Subclones of a rat parathyroid cell line (PT-r): regulation of growth and production of parathyroid hormone-related peptide (PTHRP).
Four subclones from a rat parathyroid cell line (PT-r cell) have been isolated, and morphological and functional characteristics have been examined. Subclones 1 and 2 display a polygonal shape, show growth and secretory responses to calcium (half-maximal suppressions at 1.2 and 1.7 mM, respectively), and respond to secretin with cAMP production (14.5-fold and 16.9-fold over basal) and hormone secretion (41 and 58% over basal). Subclone 4 is elongated in form and does not respond to calcium or secretin. Subclone 3 shows mixed morphology, elongated and polygonal shapes, with moderate response to calcium (half-maximal suppression at 1.7 mM) and secretin (cAMP, 3.2-fold increase and hormone secretion, 50% increase over basal). The clones were tested for content of messenger RNA (mRNA) representing parathyroid hormone (PTH) and parathyroid hormone-related peptide (PTHRP). Only PTHRP mRNA was found. The peptide released is virtually all PTHRP. PTH mRNA was not detected even with a sensitive RNA probe. The amount of mRNA for PTHRP closely paralleled the amount of PTH-like bioactivity released into the medium from each clone (144.7 +/- 12.1, 110.0 +/- 12.9, 68.0 +/- 5.6, and 39.9 +/- 2.4 pgEq of rat PTH-(-34) per 10(7) cells per 12 h in a medium with 0.7 mM ionized calcium, from subclones 1, 2, 3, and 4, respectively). Culture conditions, low-density passage (less than 1:50 split ratio) or high-density passage (greater than 1:10 split ratio), affected morphology and function of the clones 1 and 2. They became elongated and functionally dedifferentiated like subclone 4 and 3 months of high-density culture.(ABSTRACT TRUNCATED AT 250 WORDS)
Sakaguchi
K
K
Metabolic Diseases Branch, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892.
Ikeda
K
K
Curcio
F
F
Aurbach
G D
GD
Brandi
M L
ML
D000818:Animals; D015152:Blotting, Northern; D002455:Cell Division / Q000502:physiology; D002460:Cell Line; D002875:Chromosomes; D002999:Clone Cells; D000242:Cyclic AMP / Q000378:metabolism; D010280:Parathyroid Glands / Q000166:cytology / Q000378:metabolism*; D010281:Parathyroid Hormone / Q000235:genetics / Q000378:metabolism; D044162:Parathyroid Hormone-Related Protein; D011061:Poly A / Q000302:isolation & purification; D014176:Protein Biosynthesis; D011506:Proteins / Q000235:genetics; D012313:RNA / Q000302:isolation & purification; D012333:RNA, Messenger / Q000096:biosynthesis; D051381:Rats; D012633:Secretin / Q000494:pharmacology; D013936:Thymidine / Q000378:metabolism; D014316:Tritium
D010281:Parathyroid Hormone; D044162:Parathyroid Hormone-Related Protein; D011506:Proteins; D012333:RNA, Messenger; D014316:Tritium; D012633:Secretin; D011061:Poly A; D012313:RNA; D000242:Cyclic AMP; D013936:Thymidine