PTH regulates fetal blood calcium and skeletal mineralization independently of PTHrP.

PTH and PTHrP both act in the regulation of fetal mineral metabolism. PTHrP regulates placental calcium transfer, fetal blood calcium, and differentiation of the cartilaginous growth plate into endochondral bone. PTH has been shown to influence fetal blood calcium, but its role in skeletal formation remains undefined. We compared skeletal morphology, mineralization characteristics, and gene expression in growth plates of fetal mice that lack parathyroids and PTH (Hoxa3 null) with the effects of loss of PTHrP (Pthrp null), loss of PTH/PTHrP receptor (Pthr1 null), and loss of both PTH and PTHrP (Hoxa3 null x Pthrp null). Loss of PTH alone does not affect morphology or gene expression in the skeletal growth plates, but skeletal mineralization and blood calcium are significantly reduced. In double-mutant fetuses (Hoxa3 null/Pthrp null), combined loss of PTH and PTHrP caused fetal growth restriction, limb shortening, greater reduction of fetal blood calcium, and reduced mineralization. These findings suggest that 1) PTH may play a more dominant role than PTHrP in regulating fetal blood calcium; 2) blood calcium and PTH levels are rate-limiting determinants of skeletal mineral accretion; and 3) lack of both PTH and PTHrP will cause fetal growth restriction.

Reduction of blood pressure variability by amlodipine in baroreceptor denervated rats.

To determine the effect of the calcium blocker amlodipine on the variability of mean arterial pressure (MAP), amlodipine besylate was acutely administered to sino-aortic baroreceptor-denervated (SAD) rats (0, 1, 3, 10 mg/kg s.c.), and chronically administered to SAD and sham-denervated rats (0, 50, 150, 500, and 1,500 mg x kg(-1) feed, 4 days per dose). Acute amlodipine administration caused significant dose-dependent reductions of the mean MAP level and short-term MAP variability at the 3 and 10 mg/kg dose levels, respectively. Chronic administration produced dose-dependent reductions in short-term MAP variability, becoming significant at the 150 and 500 mg x kg(-1) feed dose level in SAD and Sham groups, respectively. Day-night differences in blood pressure were significantly attenuated or reversed at the 500 and 1,500 mg x kg(-1) feed dose levels. Amlodipine had little or no effect upon the 24 h MAP level, long-term MAP variability, and only modestly reduced the MAP response to hexamethonium. These results demonstrate that amlodipine can reduce MAP variability independent of changes in the mean blood pressure level.

Direct and indirect methods used to study arterial blood pressure.

A number of different approaches exist for assessing blood pressure in experimental animals. Here, we briefly consider the traditional indirect (rodent tail-cuff) and direct (saline-filled catheter) methods of blood pressure measurement before going on to describe our experience with blood pressure telemetry in rabbits, rats, and mice. Blood pressure telemetry offers the ability to obtain a high-fidelity recording of blood pressure continuously, for relatively long periods of time, in conscious, freely moving animals, without the limitations of restraint or anaesthesia. Since some drift in telemeter offset and sensitivity are inevitable, recalibration of the telemeter devices immediately before implantation and following explantation is essential to ensure and document the accuracy of the blood pressure measurements. For long-term implantations, verification of the calibration can be performed in vivo, at least in the case of large animals, such as rabbits. Telemetry devices suitable for small animals, such as mice, are also available now, which will facilitate the accurate characterization of blood pressure in transgenic animals. Telemeter implantation methods in mice are presently difficult, with relatively low success rates being reported. However, validation of new methods, such as the insertion of the catheter tip via the carotid artery, may make the technique more widely accessible in the near future.

Contribution of baroreceptors and chemoreceptors to ventricular hypertrophy produced by sino-aortic denervation in rats.

1. To test whether sino-aortic denervation (SAD)-induced right ventricular hypertrophy (RVH) is a consequence of baroreceptor or chemoreceptor denervation, we compared the effects of aortic denervation (AD), carotid denervation (CD), SAD and a SAD procedure modified to spare the carotid chemoreceptors (mSAD), 6 weeks after denervation surgery in rats. A sham surgery group served as the control. 2. The blood pressure (BP) level was unaffected by AD, CD or SAD, but increased (9 %) following mSAD. The mean heart rate level was not affected. Short-term BP variability was elevated following AD (81 %), SAD (144 %) and mSAD (146 %), but not after CD. Baroreflex heart rate responses to phenylephrine were attenuated in all denervation groups. 3. Significant RVH occurred only following CD and SAD. These procedures also produced high mortality (CD and SAD) and significant increases in right ventricular pressures and haematocrit (CD). 4. Significant left ventricular hypertrophy occurred following CD, SAD and mSAD. Normalized left ventricular weight was significantly correlated with indices of BP variability. 5. These results suggest that SAD-induced RVH is a consequence of chemoreceptor, not baroreceptor, denervation. Our results also demonstrate that a mSAD procedure designed to spare the carotid chemoreceptors produced profound baroreflex dysfunction and significant left, but not right, ventricular hypertrophy.

Effect of denervation of the rat superior mesenteric arterial bed on the distribution of resistance among the first four orders of branching.

The rat superior mesenteric artery with its first four orders of branching has been examined as an isolated perfused preparation. Lumen diameter and proportional resistance at each level have been determined and compared under conditions of constant pressure following methoxamine-induced constriction and acetylcholine-induced relaxation in intact and denervated preparations. A comparison was also made between intact preparations under conditions of constant pressure and constant flow. Lumen diameter and segment length were measured by analysis of video-taped images. The distribution of resistance among the first four orders of branching was estimated from the measured values using the Poiseuille formula. Intact first and fourth order branches had a proportionately low resistance, with the remaining resistance being fairly evenly distributed between the second and third order branches. No change in the pattern occurred as a result of either methoxamine or acetylcholine treatment. The pattern of distribution of resistance changed following denervation, so that proportional resistance was high in the first order branches and fell off from the second to fourth order branches. With methoxamine, the first order branches of denervates had a significantly higher resistance than in intact tissues, with the distribution of resistance in the second to fourth order branches following the basal resistance pattern. Reduction in the proportion of resistance with acetylcholine was similar in both intact and denervated preparations. Distribution of resistance did not differ between constant flow and constant pressure intact preparations.

Comparison of patients with idiopathic calcium phosphate and calcium oxalate stones.

Our primary objective was to test the hypothesis that a defect in acidification is more common in patients who have idiopathic calcium phosphate kidney stones than in those whose stones are formed mainly of calcium oxalate. Additionally, other risk factors might differ for these 2 stone types. Urine pH was measured serially over 24 hours, and along with ammonium and titratable acid, it was measured before and serially after ingestion of ammonium chloride in 3 groups of subjects: 24 patients with predominantly calcium phosphate stones, 30 patients with calcium oxalate stones, and 15 health non-stone-formers. Twenty-six parameters potentially related to stone formation and acidification were assayed on urines collected over 24 hours, and 15 parameters on blood. The data base was a computerized list of 5900 analyses of stones from patients living in Newfoundland. Patients not known by their physician to have had urinary tract infection, anatomical abnormality, hyperparathyroidism, or renal tubular acidosis were asked to participate in the study. Differences between means were considered significant if p values were less than 0.05 for F by analysis of variance and also less than 0.01 by t-test. In all patients with calcium oxalate stones and all non-stone-formers, urine acidified to pH less than 5.25, but in 8 of the 23 phosphate stone formers who completed the ammonium chloride study urine failed to acidify to pH less than 5.25. As all 8 had normal values for venous pH, total CO2, and chloride, they were considered to have incomplete renal tubular acidosis (IRTA). The 8 phosphate stone formers with IRTA had greater mean values for urine pH on all 9 specimens collected serially over 24 hours (all means greater than 6.2), and after administration of ammonium chloride (p less than 0.01), as well as lower mean values for urine titratable acid excretion (p less than 0.01), both after administration of ammonium chloride and in 24-hour urine samples, compared with the remaining phosphate stone formers whose urine acidified and the oxalate and non-stone-forming control groups. Nearly all the phosphate stone formers had 1 or more risk factors for stone formation, but with frequencies not significantly higher than those found in the oxalate group. Hypercalciuria and hypocitruria were the commonest, but increased oxalate or urate also occurred. Thus, idiopathic calcium phosphate stone formation can be associated with 1 or more of several risk factors, and, with the possible exception of those with IRTA, treatment should be similar to that given to patients with calcium oxalate stones.