Source: KANSAS STATE UNIV submitted to NRP
MOLECULAR MECHANISMS REGULATING SKELETAL MUSCLE GROWTH AND DIFFERENTIATION
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
COMPLETE
Funding Source
HATCH
Reporting Frequency
Annual
Accession No.
1006677
Grant No.
(N/A)
Cumulative Award Amt.
(N/A)
Proposal No.
(N/A)
Multistate No.
NC-_old1184
Project Start Date
Oct 1, 2015
Project End Date
Sep 30, 2020
Grant Year
(N/A)
Program Code
[(N/A)]- (N/A)
Recipient Organization
KANSAS STATE UNIV
(N/A)
MANHATTAN,KS 66506
Performing Department
Biochemistry and Molecular Biophysics
Non Technical Summary
As demand for pork rises, litter size has increased in swine herds to satisfy the demand. However as litter size increases, piglet birth weight has declined (Tung et al., 2012). Because of their smaller size, these piglets do not grow as fast as their contemporaries and are unable to go to slaughter. These pigs also have prewean mortality greater than 60% due to low viability, or being predisposed to being weaker (Wise et al., 1997). These piglets are competing with each other in the uterus and also with adolescent gilts that are still growing themselves (Gatford et al., 2010). Growth restricted piglets that do make it to market have reduced body weights and greater fat deposits (Tung et al., 2012). Therefore, the economic benefit of greater numbers of piglets born per sow is currently constrained because light birth weight pigs have a greater production cost than their heavier birth weight littermates. This is a result of decreased survivability (Panzardi et al, 2013), poorer growth rates, (Douglas et al, 2013), reduced carcass lean to fat ratios, and decreased feed efficiency throughout their life. Currently producers are trying to help small pigs with nurse sows or supplementation. Interventions to increase fetal growth is needed more for primiparous than multiparous sows (Gatford et al., 2009). Hence, the aim of this project is to demonstrate the molecular mechanisms responsible for the poorer performance of low birth weight piglets and explore possible countermeasures to increase their performance and survivability.
Animal Health Component
30%
Research Effort Categories
Basic
70%
Applied
30%
Developmental
(N/A)
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
3053510102050%
3053520102050%
Knowledge Area
305 - Animal Physiological Processes;

Subject Of Investigation
3510 - Swine, live animal; 3520 - Meat, swine;

Field Of Science
1020 - Physiology;
Goals / Objectives
Characterize the cellular and molecular basis of myogenesis. Characterize mechanisms of protein assembly and degradation in skeletal muscle.
Project Methods
A combination of genetic, molecular and biochemical approaches is being utilized to delineate mechanisms that contribute to the formation and/or maintenance of muscle tissue. In the lab exists a suite of genetic mutants that perturb muscle structure and/or function. Each of these mutations has been mapped to a single gene, confirmed using RNAi knockdown and/or transgenic rescue approaches, and grouped into phenotypic classes. One class of genes is required for the formation and assembly of the extracellular matrix (ECM) between myofibers and their corresponding attachment sites. Additional enhancer screens have identified genes encoding for ECM or crosslinking proteins that cooperatively function to maintain stable muscle-tendon junctions. A second class of mutants exhibit degenerative muscle phenotypes resulting from defects in cellular proteostasis. Genetic combined with yeast two-hybrid or mass spectrometry approaches have revealed that two independent mutants are defective in either the ubiquitin-proteasome or autophagic clearance pathways for protein turnover. Specific goals in the remainder of the funding period include: (1) characterization of crosslinking proteins in both the development and maintenance of the ECM environment residing at the muscle-tendon interface; (2) analysis of ubiquitin signatures on select proteins that prevent muscle degeneration and/or atrophy; and (3) delineating the biochemical complexes and genetic pathways required for the degradation of abnormal protein aggregates.

Progress 10/01/15 to 09/30/20

Outputs
Target Audience:Through this project, I estimate that we reached the following audiences: 1. Fellow scientists including muscle biologists and meat scientists that were attending sectional, regional, and national meetings. This group was also reached through peer-reviewed publications. 2. Swine and beef producers through extension related presentations and publications. 3. Pharmaceutical and industry related companies. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? Nothing Reported How have the results been disseminated to communities of interest?Journal articles What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? (1) Characterize the cellular and molecular basis of myogenesis. A common theme is the re-utilization of genes during development and again once muscle tissue maintenance ensues. A number of genes that control muscle development and homeostasis can easily be manipulated using genetics and/or biochemical approaches. We have continued to identify and characterize new genes, including the upstream signaling and downstream effectors that govern muscle formation and growth. (2) Characterize mechanisms of protein assembly and degradation in skeletal muscle. A balance between protein synthesis and degradation is crucial in muscle mass homeostasis and is mainly regulated by the ubiquitin-proteasome and autophagosome-lysosome pathways. To better understand how this balance is achieved, we have identified muscle proteins that are turned over by the proteasome in normal physiological conditions and whose degradation may be compromised in atrophy or disease conditions. In a separate project, we are in the process of characterizing target proteins that are subjected to autophagic disposal during normal muscle contraction.

Publications

  • Type: Journal Articles Status: Published Year Published: 2020 Citation: Muthukrishnan S, Mun S, Noh MY, Geisbrecht ER, Arakane Y. (2020) Insect Cuticular Chitin Contributes to Form and Function. Curr Pharm Des. May 23. doi: 10.2174/1381612826666200523175409. Online ahead of print. PMID: 32445445
  • Type: Journal Articles Status: Published Year Published: 2020 Citation: Brooks D, Naeem F, Stetsiv M, Goetting SC, Bawa S, Green N, Clark C, Bashirullah A, Geisbrecht ER. (2020) Drosophila NUAK functions with Starvin/BAG3 in autophagic protein turnover. PLOS Genetics. 16(4):e1008700. PMID: 32320396
  • Type: Journal Articles Status: Published Year Published: 2020 Citation: Bawa S, Brooks DS, Neville KE, Tipping M, Sagar MA, Kollhoff JA, Chawla G, Geisbrecht BV, Tennessen JM, Eliceiri KW, Geisbrecht ER. (2020) Drosophila TRIM32 cooperates with glycolytic enzymes to promote cell growth. Elife. Mar 30;9:e52358. PMID: 32223900


Progress 10/01/18 to 09/30/19

Outputs
Target Audience:1. Fellow scientists including muscle biologists and meat scientists that were attending sectional, regional, and national meetings. This group was also reached through peer-reviewed publications. 2. Swine and beef producers through extension related presentations and publications. 3. Poultry producers through individual presentations, radio interviews, and popular press releases. 4. Pharmaceutical and industry related companies. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Undergraduate Students: 7 Graduate Students: 3 How have the results been disseminated to communities of interest? Results were presented to fellow scientists attending the American Society of Animal Science Annual Conference and through peer-reviewed publications. Results were presented to industry sponsors. Results were presented at the Midwest and National conferences as well as peer-reviewed publications. What do you plan to do during the next reporting period to accomplish the goals?The project will be contnued under Dr. Geisbrecht

Impacts
What was accomplished under these goals? (1)Characterize the cellular and molecular basis of myogenesis. TRIM32 cooperates in a pathway with cell death proteins to regulate caspase activity and to facilitate muscle cell death. Loss of TRIM32 directly alters the localization of metabolic proteins and promotes muscle atrophy. (2)Characterize mechanisms of protein assembly and degradation in skeletal muscle. Histological measurement of poultry pectoralis major muscle from embryos injected with 2.5 mM of nicotinamide riboside at day 10 of incubation, indicate there was no effect on muscle fiber cross-sectional area. Therefore, it is hypothesized improvements in pectoralis major muscle weight may have been due to increases in muscle fiber number during myogenesis. Injecting nicotinamide riboside into the yolk of avian embryos at day 10 of incubation did not increase muscle mitochondria content. Damaged muscle tissue results in activation of the innate immune system which further drives muscle degeneration.

Publications

  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Coble, K., J. DeRouchey, J. M. Gonzalez, F. Wu, S. Dritz, J. Woodworth, M. Tokach, and R. Goodband. 2018. Effect of diet type and added copper on growth performance, carcass characteristics, energy digestibility, gut morphology, and mucosal mRNA expression of finishing pigs. J. Anim. Sci. 96:3288-3301. doi:10.1093/jas/sky196
  • Type: Journal Articles Status: Accepted Year Published: 2018 Citation: Montgomery, S. R., L. K. Mamedova, M. Zachut, G. Kra, S. H�ussler, H. Sauerwein, M. Vaughn, J. Gonzalez, and B. J. Bradford. 2018. Sodium salicylate may decrease glucose turnover rate in periparturient dairy cows through enhanced liver insulin sensitivity. J. Dairy Sci. In Press. https://doi.org/10.3168/jds.2018-15312
  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Vierck, K. R., T. G. OQuinn, J. A. Noel, T. A. Houser, E. A. E. Boyle, and J. M. Gonzalez. 2018. Effects of marbling texture on muscle fiber and collagen characteristics. Meat Mus. Bio. 2:75-82. doi:10.22175/mmb2017.10.0054
  • Type: Journal Articles Status: Under Review Year Published: 2019 Citation: Bawa S, Brooks D, Geisbrecht B, Tennessen J, and Geisbrecht ER. Drosophila TRIM32 cooperates with glycolytic enzymes in the metabolic maintenance of muscle tissue. Elife. In revision


Progress 10/01/17 to 09/30/18

Outputs
Target Audience:Through this project, I estimate that we reached the following audiences: 1. Fellow scientists including muscle biologists and meat scientists that were attending sectional, regional, and national meetings. This group was also reached through peer-reviewed publications. 2. Swine and beef producers through extension related presentations and publications. 3. Poultry producers through individual presentations, radio interviews, and popular press releases. 3. Pharmaceutical and industry related companies. Changes/Problems:Dr. Gonzalez is leaving K-State. We are in the process of determining if the Co-PD Dr. Geisbrecht will take this project over. What opportunities for training and professional development has the project provided?Undergraduate Students: 13 Graduate Students: 5 (Two Ph.D. students moved onto post-doctoral positions) How have the results been disseminated to communities of interest? Results were presented to fellow scientists attending the American Scociety of Animal Science Annual Conference and through peer-reviewed publications. Results were presented to industry sponsors. Results were presented at the Midwest and National conferences as well as peer-reviewed publications. What do you plan to do during the next reporting period to accomplish the goals?Dr. Gonzalez is leaving K-State. We are in the process of determining if the Co-PD Dr. Geisbrecht will take this project over.

Impacts
What was accomplished under these goals? Objective #1: Characterize the signal transduction pathways that regulate skeletal muscle growth and metabolism including the influence of endogenous growth factors and various production practices. TRIM32 cooperates in a pathway with cell death proteins to regulate caspase activity and to facilitate muscle cell death. Loss of TRIM32 directly alters the localization of metabolic proteins and promotes muscle atrophy. Objective #2: Characterize the cellular and molecular basis of myogenesis. Histological measurement of poultry pectoralis major muscle from embryos injected with 2.5 mM of nicotinamide riboside at day 10 of incubation, indicate there was no effect on muscle fiber cross-sectional area. Therefore, it is hypothesized improvements in pectoralis major muscle weight may have been due to increases in muscle fiber number during myogenesis. Injecting nicotinamide riboside into the yolk of avian embryos at day 10 of incubation did not increase muscle mitochondria content.

Publications

  • Type: Conference Papers and Presentations Status: Published Year Published: 2018 Citation: Davis, S. R., O. S. Khatri, and J. M. Gonzalez. 2018. Increasing the concentration of in ovo injected nicotinamide riboside has a positive influence on avian myogenesis. J. Anim. Sci. 96(E-Suppl. 2): (Abstr.).
  • Type: Conference Papers and Presentations Status: Published Year Published: 2018 Citation: Davis, S. R., O. S. Khatri, K. J. Phelps, and J. M. Gonzalez. 2018. The effects of in ovo injection of nicotinamide riboside on avian myogenesis. J. Anim. Sci. 96(E-Suppl. 2): (Abstr.).
  • Type: Conference Papers and Presentations Status: Accepted Year Published: 2018 Citation: Gonzalez, J. M., J. S. Drouillard, T. G. O⿿Quinn, and K. J. Phelps. 2018. The influence of muscle fiber structure on meat palatability. J. Anim. Sci. 96(E-Suppl. 2): (Abstr.).
  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Coble, K., J. DeRouchey, J. M. Gonzalez, F. Wu, S. Dritz, J. Woodworth, M. Tokach, and R. Goodband. 2018. Effect of diet type and added copper on growth performance, carcass characteristics, energy digestibility, gut morphology, and mucosal mRNA expression of finishing pigs. J. Anim. Sci. 96:3288-3301. doi:10.1093/jas/sky196
  • Type: Journal Articles Status: Awaiting Publication Year Published: 2019 Citation: Montgomery, S. R., L. K. Mamedova, M. Zachut, G. Kra, S. Häussler, H. Sauerwein, M. Vaughn, J. Gonzalez, and B. J. Bradford. 2018. Sodium salicylate may decrease glucose turnover rate in periparturient dairy cows through enhanced liver insulin sensitivity. J. Dairy Sci. In Press. https://doi.org/10.3168/jds.2018-15312
  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Vierck, K. R., T. G. O⿿Quinn, J. A. Noel, T. A. Houser, E. A. E. Boyle, and J. M. Gonzalez. 2018. Effects of marbling texture on muscle fiber and collagen characteristics. Meat Mus. Bio. 2:75-82. doi:10.22175/mmb2017.10.0054
  • Type: Conference Papers and Presentations Status: Published Year Published: 2018 Citation: Didier, K. D., S. M. Hammer, K. J. Phelps, J. M. Gonzalez, and T. J. Barstow. 2017. Near-infrared spectroscopy derived total heme vs. assay derived total heme. American College of Sports Medicne.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2018 Citation: Pearl, K. A., H. F. Speer, O. Khatri, S. Davis, M. D. Miesner, J. M. Gonzalez, and E. C. Titgemeyer. 2018. The effect of supplemental leucine on regulatory signaling in muscle of growing steers. J. Anim. Sci. 96(E-Suppl. 2): (Abstr.).


Progress 10/01/16 to 09/30/17

Outputs
Target Audience:Through this project, I estimate that we reached the following audiences: 1. Fellow scientists including muscle biologists and meat scientists that were attending sectional, regional, and national meetings. This group was also reached through peer-reviewed publications. 2. Swine and beef producers through extension related presentations and publications. 3. Pharmaceutical and industry related companies. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? Undergraduate Students: 7 Graduate Students: 4 Postdoctoral Associate: 1 How have the results been disseminated to communities of interest? Results were presented to fellow scientists attending the American Scociety of Animal Science Annual Conference and through peer-reviewed publications. Results were presented to industry sponsors. What do you plan to do during the next reporting period to accomplish the goals? Studies will continued to be conducted to examine the effect of nicotinamide riboside on avian myogenesis. Specifically, we will investigate mode of action. Studies will be conducted to examine the effect of in ovo nicotinamide riboside injection on avian postnatal muscle growth. Continue studies that examine the ability of nicotinamide riboside to increase NAD+ levels in the muscle of weaned pigs.

Impacts
What was accomplished under these goals? Objective #1: None. Objective #2: There were treatment × injection location interactions for chick weight and Pectoralis major weight and length (P < 0.02) when 0.0 or 2.5 mM nictotinamide riboside (NR) was injected into the yolk or albumen of the developing chick embryo. In all measures, there were no differences between 0.0 mM NR and 2.5 mM NR chicks when treatments were injected into the albumen (P > 0.14). However, when treatments were injected into the yolk, 2.5 NR chicks tended to weigh more (P = 0.07) and their Pectoralis major muscles weighed more and were longer than 0.0 NR chicks (P < 0.01). When 0.0, 2.5, 5.0, and 10.0 mM NR was injected into the yolk of day-10 embryos, Pectoralis major muscle weight, thickness, and length increased in a quadratic fashion, with no improvement in the reponse between the 5.0 and 10.0 nM treatments, when embryos were harvested at day 19 of incubtion. Data indicated supplementing porcine plasma to avian myoblasts did not affect cell proliferation, differentiation, or myotube width.

Publications

  • Type: Conference Papers and Presentations Status: Published Year Published: 2017 Citation: Hussein, A. H., E. D. Batista, M. A. Vaughn, S. R. Davis, E. F. Schwandt, E. J. McCoy, J. C. Simroth, C. D. Reinhardt, D. U. Thomson, M. D. Miesner, D. D. Burnett, J. M. Gonzalez, and E. C. Titgemeyer. 2017. Effects of ractopamine hydrochloride on lysine utilization by growing cattle. J. Anim. Sci. 95(E-Suppl. 4):275. (Abstr.)
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Lu,Y., J. S. Bradley, S. R. McCoski, J. M. Gonzalez, A. D. Ealy, and S. E. Johnson. 2017. Reduced skeletal muscle fiber size following caloric restriction is associated with calpain-mediated proteolysis and attenuation of IGF-1 signaling. Am. J. Physiol. Regul. Integr. Comp. Physiol. 312:R806-R815. doi:10.1152/ajpregu.00400.2016


Progress 10/01/15 to 09/30/16

Outputs
Target Audience:Through this project, I estimate that we reached the following audiences: 1. Fellow scientists including muscle biologists and meat scientists that were attending sectional, regional, and national meetings. This group was also reached through peer-reviewed publications. 2. Swine and beef producers through extension related presentations and publications. 3. Pharmaceutical and industry related companies. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Undergraduate Students: 7 Graduate Students: 3 Postdoctoral Associate: 1 How have the results been disseminated to communities of interest?Results have been presented to fellow muscle biologists at the NC1184 yearly meeting at Kansas State University. Results were presented to fellow scientists attending the Recipricol Meats Conference and through peer-reviewed publications. Results were presented to industry sponsors. What do you plan to do during the next reporting period to accomplish the goals?Studies will be conducted to examine the effect of nicotinamide riboside on avian myogenesis. Studies will be conducted to examine the ability of nicotinamide riboside to increase NAD+ levels in the muscle of weaned pigs. Studies will be conducted to examine the effect of porccine plasma on avain myogenesis.

Impacts
What was accomplished under these goals? Objective #1: Supplementation of market pig satellite cells with 5 mM of nicotinamide riboside reduced cell proliferation. Objective #2: We have detected a delayed trajectory of myogenesis when comparing the muscle developmental events of small, medium, and large pig fetuses. Fetal myblasts isolated small and medium sized fetuses at d-60 of gestation had a greater poliferation rate than myoblasts from large fetuses. Myoblast isolated from large fetuses had a greater differentiation capacity than small and medium, and medium myoblasts had a greater differentation capcaity than small myoblasts. When protein synthesis assays were conducted, small myoblasts had a greater myotube diameter compared to medium and large, and medium myoblasts produced greater myotube diameters compared to large. Supplementing d-60 of gestation porcine myoblasts with 10% procine plasma increased proliferation and reduced differentation and protein synthesis, while the opposite occurred in myoblasts from neonate piglets.

Publications

  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Burnett, D. D., C. B. Paulk, M. D. Tokach, J. L. Nelssen, M. A. Vaughn, K. J. Phelps, S. S. Dritz, J. M. DeRouchey, R.D. Goodband, K. D. Haydon and J. M. Gonzalez. 2016. Effects of Added Zinc on Skeletal Muscle Morphometrics and Gene Expression of Finishing Pigs Fed Ractopamine-HCl. Anim. Bio. 27:17-29. doi:10.1080/10495398.2015.1069301
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Noel, J. A., R. M. Broxterman, G. M. McCoy, J. C. Craig, K. J. Phelps, D. D. Burnett, M. A. Vaughn, T. J. Barstow, T. G. OQuinn, J. C. Woodworth, J. M. DeRouchey, T. G. Rozell, and J. M. Gonzalez. 2016. Use of electromyography to detect muscle exhaustion in finishing barrows fed ractopamine-HCl. J. Anim. Sci. 94:2344-2356. doi:10.2527/jas2016-0398
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Vaughn, M. A., C. Serrano, D. D. Burnett, D. Davis, J. Woodworth, and J. M. Gonzalez. 2016. Influence of porcine plasma supplementation on gestating sow serum IGF-1 concentration and litter weights. Prof. Ag. Workers J. 4:6.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Feldpausch, J. A., J. Jourquin, J. R. Bergstrom, C. D. Bokenkroger, J. L. Nelssen, M. J. Ritter, D. L. Davis, and J. M. Gonzalez, Birth weight threshold for identifying piglets at-risk for preweaning mortality. J. Anim. Sci. 94(E-Suppl. 2):33. (Abstr.)
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Vaughn, M. A., J. M. Morton, J. A. Feldpausch, K. J. Phelps, J. A. Noel, D. L. Davis, J. L. Nelssen, and J. M. Gonzalez. 2016. Impact of fetal size on myogenesis and Pax7+ progenitor cell populations. 69th Recip. Meat Conf. Proc., San Angelo, TX.