Cl^– concentration step. These data suggest that the macula densa cells sense changes in luminal NaCl concentration via coupled uptake of Na⁺ and Cl^–. The transport pathways for NaCl transport in macula densa cells are probably identical to those in the thick ascending limb: the (Na⁺+K⁺)-ATPase in the basolateral membrane drives Na⁺ and Cl^– uptake via the luminal Na⁺2Cl^–K⁺ cotransport, Cl^– leaves the cell via basolateral Cl^– channels and K⁺ recycles across the apical membrane via K⁺ channels. Changes in intracellular Cl^– activity as a result of altered luminal NaCl uptake, and thus voltage changes of the basolateral membrane are probably the first signal in the tubuloglomerular feedback regulation.This study was supported by Deutsche Forschungsgemeinschaft Gr. 480/9     

Quantitative localization of Na-K pump sites in the frog sacculus

Summary The Na-K pump site distribution within the macula, perimacula, and wall epithelia of the sacculus in the frog inner ear was examined with quantitative [³H]ouabain autoradiography. Excised tissue was incubated for 10–30 min (23 ° C) in micromolar concentrations of high specific activity [³H]ouabain (14–70 Ci mT^–1, 5–15 Ci mmor^–1), washed for 30 min (4 ° C), then rapidly frozen (–175 ° C) and processed for light and electron microscope autoradiography. Control experiments based on (1) high K⁺ (50 mm) in the incubation and (2) low specific activity [³H]Jouabain (1 mM, 0.013–0.025 Ci mmol^–1) indicated negligible nonspecific binding of the [³H]ouabain.Measurable levels of specific [³H]ouabain binding occurred in all saccular regions examined. Binding was localized to the basolateral cell membranes with no detectable binding to the apical membranes. [³H]ouabain binding across the apical-basal axis of the saccule macular epithelium was nonuniform. Binding was low in the apical region, rose to a peak in the middle two-thirds, and then fell again close to the basement membrane. Electron microscope autoradiography suggested that this peak was due to ouabain binding to nerve terminals. Denervation of the sacculus eliminated the peak in [³H]ouabain binding and quantitative grain density analysis revealed that 45% of the Na-K pumps within the saccule macula were located on the nerve terminals.Na-K pump site density per unit volume was estimated by quantitative grain density analysis and the following values were obtained (sites m^–3 × 10³, means ± S.E.M.): saccule macula, 1.9 ± 0.2; saccule perimacula, 1.1 ± 0.1; saccule wall, 2.3 ± 0.3. Stereological analysis of conventionally fixed tissue was used to estimate overall plasma membrane surface area per unit volume (S _v). Na-K pump site densities per unit membrane area for the various regions were calculated by combining the autoradiographical and Stereological data. The following values were obtained (sites m^–2 ± 25%): saccule macula, 2500; saccule perimacula, 2500. Values for individual cells within the macula (sites m^–2 ± 25%) were: hair cells, 3000; nerve terminals, 3000; supporting cells, 1500.     

Increased activity of the Na-K-ATPase in red cell-ghosts of patients with Cushing's Syndrome: Possible significance for the pathogenesis of glucocorticoid-induced hypertension

Summary The Na-K-ATPase activity of erythrocyte ghosts was increased in 6 patients with Cushing's syndrome compared with 28 control subjects (0.986±0.291 versus 0.259±0.1 µM P_i·h^–1·mg^–1,p<0.001). Ouabain insensitive Mg-ATPase activity was similar in both groups. These data support the concept of an activation of the Na-pump in patients with glucocorticoid excess.     

Heterogeniety in regulation of glomerular function

The aim was to study differences in filtration driving forces and glomerular filtration rates between superficial and deep nephrons when urine flow rate was altered at the macula densa region. In young rats stop-flow pressures and single nephron glomerular filtration rates (SNGFR) were measured in the superficial proximal tubules and in the loops of Henle in the papilla. SNGFR was also measured with a modified Hanssen technique. The stop-flow pressures of superficial nephrons amounted to 30.9±0.8 mmHg (mean ± SE) and those of juxtamedullary nephrons to 52.2±1.6 mmHg. In the stop-flow condition the net driving filtration forces were calculated to be about 19 mmHg and 50 mmHg for the superficial and deep glomeruli, respectively. In free flow conditions both net driving forces were calculated to be 19 mmHg. The micropuncture technique gave a SNGFR value for superficial nephrons of 29.6±2.9 and for deep nephrons of 84.1±8.5 nl±min^-1 g^-1 kidney weight (KW). With a modified Hanssen technique the corresponding values were 25.8±3.3 and 27.7±2.9 nl. min^-1.g^-1KW. The tubuloglomerular feedback mechanism is considered to have a powerful regulatory influence on the glomerular filtration rate of deep nephrons.     

Thermogenesis due to noradrenaline in muscles under different rates of perfusion

Vascularly isolated hind legs of cold acclimated rats were perfused with arterial blood either without noradrenaline (NA) or with a constant concentration of NA (10 ng·ml^–1) at different perfusion rates ranging from 2 to 14l·g^–1·min^–1. The oxygen consumption of the leg during perfusion both with or without NA was linearly related to the perfusion rate. The linear increase of leg oxygen consumption with respect to the perfusion was steeper after NA, which indicates that the same arterial concentration of NA may produce a greater thermogenic effect at higher blood flow rates (the difference between resting metabolic rate and the thermogenesis stimulated by NA, was 8.20 l O₂·g^–1·h^–1 at a blood flow of 3l·g^–1·min^–1, compared with 45.02 l O₂·g^–1·h^–1 at a blood flow of 14 l·g^–1·min^–1). These data confirm the important role of the extravascular influx rate of NA in the control of thermogenesis due to NA in muscles.     

Tubular transport processes in proximal tubules of hypothyroid rats. Lack of relationship between thyroidal dependent rise of isotonic fluid reabsorption and Na+−K+-ATPase activity

Hypothyroid rats reconstituted with 10 g/kg b.w. per day of tri-iodothironine (T₃) for 4 days resulted in normal free T₃ and TSH levels. FT₃ levels were: 0.53±0.3 pg/ml in hypothyroid rats; 2.78±1.21 pg/ml in hormone reconstituted rats and 2.90±0.90 pg/ml in euthyroid rats. TSH levels were 3,508±513 g/ml in hypothyroid rats; 1,008±204 g/ml in reconstituted rats and 270±184 ng/ml in euthyroid rats.When hypothyroid rats were reconstituted with 50 g T₃/kg b.w. per day, TSH levels were nearly normal after 4 days (1,157±621 ng/ml). However FT₃ levels after 1–4 days were always higher than in euthyroid rats.Hypothyroid rats show a decrease in isotonic fluid reabsorption (J _v) in the proximal tubule (1.50±0.08 versus 4.96±0.23 10^–2 nl·mm^–1·s^–1 in euthyroid animals). 1 day after T₃ (10 g/kg b.w./day) injectionJ _v was increased significantly to 2.05±0.20 10^–2 nl·mm^–1·s^–1 and continued to increase during 4 days of T₃ reconstitution.When 50 g T₃/kg b.w./day was used,J _v increased to 2.75±0.07 after 1 day and to 3.10±0.42 10^–2 nl·mm^–1·s^–1 after 4 days.J _v was never reaching a value close to that of euthyroid rats because the tubular radius in hypothyroid rats (14.7±1.8 m) is less than that of euthyroid rats (19.2±0.5 m). The radius in hypothyroid rats treated with T₃ was unchanged over a 4 day course with either high or low doses of T₃.Na⁺–K⁺-ATPase activity was found to be 2.91±0.16 M P_i/h×mg protein in homogenates of kidney cortex from hypothyroid rats. Treatment of hypothyroid rats with 10 g or 50 g of T₃ resulted in an initial decrease in ATPase activity, followed by an increase to base level in hypothyroid rats with 10 g and a significantly higher level with 50 g. This decrease in ATPase activity was contrasted to the increase inJ _v.These data indicate that there is a dissociation between the effects of physiological doses of thyroid hormones on proximal tubular reabsorption and the effects of T₃ on Na⁺–K⁺-ATPase activity of kidney cortex. This leads to question the relationship between sodium transport and ATPase activity under physiological doses of thyroid hormones. An early effect of physiological doses of thyroid hormones on brush border Na⁺ permeability is suggested.     

Furosemide stimulates renin expression in the kidneys of salt-supplemented rats

This study was conducted to obtain information about a possible influence of salt transport by the thick ascending limb of Henle (TALH) and the macula densa on the expression of renin in the kidney. To this end, adult male rats were subcutaneously infused with furosemide (12 mg/24 h), an established inhibitor of TALH and macula densa salt transport, or with vehicle for 6 days. The animals had free access to chow, water and salt water (0.8% NaCl, 0.1% KCl) to maintain salt and water balance. Chronic furosemide treatment led to a 20-fold increase in urine flow rates and 50% increase in kidney weights, while urine osmolality decreased by 60% and body weight gain decreased by 40% in the furosemide-treated animals. Plasma renin activities increased from 2.9±0.5 ng angiotensin I h^–1 ml^–1 in controls to 10.6±2.2 ng angiotensin I h^–1 ml^–1 in furosemide-treated rats. In parallel, kidney areas immunoreactive for renin increased by 80% and the renal content of renin mRNA increased by 120% in the animals receiving furosemide. Under the assumption that the effects seen on renal renin expression were primarily due to the inhibition of TALH and macula densa function by furosemide, our findings suggest that salt transport across the TALH and macula densa exerts a negative control function not only on the secretion but also on the expression of renin in the kidney.     

Characteristics of the transport of oxalate and other ions across rabbit proximal colon

In order to characterize oxalate handling by the P2 segment of the rabbit proximal colon, the fluxes of [¹⁴C]oxalate, ²²Na⁺, and ³⁶Cl^– were measured in vitro using conventional short-circuiting techniques. In standard buffer the proximal colon exhibited net secretion of Na⁺ (–2.31±0.64 equiv cm^–2 h^–1), negligible net Cl^– transport, and net secretion of oxalate (–12.7±1.6 pmol cm^–2 h^–1). Replacement of buffer Na⁺ or Cl^– abolished net oxalate secretion, while HCO ₃ ^– -free media revealed a net absorption of oxalate (19.3±4.2 pmol cm^–2 h^–1) and stimulated NaCl absorption. Mucosal amiloride and dimethylamiloride (1 mM) significantly reduced the unidirectional fluxes of oxalate and enhanced sodium secretion by decreasing J _ms ^Na . The anion exchange inhibitor 4,4-diisothiocyanatostilbene-2,2-disulfonic acid (DIDS; 0.1 mM, both sides) reduced the unidirectional fluxes of oxalate and chloride. Serosal epinephrine (50 M) stimulated oxalate absorption (21.3±6.3 pmol cm^–2 h^–1) and sodium absorption (5.71±1.20 equiv cm^–2 h^–1), whereas dibutyryl-cAMP enhanced oxalate secretion (–43.4±6.9 pmol cm^–2 h^–1) and stimulated chloride secretion (–7.27 ±0.64 equiv cm^–2 h^–1). These results indicate that the P2 segment of the proximal colon possesses (a) secretory as well as absorptive capacities, (b) oxalate fluxes that are mediated by pathways involving Na⁺, Cl^–, HCO ₃ ^– transport and (c) a net oxalate flux that is sensitive to absorptive and secretory stimuli.     

Lipoprotein lipase activity in skeletal muscle and adipose tissue of marathon runners after simple and complex carbohydrate-rich diets

Summary A comparison of the influence of simple and complex carbohydrate (CHO) consumption on adipose tissue- and skeletal muscle-lipoprotein lipase activity (AT-LPLA, SM-LPLA) was examined. Twenty male marathon runners were divided into two equal dietary groups: simple-CHO and complex-CHO. Half of the subjects in each group consumed either a low-CHO (15% energy [E] intake), or a mixed diet (50% CHO) for 3 days. Immediately following this dietary period, the subjects consumed a CHO-rich diet (70% E intake) predominant in simple-CHO or in complex-CHO for an additional 3 days. Thereafter, all subjects returned to a normal mixed diet. Skeletal muscle biopsies, adipose tissue aspirations, and venous blood samples were obtained prior to dietary manipulation (PRE), upon completion of the 6 day diet (POST I), and 2 weeks after returning to a normal diet (POST II). The samples were analysed for AT-LPLA, SM-LPLA, serum insulin, and free fatty acids (FFA), and blood glucose, and lactate. SM-LPLA fell 71% from PRE values of 0.39±0.30 mol · g^–1 · h^–1 to POST I values of 0.11 ±0.09 mol · g^–1 · h^–1 (means±SD) (p<0.05), after a complex-CHO diet. However, the simple-CHO diet did not alter SM-LPLA. AT-LPLA similarly decreased (p<0.05) after the complex-CHO diet, and no significant decrease was noted after the simple-CHO diet. Serum FFA decreased significantly (p<0.05) after a simple-CHO diet (0.82±0.13 to 0.65±0.10 mmol l^–1) but were unchanged after a complex-CHO diet. Blood glucose and lactate, and serum insulin were not altered following a CHO-rich diet.     

Vascular fluid shifts and endocrine responses to exercise in the heat

Summary This study examines the relationships between vascular changes and endocrine responses to prolonged exercise in the heat, associated with dehydration and rehydration by fluids of different osmolarity. Five subjects were exposed, in a 34 C environment for 4 h of intermittent exercise on a cycle ergometer at 85±12 Watts (SD). Fluid regulatory hormones and cortisol were analysed in 3 experimental sessions: one without any fluid supplement (NO FLUID), and two with progressive rehydration, either by spring water (WATER) or isotonic solution (ISO), given after 70 min of exercise. Results were expressed in terms of differences between the mean values observed at the end of the exercise and the first hour values taken as references.Dehydration (NO FLUID) elicited a 4.0±0.8% (SE) decrease in plasma volume (PV) and an increase in osmolarity (8.4±3.1 mosmol · l^–1). Concomitantly, plasma aldosterone (PA), renin activity (PRA), arginin vasopressin (AVP) and cortisol (PC) levels increased greatly in response to exercise in the heat (PA: 37.2+-10.8 ng. 100 ml^–1; PRA: 13.4±2.5 ng · ml^–1 · h^–1; AVP: 3.8±1.3 pg · ml^–1; PC: 12.2±2.7 g · 100 ml^–1). Rehydration with water led to decreased osmolarity (–8.2±2.1 mosmol · l^–1) with no significant changes in PV. With ISO, PV increased by 6.0±1.3% and the decrease in osmolarity was –5.8±1.8 mosmol · l^–1. With both modes of rehydration, the increases in PRA, AVP and cortisol were blunted; only ISO prevented the rise in PA.These data indicate that prolonged exercise in moderate heat is extremely effective in increasing cortisol and fluid-electrolyte regulatory hormones in dehydrated subjects. Progressive rehydration with water or isotonic solution, in the absence of osmotic and volemic stimuli, prevents the hormonal increases.     

Gastric emptying and serum insulin levels after intake of glucose-polymer solutions

Summary To examine the gastric emptying characteristics of four drinks varying in carbohydrate composition and concentration, five men ingested 600 ml of one of the different drinks on four separate occasions. All drinks contained Na⁺ 71 mmol · l^–1, Cl^– 60 mmol · l^–1, Mg⁺² 5 mmol · l^–1 and citrate 7 mmol · l^–1; the carbohydrate component was either 3% glucose, 3% glucose-polymer (GP), 5% GP or 10% GP. With ^99mTc-diethylene-triaminepenta-acetic acid (DTPA) as a marker, a scintillation camera and computer were used to measure the rate of gastric emptying. The half-emptying times (T _1/2) were inversely related to the glucose content of the solutions. The T _1/2 for 3% PG was 22.4±4.4 min (mean±SE) and for 10% GP 50±3.3 min (p< 0.005). There was no significant difference in T_1/2 between the 3% glucose and 3% GP solutions. The increments in blood glucose (highest blood levels from 7.4±0.3 mmol · l^–1 to 8.9±0.8 mmol · l^–1), serum insulin (from 28±6 mU · l^–1 to 77±13 mU · l^–1) and C-peptide (from 3.6±0.4 g · l^–1 to 5.8±0.9 g · l^–1) were related to the amount of carbohydrate ingested. In all cases the serum insulin levels were high enough to inhibit the liberation of free fatty acids from the adipose tissue. It is concluded that the amount of carbohydrate in glucosyl units in the solution is a major determinant of gastric emptying. Furthermore, the inhibitory effect of glucose is not modified by replacing monomer glucose with glucose polymer or adding NaCl (about 70 mmol · l^–1) in the solution.     

Vasoactive intestinal peptide stimulation of renal adenylate cyclase and antagonism by (4Cl-d-Phe6Leu17)VIP

The effect of vasoactive intestinal peptide (VIP) and related peptides [glucagon, secretin, PHI 1-27 (peptide with N-terminal histidine and C-terminal isoleucine)] on renal adenylate cyclase (AC) has been determined in several species. The largest stimulation (4.1±0.5-fold basal) of AC by 1 mol · l^–1 VIP was observed in feline cortical plasma membranes. In rabbit and guinea-pig, VIP increased AC activity 1.5±0.3- and 1.8±0.3-fold respectively but glucagon had no such action. Conversely in the rat glucagon stimulated AC some 3-fold over basal activity whereas VIP had little effect. In dog, cat and mouse both peptides were effective in increasing AC activity. For cat, half-maximal stimulation of cortical plasma membrane AC by VIP was seen at 27.0±9.0 nmol · l^–1 (SEN=9 animals). VIP also increased AC activity in both outer (red) and inner (white) medulla. In feline cortical membranes VIP and PTH (parathyroid hormone) when added in combination were fully additive. However for VIP and glucagon in combination there was no cumulative increase in AC activity, indeed the resultant activity was less than that attained by VIP alone. The VIP analogue (4Cl-d-Phe⁶Leu¹⁷)VIP at 10 mol · l^–1 produced a right shift in the VIP-dose response curve and increased the EC₅₀ from 17.2±5.8 nmol · l^–1 to 132.0±22.2 nmol · l^–1 VIP (SEN=4). There was no reduction in the maximum response elicited by VIP consistent with a competitive type of antagonism by this analogue. PHI-stimulated AC was also reduced by (4Cl-d-Phe⁶-Leu¹⁷)VIP resulting in a similar right shift in the dose response curve. However, this analogue of VIP had no effect on glucagon- or secretin-stimulated AC, indicated by no change in EC₅₀ values.