Abstract
Classical stimulus-secretion theory suggests that each individual cell responds to a given stimulus. We tested this theory by determining the response of single bovine parathyroid cells to calcium with the reverse hemolytic plaque assay (RHPA), an assay that measures hormone release from individual cells. As calcium concentrations decreased, the amount of parathyroid hormone (PTH) released per cell increased, and cells were recruited to release PTH. To confirm that adequate stores of PTH were present, immunocytochemistry and in situ hybridization were performed. To test if cells that did not release PTH were capable of secretion, we performed a sequential RHPA; 47.8% of cells did not release PTH after the first stimulus. After the second exposure to low concentrations of calcium, 26.5% of these "nonsecretory" cells were able to release PTH. We conclude that parathyroid cells are homogeneous for PTH content and synthetic capability. Parathyroid cells respond to changes in extracellular calcium heterogeneously in that more PTH per cell is released, and individual parathyroid cells are "recruited" to release PTH at low calcium concentrations. In addition, parathyroid cells can be induced to secrete suggesting that cells are viable but in a depressed secretory state. Parathyroid cells may exist in an "on" or "off" secretory state.
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- BLACK L., BERENBAUM M. C. FACTORS AFFECTING THE DYE EXCLUSION TEST FOR CELL VIABILITY. Exp Cell Res. 1964 Jun;35:9–13. doi: 10.1016/0014-4827(64)90066-7. [DOI] [PubMed] [Google Scholar]
- Banerjee D. K., Ornberg R. L., Youdim M. B., Heldman E., Pollard H. B. Endothelial cells from bovine adrenal medulla develop capillary-like growth patterns in culture. Proc Natl Acad Sci U S A. 1985 Jul;82(14):4702–4706. doi: 10.1073/pnas.82.14.4702. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Boockfor F. R., Frawley L. S. Functional variations among prolactin cells from different pituitary regions. Endocrinology. 1987 Mar;120(3):874–879. doi: 10.1210/endo-120-3-874. [DOI] [PubMed] [Google Scholar]
- Brandi M. L., Ornberg R. L., Sakaguchi K., Curcio F., Fattorossi A., Lelkes P. I., Matsui T., Zimering M., Aurbach G. D. Establishment and characterization of a clonal line of parathyroid endothelial cells. FASEB J. 1990 Oct;4(13):3152–3158. doi: 10.1096/fasebj.4.13.1698682. [DOI] [PubMed] [Google Scholar]
- Brown E. M., Katz C., Butters R., Kifor O. Polyarginine, polylysine, and protamine mimic the effects of high extracellular calcium concentrations on dispersed bovine parathyroid cells. J Bone Miner Res. 1991 Nov;6(11):1217–1225. doi: 10.1002/jbmr.5650061112. [DOI] [PubMed] [Google Scholar]
- Calvo M. S., Eastell R., Offord K. P., Bergstralh E. J., Burritt M. F. Circadian variation in ionized calcium and intact parathyroid hormone: evidence for sex differences in calcium homeostasis. J Clin Endocrinol Metab. 1991 Jan;72(1):69–76. doi: 10.1210/jcem-72-1-69. [DOI] [PubMed] [Google Scholar]
- Conn P. M., Rogers D. C., Seay S. G., Staley D. Activation of luteinizing hormone release from pituitary cells by polycations. Endocrinology. 1984 Nov;115(5):1913–1917. doi: 10.1210/endo-115-5-1913. [DOI] [PubMed] [Google Scholar]
- Edidin M. A rapid, quantitative fluorescence assay for cell damage by cytotoxic antibodies. J Immunol. 1970 May;104(5):1303–1306. [PubMed] [Google Scholar]
- Fitzpatrick L. A., Leong D. A. Individual parathyroid cells are more sensitive to calcium than a parathyroid cell population. Endocrinology. 1990 Mar;126(3):1720–1727. doi: 10.1210/endo-126-3-1720. [DOI] [PubMed] [Google Scholar]
- Frawley L. S., Neill J. D. A reverse hemolytic plaque assay for microscopic visualization of growth hormone release from individual cells: evidence for somatotrope heterogeneity. Neuroendocrinology. 1984 Nov;39(5):484–487. doi: 10.1159/000124025. [DOI] [PubMed] [Google Scholar]
- Jia L. G., Canny B. J., Leong D. A. Paracrine communication regulates adrenocorticotropin secretion. Endocrinology. 1992 Jan;130(1):534–539. doi: 10.1210/endo.130.1.1309348. [DOI] [PubMed] [Google Scholar]
- Jia L. G., Canny B. J., Orth D. N., Leong D. A. Distinct classes of corticotropes mediate corticotropin-releasing hormone- and arginine vasopressin-stimulated adrenocorticotropin release. Endocrinology. 1991 Jan;128(1):197–203. doi: 10.1210/endo-128-1-197. [DOI] [PubMed] [Google Scholar]
- Jones K. H., Senft J. A. An improved method to determine cell viability by simultaneous staining with fluorescein diacetate-propidium iodide. J Histochem Cytochem. 1985 Jan;33(1):77–79. doi: 10.1177/33.1.2578146. [DOI] [PubMed] [Google Scholar]
- Kineman R. D., Faught W. J., Frawley L. S. Bovine pituitary cells exhibit a unique form of somatotrope secretory heterogeneity. Endocrinology. 1990 Nov;127(5):2229–2235. doi: 10.1210/endo-127-5-2229. [DOI] [PubMed] [Google Scholar]
- Kitamura N., Shigeno C., Shiomi K., Lee K., Ohta S., Sone T., Katsushima S., Tadamura E., Kousaka T., Yamamoto I. Episodic fluctuation in serum intact parathyroid hormone concentration in men. J Clin Endocrinol Metab. 1990 Jan;70(1):252–263. doi: 10.1210/jcem-70-1-252. [DOI] [PubMed] [Google Scholar]
- Klee G. G., Preissner C. M., Schryver P. G., Taylor R. L., Kao P. C. Multisite immunochemiluminometric assay for simultaneously measuring whole-molecule and amino-terminal fragments of human parathyrin. Clin Chem. 1992 May;38(5):628–635. [PubMed] [Google Scholar]
- Krishan A. Rapid flow cytofluorometric analysis of mammalian cell cycle by propidium iodide staining. J Cell Biol. 1975 Jul;66(1):188–193. doi: 10.1083/jcb.66.1.188. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Leong D. A. A complex mechanism of facilitation in pituitary ACTH cells: recent single-cell studies. J Exp Biol. 1988 Sep;139:151–168. doi: 10.1242/jeb.139.1.151. [DOI] [PubMed] [Google Scholar]
- Logue F. C., Fraser W. D., O'Reilly D. S., Cameron D. A., Kelly A. J., Beastall G. H. The circadian rhythm of intact parathyroid hormone-(1-84): temporal correlation with prolactin secretion in normal men. J Clin Endocrinol Metab. 1990 Dec;71(6):1556–1560. doi: 10.1210/jcem-71-6-1556. [DOI] [PubMed] [Google Scholar]
- Mayer G. P., Habener J. F., Potts J. T., Jr Parathyroid hormone secretion in vivo. Demonstration of a calcium-independent nonsuppressible component of secretion. J Clin Invest. 1976 Mar;57(3):678–683. doi: 10.1172/JCI108324. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Poenie M., Alderton J., Tsien R. Y., Steinhardt R. A. Changes of free calcium levels with stages of the cell division cycle. Nature. 1985 May 9;315(6015):147–149. doi: 10.1038/315147a0. [DOI] [PubMed] [Google Scholar]
- Porter T. E., Wiles C. D., Frawley L. S. Evidence for bidirectional interconversion of mammotropes and somatotropes: rapid reversion of acidophilic cell types to pregestational proportions after weaning. Endocrinology. 1991 Sep;129(3):1215–1220. doi: 10.1210/endo-129-3-1215. [DOI] [PubMed] [Google Scholar]
- Roth S. I., Raisz L. G. The course and reversibility of the calcium effect on the ultrastructure of the rat parathyroid gland in organ culture. Lab Invest. 1966 Jul;15(7):1187–1211. [PubMed] [Google Scholar]
- Rotman B., Papermaster B. W. Membrane properties of living mammalian cells as studied by enzymatic hydrolysis of fluorogenic esters. Proc Natl Acad Sci U S A. 1966 Jan;55(1):134–141. doi: 10.1073/pnas.55.1.134. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Shannon W. A., Jr, Roth S. I. An ultrastructural study of acid phosphatase activity in normal, adenomatous and hyperplastic (chief cell type) human parathyroid glands. Am J Pathol. 1974 Dec;77(3):493–506. [PMC free article] [PubMed] [Google Scholar]
- Smith P. F., Luque E. H., Neill J. D. Detection and measurement of secretion from individual neuroendocrine cells using a reverse hemolytic plaque assay. Methods Enzymol. 1986;124:443–465. doi: 10.1016/0076-6879(86)24034-3. [DOI] [PubMed] [Google Scholar]
- Uhl G. R. In situ hybridization: quantitation using radiolabeled hybridization probes. Methods Enzymol. 1989;168:741–752. doi: 10.1016/0076-6879(89)68055-x. [DOI] [PubMed] [Google Scholar]




