title: Expression of neuropeptide receptor mRNA during osteoblastic differentiation of mouse iPS cells.
abstract: Various studies have shown a relationship between nerves and bones. Recent evidence suggests that both sensory and sympathetic nerves affect bone metabolism; however, little is known about how neuropeptides are involved in the differentiation of pluripotent stem cells into osteoblastic (OB) cells. To evaluate the putative effects of neuropeptides during the differentiation of mouse induced pluripotent stem (iPS) cells into calcified tissue-forming OB cells, we investigated the expression patterns of neuropeptide receptors at each differentiation stage. Mouse iPS cells were seeded onto feeder cells and then transferred to low-attachment culture dishes to form embryoid bodies (EBs). EBs were cultured for 4 weeks in osteoblastic differentiation medium. The expression of α1-adrenergic receptor (AR), α2-AR, β2-AR, neuropeptide Y1 receptor (NPY1-R), neuropeptide Y2 receptor (NPY2-R), calcitonin gene-related protein receptor (CGRP-R), and neurokinin 1-R (NK1-R) was assessed by reverse transcription-polymerase chain reaction (RT-PCR) and real-time PCR. Among these neuropeptide receptors, CGRP-R and β2-AR were expressed at all stages of cell differentiation, including the iPS cell stage, with peak expression occurring at the early osteoblastic differentiation stage. Another sensory nervous system receptor, NK1-R, was expressed mainly in the late osteoblastic differentiation stage. Furthermore, CGRP-R mRNA showed an additional small peak corresponding to EBs cultured for 3 days, suggesting that EBs may be affected by serum CGRP. These data suggest that the sensory nervous system receptor CGRP-R and the sympathetic nervous system receptor β2-AR may be involved in the differentiation of iPS cells into the osteoblastic lineage. It follows from these findings that CGRP and β2-AR may regulate cell differentiation in the iPS and EB stages, and that each neuropeptide has an optimal period of influence during the differentiation process.
authors.affiliation: Division of Developmental Stomatognathic Function Science, Kyushu Dental University, Kitakyushu 803-8580, Japan.
authors.forename: Satomi
authors.initials: S
authors.lastname: Nagao
authors.affiliation: Division of Oral Anatomy and Histology, Graduate School of Medical and Dental Sciences, Kagoshima University, Kagoshima 890-8544, Japan. Electronic address: tgoto@dent.kagoshima-u.ac.jp.
authors.forename: Tetsuya
authors.initials: T
authors.lastname: Goto
authors.affiliation: Anatomy, Kyushu Dental University, Kitakyushu 803-8580, Japan.
authors.forename: Shinji
authors.initials: S
authors.lastname: Kataoka
authors.affiliation: Oral Histology and Neurobiology, Kyushu Dental University, Kitakyushu 803-8580, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Toyono
authors.affiliation: Anatomy, Kyushu Dental University, Kitakyushu 803-8580, Japan.
authors.forename: Takaaki
authors.initials: T
authors.lastname: Joujima
authors.affiliation: Division of Molecular and Regenerative Prosthodontics, Tohoku University Graduate School of Dentistry, Sendai-city 980-8575, Japan.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Egusa
authors.affiliation: Division of Fixed Prosthodontics, Osaka University Graduate School of Dentistry, Suita, Osaka 565-0871, Japan.
authors.forename: Hirofumi
authors.initials: H
authors.lastname: Yatani
authors.affiliation: Anatomy, Kyushu Dental University, Kitakyushu 803-8580, Japan.
authors.forename: Shigeru
authors.initials: S
authors.lastname: Kobayashi
authors.affiliation: Division of Developmental Stomatognathic Function Science, Kyushu Dental University, Kitakyushu 803-8580, Japan.
authors.forename: Kenshi
authors.initials: K
authors.lastname: Maki
mesh_terms: D000818:Animals; D002454:Cell Differentiation; D002478:Cells, Cultured; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D010006:Osteoblasts / Q000166:cytology* / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism*; D011941:Receptors, Adrenergic / Q000378:metabolism*; D018340:Receptors, Adrenergic, alpha-1 / Q000378:metabolism; D018341:Receptors, Adrenergic, alpha-2 / Q000378:metabolism; D018343:Receptors, Adrenergic, beta-2 / Q000378:metabolism; D018015:Receptors, Calcitonin Gene-Related Peptide / Q000378:metabolism; D018040:Receptors, Neurokinin-1 / Q000378:metabolism; D018013:Receptors, Neuropeptide / Q000378:metabolism*; D017476:Receptors, Neuropeptide Y / Q000378:metabolism
chemical_list: D012333:RNA, Messenger; D011941:Receptors, Adrenergic; D018340:Receptors, Adrenergic, alpha-1; D018341:Receptors, Adrenergic, alpha-2; D018343:Receptors, Adrenergic, beta-2; D018015:Receptors, Calcitonin Gene-Related Peptide; D018040:Receptors, Neurokinin-1; D018013:Receptors, Neuropeptide; D017476:Receptors, Neuropeptide Y; C097212:neuropeptide Y-Y1 receptor; C095889:neuropeptide Y2 receptor
keywords: Calcitonin gene-related peptide receptor (CGRP-R); Induced pluripotent stem (iPS) cells; Osteoblastic (OB) cells; Sensory nervous system receptors; Sympathetic nervous system receptors; β2-adrenergic receptor (β2-AR)