Advertisement

Osteoporosis International

, Volume 27, Issue 10, pp 3091–3101 | Cite as

Room temperature housing results in premature cancellous bone loss in growing female mice: implications for the mouse as a preclinical model for age-related bone loss

  • U. T. Iwaniec
  • K. A. Philbrick
  • C. P. Wong
  • J. L. Gordon
  • A. M. Kahler-Quesada
  • D. A. Olson
  • A. J. Branscum
  • J. L. Sargent
  • V. E. DeMambro
  • C. J. Rosen
  • R. T. Turner
Original Article

Abstract

Summary

Room temperature housing (22 °C) results in premature cancellous bone loss in female mice. The bone loss was prevented by housing mice at thermoneutral temperature (32 °C). Thermogenesis differs markedly between mice and humans and mild cold stress induced by standard room temperature housing may introduce an unrecognized confounding variable into preclinical studies.

Introduction

Female mice are often used as preclinical models for osteoporosis but, in contrast to humans, mice exhibit cancellous bone loss during growth. Mice are routinely housed at room temperature (18–23 °C), a strategy that exaggerates physiological differences in thermoregulation between mice (obligatory daily heterotherms) and humans (homeotherms). The purpose of this investigation was to assess whether housing female mice at thermoneutral (temperature range where the basal rate of energy production is at equilibrium with heat loss) alters bone growth, turnover and microarchitecture.

Methods

Growing (4-week-old) female C57BL/6J and C3H/HeJ mice were housed at either 22 or 32 °C for up to 18 weeks.

Results

C57BL/6J mice housed at 22 °C experienced a 62 % cancellous bone loss from the distal femur metaphysis during the interval from 8 to 18 weeks of age and lesser bone loss from the distal femur epiphysis, whereas cancellous and cortical bone mass in 32 °C-housed mice were unchanged or increased. The impact of thermoneutral housing on cancellous bone was not limited to C57BL/6J mice as C3H/HeJ mice exhibited a similar skeletal response. The beneficial effects of thermoneutral housing on cancellous bone were associated with decreased Ucp1 gene expression in brown adipose tissue, increased bone marrow adiposity, higher rates of bone formation, higher expression levels of osteogenic genes and locally decreased bone resorption.

Conclusions

Housing female mice at 22 °C resulted in premature cancellous bone loss. Failure to account for species differences in thermoregulation may seriously confound interpretation of studies utilizing mice as preclinical models for osteoporosis.

Keywords

Animal models Osteoporosis Sympathetic signaling Thermogenesis 

Notes

Compliance with ethical standards

The experimental protocols were approved by the Institutional Animal Care and Use Committee, and the mice were maintained in accordance with the NIH Guide for the Care and the Use of Laboratory Animals.

Funding

This study received financial support from NIH AR060913, NASA NNX12AL24, and USDA 38420–17804.

Conflicts of interest

None.

Supplementary material

198_2016_3634_MOESM1_ESM.xlsx (14 kb)
ESM 1 (XLSX 13 kb)

References

  1. 1.
    Iwaniec UT, Turner RT (2013) Animal models for osteoporosis. In: Marcus R, Feldman D, Dempster D, Luckey M, Cauley JA (eds) Osteoporosis. Elsevier Academic press, New York, pp 939–961CrossRefGoogle Scholar
  2. 2.
    Guglielmi G, De Serio A, Fusilli S, Scillitani A, Chiodini I, Torlontano M, Cammisa M (2000) Age-related changes assessed by peripheral QCT in healthy Italian women. Eur Radiol 10:609–614CrossRefPubMedGoogle Scholar
  3. 3.
    Lee EY, Kim D, Kim KM, Kim KJ, Choi HS, Rhee Y, Lim SK (2012) Age-related bone mineral density patterns in Koreans (KNHANES IV). J Clin Endocrinol Metab 97:3310–3318CrossRefPubMedGoogle Scholar
  4. 4.
    Glatt V, Canalis E, Stadmeyer L, Bouxsein ML (2007) Age-related changes in trabecular architecture differ in female and male C57BL/6J mice. J Bone Mineral Res: Off J Am Soc Bone Mineral Res 22:1197–1207CrossRefGoogle Scholar
  5. 5.
    Rickard DJ, Iwaniec UT, Evans G et al (2008) Bone growth and turnover in progesterone receptor knockout mice. Endocrinology 149:2383–2390CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Frost HM (1999) Changing views about ‘Osteoporoses’ (a 1998 overview). Osteoporos Int: J Established Res Coop Between Eur Found Osteoporos Natl Osteoporos Found USA 10:345–352CrossRefGoogle Scholar
  7. 7.
    Hadjidakis DJ, Androulakis II (2006) Bone remodeling. Ann N Y Acad Sci 1092:385–396CrossRefPubMedGoogle Scholar
  8. 8.
    Seeman E (2009) Bone modeling and remodeling. Crit Rev Eukaryot Gene Expr 19:219–233CrossRefPubMedGoogle Scholar
  9. 9.
    Turner RT (1994) Cancellous bone turnover in growing rats: time-dependent changes in association between calcein label and osteoblasts. J Bone Miner Res: Off J Am Soc Bone Miner Res 9:1419–1424CrossRefGoogle Scholar
  10. 10.
    Turner RT, Evans GL, Wakley GK (1993) Mechanism of action of estrogen on cancellous bone balance in tibiae of ovariectomized growing rats: inhibition of indices of formation and resorption. J Bone Miner Res: Off J Am Soc Bone Miner Res 8:359–366CrossRefGoogle Scholar
  11. 11.
    Westerlind KC, Wronski TJ, Ritman EL, Luo ZP, An KN, Bell NH, Turner RT (1997) Estrogen regulates the rate of bone turnover but bone balance in ovariectomized rats is modulated by prevailing mechanical strain. Proc Natl Acad Sci U S A 94:4199–4204CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Turner RT, Philbrick KA, Wong CP, Olson DA, Branscum AJ, Iwaniec UT (2014) Morbid obesity attenuates the skeletal abnormalities associated with leptin deficiency in mice. J Endocrinol 223:M1–15CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Serrat MA, King D, Lovejoy CO (2008) Temperature regulates limb length in homeotherms by directly modulating cartilage growth. Proc Natl Acad Sci U S A 105:19348–19353CrossRefPubMedPubMedCentralGoogle Scholar
  14. 14.
    Iwaniec UT, Wronski TJ, Turner RT (2008) Histological analysis of bone. Methods Mol Biol 447:325–341CrossRefPubMedGoogle Scholar
  15. 15.
    Akhter MP, Iwaniec UT, Covey MA, Cullen DM, Kimmel DB, Recker RR (2000) Genetic variations in bone density, histomorphometry, and strength in mice. Calcif Tissue Int 67:337–344CrossRefPubMedGoogle Scholar
  16. 16.
    Turner RT, Kalra SP, Wong CP, Philbrick KA, Lindenmaier LB, Boghossian S, Iwaniec UT (2013) Peripheral leptin regulates bone formation. J Bone Miner Res: Off J Am Soc Bone Miner Res 28:22–34CrossRefGoogle Scholar
  17. 17.
    Menagh PJ, Turner RT, Jump DB, Wong CP, Lowry MB, Yakar S, Rosen CJ, Iwaniec UT (2010) Growth hormone regulates the balance between bone formation and bone marrow adiposity. J Bone Miner Res: Off J Am Soc Bone Miner Res 25:757–768Google Scholar
  18. 18.
    Dempster DW, Compston JE, Drezner MK, Glorieux FH, Kanis JA, Malluche H, Meunier PJ, Ott SM, Recker RR, Parfitt AM (2013) Standardized nomenclature, symbols, and units for bone histomorphometry: a 2012 update of the report of the ASBMR Histomorphometry Nomenclature Committee. J Bone Miner Res: Off J Am Soc Bone Miner Res 28:2–17CrossRefGoogle Scholar
  19. 19.
    Benjamini Y, Hochberg Y (1995) Controlling the false discovery rate: a practical and powerful approach to multiple testing. J R Stat Soc B 57:289–300Google Scholar
  20. 20.
    Team RC (2015) R: A language and environment for statistical computing. R Foundation for Statistical Computing, ViennaGoogle Scholar
  21. 21.
    Turner RT, Iwaniec UT, Andrade JE, Branscum AJ, Neese SL, Olson DA, Wagner L, Wang VC, Schantz SL, Helferich WG (2013) Genistein administered as a once-daily oral supplement had no beneficial effect on the tibia in rat models for postmenopausal bone loss. Menopause (New York, NY) 20:677–686CrossRefGoogle Scholar
  22. 22.
    Devlin MJ, Cloutier AM, Thomas NA, Panus DA, Lotinun S, Pinz I, Baron R, Rosen CJ, Bouxsein ML (2010) Caloric restriction leads to high marrow adiposity and low bone mass in growing mice. J Bone Miner Res: Off J Am Soc Bone Miner Res 25:2078–2088CrossRefGoogle Scholar
  23. 23.
    Turner RT, Iwaniec UT (2011) Low dose parathyroid hormone maintains normal bone formation in adult male rats during rapid weight loss. Bone 48:726–732CrossRefPubMedPubMedCentralGoogle Scholar
  24. 24.
    Brown RT, Baust JG (1980) Time course of peripheral heterothermy in a homeotherm. Am J Phys 239:R126–129Google Scholar
  25. 25.
    Swoap SJ, Gutilla MJ (2009) Cardiovascular changes during daily torpor in the laboratory mouse. Am J Physiol Regul integr Comp Physiol 297:R769–774CrossRefPubMedPubMedCentralGoogle Scholar
  26. 26.
    Tracy CR (1977) Minimum size of mammalian homeotherms: role of the thermal environment. Science (New York, NY) 198:1034–1035CrossRefGoogle Scholar
  27. 27.
    Eng JW, Reed CB, Kokolus KM, Pitoniak R, Utley A, Bucsek MJ, Ma WW, Repasky EA, Hylander BL (2015) Housing temperature-induced stress drives therapeutic resistance in murine tumour models through beta2-adrenergic receptor activation. Nat Commun 6:6426CrossRefPubMedPubMedCentralGoogle Scholar
  28. 28.
    Kokolus KM, Capitano ML, Lee CT et al (2013) Baseline tumor growth and immune control in laboratory mice are significantly influenced by subthermoneutral housing temperature. Proc Natl Acad Sci U S A 110:20176–20181CrossRefPubMedPubMedCentralGoogle Scholar
  29. 29.
    Rosania K (2014) Chilly mice confound cancer studies. Lab Anim 43:3CrossRefGoogle Scholar
  30. 30.
    Stemmer K, Kotzbeck P, Zani F, Bauer M, Neff C, Muller TD, Pfluger PT, Seeley RJ, Divanovic S (2015) Thermoneutral housing is a critical factor for immune function and diet-induced obesity in C57BL/6 nude mice. Int J Obes (2005) 39:791–797CrossRefGoogle Scholar
  31. 31.
    Bechtold DA, Sidibe A, Saer BR et al (2012) A role for the melatonin-related receptor GPR50 in leptin signaling, adaptive thermogenesis, and torpor. Curr Biol: CB 22:70–77CrossRefPubMedGoogle Scholar
  32. 32.
    Gavrilova O, Leon LR, Marcus-Samuels B, Mason MM, Castle AL, Refetoff S, Vinson C, Reitman ML (1999) Torpor in mice is induced by both leptin-dependent and -independent mechanisms. Proc Natl Acad Sci U S A 96:14623–14628CrossRefPubMedPubMedCentralGoogle Scholar
  33. 33.
    Swoap SJ, Weinshenker D (2008) Norepinephrine controls both torpor initiation and emergence via distinct mechanisms in the mouse. PLoS ONE 3, e4038CrossRefPubMedPubMedCentralGoogle Scholar
  34. 34.
    Swoap SJ, Gutilla MJ, Liles LC, Smith RO, Weinshenker D (2006) The full expression of fasting-induced torpor requires beta 3-adrenergic receptor signaling. J Neurosci: Off J Soc Neurosci 26:241–245CrossRefGoogle Scholar
  35. 35.
    Nguyen KD, Qiu Y, Cui X, Goh YP, Mwangi J, David T, Mukundan L, Brombacher F, Locksley RM, Chawla A (2011) Alternatively activated macrophages produce catecholamines to sustain adaptive thermogenesis. Nature 480:104–108CrossRefPubMedPubMedCentralGoogle Scholar
  36. 36.
    Denjean F, Lachuer J, Geloen A, Cohen-Adad F, Moulin C, Barre H, Duchamp C (1999) Differential regulation of uncoupling protein-1, −2 and −3 gene expression by sympathetic innervation in brown adipose tissue of thermoneutral or cold-exposed rats. FEBS Lett 444:181–185CrossRefPubMedGoogle Scholar
  37. 37.
    Vosselman MJ, van Marken Lichtenbelt WD, Schrauwen P (2013) Energy dissipation in brown adipose tissue: from mice to men. Mol Cell Endocrinol 379:43–50CrossRefPubMedGoogle Scholar
  38. 38.
    Gat-Yablonski G, Phillip M (2008) Leptin and regulation of linear growth. Curr Opin Clin Nutr Metab Care 11:303–308CrossRefPubMedGoogle Scholar
  39. 39.
    Hamrick MW, Pennington C, Newton D, Xie D, Isales C (2004) Leptin deficiency produces contrasting phenotypes in bones of the limb and spine. Bone 34:376–383CrossRefPubMedGoogle Scholar
  40. 40.
    Hill EL, Elde R (1991) Distribution of CGRP-, VIP-, D beta H-, SP-, and NPY-immunoreactive nerves in the periosteum of the rat. Cell Tissue Res 264:469–480CrossRefPubMedGoogle Scholar
  41. 41.
    Hohmann EL, Elde RP, Rysavy JA, Einzig S, Gebhard RL (1986) Innervation of periosteum and bone by sympathetic vasoactive intestinal peptide-containing nerve fibers. Science (New York, NY) 232:868–871CrossRefGoogle Scholar
  42. 42.
    Gordon CJ, Aydin C, Repasky EA, Kokolus KM, Dheyongera G, Johnstone AF (2014) Behaviorally mediated, warm adaptation: a physiological strategy when mice behaviorally thermoregulate. J Therm Biol 44:41–46CrossRefPubMedGoogle Scholar

Copyright information

© International Osteoporosis Foundation and National Osteoporosis Foundation 2016

Authors and Affiliations

  • U. T. Iwaniec
    • 1
    • 2
  • K. A. Philbrick
    • 1
  • C. P. Wong
    • 1
  • J. L. Gordon
    • 1
  • A. M. Kahler-Quesada
    • 1
  • D. A. Olson
    • 1
  • A. J. Branscum
    • 3
  • J. L. Sargent
    • 4
  • V. E. DeMambro
    • 5
  • C. J. Rosen
    • 5
  • R. T. Turner
    • 1
    • 2
  1. 1.Skeletal Biology Laboratory, School of Biological and Population Health SciencesOregon State UniversityCorvallisUSA
  2. 2.Center for Healthy Aging ResearchOregon State UniversityCorvallisUSA
  3. 3.Biostatistics Program, School of Biological and Population Health SciencesOregon State UniversityCorvallisUSA
  4. 4.College of Veterinary MedicineOregon State UniversityCorvallisUSA
  5. 5.Maine Medical Center Research InstituteScarboroughUSA

Personalised recommendations