The expression of PHOSPHO1, nSMase2 and TNAP is coordinately regulated by continuous PTH exposure in mineralising osteoblast cultures

Dean Houston; Katherine Myers; Victoria MacRae; Katherine Staines; Colin Farquharson

doi:10.1007/s00223-016-0176-9

The expression of PHOSPHO1, nSMase2 and TNAP is coordinately regulated by continuous PTH exposure in mineralising osteoblast cultures

Dean Houston, Katherine Myers, Victoria MacRae, Katherine Staines, Colin Farquharson

Royal (Dick) School of Veterinary Studies

Research output: Contribution to journal › Article › peer-review

Abstract

Sustained exposure to high levels of parathyroid hormone (PTH), as observed in hyperparathyroidism, are catabolic to bone. The increase in the RANKL/OPG ratio in response to continuous PTH, resulting in increased osteoclastogenesis is well established. However, the effects of prolonged PTH exposure on key regulators of skeletal mineralisation has yet to be investigated. This study sought to examine the temporal expression of PHOSPHO1, TNAP and nSMase2 in mineralising osteoblast-like cell cultures and to investigate the effects of continuous PTH exposure on the expression of these genes in vitro. PHOSPHO1, nSMase2 and TNAP expression in cultured MC3T3-C14 cells significantly increased from day 0 to day 10. PTH induced a rapid downregulation of Phospho1 and Smpd3 gene expression in MC3T3-C14 cells and cultured hemi-calvariae. Alpl was differentially regulated by PTH, displaying upregulation in cultured MC3T3-C14 cells and downregulation in hemi-calvariae. The effects of PTH on Phospho1 expression were mimicked with the cAMP agonist forskolin, and blocked by the PKA inhibitor PKI (5-24), highlighting a role for the cAMP/PKA pathway in this regulation. The potent down-regulation of Phospho1 and Smpd3 in osteoblasts in response to continuous PTH may provide a novel explanation for the catabolic effects on the skeleton of such an exposure. Furthermore, our findings support the hypothesis that PHOSPHO1, nSMase2 and TNAP function cooperatively in the initiation of skeletal mineralisation.

Original language	English
Pages (from-to)	510-524
Journal	Calcified Tissue International and Musculoskeletal Research
Volume	99
Issue number	5
Early online date	21 Jul 2016
DOIs	https://doi.org/10.1007/s00223-016-0176-9
Publication status	Published - Nov 2016

Keywords / Materials (for Non-textual outputs)

Parathyroid hormone
osteoblast
mineralisation regulators
hyperparathyroidism
PHOSPHO1

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10.1007/s00223-016-0176-9

The Expression of PHOSPHO1, nSMase2 and TNAP is Coordinately Regulated by Continuous PTH Exposure in Mineralising Osteoblast Cultures
© The Author(s) 2016 Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.
Final published version, 1.83 MBLicence: Creative Commons: Attribution (CC-BY)

Subchondral bone sclerosis in OA is driven by defective mineralisation and osteocyte formation
Farquharson, C. (Principal Investigator)
UK-based charities
1/01/14 → 31/12/16
Project: Research
Eastbio Doctoral Training Partnership
Farquharson, C. (Principal Investigator) & MacRae, V. (Co-investigator)
Biotechnology and Biological Sciences Research Council
1/10/12 → 30/09/18
Project: Research
Control of development and reproductive traits
Burdon, T. (Principal Investigator), Argyle, D. (Co-investigator), Ashworth, C. (Co-investigator), Beard, P. (Co-investigator), Brunton, P. (Co-investigator), Burt, D. (Co-investigator), Clinton, M. (Co-investigator), Dunn, I. (Co-investigator), Farquharson, C. (Co-investigator), Headon, D. (Co-investigator), Hocking, P. (Co-investigator), Hohenstein, P. (Co-investigator), Hume, D. (Co-investigator), Jackson, I. (Co-investigator), McColl, B. (Co-investigator), McGrew, M. (Co-investigator), McLachlan, G. (Co-investigator), Sang, H. (Co-investigator), Summers, K. (Co-investigator) & Whitelaw, B. (Co-investigator)
Biotechnology and Biological Sciences Research Council
1/04/12 → 31/03/17
Project: Research

PLA2 and ENPP6 may act in concert to generate phosphocholine from the matrix vesicle membrane during skeletal mineralization
Stewart, A., Leong, D. & Farquharson, C., Jan 2018, In: The FASEB Journal. 32, 1, p. 20-25
Research output: Contribution to journal › Article › peer-review

Cite this

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title = "The expression of PHOSPHO1, nSMase2 and TNAP is coordinately regulated by continuous PTH exposure in mineralising osteoblast cultures",

abstract = "Sustained exposure to high levels of parathyroid hormone (PTH), as observed in hyperparathyroidism, are catabolic to bone. The increase in the RANKL/OPG ratio in response to continuous PTH, resulting in increased osteoclastogenesis is well established. However, the effects of prolonged PTH exposure on key regulators of skeletal mineralisation has yet to be investigated. This study sought to examine the temporal expression of PHOSPHO1, TNAP and nSMase2 in mineralising osteoblast-like cell cultures and to investigate the effects of continuous PTH exposure on the expression of these genes in vitro. PHOSPHO1, nSMase2 and TNAP expression in cultured MC3T3-C14 cells significantly increased from day 0 to day 10. PTH induced a rapid downregulation of Phospho1 and Smpd3 gene expression in MC3T3-C14 cells and cultured hemi-calvariae. Alpl was differentially regulated by PTH, displaying upregulation in cultured MC3T3-C14 cells and downregulation in hemi-calvariae. The effects of PTH on Phospho1 expression were mimicked with the cAMP agonist forskolin, and blocked by the PKA inhibitor PKI (5-24), highlighting a role for the cAMP/PKA pathway in this regulation. The potent down-regulation of Phospho1 and Smpd3 in osteoblasts in response to continuous PTH may provide a novel explanation for the catabolic effects on the skeleton of such an exposure. Furthermore, our findings support the hypothesis that PHOSPHO1, nSMase2 and TNAP function cooperatively in the initiation of skeletal mineralisation.",

keywords = "Parathyroid hormone, osteoblast, mineralisation regulators, hyperparathyroidism, PHOSPHO1",

author = "Dean Houston and Katherine Myers and Victoria MacRae and Katherine Staines and Colin Farquharson",

year = "2016",

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doi = "10.1007/s00223-016-0176-9",

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journal = "Calcified Tissue International and Musculoskeletal Research",

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AU - Houston, Dean

AU - Myers, Katherine

AU - MacRae, Victoria

AU - Staines, Katherine

AU - Farquharson, Colin

PY - 2016/11

Y1 - 2016/11

N2 - Sustained exposure to high levels of parathyroid hormone (PTH), as observed in hyperparathyroidism, are catabolic to bone. The increase in the RANKL/OPG ratio in response to continuous PTH, resulting in increased osteoclastogenesis is well established. However, the effects of prolonged PTH exposure on key regulators of skeletal mineralisation has yet to be investigated. This study sought to examine the temporal expression of PHOSPHO1, TNAP and nSMase2 in mineralising osteoblast-like cell cultures and to investigate the effects of continuous PTH exposure on the expression of these genes in vitro. PHOSPHO1, nSMase2 and TNAP expression in cultured MC3T3-C14 cells significantly increased from day 0 to day 10. PTH induced a rapid downregulation of Phospho1 and Smpd3 gene expression in MC3T3-C14 cells and cultured hemi-calvariae. Alpl was differentially regulated by PTH, displaying upregulation in cultured MC3T3-C14 cells and downregulation in hemi-calvariae. The effects of PTH on Phospho1 expression were mimicked with the cAMP agonist forskolin, and blocked by the PKA inhibitor PKI (5-24), highlighting a role for the cAMP/PKA pathway in this regulation. The potent down-regulation of Phospho1 and Smpd3 in osteoblasts in response to continuous PTH may provide a novel explanation for the catabolic effects on the skeleton of such an exposure. Furthermore, our findings support the hypothesis that PHOSPHO1, nSMase2 and TNAP function cooperatively in the initiation of skeletal mineralisation.

AB - Sustained exposure to high levels of parathyroid hormone (PTH), as observed in hyperparathyroidism, are catabolic to bone. The increase in the RANKL/OPG ratio in response to continuous PTH, resulting in increased osteoclastogenesis is well established. However, the effects of prolonged PTH exposure on key regulators of skeletal mineralisation has yet to be investigated. This study sought to examine the temporal expression of PHOSPHO1, TNAP and nSMase2 in mineralising osteoblast-like cell cultures and to investigate the effects of continuous PTH exposure on the expression of these genes in vitro. PHOSPHO1, nSMase2 and TNAP expression in cultured MC3T3-C14 cells significantly increased from day 0 to day 10. PTH induced a rapid downregulation of Phospho1 and Smpd3 gene expression in MC3T3-C14 cells and cultured hemi-calvariae. Alpl was differentially regulated by PTH, displaying upregulation in cultured MC3T3-C14 cells and downregulation in hemi-calvariae. The effects of PTH on Phospho1 expression were mimicked with the cAMP agonist forskolin, and blocked by the PKA inhibitor PKI (5-24), highlighting a role for the cAMP/PKA pathway in this regulation. The potent down-regulation of Phospho1 and Smpd3 in osteoblasts in response to continuous PTH may provide a novel explanation for the catabolic effects on the skeleton of such an exposure. Furthermore, our findings support the hypothesis that PHOSPHO1, nSMase2 and TNAP function cooperatively in the initiation of skeletal mineralisation.

KW - Parathyroid hormone

KW - osteoblast

KW - mineralisation regulators

KW - hyperparathyroidism

KW - PHOSPHO1

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DO - 10.1007/s00223-016-0176-9

M3 - Article

SN - 0171-967X

VL - 99

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EP - 524

JO - Calcified Tissue International and Musculoskeletal Research

JF - Calcified Tissue International and Musculoskeletal Research

IS - 5

ER -

The expression of PHOSPHO1, nSMase2 and TNAP is coordinately regulated by continuous PTH exposure in mineralising osteoblast cultures

Abstract

Keywords / Materials (for Non-textual outputs)

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Projects

Subchondral bone sclerosis in OA is driven by defective mineralisation and osteocyte formation

Eastbio Doctoral Training Partnership

Control of development and reproductive traits

Research output

PLA2 and ENPP6 may act in concert to generate phosphocholine from the matrix vesicle membrane during skeletal mineralization

Cite this