Ammonia elicits a different myogenic response in avian and murine myotubes

Rachel A. Stern, Srinivasan Dasarathy, Paul E. Mozdziak

Research output: Contribution to journalArticle

Abstract

Increased myostatin expression, resulting in muscle loss, has been associated with hyperammonemia in mammalian models of cirrhosis. However, there is evidence that hyperammonemia in avian embryos results in a reduction of myostatin expression, suggesting a proliferative myogenic environment. The present in vitro study examines species differences in myotube and liver cell response to ammonia using avian and murine-derived cells. Primary myoblasts and liver cells were isolated from embryonic day 15 and 17 chick embryos to be compared with mouse myoblasts (C2C12) and liver (AML12) cells. Cells were exposed to varying concentrations of ammonium acetate (AA; 2.5, 5, or 10 mM) to determine the effects of ammonia on the cells. Relative expression of myostatin mRNA, determined by quantitative real-time PCR, was significantly increased in AA (10 mM) treated C2C12 myotubes compared to both ages of chick embryonic myotube cultures after 48 h (P < 0.02). Western blot analysis of myostatin protein confirmed an increase in myostatin expression in AA-treated C2C12 myotubes compared to the sodium acetate (SA) controls, while myostatin expression was decreased in the chick embryonic myotube cultures when treated with AA. Myotube diameter was significantly decreased in AA-treated C2C12 myotubes compared to controls, while avian myotube diameter increased with AA treatment (P < 0.001). There were no significant differences between avian and murine liver cell viability, assessed using 2′, 7′- bis-(2-carboxyethyl)-5-(and-6-)-carboxyfluorescein, acetoxymethyl ester, when treated with AA. However, after 24 h, AA-treated avian myotubes showed a significant increase in cell viability compared to the C2C12 myotubes (P < 0.05). Overall, it appears that there is a positive myogenic response to hyperammonemia in avian myotubes compared to murine myotubes, which supports a proliferative myogenic environment.

Original languageEnglish (US)
Pages (from-to)1-12
Number of pages12
JournalIn Vitro Cellular and Developmental Biology - Animal
DOIs
StateAccepted/In press - Aug 29 2016
Externally publishedYes

Fingerprint

Skeletal Muscle Fibers
Ammonia
Myostatin
Liver
Hyperammonemia
Myoblasts
Cell Survival
Sodium Acetate
Chick Embryo
Ammonium Compounds
Real-Time Polymerase Chain Reaction
Esters
Acetates
Fibrosis
Embryonic Structures
Western Blotting
Muscles
Messenger RNA
Proteins
In Vitro Techniques

Keywords

  • Cirrhosis
  • Liver
  • Muscle
  • Myoblast
  • Myogenesis

ASJC Scopus subject areas

  • Developmental Biology
  • Cell Biology

Cite this

Ammonia elicits a different myogenic response in avian and murine myotubes. / Stern, Rachel A.; Dasarathy, Srinivasan; Mozdziak, Paul E.

In: In Vitro Cellular and Developmental Biology - Animal, 29.08.2016, p. 1-12.

Research output: Contribution to journalArticle

Stern, Rachel A.; Dasarathy, Srinivasan; Mozdziak, Paul E. / Ammonia elicits a different myogenic response in avian and murine myotubes.

In: In Vitro Cellular and Developmental Biology - Animal, 29.08.2016, p. 1-12.

Research output: Contribution to journalArticle

@article{55230396543c4857a80035f46ef6e518,
title = "Ammonia elicits a different myogenic response in avian and murine myotubes",
abstract = "Increased myostatin expression, resulting in muscle loss, has been associated with hyperammonemia in mammalian models of cirrhosis. However, there is evidence that hyperammonemia in avian embryos results in a reduction of myostatin expression, suggesting a proliferative myogenic environment. The present in vitro study examines species differences in myotube and liver cell response to ammonia using avian and murine-derived cells. Primary myoblasts and liver cells were isolated from embryonic day 15 and 17 chick embryos to be compared with mouse myoblasts (C2C12) and liver (AML12) cells. Cells were exposed to varying concentrations of ammonium acetate (AA; 2.5, 5, or 10 mM) to determine the effects of ammonia on the cells. Relative expression of myostatin mRNA, determined by quantitative real-time PCR, was significantly increased in AA (10 mM) treated C2C12 myotubes compared to both ages of chick embryonic myotube cultures after 48 h (P < 0.02). Western blot analysis of myostatin protein confirmed an increase in myostatin expression in AA-treated C2C12 myotubes compared to the sodium acetate (SA) controls, while myostatin expression was decreased in the chick embryonic myotube cultures when treated with AA. Myotube diameter was significantly decreased in AA-treated C2C12 myotubes compared to controls, while avian myotube diameter increased with AA treatment (P < 0.001). There were no significant differences between avian and murine liver cell viability, assessed using 2′, 7′- bis-(2-carboxyethyl)-5-(and-6-)-carboxyfluorescein, acetoxymethyl ester, when treated with AA. However, after 24 h, AA-treated avian myotubes showed a significant increase in cell viability compared to the C2C12 myotubes (P < 0.05). Overall, it appears that there is a positive myogenic response to hyperammonemia in avian myotubes compared to murine myotubes, which supports a proliferative myogenic environment.",
keywords = "Cirrhosis, Liver, Muscle, Myoblast, Myogenesis",
author = "Stern, {Rachel A.} and Srinivasan Dasarathy and Mozdziak, {Paul E.}",
year = "2016",
month = "8",
doi = "10.1007/s11626-016-0088-z",
pages = "1--12",
journal = "In Vitro Cellular and Developmental Biology - Animal",
issn = "1071-2690",
publisher = "Springer New York",

}

TY - JOUR

T1 - Ammonia elicits a different myogenic response in avian and murine myotubes

AU - Stern,Rachel A.

AU - Dasarathy,Srinivasan

AU - Mozdziak,Paul E.

PY - 2016/8/29

Y1 - 2016/8/29

N2 - Increased myostatin expression, resulting in muscle loss, has been associated with hyperammonemia in mammalian models of cirrhosis. However, there is evidence that hyperammonemia in avian embryos results in a reduction of myostatin expression, suggesting a proliferative myogenic environment. The present in vitro study examines species differences in myotube and liver cell response to ammonia using avian and murine-derived cells. Primary myoblasts and liver cells were isolated from embryonic day 15 and 17 chick embryos to be compared with mouse myoblasts (C2C12) and liver (AML12) cells. Cells were exposed to varying concentrations of ammonium acetate (AA; 2.5, 5, or 10 mM) to determine the effects of ammonia on the cells. Relative expression of myostatin mRNA, determined by quantitative real-time PCR, was significantly increased in AA (10 mM) treated C2C12 myotubes compared to both ages of chick embryonic myotube cultures after 48 h (P < 0.02). Western blot analysis of myostatin protein confirmed an increase in myostatin expression in AA-treated C2C12 myotubes compared to the sodium acetate (SA) controls, while myostatin expression was decreased in the chick embryonic myotube cultures when treated with AA. Myotube diameter was significantly decreased in AA-treated C2C12 myotubes compared to controls, while avian myotube diameter increased with AA treatment (P < 0.001). There were no significant differences between avian and murine liver cell viability, assessed using 2′, 7′- bis-(2-carboxyethyl)-5-(and-6-)-carboxyfluorescein, acetoxymethyl ester, when treated with AA. However, after 24 h, AA-treated avian myotubes showed a significant increase in cell viability compared to the C2C12 myotubes (P < 0.05). Overall, it appears that there is a positive myogenic response to hyperammonemia in avian myotubes compared to murine myotubes, which supports a proliferative myogenic environment.

AB - Increased myostatin expression, resulting in muscle loss, has been associated with hyperammonemia in mammalian models of cirrhosis. However, there is evidence that hyperammonemia in avian embryos results in a reduction of myostatin expression, suggesting a proliferative myogenic environment. The present in vitro study examines species differences in myotube and liver cell response to ammonia using avian and murine-derived cells. Primary myoblasts and liver cells were isolated from embryonic day 15 and 17 chick embryos to be compared with mouse myoblasts (C2C12) and liver (AML12) cells. Cells were exposed to varying concentrations of ammonium acetate (AA; 2.5, 5, or 10 mM) to determine the effects of ammonia on the cells. Relative expression of myostatin mRNA, determined by quantitative real-time PCR, was significantly increased in AA (10 mM) treated C2C12 myotubes compared to both ages of chick embryonic myotube cultures after 48 h (P < 0.02). Western blot analysis of myostatin protein confirmed an increase in myostatin expression in AA-treated C2C12 myotubes compared to the sodium acetate (SA) controls, while myostatin expression was decreased in the chick embryonic myotube cultures when treated with AA. Myotube diameter was significantly decreased in AA-treated C2C12 myotubes compared to controls, while avian myotube diameter increased with AA treatment (P < 0.001). There were no significant differences between avian and murine liver cell viability, assessed using 2′, 7′- bis-(2-carboxyethyl)-5-(and-6-)-carboxyfluorescein, acetoxymethyl ester, when treated with AA. However, after 24 h, AA-treated avian myotubes showed a significant increase in cell viability compared to the C2C12 myotubes (P < 0.05). Overall, it appears that there is a positive myogenic response to hyperammonemia in avian myotubes compared to murine myotubes, which supports a proliferative myogenic environment.

KW - Cirrhosis

KW - Liver

KW - Muscle

KW - Myoblast

KW - Myogenesis

UR - http://www.scopus.com/inward/record.url?scp=84984622681&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84984622681&partnerID=8YFLogxK

U2 - 10.1007/s11626-016-0088-z

DO - 10.1007/s11626-016-0088-z

M3 - Article

SP - 1

EP - 12

JO - In Vitro Cellular and Developmental Biology - Animal

T2 - In Vitro Cellular and Developmental Biology - Animal

JF - In Vitro Cellular and Developmental Biology - Animal

SN - 1071-2690

ER -