Abstract
A number of studies have indicated that microgravity induces osteopenia and modulates functions of mammalian cells. However, the molecular mechanisms underlying these effects of microgravity are still unknown. Rat osteoblasts were cultured for 4 and 5 days during Shuttle-Spacelab flight, and fixed by guanidine isothiocyanate solution on board after treatment with 1α, 25 (OH)2 vitamin D3. The mRNA levels for platelet-derived growth factor (PDGF)-β receptor, epidermal growth factor (EGF) receptor, the growth factor receptor adaptor protein Shc, and c-fos were determined using the method of quantitative reverse transcription-polymerase chain reaction. The mRNA levels for EGF receptor were not altered by microgravity. However, the mRNA levels for PDGF-β receptor, Shc, and c-fos were decreased to 62, 55 and 25% on the 4th day of flight, and 47, 40, and 43% on the 5th day, respectively, as compared to the corresponding ground controls. Expression of the growth factor receptor and the receptor adaptor protein was modulated in rat osteoblasts during spaceflight. Data suggest that signal transduction via growth factor receptors in rat osteoblasts is impaired by microgravity. Dysfunction of osteoblasts might be involved in spaceflight-induced osteopenia.
Similar content being viewed by others
References
Morey-Holton ER, Arnaud SB: Skeletal responses to spaceflight. Adv Space Biol Med 1: 37–69, 1991
De Groot RP, Rijken PJ, Den Hertog J, Boonstra J, Verkleij AJ, De Laat SW, Kruijer W: Microgravity decreases c-fos induction and serum response element activity. J Cell Sci 97: 33–38, 1990
De Groot RP, Rijken PJ, Den Hertog J, Boonstra J, Verkleij AJ, De Laat SW, Kruijer W: Nuclear responses to protein kinase C signal transduction are sensitive to gravity changes. Exp Cell Res 197: 87–90, 1991
Rijken PJ, De Groot RP, Van Belzen N, De Laat SW, Boonstra J, Verkleij AJ: Inhibition of EGF-induced signal transduction by microgravity is independent of EGF receptor redistribution in the plasma membrane of Human A431 cells. Exp Cell Res 204: 373–377, 1993
Siddhanti SR, Quarles D: Molecular to pharmacologic control of osteoblast proliferation and differentiation. J Cell Biochem 55: 310–320, 1994
Batzer AG, Rotin D, Urena JM, Skolnik EK, Schlessinger J: Hierarchy of binding sites for Grb2 and Shc on the epidermal growth factor receptor. Mol Cell Biol 14: 5192–5201, 1994
Yokote K, Mori S, Hausen K, Mcglade J, Pawson T, Heldin C-H, Claesson-Welsh L: Direct interaction between Shc and the platelet-derived growth factor fl-receptor. J Biol Chem 269: 15337–15343, 1994
Arcellana-Panlilio MY, Schultz GA: Analysis of Messenger RNA. Meth Enzymol 225: 303–328, 1993
Singer-Sam J, Riggs AD: Quantitative analysis of messenger RNA levels: Reverse transcription-polymerase chain reaction single nucleotide primer extension assay. Meth Enzymol 225: 344–351, 1993
Robinson MO, Simon MI: Determining transcript number using the polymerase chain reaction: Pgk-2 mP2, and PGK-2 transgene mRNA levels during spermatogenesis. Nuc Acid Res 19: 1557–1562, 1991
Hirata S, Osada T, Hirai M, Hagihara K, Kato J: Expression of estrogen receptor in the rat brain. J Steroid Biochem Mol Biol 41: 583–587, 1992
Kageyama Y, Koide Y, Miyamoto S, Inoue T, Yoshida T: The biased V gene usage in the synovial fluid of patients with rheumatoid arthritis. Eur J Immunol 24: 1122–1129, 1994
Lee BY, Huynh T, Prichard LE, McGuire J, Polan ML: Gonadal steroids modulate interleukin-1 receptor antagonist mRNA expression in cultured human monocytes. Biochem Biophys Res Comm 209: 279–285, 1995
Akiyama H, Kanai S, Hirano M, Kumei Y, Shimokawa Y, Katano H, Hara E, Song S, Mukai C, Nagaoka S: An improved quantitative RTPCR fluorescent method for analysis of gene transcripts in STS-65 space shuttle experiments. J Biotechnol 47: 325–333, 1996
Kumei Y, Shimokawa Y, Katano H, Hara E, Akiyama H, Hirano M, Mukai C, Nagaoka S, Whitson PA, Sams CF: Spaceflight modulates insulin-like growth factor binding proteins and glucocorticoid receptor. J Appl Physiol 85: 139–149, 1998
Kumei Y, Shimokawa Y, Katano H, Hara E, Akiyama H, Hirano M, Mukai C, Nagaoka S, Whitson PA, Sams CF: Microgravity induces prostaglandin E2 and interleukin-6 production in normal rat osteoblasts: Role in bone demineralization. J Biotechnol 47: 313–324, 1996
Haussler MR, Jurutka PW, Hsieh J-C, Thompson PD, Selznick SH, Haussler CA, Whitfield GK: New understanding of the molecular mechanism of receptor-mediated genomic actions of the vitamin D hormone. Bone 17: 33–38, 1995
Wang WW, Howells RD: Sequence of the 5′-flanking region of the rat c-fos proto-oncogene. Gene 143: 261–264, 1994
Schule R, Rangarajan P, Kliewer S, Ransone LJ, Bolado J, Verma IM, Evans RM: Functional antagonism between oncoprotein c-jun and the glucocorticoid receptor. Cell 62: 1217–1226, 1990
Diamond MI, Miner JN, Yoshinaga SK, Yamamoto KR: Transcription factor interactions: selectors of positive or negative regulation from a single DNA element. Science 249: 1266–1272, 1990
Yin J, Howells RD: Glococorticoid-mediated down regulation of c-fos mRNA in C6 glioma cells: Lack of correlation with proenkephalin mRNA. Mol Brain Res 12: 187–194, 1992
Hughes-Fulford M, Tjandrawinata R, Fitzgerald J, Gasuad K, Gilbertson V: Effects of microgravity on osteoblast growth. Gravit Space Biol Bull 11: 51–60, 1998
Hunter T: Oncoprotein networks. Cell 88: 333–346, 1997
Rani CSS, Wang F, Fuior E, Berger A, Wu J, Sturgill TW, Beitner-Johnson D, LeRoith D, Varticovski L, Spiegel S: Divergence in signal transduction pathways of platelet-derived growth factor (PDGF) and epidermal growth factor (EGF) receptors. J Biol Chem 272: 10777–10783, 1997
Ballagi AE, Ishizaki A, Nehlin J-O, Funa K: Isolation and characterization of the mouse PDGF-′ receptor promoter. Biochem Biophys Res Comm 210: 165–173, 1995
Turner RT, Evans GL, Cavolina JM, Halloran B, Morey-Holton E: Programmed administration of parathyroid hormone increases bone formation and reduces bone loss in hindlimb-unloaded ovariectomized rats. Endocrinol 139: 4086–4091, 1998
Chow JW, Fox S, Jagger CJ, Chambers TJ: Role of parathyroid hormone in mechanical responsiveness of rat bone. Am J Physiol 274: E146–E154, 1998
Herrlich A, Daub H, Knebel A, Herrlich P, Ullrich A, Schultz G, Gudermann T: Ligand-independent activation of platelet-derived growth factor receptor is a necessary intermediate in lysophosphatidic, acid-stimulated mitogenic activity in L cells. Proc Natl Acad Sci USA 95: 8985–8990, 1998
Sambrrok J, Fritsch EF, Maniatis T: Molecular Cloning (2nd ed). Cold Spring Harbor Laboratory Press, New York, 1989
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Akiyama, H., Kanai, S., Hirano, M. et al. Expression of PDGF-β receptor, EGF receptor, and receptor adaptor protein Shc in rat osteoblasts during spaceflight. Mol Cell Biochem 202, 63–71 (1999). https://doi.org/10.1023/A:1007097511914
Issue Date:
DOI: https://doi.org/10.1023/A:1007097511914