Paul H. Brand

Body fluid expansion is not essential for salt-induced hypertension in SS/Jr rats

To evaluate the importance of volume in the development of hypertension in inbred Dahl salt-sensitive rats (SS/Jr), we measured the changes in blood pressure (BP) that occurred with oral intake of food (salt) and water in rats whose body weight was permitted to increase versus those in which body weight was maintained constant with a servo-control system. We hypothesized that if volume expansion is essential in the development of hypertension, then BP would not increase if body weight was held constant. We found that oral presentation of chow containing 4% salt to SS/Jr rats caused BP to increase 32.2 ± 2.9 mmHg over 4 days when body weight was controlled at its initial value. Plasma sodium increased from 142.0 to 145.2 meq/l during 4 days of high salt. Neither plasma volume, hematocrit, nor central venous pressure changed significantly on the high-salt diet. In contrast, the inbred Dahl salt-resistant rats (SR/Jr) did not increase their BP during body weight control when given 4% salt. This demonstrates that volume expansion is not an obligatory step in the pressure response to increased salt in SS/Jr rats. Our results obtained with oral presentation of salt, in contrast to intravenous, represent a physiological evaluation of the significance of volume changes in response to dietary salt because no potential regulatory reflexes have been bypassed.

Spontaneous changes in arterial blood pressure and renal interstitial hydrostatic pressure in conscious rats.

1. Previous work has demonstrated a positive relationship between experimentally induced changes in arterial pressure (AP) and renal interstitial hydrostatic pressure (RIHP). The purpose of the present study was to test the hypothesis that RIHP is positively correlated with the normal changes in AP that occur spontaneously in conscious rats. 2. Rats were chronically instrumented for the recording of AP (via an aortic catheter) and RIHP. RIHP was measured by implanting a Millar microtransducer, whose tip had been encapsulated in a 35 microns pore polyethylene matrix (5 mm long, 2 mm o.d.), approximately 5 mm below the renal cortical surface. 3. A total of 56 h of simultaneous analog recording of AP and RIHP was obtained from ten rats. Each 1 h segment was digitized and evaluated at frequencies of 1, 0.1, 0.02 and 0.01 Hz. 4. In forty‐nine out of fifty‐six of these 1 h recordings taken at 1 Hz, there were significant positive linear correlations between AP and RIHP (mean r = 0.32) with a mean slope of 0.11 mmHg RIHP/1 mmHg AP. Low‐pass filtering to 0.01 Hz significantly increased the r value to 0.48. 5. These results demonstrate that spontaneous changes in AP and RIHP are positively correlated. The spontaneous coupling of AP and RIHP may be of importance in the regulation of salt and water excretion by the pressure diuresis mechanism.

A GRAVIMETRIC METHOD FOR THE MEASUREMENT OF TOTAL SPONTANEOUS ACTIVITY IN RATS

Currently available methods for the measurement of spontaneous activity of laboratory animals require expensive, specialized equipment and may not be suitable for use in low light conditions with nocturnal species. We developed a gravimetric method that uses common laboratory equipment to quantify the total spontaneous activity of rats and is suitable for use in the dark. The rat in its home cage is placed on a top-loading electronic balance interfaced to a computer. Movements are recorded by the balance as changes in weight and transmitted to the computer at 10 Hz. Data are analyzed on-line to derive the absolute value of the difference in weight between consecutive samples, and the one-second average of the absolute values is calculated. The averages are written to file for off-line analysis and summed over the desired observation period to provide a measure of total spontaneous activity. The results of in vitro experiments demonstrated that: 1) recorded weight changes were not influenced by position of the weight on the bottom of the cage, 2) values recorded from a series of weight changes were not significantly different from the calculated values, 3) the constantly decreasing force exerted by a swinging pendulum placed on the balance was accurately recorded, 4) the measurement of activity was not influenced by the evaporation of a fluid such as urine, and 5) the method can detect differences in the activity of sleeping and waking rats over a 10-min period, as well as during 4-hr intervals recorded during active (night-time) and inactive (daytime) periods. These results demonstrate that this method provides an inexpensive, accurate, and noninvasive method to quantitate the spontaneous activity of small animals.

Phenotypic variation in sensorimotor performance among eleven inbred rat strains

As a first step toward identifying the genes that determine sensorimotor ability (motor coordination) we subjected 11 inbred strains of rats to three different tests for this trait. Rats were tested at 13 wk of age to determine how long they could remain on 1) a rotating cylinder as the velocity of rotation increased every 5 s (1-direction rotation test), 2) a rotating cylinder that reversed direction every 5 s and increased velocity every 10 s (2-direction rotation test), and 3) a platform that was tilted 2 degrees every 5 s from 22 to 47 degrees (tilt test). On all three tests, rats of the PVG strain demonstrated the greatest sensorimotor ability. In contrast, rats of the MNS strain were most often represented among the group of strains that demonstrated the lowest performance on all tests. Considering all three tests, there was a 3- to 13-fold range in sensorimotor performance between the highest and lowest strains. This large divergence between the highest and lowest strains provides a genetic model that can be used to identify intermediate phenotypes and quantitative trait loci that contribute to sensorimotor ability.As a first step toward identifying the genes that determine sensorimotor ability (motor coordination) we subjected 11 inbred strains of rats to three different tests for this trait. Rats were tested at 13 wk of age to determine how long they could remain on 1) a rotating cylinder as the velocity of rotation increased every 5 s (1-direction rotation test), 2) a rotating cylinder that reversed direction every 5 s and increased velocity every 10 s (2-direction rotation test), and 3) a platform that was tilted 2° every 5 s from 22 to 47° (tilt test). On all three tests, rats of the PVG strain demonstrated the greatest sensorimotor ability. In contrast, rats of the MNS strain were most often represented among the group of strains that demonstrated the lowest performance on all tests. Considering all three tests, there was a 3- to 13-fold range in sensorimotor performance between the highest and lowest strains. This large divergence between the highest and lowest strains provides a genetic model that can be used to identify intermediate phenotypes and quantitative trait loci that contribute to sensorimotor ability.

Improved microdialysis technique

Abstract A simple, inexpensive method is described for dialysis of microliter amounts of aqueous samples against large volumes of solution with complete recovery of the fluid dialyzed. An example is given of application of the method to separation of [3H]inulin from a monosaccharide.

Phenotypic variation in strength among eleven inbred strains of rats

Abstract As a first step toward the long-range goal of identifying the genes that determine strength, we subjected 11 inbred strains of rats to three tests of muscular strength. The tests consisted of measuring (1) the force exerted by the rat as it was pulled by the base of the tail off a grid on the pan of a top-loading electronic balance (scale test); (2) the length of time the rat hung from a 2.5-mm-diameter U-shaped wire (wire-hanging test); and (3) the length of time the rat hung from a vertically oriented grid consisting of 4-mm-diameter rods (grid-hanging test). Six rats of each gender from each strain were tested at 12 weeks of age, once/day for 5 consecutive days. For the two tests that required use of all four limbs (the scale and grid-hanging tests), one strain performed best (DA). In contrast, on the test that required primarily the use of the front limbs (wire-hanging test), the DA was the lowest performing strain and the F344 rats the best. This differential ranking suggests that the tests selected for variance in the morphological distribution of strength among the strains. There was a 1.5- to 5.2-fold divergence observed between the males of the highest and lowest strains on the scale test and grid hanging tests. This large divergence provides the opportunity to search for intermediate phenotypes and quantitative trait loci that contribute to the different performances of the strains on these strength tests.

State-Dependent Expression of Pressure Diuresis in Conscious Rats

In 1967, Guyton and Coleman modeled pressure diuresis as the underlying, essential, long-term mechanism that regulates arterial pressure when sodium intake changes. Other mechanisms that influence renal function interact with pressure diuresis to achieve sodium balance and determine the blood pressure. Increases in sodium intake suppress sodium conserving mechanisms and activate natriuretic mechanisms; decreases in sodium intake have the opposite effect. If the Guyton-Coleman model is correct, then pressure diuresis should be more readily detected in animals on a high-salt diet than in animals on a low-salt diet. We measured spontaneous changes in arterial pressure and urine flow in conscious rats fed low-salt (0. 4% NaCl) and high-salt (8.0% NaCl) chow. For 10 rats fed a high-salt diet, arterial pressure and urine flow were positively correlated in 19 of 32 (59%) trials. In 10 rats fed a low-salt diet, a positive correlation was observed in 10 of 33 (30%) trials. Chi-square analysis revealed that differences in Na+ content of the diet were significantly associated with the probability of a positive relationship between blood pressure and urine flow. These results support the hypothesis that the expression of pressure diuresis across time is dependent on the state of sodium balance.

Lactate Oxidation by Three Segments of the Rabbit Proximal Tubule

Abstract Oxidation of [U14C]lactate to 14CO2 was measured in vitro, in nonperfused anatomically defined segments of rabbit proximal tubule (S1, proximal convoluted, and S2 and S3, proximal straight tubules). The rate of lactate oxidation was similar in S2 and S3 segments, and within the range of lactate oxidation rates measured in vivo. In contrast, the oxidation rate of S1 segments was significantly lower than that of S2 or S3. In proximal straight tubules, lactate oxidation was inhibited by incubation at 0°C, or by application of 1 mM ouabain. To determine if the rate of transepithelial transport affected the rate of lactate oxidation, lactate oxidation was measured in proximal straight tubules after the lumen had been opened by perfusion with Ringers containing 10 mM polyethylene glycol. No difference in lactate oxidation rate was observed between tubules with patent lumina and nonperfused tubules. These results suggest that the various segments of the renal proximal tubule have different metabolic characteristics, and that the rate of substrate oxidation is related to the activity of the Na+, K+-ATPase.

Gravimetric Method for the Dynamic Measurement of Urine Flow

Abstract The rate of urine formation is a primary index of renal function, but no techniques are currently available to accurately measure low rates of urine flow on a continuous basis, such as are normally found in rats. We developed a gravimetric method for the dynamic measurement of urine flow in anesthetized rats. Catheters were inserted directly into the ureters close to the renal pelves, and a siphon was created to collect all of the urine formed as rapidly as it was produced. Urine flow was determined by measuring the weight of the urine using a direct-reading analytical balance interfaced to a computer. Basal urine flow was measured at 2-sec intervals for 30 to 60 min. The dynamic response of urine flow to a rapid decrease in arterial pressure produced by a bolus intravenous injection of acetylcholine (0.5 μg) was also measured. Intrinsic drift, evaporative losses, and the responsiveness of the system to several fixed pump flows in the low physiologic range were evaluated in vitro. The gravimetric method described was able to continuously measure basal urine flows that averaged 37.3 ± 12.4 μl/min. Error due to drift and evaporation was negligible, totaling less than 1% of the measured urine flow. Acetylcholine-induced declines in arterial pressure were followed within 8 sec by a decline in urine flow. These data demonstrate that this new gravimetric method provides a simple, inexpensive, dynamic measurement of urine flow in the μl/min range.