Progress 10/22/13 to 09/30/18
Outputs
Target Audience:The target audience comprises scientists working in the field of muscle growth and differentiation as well as the broader areas of meat science, and meat animal growth and development. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?This project has resulted in training of 4 graduate students and 2 undergraduate student researchers during the previous 5 years How have the results been disseminated to communities of interest?Publications in the peer-reviewed literature and presentations at annual meetings (Plant and Animal Genome and NC1184) What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts
What was accomplished under these goals?
The Michigan station completed the analysis of NGS sequence of several chicken lines, particularly developmental mutants: stumpy,diplopodia3, diplopodia4, talpid2, limbless and eudiplopodia. Candidate genes and SNP were identified and are being pursued further. A likely candidate gene for talpid2 has been identified as the C2CD3 gene and a likely causal allele, a 19 bp indel, has been found in talpid2 C2CD3. In addition, we have been examining the latest chicken genome sequence assembly (galGal4) and exploring approaches to fill gaps and obtain missing segments of the assembly (particularly on microchromosomes). Optical map data have recently been obtained for the UCD001 reference bird (#256) and Moleculo sequence analysis is in progress. These efforts parallel work at the Wash. U. Genome Center that is doing targeted gap filling and PacBio sequencing. We have been examining the latest chicken genome sequence assembly (galGal4) and exploring approaches to fill gaps and obtain missing segments of the assembly (particularly on microchromosomes). Optical map and Moleculo sequence data have been obtained for the UCD001 reference bird (#256). For single gene traits, this provides a short list of possible candidate SNP that may include the causal allele. For complex traits, this provides data for genome-wide association analysis. All of these efforts are compromised by the incomplete nature of the reference genome sequence and the annotation that links it with genes and transcripts. New methodologies will substantially improve the reference sequence and better annotation will make that reference more useful to all users. Dr. Strasburg transitioned in as the Station rep in 2015 as Dr. Dodgson retired. His work in collaboration with Dr. Kent Reed at Minnesota and Dr. Sandra Velleman at Ohio State focused on effects of thermal challenge on cultured satellite cells from slow-growing and fast growing turkey lines. Satellite cells were cultured at various temperatures between 33 and 43°C and compared to cells cultured at the control temperature of 38 °C to ascertain temperature effects on lipid accumulation and expression of three different adipogenic genes: CCAAT/enhancer-binding protein-β (C/EBPbeta), proliferator-activated receptor-gamma (PPARgamma), and stearoyl-CoA desaturase (SCD). During satellite cell proliferation (increase in number), the amount of quantifiable lipid in both F and RBC2 satellite cells increased at temperatures above 38 °C and decreased at temperatures below 38 °C. Above 38 °C, RBC2 satellite cells had more lipid compared to the F line, whereas there were few differences between lines below 38 °C. At 72 h of proliferation, expression of C/EBPbeta, PPARgamma and SCD decreased as temperatures increased from 33 to 43 °C in both cell lines. During differentiation expression of C/EBPβ increased as temperatures increased from 33 to 43 °C in both cell lines. In F line satellite cells, PPARgamma expression decreased with increasing temperatures during differentiation, whereas there was no linear trend in RBC2 cells. During differentiation, expression of SCD increased as temperatures increased in RBC2 cells, and there was no linear trend within the F line.Results suggest that environmental temperature can affect breast muscle satellite cellular fate; however, selection for body weight had little impact on these cellular responses. Isolated skeletal muscle satellite cells of 7 wk old male turkeys (Meleagris gallopavo) were differentiated in culture at 38° C or thermally challenged at 33° C or 43° C. Included in this experiment were cells from two breeding lines; the F-line (16 wk body weight-selected) and RBC2 (it's randombred control). After for 48 h of differentiation, cells were harvested and total RNA was isolated for RNAseq analysis. Analysis of 39.9 Gb of sequence found 89% mapped to the turkey genome (UMD5.0, annotation 101) with average expression of 18917 genes per library. In the cultured satellite cells, slow/cardiac muscle isoforms are generally present in greater abundance than fast skeletal isoforms. Statistically significant differences in gene expression were observed among treatments and between turkey lines, with a greater number of genes affected in the F-line cells following cold treatment whereas more DE genes were observed in the RBC2 cells following heat treatment. Many of the most significant pathways involved signaling, consistent with ongoing cellular differentiation. Regulation of Ca2+ homeostasis appears to be significantly affected by temperature treatment, particularly cold treatment. Satellite cell differentiation is directly influenced by temperature at the level of gene transcription with greater effects attributed to selection for fast growth. At lower temperature, muscle-associated genes in the satellite cells were among the genes with the greatest down regulation consistent with slower differentiation and smaller myotubes. Fewer expression differences were observed in the differentiating cells than previously observed for proliferating cells. This suggests the impact of temperature on satellite cells occurs primarily at early points in satellite cell activation. Our most recent studies were designed to investigate changes in gene expression in the breast muscle as a function of temperature exposure of young turkey poults from a randombred turkey line representing the turkey of the 1960s, and a turkey line bred for 16-week body weight. Newly hatched turkey poults were brooded for 3 days at one of 3 temperatures: control (35°C), cold (31°C), or hot (39°C). Samples of the pectoralis major were harvested and subjected to RNA deep sequencing. Significant differential gene expression was observed in both growth-selected and randombred birds at both temperature extremes when compared to control-brooded poults. Broadly speaking, in this study the slower growing RBC2 birds responded to thermal stress almost exclusively with changes in expression of lipid-related genes. Both temperature treatments suggested a reduction in lipid storage, transport, or synthesis, consistent with changes in energy metabolism required to maintain body temperature. In contrast, the fast-growing F-line responded to thermal stress through changes in genes (ubiquitination and gene expression modulators) that would influence the levels of other gene products downstream. The F-line showed changed in genes that are predicted to result in reduced muscle growth. The temperature differences among treatment groups in this study were only 4°C warmer or colder relative to the standard brooding temperature. Temperature extremes during transportation of hatchlings from hatchery to grow-out facility may often be much greater, which in turn, will likely have greater effects on gene expression. This is the first to study to examine the effects of thermal challenge on gene expression in turkeys over the first 3 d following hatch.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2018
Citation:
Barnes, N.E., Mendoza, K.M., Strasburg, G.M., Velleman, S.G., and Reed, K.M. 2018. Thermal challenge alters the transcriptional profile of the breast muscle in turkey poults. Poultry Science pey401, https://doi.org/10.3382/ps/pey401
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2018
Citation:
Strasburg, G.M., and Aljarbou, W.A. 2018. Quantification of Total and Phosphorylated Pyruvate Dehydrogenase in Normal and Pale, Soft, Exudative Turkey Muscle. Presentation W831, Plant and Animal Genome Meeting, January 13, 2018, San Diego, CA.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2017
Citation:
Strasburg, G.M. 2017. Influence of Post-hatch Thermal Challenge on Turkey Breast Muscle Gene Expression. Presentation given at the annual meeting of the NC1184 USDA Multi-state Project Meeting, October 20, 2017, Gainesville,
FL
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Progress 10/01/16 to 09/30/17
Outputs
Target Audience:The target audience comprises scientists working in the field of muscle growth and differentiation as well as the broader areas of meat science, and meat animal growth and development. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?This project supported two graduate students: Galen George, who graduated with his M.S. in Food Science from Michigan State University in December 2016, and Natalie Barnes who is working on an M.S. at the University of Minnesota under the guidance of my collaborator, Dr. Kent Reed. In addition, Dr. Kristelle Mendoza has been working on this project at the University of Minnesota. How have the results been disseminated to communities of interest?Results of our work have been presented at the Plant and Animal Genome meeting and at the NC1170 USDA Multistate meeting. In addition, one manuscript was published in Poultry Science, one was published in BMC Genomics, and one has been submitted. What do you plan to do during the next reporting period to accomplish the goals?Together with the Minnesota and Ohio groups, we will continue analysis of RNA-Seq data to analyze changes in gene expression in birds subjected to posthatch thermal challenge. We will also be testing the effects of prehatch thermal challenge of developing embroyos on muscle growth and development of thermotolerance.
Impacts
What was accomplished under these goals?
The Michigan station is collaborating with the Minnesota and Ohio stations to define the effects of an acute thermal change on turkey satellite cell growth and development. Both in ovo and post-hatch thermal challenges will be used to isolate those times and tissue events most sensitive to increases in temperature. Finally, these investigators will attempt to develop embryonic thermal manipulation approaches to mitigate the damaging effects of post-hatch thermal challenges on long-term muscle growth and development in turkeys. Isolated skeletal muscle satellite cells of 7 wk old male turkeys (Meleagris gallopavo) were differentiated in culture at 38° C or thermally challenged at 33° C or 43° C. Included in this experiment were cells from two breeding lines; the F-line (16 wk body weight-selected) and RBC2 (it's randombred control). After for 48 h of differentiation, cells were harvested and total RNA was isolated for RNAseq analysis. Analysis of 39.9 Gb of sequence found 89% mapped to the turkey genome (UMD5.0, annotation 101) with average expression of 18917 genes per library. In the cultured satellite cells, slow/cardiac muscle isoforms are generally present in greater abundance than fast skeletal isoforms. Statistically significant differences in gene expression were observed among treatments and between turkey lines, with a greater number of genes affected in the F-line cells following cold treatment whereas more DE genes were observed in the RBC2 cells following heat treatment. Many of the most significant pathways involved signaling, consistent with ongoing cellular differentiation. Regulation of Ca2+ homeostasis appears to be significantly affected by temperature treatment, particularly cold treatment. Satellite cell differentiation is directly influenced by temperature at the level of gene transcription with greater effects attributed to selection for fast growth. At lower temperature, muscle-associated genes in the satellite cells were among the genes with the greatest down regulation consistent with slower differentiation and smaller myotubes. Fewer expression differences were observed in the differentiating cells than previously observed for proliferating cells. This suggests the impact of temperature on satellite cells occurs primarily at early points in satellite cell activation. Exposure of newly hatched turkey poults to hot or cold thermal stress often results in detrimental effects on breast muscle growth and development. Typical changes include increased lipid deposition and damage to muscle ultrastructure, leading to inferior meat quality with consequent economic losses to producers and processors. Likewise, exposure of market-weight turkeys to acute heat stress immediately prior to harvest, frequently results in a high incidence of inferior meat quality characterized by pale color, reduced marinade uptake and water-holding capacity, and poor protein functionality. Thermal manipulation of embryonic development has met with some success as a strategy to improve thermotolerance of broilers. We hypothesized that exposure of turkey eggs to a mild heat challenge at a critical developmental stage would alter muscle growth and development, thereby setting the stage for improved thermotolerance of the growing bird. Eggs from RBC2 (slow-growing) and F (fast-growing) turkey lines were exposed to a control temperature of 38C throughout 28 days of incubation or 12h of 39.5C between days 21-25. Following hatch, birds were brooded at temperatures of 31C, 35C (control), or 39C for 3d, followed by brooding at 35C until 14d of age when the birds were sacrificed. Breast muscle (P. major) was collected for analyses including muscle weight and fiber diameter. Results from this preliminary study suggest that mild thermal manipulation of turkey eggs results in changes in myogenesis, muscle growth, and development that may be associated with altered thermotolerance in the growing bird.
Publications
- Type:
Conference Papers and Presentations
Status:
Other
Year Published:
2016
Citation:
Strasburg, G.M. 2016. Influence of thermal challenge on turkey meat quality. Oral presentation at the NC1184 USDA Annual Multistate Project Meeting. October 21, 2016, Manhattan, Kansas
- Type:
Journal Articles
Status:
Published
Year Published:
2017
Citation:
Clark, D,L,; Strasburg, G,M,; Reed, K.M.; Velleman, S.G. 2017. Influence of temperature and growth selection on turkey pectoralis major muscle satellite cell adipogenic gene expression and lipid accumulation. Poultry Science 96:1015-1027.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2017
Citation:
Barnes, N.E.; Strasburg, G.M.; Velleman, S.G.; Reed, K.M. 2017. Transcriptional Response to Thermal Stress in Turkey Muscle. Poster 1134. Plant and Animal Genome Meeting, January 14 18, 2017. San Diego, CA
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2017
Citation:
Reed, K.M.; Velleman, S.G.; Strasburg, G.M. 2017 Temperature effects on differential gene expression in turkey satellite cells during proliferation and differentiation. Presentation W765, Plant and Animal Genome Meeting, January 14 18, 2017. San Diego, CA
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2017
Citation:
Strasburg, G.M.; Clark, D.L.; George, G.; Reed, K.M.; Velleman, S.G. 2017. Effect of embryonic and post-hatch thermal challenge on turkey muscle development. Presentation W766, Plant and Animal Genome Meeting, January 14 18, 2017. San Diego, CA
- Type:
Conference Papers and Presentations
Status:
Submitted
Year Published:
2017
Citation:
Reed, K.M.; Mendoza, K.M.; Strasburg, G.M.; Velleman, S.G. 2017. Response of turkey muscle satellite cells to thermal challenge. II. Transcriptome effects in differentiating cells. Submitted to Frontiers in Avian Physiology
- Type:
Journal Articles
Status:
Published
Year Published:
2017
Citation:
Reed, K.M.; Mendoza, K.M.; Abrahante, J.E.; Barnes, N.E.; Velleman, S.G., Strasburg, G.M. 2017. Response of turkey muscle satellite cells to thermal challenge. I. transcriptome effects in proliferating cells. BMC Genomics 18(1):352. doi: 10.1186/s12864-017-3740-4
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Progress 10/01/15 to 09/30/16
Outputs
Target Audience:The target audience comprises researchers and industry scientists working in the field of poultry genetics, as well as poultry muscle biology, growth, and development. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?This project has supported a graduate students at Michigan State University. Galen George is a M.S. candidate. In addition, it has supported a postdoctoral associate, Dr. Daniel Clark at Ohio State University, who is working under the direction of Dr. Sandy Velleman. How have the results been disseminated to communities of interest?Results of satellite cell culture work were communicated at the Plant and Animal Genome Meeting in San Diego, CA in January 2016, the Reciprocal Meat Conference of the American Meat Science Organization in San Angelo, TX in June 2016, and the Annual Meeting of the Poultry Science Association in New Orleans, LA in July 2016. What do you plan to do during the next reporting period to accomplish the goals?We are continuing studies on the effects of thermal challenge on differentiating muscle satellite cells from two different turkey genetic lines (RBC2 and F) from Ohio State University. In addition, we are analyizing results of the exposure of posthatch turkeys from the two genetic lines to different brooding temperatures on gene expression during the first 3 days, as well as muscle ultrastructure and meat quality characteristics at 16 weeks of age. In the next phase of the study, we will analyze the effect of embryonic thermal challenge on development of thermotolerance in posthatch RBC2 and F-line turkeys. Eggs will be exposed to brief thermal challenge late in development, and posthatch poults will be exposed to different brooding thermal challenge conditions. Muscle tissue from market weight birds will be analyzed for muscle ultrastructure and meat quality characteristics.
Impacts
What was accomplished under these goals?
The importance of muscle as a food is exemplified by the turkey meat processing industry. In the USA, there has been a continual increase in consumption of turkey meat from 4.9 pounds of boneless equivalent in 1960 to 14.4 pounds in 2002. Breast meat yield is the primary profit center for the commercial poultry industry. A predominant factor leading to this increase in consumer consumption is that poultry breast meat is regarded as the ideal lean meat for a healthy diet. This is coupled to its affordability and ease of preparation. A 1% increase in breast yield will amount to at least $75 million increase in revenues to the poultry industry. Commercial geneticists have placed a tremendous amount of selection pressure on breast yield and conformation, but not the mechanisms regulating the growth of muscle. The ability to regulate the growth and development of muscle depends on understanding the cellular regulatory mechanisms and cellular interactions that occur. Changes in muscle fiber size, extracellular areas available between the individual fibers and fiber bundles, and increased fat deposition will alter meat quality as have been illustrated by problems like pale, soft, and exudative turkey and pork. The results from this study will provide information on the role of specific genes whose function is currently poorly understood, on the growth and development of muscle. Results from these experiments will also enable breeders to develop strategies for genetic improvement of animals which will yield higher quality meat at low cost. During the past year, we have focused on objective 3, which is to elucidate genetic mechanisms that underlie economic traits and develop new methods to apply that knowledge to poultry breeding practices. Our focus this year was on the effects of thermal challenge on adipogenic properties (tendency to produce fat deposits) of satellite cells from turkey skeletal muscle from two genetic lines. The lines were a slow growing line (RBC2) and fast growing line (F line) derived from the RBC2 line with selection only for 16-week breast muscle mass. Satellite cells were cultured at various temperatures between 33 and 43 °C and compared to cells cultured at the control temperature of 38 °C to ascertain temperature effects on lipid accumulation and expression of three different adipogenic genes: CCAAT/enhancer-binding protein-β (C/EBPbeta), proliferator-activated receptor-gamma (PPARgamma), and stearoyl-CoA desaturase (SCD). During satellite cell proliferation (increase in number), the amount of quantifiable lipid in both F and RBC2 satellite cells increased at temperatures above 38 °C and decreased at temperatures below 38 °C. Above 38 °C, RBC2 satellite cells had more lipid compared to the F line, whereas there were few differences between lines below 38 °C. At 72 h of proliferation, expression of C/EBPbeta, PPARgamma and SCD decreased as temperatures increased from 33 to 43 °C in both cell lines. During differentiation expression of C/EBPβ increased as temperatures increased from 33 to 43 °C in both cell lines. In F line satellite cells, PPARgamma expression decreased with increasing temperatures during differentiation, whereas there was no linear trend in RBC2 cells. During differentiation, expression of SCD increased as temperatures increased in RBC2 cells, and there was no linear trend within the F line. Results from the current study suggest that environmental temperature can affect breast muscle satellite cellular fate; however, selection for body weight had little impact on these cellular responses.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2016
Citation:
Clark DL, Coy CS, Strasburg GM, Reed KM, Velleman SG. 2016. Temperature effect on proliferation and differentiation of satellite cells from turkeys with different growth rates. Poultry Science 95:934-47.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2016
Citation:
Clark DL, Strasburg GM, Reed KM, Velleman SG. 2016. Effects of temperature and growth selection on adipogenic potential of turkey pectoralis major muscle satellite cells. Book of Abstracts, 105th Annual Meeting Poultry Science Association. Poultry Science 95: (E-Suppl. 1), Abstract 189.
- Type:
Conference Papers and Presentations
Status:
Other
Year Published:
2016
Citation:
Strasburg G.M. Effects of Thermal Challenge on Turkey Muscle Development and Meat Quality. Oral Presentation at the Reciprocal Meat Conference, San Angelo, TX June 21, 2016
- Type:
Conference Papers and Presentations
Status:
Other
Year Published:
2016
Citation:
Fields B, Strasburg GM, Brackenridge A, Howard S, Hueser, K. 2016. All Species Are Getting Bigger - How Do We Handle and Is the Trend Here to Stay? Oral Presentation at the Reciprocal Meat Conference, San Angelo, TX June 21, 2016
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2016
Citation:
Strasburg GM, Clark DL, Coy CS, Velleman SG, George G, and Reed KM. 2016. Effects of Thermal Challenge on Turkey Muscle Development and Meat Quality. Plant and Animal Genome Meeting Abstracts, W762. Available from: https://pag.confex.com/pag/xxiv/webprogram/Paper18995.html
- Type:
Journal Articles
Status:
Published
Year Published:
2016
Citation:
Reed KM, Mendoza KM, Coy CS, Strasburg GM, and Velleman SG 2016. Temperature Effects on Gene Expression in Turkey Satellite Cells. Plant and Animal Genome Meeting Abstracts, W779. Available from: https://pag.confex.com/pag/xxiv/webprogram/Paper18923.html
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Progress 10/01/14 to 09/30/15
Outputs
Target Audience:Animal scientists. Poultry breeders. Poultry industry representatives. 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?
Nothing Reported
What do you plan to do during the next reporting period to accomplish the goals?Complete a new chicken genome sequence assembly (Galgal5). Complete manual annotation of chicken genes in the current chicken genome sequence assembly for use in annotating the new assembly.
Impacts
What was accomplished under these goals?
We have been examining the latest chicken genome sequence assembly (galGal4) and exploring approaches to fill gaps and obtain missing segments of the assembly (particularly on microchromosomes). Optical map and Moleculo sequence data have been obtained for the UCD001 reference bird (#256). These efforts parallel work at the Wash. U. Genome Center that is doing targeted gap filling and PacBio sequencing. In addition, a full manual annotation of the transcripts in the latest galGal4 reference sequence assembly is nearly complete. NGS sequence analysis is now feasible for numerous individual chicken lines. For single gene traits, this provides a short list of possible candidate SNP that may include the causal allele. For complex traits, this provides data for genome-wide association analysis. All of these efforts are compromised by the incomplete nature of the reference genome sequence and the annotation that links it with genes and transcripts. New methodologies will substantially improve the reference sequence and better annotation will make that reference more useful to all users.
Publications
|
Progress 10/22/13 to 09/30/14
Outputs
Target Audience: Animal scientists. Poultry breeders. Poultry industry representatives. 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? Publications What do you plan to do during the next reporting period to accomplish the goals? Continued manual annotation of the genome sequence and additional gap filling and expansion of the chicken genome reference sequence are planned. Generation and release of a new chicken genome reference sequence, galGal5, is anticipated.
Impacts
What was accomplished under these goals?
We completed the analysis of NGS sequence of several chicken lines, particularly developmental mutants: stumpy, diplopodia3, diplopodia4, talpid2, limbless and eudiplopodia. Candidate genes and SNP were identified and are being pursued further. A likely candidate gene for talpid2 has been identified as the C2CD3 gene and a likely causal allele, a 19 bp indel, has been found in talpid2 C2CD3. In addition, we have been examining the latest chicken genome sequence assembly (galGal4) and exploring approaches to fill gaps and obtain missing segments of the assembly (particularly on microchromosomes). Optical map data have recently been obtained for the UCD001 reference bird (#256) and Moleculo sequence analysis is in progress. These efforts parallel work at the Wash. U. Genome Center that is doing targeted gap filling and PacBio sequencing. In addition, a full manual annotation of the transcripts in the latest galGal4 reference sequence assembly is currently in progress.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2014
Citation:
Chang, C.-F., E.N. Schock, E.A. OHare, J. Dodgson, H.H. Cheng, W.M. Muir, R.E. Edelmann, M.E. Delany and S.A. Brugmann. 2014. The cellular and molecular etiology of the craniofacial defects in the avian ciliopathic mutant, talpid2. Development, 141:3003-3012.
- Type:
Book Chapters
Status:
Published
Year Published:
2014
Citation:
Dodgson, J.B. 2014. Genomics of food animals. In N. Van Alfen (ed.), Encyclopedia of Agriculture and Food Systems, Elsevier, Oxford, UK, pp. 433-453.
- Type:
Book Chapters
Status:
Published
Year Published:
2014
Citation:
Dodgson, J.B. 2014. Chapter 1: Avian Genomics. In C. Scanes, (ed.), Sturckies Avian Physiology, 6th ed., Elsevier, London, UK, pp. 3-13.
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