Parathyroid hormone secretion in vivo. Demonstration of a calcium-independent nonsuppressible component of secretion.

The response of normal bovine parathyroid glands to hypercalcemia was assessed in vivo by radioimmunoassay of immunoreactive parathyroid hormone concentrations in parathyroid effluent blood obtained by surgical cannulation of both anesthetized and nonanesthetized calves. Hypercalcemia was induced for periods of 0.3-35 h by intravenous infusion of a solution of calcium chloride. Assessment of immunoreactivity in effluent and peripheral blood included measurements of selected samples by use of a radioimmunoassay specific for a site residing in the biologically active portion of the hormone molecule. In all instances, the concentration of immunoreactive parathyroid hormone in hypercalcemic venous effluent from a superior parathyroid gland exceeded that of the peripheral blood. Failure of hypercalcemia to suppress completely secretion by normal parathyroids indicates that a portion of parathyroid hormone secretion occurs independent of blood calcium concentration. Consequently, continued parathyroid hormone secretion despite hypercalcemia can no longer be regarded as a unique feature of parathyroid neoplasia.

Sigmoidal relationship between parathyroid hormone secretion rate and plasma calcium concentration in calves.

Parathyroid hormone (PTH) secretion rate was measured in 16 anesthetized calves by using a technique involving radioimmunoassay of parathyroid venous blood which was collected during timed intervals and measured volumetrically. The calves ranged in age from 2-14 weeks. Plasma calcium concentration was altered by infusion of solutions of CaCl2 or disodium ethylenediamine tetracetate (Na2 EDTA) into the jugular vein. When plasma calcium concentrations exceeded 10.5 mg/100 ml, a basal, non-suppressible secretion rate of 0.3 ng/kg/min was maintained despite the induction of hypercalcemia. Slight changes in secretion rate were observed in response to changes of plasma calcium in the range between 9 and 10.5 mg/100 ml. Below 9 mg/100 ml, a small decrease in plasma calcium concentration evoked a pronounced increase in secretion rate. A maximal secretion rate of about 5.5 ng/kg/min was attained at a plasma calcium concentration of approximately 7.5 mg/100 ml and it was not increased by more severe hypocalcemia. These observations confirm the sigmoidal relationship between PTH secretion rate and plasma calcium concentration which was previously suggested by measurement of PTH concentration in peripheral plasma of hypocalcemic, parturient cows.

Comparison of the effects of calcium and magnesium on parathyroid hormone secretion rate in calves.

Parathyroid hormone (PTH) secretion rate was measured in seven calves by using a technique which involved RIA of parathyroid venous blood collected during timed intervals and measured volumetrically. Infusion of solutions of NA2EDTA and MgCl2 into the jugular vein was used to alter plasma calcium and magnesium concentrations. In four calves, elevation of plasma magnesium concentration rapidly decreased the PTH secretion rate that had been stimulated by induced hypocalcemia. In three calves, equimolar and opposite changes in plasma calcium and magnesium concentrations were induced by simultaneous infusions of Na2EDTA and MgCl2. Despite the equimolar increase in plasma magnesium concentration, PTH secretion was increased in response to the decline in plasma calcium concentration. In three experiments, the concentration of each cation was kept constant during periods in which the concentration of the other cation was varied. The effect of variations in plasma magnesium concentration on PTH secretion rate was approximately 1/3-1/2 as great as that observed with changes in plasma calcium concentration. These observations indicate that the effect of magnesium on PTH secretion rate is similar to that of calcium, but not equipotent.

Diminished prepartal plasma calcitonin concentration on cows developing parturient hypocalcemia.

Immunoreactive calcitonin and calcium concentrations were determined on 581 plasma samples collected during 23 studies on 20 cows. Sample collections in each study was begun approximately 1 month prior to parturition and continued for about 1 month after parturition. The cows were grouped according to the degree of hypocalcemia encountered at parturition. The parturient paresis group consisted of 10 cows which developed severe hypocalcemia (3.91 plus or minus 0.22 mg/100 ml, mean plus or minus se) accompanied by paresis; the nonparetic hypocalcemic group consisted of 5 cows which developed severe hypocalcemia (5.70 plus or minus 0.03 mg/100 ml) but not paresis; and the control group consisted of 8 cows which experienced only a mild hypocalcemia (8.50 plus or minus 0.27 mg/100 ml) at parturition. In the prepartal period prior to the onset of hypocalcemia, the respective mean plasma calcium concentrations (plus or minus se) of the 3 groups were 10.1 plus or minus 0.11, 9.95 plus or minus 0.20, and 10.2 plus or minus 0.17 mg/100 ml. The development of severe hypocalcemia in the parturient paresis and nonparetic hypocalcemic groups was not accompanied by an increase in plasma calcitonin concentration. Furthermore, plasma calcitonin concentraion of these 2 groups was less than that of control cows during the parturient period as well as during the month before and the month after parturition. The plasma calcium nadir at parturition was positively related to the mean prepartal (encompassing the period from 30 days until 60 h before parturition) plasma calcitonin concentration (r = 0.57, t= 3.14, p less than 0.005); i.e., the lower the prepartal plasma calcitonin concentration the more severe the hypocalcemia at parturtion. These observations suggest that the development of hypocalcemia at parturition is not due to an increased secretion of calcitonin, but instead they suggest that parturient hypocalcemia may be associated with a diminished prepartal secretion of calcitonin.

Metabolism of amino- and carboxyl-sequence immunoreactive parathyroid hormone in the bovine: evidence for peripheral cleavage of hormone.

Radioimmunoassays that detect specifically peptide sequences within either the biologically active amino region (N-assay) or inactive carboxyl region (C-assay) of parathyroid hormone (PTH) were used to evaluate the metabolism of PTH during and after infusion and injection of homogeneous (containing less than 0.1% hormonal fragments) intact bovine PTH (bPTH) into calves. During continuous infusions of hormone, when constant blood levels of immunoreactive PTH were reached, a dissociation between the concentrations of amino versus carboxyl immunoreactivity was observed; concentrations of hormone measured by the C-assay rose to a level of approximately three times higher than that measured by the N-assay. Analysis by gel filtration of immunoreactive PTH in plasma samples from calves after injection of hormone showed the rapid disappearance of intact hormone (N- and C-assays) and the appearance of a large fragment detected by the C-assay but not by the N-assay. The hormonal fragment lacked antigenic determinants within the amino peptide sequence required for biologic activity. No additional fragments of PTH were detected by gel filtration using the N- and C-assays. No detectable conversion of intact PTH to hormonal fragments occurred during incubation in vitro in bovine serum. The results are consistent with the concept that PTH is metabolized after entry into the circulation at peripheral sites located outside the vascular space, resulting in the rapid disappearance from blood of intact hormone and the appearance of a biologically inactive hormonal fragment(s). These studies done in calves agree with earlier studies done in dogs and man and point to the existence in mammals of common pathways for the peripheral metabolism of PTH.

Dietary calcium, calcium kinetics and plasma parathyroid hormone concentration in cows.

Kinetic analysis of radiocalcium data was combined with radioimmunoassays of plasma parathyroid hormone in nonpregnant nonlactating cows 4.5 to 7.8 years of age whose calcium intake was varied from 0.05% to 1.4% of the diet. Calcium intake had no significant effect on plasma concentration of calcium or parathyroid hormone. It had a slight but significant effect on the plasma phosphorus concentration. As calcium intake increased, calcium absorption increased but calcium removal from bone decreased so that total calcium transport into the exchangeable pool remained fairly constant. A decline in calcium clearance with age was accompanied by a fall in plasma parathyroid hormone concentration, and a decrease in calcium transport into the pool. These observations are consistent with feedback controls between plasma calcium, parathyroid hormone and calcium transport operating to maintain plasma calcium homeostasis as calcium clearance declined with advancing age.

Parathyroid hormone: secretion and metabolism in vivo.

Gel filtration and radioimmunoassay were used to determine the molecular size and immunochemical reactivity of parathyroid hormone present in gland extracts, in the general peripheral circulation, and in parathyroid effluent blood (obtained by venous catheterization) from patients with hyperparathyroidism, as well as from calves and from cattle. Hormone secreted in vivo from normal bovine parathyroid glands and from human parathyroid adenomas is similar in size to the 84-amino-acid peptide (molecular weight of 9500) extracted from the parathyroids. However, much of the immunoreactive parathyroid hormone present in the peripheral circulation of man and cattle is smaller than the extracted or secreted hormone; it elutes from gel columns at a position corresponding to a molecular weight of about 7000. The immunological characteristics of extracted and secreted hormone are identical, while hormone in the general circulation is immunologically dissimilar to extracted and secreted hormone. The results indicate that parathyroid hormone secreted from the parathyroids in man and cattle is at least as large as the molecule extracted from normal bovine glands. However, once secreted into the circulation the hormone is cleaved, and one or more fragments, immunologically dissimilar to the originally secreted hormone, constitute the dominant form of circulating immunoreactive hormone.

Model for parathyroid hormone secretion and metabolism in calves.

A model for parathyroid gland response to differing calcium and magnesium concentrations is proposed based on direct determinations of parathyroid hormone (PTH) secretion rate. A proportional control seems adequate for physiological variations of calcium and magnesium levels. At extreme hypocalcemia, nonlinearities are observed: a model involving the depletion of a storage compartment is proposed, the size of which is calculated. PTH distribution was studied in another group of animals; sampling was made from the saphenous artery and the thoracic duct. Concentrations of plasma and lymph-intact PTH and carboxy-terminal fragments were determined by means of two different radioimmunoassays. The analysis of the results leads to the formulation of a five-compartment model, which shows that the metabolism of intact PTH is only partly due to the formation of carboxy-terminal fragments, most being directly secreted or catabolized. Linking these models for secretion and distribution of PTH, simulation studies were undertaken and compared with published data. This model is then discussed in comparison with previous work concerning other polypeptide hormones.

Parathyroid gland response to epinephrine: a rate-sensitivity mechanism.

A model for parathyroid gland response to an epinephrine infusion is proposed. It is based on direct measurements of the parathyroid hormone (PTH) secretion rate as measured directly after cannulation of the venous drainage of single superior parathyroid glands of calves. When epinephrine was infused into a jugular vein, a large increase of PTH secretion rate was observed at the beginning of the infusion. The PTH secretion rate thereafter fell, but it was still significantly above the control rate. At the end of the epinephrine infusion a sharp decrease was observed when the infusion was terminated. These observations are consistent with a rate-sensitive control mechanism. Calculation of the parameters yielded a model, which was then linked with a kinetic model for PTH distribution. Simulation studies were undertaken to predict plasma PTH concentration in response to an epinephrine infusion. Good agreement was found between predicted results and previously published work. The role of this rate-sensitive mechanism in the parathyroid gland is discussed.