The effect of denervation on protein synthesis and degradation in adult rat diaphragm muscle

Heather M. Argadine, Nathan J. Hellyer, Carlos Bernardo Mantilla, Wen Zhi Zhan, Gary C Sieck

Research output: Contribution to journalArticle

38 Citations (Scopus)

Abstract

Previous studies showed that unilateral denervation (DNV) of the rat diaphragm muscle (DIAm) results in loss of myosin heavy chain protein by 1 day after DNV. We hypothesize that DNV decreases net protein balance as a result of activation of the ubiquitin-proteasome pathway. In DIAm strips, protein synthesis was measured by incorporation of 3H-Tyr, and protein degradation was measured by Tyr release at 1, 3, 5, 7, and 14 days after DNV. Total protein ubiquitination, caspase-3 expression/activity, and actin fragmentation were analyzed by Western analysis. We found that, at 3 days after DNV, protein synthesis increased by 77% relative to sham controls. Protein synthesis remained elevated at 5 (85%), 7 (53%), and 14 days (123%) after DNV. At 5 days after DNV, protein degradation increased by 43% relative to sham controls and remained elevated at 7 (49%) and 14 days (74%) after DNV. Thus, by 5 days after DNV, net protein balance decreased by 43% compared with sham controls and was decreased compared with sham at 7 (49%) and 14 days (72%) after DNV. Protein ubiquitination increased at 5 days after DNV and remained elevated. DNV had no effect on caspase-3 activity or actin fragmentation, suggesting that the ubiquitin-proteasome pathway rather than caspase-3 activation is important in the DIAm response to DNV. Early loss of contractile proteins, such as myosin heavy chain, is likely the result of selective protein degradation rather than generalized protein breakdown. Future studies should evaluate this selective effect of DNV.

Original languageEnglish (US)
Pages (from-to)438-444
Number of pages7
JournalJournal of Applied Physiology
Volume107
Issue number2
DOIs
StatePublished - Aug 2009

Fingerprint

Denervation
Diaphragm
Proteolysis
Muscles
Proteins
Caspase 3
Myosin Heavy Chains
Ubiquitination
Proteasome Endopeptidase Complex
Ubiquitin
Actins
Contractile Proteins
Muscle Proteins

Keywords

  • Innervation
  • Protein balance
  • Skeletal muscle

ASJC Scopus subject areas

  • Physiology
  • Physiology (medical)

Cite this

The effect of denervation on protein synthesis and degradation in adult rat diaphragm muscle. / Argadine, Heather M.; Hellyer, Nathan J.; Mantilla, Carlos Bernardo; Zhan, Wen Zhi; Sieck, Gary C.

In: Journal of Applied Physiology, Vol. 107, No. 2, 08.2009, p. 438-444.

Research output: Contribution to journalArticle

Argadine, Heather M. ; Hellyer, Nathan J. ; Mantilla, Carlos Bernardo ; Zhan, Wen Zhi ; Sieck, Gary C. / The effect of denervation on protein synthesis and degradation in adult rat diaphragm muscle. In: Journal of Applied Physiology. 2009 ; Vol. 107, No. 2. pp. 438-444.
@article{c8e2ec0f67e645bf9a05868442e4921b,
title = "The effect of denervation on protein synthesis and degradation in adult rat diaphragm muscle",
abstract = "Previous studies showed that unilateral denervation (DNV) of the rat diaphragm muscle (DIAm) results in loss of myosin heavy chain protein by 1 day after DNV. We hypothesize that DNV decreases net protein balance as a result of activation of the ubiquitin-proteasome pathway. In DIAm strips, protein synthesis was measured by incorporation of 3H-Tyr, and protein degradation was measured by Tyr release at 1, 3, 5, 7, and 14 days after DNV. Total protein ubiquitination, caspase-3 expression/activity, and actin fragmentation were analyzed by Western analysis. We found that, at 3 days after DNV, protein synthesis increased by 77{\%} relative to sham controls. Protein synthesis remained elevated at 5 (85{\%}), 7 (53{\%}), and 14 days (123{\%}) after DNV. At 5 days after DNV, protein degradation increased by 43{\%} relative to sham controls and remained elevated at 7 (49{\%}) and 14 days (74{\%}) after DNV. Thus, by 5 days after DNV, net protein balance decreased by 43{\%} compared with sham controls and was decreased compared with sham at 7 (49{\%}) and 14 days (72{\%}) after DNV. Protein ubiquitination increased at 5 days after DNV and remained elevated. DNV had no effect on caspase-3 activity or actin fragmentation, suggesting that the ubiquitin-proteasome pathway rather than caspase-3 activation is important in the DIAm response to DNV. Early loss of contractile proteins, such as myosin heavy chain, is likely the result of selective protein degradation rather than generalized protein breakdown. Future studies should evaluate this selective effect of DNV.",
keywords = "Innervation, Protein balance, Skeletal muscle",
author = "Argadine, {Heather M.} and Hellyer, {Nathan J.} and Mantilla, {Carlos Bernardo} and Zhan, {Wen Zhi} and Sieck, {Gary C}",
year = "2009",
month = "8",
doi = "10.1152/japplphysiol.91247.2008",
language = "English (US)",
volume = "107",
pages = "438--444",
journal = "Journal of Applied Physiology",
issn = "8750-7587",
publisher = "American Physiological Society",
number = "2",

}

TY - JOUR

T1 - The effect of denervation on protein synthesis and degradation in adult rat diaphragm muscle

AU - Argadine, Heather M.

AU - Hellyer, Nathan J.

AU - Mantilla, Carlos Bernardo

AU - Zhan, Wen Zhi

AU - Sieck, Gary C

PY - 2009/8

Y1 - 2009/8

N2 - Previous studies showed that unilateral denervation (DNV) of the rat diaphragm muscle (DIAm) results in loss of myosin heavy chain protein by 1 day after DNV. We hypothesize that DNV decreases net protein balance as a result of activation of the ubiquitin-proteasome pathway. In DIAm strips, protein synthesis was measured by incorporation of 3H-Tyr, and protein degradation was measured by Tyr release at 1, 3, 5, 7, and 14 days after DNV. Total protein ubiquitination, caspase-3 expression/activity, and actin fragmentation were analyzed by Western analysis. We found that, at 3 days after DNV, protein synthesis increased by 77% relative to sham controls. Protein synthesis remained elevated at 5 (85%), 7 (53%), and 14 days (123%) after DNV. At 5 days after DNV, protein degradation increased by 43% relative to sham controls and remained elevated at 7 (49%) and 14 days (74%) after DNV. Thus, by 5 days after DNV, net protein balance decreased by 43% compared with sham controls and was decreased compared with sham at 7 (49%) and 14 days (72%) after DNV. Protein ubiquitination increased at 5 days after DNV and remained elevated. DNV had no effect on caspase-3 activity or actin fragmentation, suggesting that the ubiquitin-proteasome pathway rather than caspase-3 activation is important in the DIAm response to DNV. Early loss of contractile proteins, such as myosin heavy chain, is likely the result of selective protein degradation rather than generalized protein breakdown. Future studies should evaluate this selective effect of DNV.

AB - Previous studies showed that unilateral denervation (DNV) of the rat diaphragm muscle (DIAm) results in loss of myosin heavy chain protein by 1 day after DNV. We hypothesize that DNV decreases net protein balance as a result of activation of the ubiquitin-proteasome pathway. In DIAm strips, protein synthesis was measured by incorporation of 3H-Tyr, and protein degradation was measured by Tyr release at 1, 3, 5, 7, and 14 days after DNV. Total protein ubiquitination, caspase-3 expression/activity, and actin fragmentation were analyzed by Western analysis. We found that, at 3 days after DNV, protein synthesis increased by 77% relative to sham controls. Protein synthesis remained elevated at 5 (85%), 7 (53%), and 14 days (123%) after DNV. At 5 days after DNV, protein degradation increased by 43% relative to sham controls and remained elevated at 7 (49%) and 14 days (74%) after DNV. Thus, by 5 days after DNV, net protein balance decreased by 43% compared with sham controls and was decreased compared with sham at 7 (49%) and 14 days (72%) after DNV. Protein ubiquitination increased at 5 days after DNV and remained elevated. DNV had no effect on caspase-3 activity or actin fragmentation, suggesting that the ubiquitin-proteasome pathway rather than caspase-3 activation is important in the DIAm response to DNV. Early loss of contractile proteins, such as myosin heavy chain, is likely the result of selective protein degradation rather than generalized protein breakdown. Future studies should evaluate this selective effect of DNV.

KW - Innervation

KW - Protein balance

KW - Skeletal muscle

UR - http://www.scopus.com/inward/record.url?scp=68249161970&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=68249161970&partnerID=8YFLogxK

U2 - 10.1152/japplphysiol.91247.2008

DO - 10.1152/japplphysiol.91247.2008

M3 - Article

C2 - 19520837

AN - SCOPUS:68249161970

VL - 107

SP - 438

EP - 444

JO - Journal of Applied Physiology

JF - Journal of Applied Physiology

SN - 8750-7587

IS - 2

ER -