ANNUAL REPORT
Regional Research Project NC-131
1997

PROJECT TITLE: Molecular Mechanisms Regulating Muscle Growth and Differentiation

PROJECT PERIOD: 10-1-96 TO 9-30-97

COOPERATING AGENCIES AND PRINCIPAL:

PROGRESS OF THE WORK AND PRINCIPAL ACCOMPLISHMENTS:

Objective 1: To Characterize the signal transduction pathways that regulate skeletal muscle growth and differentiation.

The Arizona station demonstrated that hepatocyte growth factor/scatter factor can stimulate activation and early division of adult satellite cells in culture. New evidence indicates that hepatocyte growth factor/scatter factor is present in uninjured adult rat skeletal muscle and that the activating factor in crushed muscle extract is hepatocyte growth factor/scatter factor. Adult rat skeletal muscle shows the presence of hepatocyte growth factor/scatter factor in the extracellular matrix surrounding muscle fibers; in addition, the receptor for hepatocyte growth factor/scatter factor, c-met, is localized to putative satellite cells. In muscle from mdx mice, hepatocyte growth factor/scatter factor and c-met are co-localized in activated satellite cells and in early myotubes in regions of muscle repair. Finally, the satellite cell activating activity of crushed muscle extract is abolished by preincubation with anti-hepatocyte growth factor antibodies. In conclusion, hepatocyte growth factor/scatter factor is present in muscle, can be released upon injury and can function in a paracrine or autocrine manner to activate quiescent satellite cells.

In another study, the Arizona station screened over 40 antibodies (monoclonals) to seven different peptide antigens. A number of antibodies were identified that recognize polypeptides migrating in the 90-100 kDa range in SDS-PAGE (predicted MW of skm-calpain is 94-kDa). The polypeptides recognized by these antibodies have been phosphorylase in several instances and gelsolin in another instance. Several antibodies recognized a polypeptide in the 25-30 kDa range that existed specifically in skeletal muscle and not in liver, kidney, or cardiac tissue (a requirement for a skm-calpain antibody). In the most recent effort, a "polyclonal" antibody obtained from France against a peptide representing amino acids Gln⁶⁴⁵ to Gln⁶⁶² (human sequence numbers) labels a 90-100 kDa polypeptide in skeletal muscle extracts but not in liver, heart, or kidney extracts. This antibody provides some promise in recognizing skm-calpain.

The Indiana station determined parameters for isolating porcine myoblasts from mixed primary cell preparations. Clones were determined to be myogenic by cell fusion and reverse transcriptase-PCR analysis of muscle specific mRNA. Populations of larger cells either detached or remained mononucleated, whereas the smaller cells readily fused to form multinucleated myotubes. Size differences between myoblasts and fibroblasts are lost after a single passage of mixed primary cultures. Therefore, porcine myoblasts can be isolated from mixed primary cultures prior to the first passage based on size alone.

The Michigan station used clonal cultures of porcine muscle satellite cells to characterize direct effects of testosterone on muscle cell proliferation and differentiation. They demonstrated the presence of nuclear androgen receptors in porcine satellite cells and satellite cell-derived myotubes. Testosterone administration increased immunoreactive androgen receptor protein and reduced (20-30%) satellite cell differentiation. Testosterone had no effect on satellite cell proliferation or responsiveness to growth factors (FGF, IGF-1, PDGF-BB, or EGF). Another study identified specific binding of beta adrenergic agonists (BAA) to cultured C₂C₁₂ muscle cells, which resulted in increased intracellular cAMP. These data indicate that androgens and BAA interact with skeletal muscle cell receptors and may influence muscle growth via direct action.

The Nebraska station conducted a study to determine the effects at differentiation on ribosome recruitment. Myoblasts had higher polyribosome percentages (indicator of increased ribosome initiation) than myotubes. Polyribosome percent decreased after the cells fused. Total RNA concentration did not change. It was concluded that changes in the rate of protein synthesis during differentiation is due to changes in the rate of initiation of polyribosomes, not to the amount of RNA that is present. Ribosomes are thought to be present in the cell as free, membrane bound, or cytoskeletal bound polysomes. Data also indicate that polysome percentage in the free fraction decreased from 24 hours prior to addition of fusion media to 24 hours after addition of fusion media. It was also observed that the percent of polysomes increased in the cytoskeletal fraction during the same time.

The New York Station used the Cornell Net Carbohydrate and Protein System (CNCPS) model to formulate a "balanced" combination of animal by-product proteins to be used as the protein supplement for optimizing amino acid composition of the absorbed pool in growing-finishing beef steers, and 2) to determine the optimum quantity of the "balanced" combination of proteins for facilitating maximum rate and efficiency of protein gain. In summary, the CNCPS model to balance quantity of amino acids absorbed from both microbial and undegraded intake protein sources of protein in growing-finishing cattle fed corn-based grain diets improves efficiency and rates of protein gain. Use of animal by-product undegraded intake protein sources in a combination indicated by the models improved apparent N digestibility, protein accretion rates, efficiency of protein gain, and skeletal muscle growth.

The Ohio Station continued work on the role of the extracellular matrix in chicken skeletal muscle development working with Low Score Normal (LSN) muscle weakness in collaboration with the South Dakota Station. A significant elevation in both decorin protein and transcript levels was reported at 20 days of embryonic development. Collagen concentration is unaffected whereas collagen crosslinking is significantly elevated at six weeks posthatch. In a second study, TGF-ß increased decorin expression. On embryonic Day 20 through 1 wk posthatch, TGF-ß2 transcript levels were elevated whereas an increase in TGF-ß1 was only noted at 1 d posthatch. The data suggests that an increase in decorin expression may be associated with a modification in a TGF-ß signal transduction pathway.

Collaboration of the South Dakota and Ohio Stations have examined normal chicken satellite cells for the synthesis of proteoglycans. Initial studies have identified a heparan sulfate proteoglycan synthesized during satellite cell proliferation and differentiation. After the induction of fusion, this heparan sulfate proteoglycan has both an intracellular and extracellular distribution. Similar to chicken, the turkey muscle matrix changes from a chondroitin sulfate-rich matrix to one containing both heparan and chondroitin sulfate proteoglycans.

The South Dakota station satellite cells were isolated from the diaphragms and limb muscles of dystrophic and normal hamsters to explore cellular mechanisms in muscular dystrophy. The results indicate that satellite cells can be successfully cultured from dystrophic and normal hamsters and rapid proliferation can be obtained with the medium developed. Clonal populations of satellite cells are now being developed to alleviate problems with fibroblast overgrowth of the cultures. In another study the responses of fast growing and slow growth myogenic cell clones to growth factors were determined. The fast growing clone was more responsive to the mitogenic effects of FGF, IGF-I, insulin, and PDGF and the proliferation and differentiation depressing effects of TGF-ß Administration of hepatocyte growth factor (HGF) to the slow growing clone increases proliferation rates, but has no effect on the fast growing clone.

The USDA-MARC station investigated the mechanism of muscle hypertrophy in callipyge lambs. Callipyge muscle weights increased at a greater rate than non-callipyge controls. At death, calpastatin activity in callipyge muscle was higher (P < .05) than non-callipyge controls. Callipyge phenotype muscle had elevated protein:DNA and RNA:DNA ratios. Fractional protein synthesis rate was decreased in callipyge muscle compared to non-callipyge. Fractional protein degradation rate was decreased in callipyge muscle. In conclusion callipyge-induced muscle hypertrophy is, in part, due to decreased muscle protein degradation.

A study to examine the ultrastructure of muscle from callipyge lambs concluded that the major factor responsible for the toughness of meat from callipyge longissimus is the stability of myofibrils and myofibril associated cytoskeleton postmortem. Another group of experiments were conducted to determine the effect of rapid prerigor freezing and postrigor calcium chloride injection on tenderness of callipyge longissimus. Results indicate that either treatment can effectively mitigate the negative effect of callipyge phenotype on longissimus tenderness. Callipyge lamb muscle subjected to the combination of prerigor liquid nitrogen freezing, postrigor calcium chloride injection, and 14 d postmortem storage was similar in (P > .05) tenderness to normal untreated lamb after 14 d of postmortem storage.

The Washington station examined the differences in variables of satellite cell strain morphology, proliferation and differentiation. It identified satellite cell strains that 1) are morphologically distinct, 2) grow at different rates [+/- growth factors], 3) differentiate to distinguishably different (and repeatable) levels [measured by fusion] and 4) express different amounts of endogenous mitogenic growth-regulating agents (IGFs and IGFBPs). Preliminary results indicate at least four non-transformed, sheep-derived satellite cell strains responded to (RIA grade) sheep-derived growth hormone, whereas at least nine other strains did not respond to GH. The Washington station is also developing an in vitro system to examine paracrine regulation between muscle and fat cells. Two different defined media have been examined; one media was best for optimizing fusion of ovine satellite cells, and the second for promoting differentiation of preadipocytes.

Objective 2: To Determine the nuclear mechanisms that control gene expression in skeletal muscle.

The Indiana station studied whether if intramuscular injection of a DNA construct for luciferase in pigs is an effective method for obtaining production of recombinant protein. Greatest activity was detected at the injection sites, and acitivity decreased as distance from the site increased. Greater doses of DNA appeared to increase the migration of luciferase to more distant sites, even though the volume of injection was the same for all doses.

The Indiana srtation also identified the all three fast MyHC transcripts were expressed in the longissimus, whereas only type IIA and type IIX transcripts were present in deep red semitendinosus muscle. Expression of the three fast adult MyHC isoforms in longissimus was spatially regulated around the typical islets of type I fibers encountered in pig skeletal muscle.

The Minnesota station completed several studies examining the role of IGF and insulin-like growth factor binding proteins (IGFBP’s) in controlling muscle growth and differentiation. They observed that compared to levels present in myogenin negative, nonfused cultures, the steady-state level of IGFBP-3 mRNA was decreased by approximately three-fold in differentiating cultures. In extensively fused cultures, the steady-state level of IGFBP-3 mRNA was increased by approximately five-fold as compared to the level in myogenin negative, nonfused cultures. The data suggests very little change in steady-state levels of IGFBP-5 mRNA during differentiation of myogenic cultures. Steady-state IGFBP-2 mRNA levels were similar early on but later steady-state IGFBP-2 mRNA levels were increased approximately three-fold as compared to nonfused cultures. No IGFBP-4 mRNA was detectable in total RNA from differentiating porcine myogenic cultures. Changes in steady-state IGFBP-3 mRNA levels during porcine myogenic cell differentiation suggest this IGFBP may play an important role in regulating the bioactivity of IGF during differentiation of porcine myogenic cells. A second study was designed to assess changes in IGFBP mRNA levels in the presence of TGF-b1 or IGF-I during differentiation of porcine myogenic cells. Observations indicate that TGF-b1 increased steady-state IGFBP-3 mRNA levels and IGFBP-3 secretion at the same time is suppressed fusion, suggests that elevated IGFBP-3 mRNA and protein may play a role in TGF-b1-induced inhibition of fusion in porcine embryonic muscle cells. A third study investigated the effects of IGF-I and Des (1-3)-IGF-I on IGFBP production by cultured porcine embryonic myoblasts and muscle-derived fibroblasts. Des (1-3)-IGF-I is an IGF-I analog which has greatly decreased affinity for binding proteins but has a similar affinity for IGF-I receptors as compared to IGF-I. Proliferation of porcine embryonic myoblasts increases in response to both IGF-I and Des (1-3)-IGF-I in a dose-dependent manner, however, Des (1-3)-IGF-I stimulates proliferation at a lower concentration than does IGF-I. The IGFBP production levels in conditioned medium. The results provide evidence that IGF-I regulates IGFBP production in primary porcine embryonic muscle cells. This suggests that altering local IGFBP production may be another mechanism by which IGF regulates myoblast proliferation and differentiation.

The Minnesota station studied the effects of the combined trenbolone acetate and estrdiol implants on IGF-I mRNA concentrations. They observed an increase in serum concentrations of IGF-I as early as 3 days after implantation. There was also an increase in IGF-I mRNA in the liver and muscle. In conclusion , increased local production of IGF-I by muscle tissue may play a role in increasing circulating IGF-I concentrations as well as an autocrine or paracrine role in stimulating muscle growth in implanted steers. After implantation, sera from steers implanted with TBA + E₂ showed an increase in IGF-I concentrations as compared to sera from nonimplanted steers. The TBA+E₂ increased the percentage of muscle satellite cells that were able to proliferate. Fusion percentage was greater in implanted versus non-implanted steer cultures. Implanted steer cultures also possessed a significantly greater number of myotube nuclei than non-implanted steer cultures. Because of the crucial role muscle satellite cells play in postnatal muscle growth, this TBA+E₂-induced satellite cell activation may play an important role in TBA+E₂-enhanced muscle growth.

The Washington station has screened a variety of growth regulators, including cytokines, for ability to promote equine satellite cell proliferation and differentiation in vitro. Some factors (ex. basic FGF), which are powerful growth-promoting agents in other satellite cell systems were not as proficient in promoting activity of equine satellite cells.

The Wisconsin station demonstrated that after 28 days of hindlimb suspension, there was significantly less myosin heavy chain Type IIA, and more myosin heavy chain Type IIX in the soleus muscles of hindlimb suspended rats compared to soleus muscles from weight bearing rats. Although there was a shift to a faster myosin heavy chain phenotype in the hindlimb suspended rat soleus muscles, there was no change in myogenin expression or MyoD expression. Semi-quantitiative RT-PCR revealed an upregulation of MyoD expression following 14 days of hindlimb suspension. It was concluded that myogenin does not play a role in the slow to fast myofiber phenotypic transition that occurs during hindlimb suspension, but MyoD may play a role in the phenotypic transition. The Wisconsin station also examined the potential relationship between hypertrophy of the rat soleus after clenbuterol and 3,3',5-Triiodo-L-Thyronine treatment (CT) or after surgical overload, and myogenin or MyoD expression. The CT treatment induced the appearance of the fast Type IIX myosin heavy chain isoform, depressed myogenin expression, and induced MyoD expression. However, functional overload did not alter myogenin or MyoD expression in CT treated or non-CT treated rats. Thus, pharmacologically and surgically induced hypertrophy involves different mechanisms because myogenin and MyoD expression levels were associated with changes in myosin heavy chain composition (especially Type IIX) rather than the changes in muscle mass.

Objective 3: To Characterize muscle proteins and their function domains involved in myofibrillar assembly and disassembly.

The Arizona station has completed the characterization of a group of monoclonal antibodies to 1/4-calpain, m-calpain, and calpastatin. Six of these monoclonal antibodies have been made available commercially through Affinity Bereagents, Inc.

The California station has constructed expression vectors containing the full length LMM domain, and mutants lacking 16, 44, 72, and 100 aa residues. They demonstrated that removal of the terminal 100 aa residues, generated a LMM-peptide that remains soluble in low ionic strength buffers. In another study, they cloned and expressed neonatal and adult histidine-tagged and untagged LMM constructs in order to study LMM subunit exchange. It was concluded that stability of a discrete length at the N-terminal end of the LMM is required to nucleate formation of the ±-helical coiled-coil and that amino acid differences at the interhelical interface in this region are inhibitory to coiled-coil formation. In the second study, it was observed that amino acid substitutions in the e and g helix position (as opposed to the a and d position) on the heptad could explain the preference of chicken myosin rods to form homodimers. The e and g interhelical interactions are known to regulate dimerization of ±-helical coiled coils such as Fos/Jun and B-Zip transcription factor members. It was observed that e and g charge distribution may contribute to the different arrangements of molecules in sarcomeric and non-sarcomeric myosin filaments.

The Iowa Station determined that enzymatic transfer of ADP-ribose from NAD to arginine residues of specific cellular proteins appears to be an important regulatory process in many tissues. Summaries of earlier studies on the role of this important covalent protein modification in developing skeletal muscle have demonstrated that: (1) both the arginine-specific ADP-ribosyl transferase and the ribosyl-arginine hydrolase, which removes ADP-ribose from proteins, are present and can be isolated from skeletal muscle; (2) transferase activity is present in myoblasts in early cultures of embryonic chick myogenic cell cultures, and this activity increases dramatically with myogeinc differentiation; (3) a specific inhibitor of arginine mono-ADP-ribosylation of proteins, meta-iodobenzylguanidine (MIBG) reversibly blocks differentiation and proliferation of myoblasts in myogenic cell cultures; (4) the muscle-specific intermediate filament (IF) protein desmin is an in vitro substrate for the muscle transferase; (5) the ADP-ribosylation of desmin blocks the ability of this protein to assemble into IFs in vitro. Other studies have identified two major sites of arginine ADP-ribosylation in desmin. Both of these arginine residues are in the head region of the desmin molecule, a region that is known to be important in regulation of assembly. These results suggest that desmin is ADP-ribosylated in muscle cells. This process may be involved in regulating assembly of IFs during the process of myofibrillogenesis in developing muscle cells. Experiments are in progress to further characterize the role of ADP-ribosylation, and specifically the role of desmin modification, in myogenic cells.

Paranemin was purified from embryonic skeletal muscle and specific monoclonal antibodies were prepared. To further characterize this protein, cDNA clones encoding the complete sequence of paranemin were obtained. Northern blot analysis revealed a single transcript of 5.3 kb, which is much smaller than the predicted size expected for a protein of 280 kDa. The sequence coded for a protein of 1,606 residues (178,161 kDa ). The derived amino acid sequence demonstrated the presence of the conserved rod domain characteristic of IF proteins, thus demonstrating that paranemin is a novel IF protein. Of the various other known IF proteins, paranemin demonstrated the highest homology to nestin and tanabin. Immunofluorescence with paranemin and vimentin antibodies demonstrated that expressed paranemin was associated with the IFs in cells expressing vimentin, but was diffusely distributed in cells without IFs. These latter results suggest that paranemin alone does not form IFs, but rather is incorporated into heteropolmeric IFs with other IF proteins.

The Nebraska station has employed capillary electrophoresis to examine the action of purified cathepsin D on hemoglobin. The study found that cathepsin D produced a characteristic "fingerprint" of hemoglobin peptides and that its action is rapid. The sensitivity of the assay could be improved by labeling hemoglobin with fluorescein derivatives.

The Wisconsin station developed a monoclonal antibody (H-4) against bovine cardiac titin and has labeled seven repetitive epitopes in the frog skeletal muscle half sarcomere. However, since the resolution of immunoelectron microscopy is limited, it is impossible to define unambiguously whether the repetitive epitopes correspond exactly with the positions of the thick filament associated proteins. Nevertheless, the common antigenic properties of the repetitive epitopes and the titin-C protein interactions shown in binding assays all support the protein ruler hypothesis for both the thick filament associated proteins and titin.

USEFULNESS OF FINDINGS

The result found in this progress report indicate the NC-131 research committee has been very active during 1996-97 in doing basic research that has relevance to animal agriculture. The in the project the overall goal of the committee is to increase the efficiency of lean meat production through the use of the basic research into the biological and molecular mechanisms that control muscle growth and differentiation in meat producing livestock. Information from this research is vital for the future development of the applied methods and techniques for improving meat production, both it efficiency and safety. This overall it very much as part of the national priorities for the research in animal agriculture.

The committee has been very productive and evidenced by the number and quality of the publication resulting from activities in the laboratories of the committee members. This report covers the second year of the renewal of the NC-131 project. This has been substantial work done on each objective which suggests that the committee is focused and committed to accomplishing the objective that they have set. The committee has demonstrated a tremendous amount of the collaboration and cooperation as demonstrated by the sharing of techniques, culture models, antibodies, molecular probes, and expertise in different areas. This has been a valuable outcome of the committee and has been wise investment of the resources.

WORK PLANNED FOR NEXT YEAR

Studies will continue as described in the project proposal. Some changes in project personnel and participation by specific stations have occurred. Dr Katherine Byrne of the Washington station became a member of the committee and Dr. Matthew Doumit became the representative for the Michigan station. Effort will be put forth to include industry representation to improve the technology transfer to the animal industry.

PUBLICATIONS FOR THE YEAR :

Published Full Length Articles

Arizona

Allen, R. E., C. J. Temm-Grove, S. M. Sheehan, and G. Rice. 1997. Skeletal muscle satellite cell cultures. Meth. Cell Biol. 52, (in press).

Tatsumi, R., J. E. Anderson, O. Halevy, and R. E. Allen. 1997. HGF/SF is present in normal adult skeletal muscle and is capable of activating satellite cells. (Submitted).

Cong, M., V. F. Thompson, D. E. Goll, and P. Antin. 1997. The bovine calpastatin gene promoter and a new N-terminal region of the protein are targets for cAMP dependent portein kinase activity. J. Biol. Chem. (in press).

Kumamoto, T., H. Ueyama, R. Sugihara, E. Kominami, D. E. Goll, and T. Tsuda. 1997. Calpain and cathepsins in the skeletal muscle of inflammatory myopathies. Europ. Neurol. 37:176-181.

California

Rosser, B. W. C., M. Wick, E. Waldbillig, and E. Bandman. 1996. Heterogeneity of myosin heavy chain expression in fast twitch fibers of mature avian pectoralis muscle. Biochemistry and Cell Biology. 74:715-728.

Wick, M., F. Tablin, and E. Bandman. 1996. The effects of anti-LMM antiobodies on the solubility of chicken skeletal muscle myosin. Journal of Food Biochemistry. 20:379-395.

Tidyman, W. E., L. A. Moore, and E. Bandman. 1997. Expression of fast myosin heavy chain transcripts in developing and dystrophic chicken skeletal muscle. Developmental Dynamics. 208:491-504.

Bandman, E., M. J. Arrizubieta, M. Wick, A. Hattori, F. Tablin, S. Zhang, and Q. Zhang. 1997. Functional analysis of the chicken sarcomeric myosin rod: Regulation of dimerization, solubility, and fibrillogenesis. Cell Structure and Function. 22:131-137.

Chen, Q., L. A. Moore, M. Wick, and E. Bandman. 1997. Identification of a genomic locus containing three myosin heavy chain genes in the chicken. Biochemica Biophysica Acta - Gene Structure and Expression. 1353:148-156.

Chao, T-H. and E. Bandman. 1997. Cloning, nucleotide sequence, and characterizationof a full length cDNA encoding the myosin heavy chain from adult chicken pectoral major muscle. Gene. (in press).

Indiana

Lefaucheur, L.M., R.K. Hoffman, C.S. Okamura, D.E. Gerrard, N.A. Rubinstein, and A.M. Kelly. 1997. Transitory expression of alpha cardiac m. yosin heavy chain in subpopulation of secondary generation muscle fibers in the pig. Dev. Dyn. (In press).

Reecy, J.M., C.A. Bidwell, G.P. Briley, and A.L. Grant. 1996. Structure and regulation of the porcine skeletal _-actin-encoding gene. Gene. 180:23-28.

Beermann, D. H., A. P. Moloney, D. E. Gerrard, T. F. Robinson, K. D. Robinson, K. D. Finnerty, and J. M. Elliot. Temporal change in skeletal muscle IGF-I mRNA abundance and nitrogen metabolism responses to abomasal casein infusion in steers. (Accepted).

Gerrard, D. E., A. L. Grant, and C. S. Okamura. Developmental expression and location of muscle tissue IGF-II mRNA in pigs. (Accepted).

Fligger, J. M., P. V. Malven, R. A. Merkel, M. E. Doumit, and A. L. Grant. Increases in IGFBP-2 expression accompany decreases in proliferation and differentiation when porcine muscle satellite cells undergo multiple passages. (Submitted).

Gerrard, D. E., C. S. Okamura, and A. L. Grant. Expression and location of IGF binding proteins-2, -4, and -5 in developing fetal tissues.(Submitted).

Lefaucheur, L., R. K. Hoffman, D. E. Gerrard, C. S. Okamura, N. Rubinstein, and A. Kelly. Evidence for three adult fast myosin heavy chain isoforms in type II skeletal muscle fibers in the pig. (Submitted).

Reecy, J. M., C. A. Bidwell, O. M. Andrisani, D. E. Gerrard, and A. L. Grant. Multiple regions of the porcine alpha-skeletal actin gene modulate muscle-specific expression in cell culture and directly injected skeletal muscle. (Submitted).

Taylor-Roth, J. L., P. V. Malven, D. E. Gerrard, S. E. Mills, and A. L. Grant. Independent effects of food intake and insulin status on insulin-like growth factor-I in young pigs. (Submitted).

Zhang, H., F. F. Depreux, D. E. Gerrard, and J. Tan. Cell image processing and cell growth modeling. (Submitted).

Iowa

Graves, D. J., T. W. Huiatt, H. Zhou, H.-Y. Huang, S. W. Sernett, R. M. Robson, and K. K. McMahon. 1997. Regulatory role of arginine-specific mono-ADP-ribosyltransferase in muscle cells. Adv. Exp. Med. Biol. 419:305-313.

Ho, C.-Y., M. H. Stromer, G. Rouse, and R. M. Robson. 1997. Effects of electrical stimulation and postmortem storage on changes in titin, nebulin, desmin, troponin-T, and muscle ustrastructure in Bos indicus crossbred cattle. J. Anim. Sci. 75:366-376.

Hemken, P. M., R. M. Bellin, S. W. Sernett, B. Becker, T. W. Huiatt, and R. M. Robson. 1997. Molecular characteristics of the novel intermediate filament protein paranemin. J. Biol. Chem. (in press).  

Wang, R., M. H. Stromer, and T. W. Huiatt. 1997. Integrin expression in developing smooth muscle cells. J. Histochem. Cytochem. (in press).

Michigan

Bergen, W. G., P. S. Dickerson-Weber, M. K. Mater, and S. A. Kramer. 1996. Approaches to gene analysis, gene transfer into cells, and gene targeting strategies. J. Anim. Sci. 74(Suppl. 2):9-19.

Doumit, M. E., D. R. Cook, and R. A. Merkel. 1996. Testosterone up-regulates androgen receptors and decreases differential of porcine myogenic satellite cells in vitro. Endrocrinology. 137(4):1385-1394.

Minnesota

Johnson, B. J., M. E. White, M. R. Hathaway, C. J. Christians, and W. R. Dayton. 1997. Effect of a combined trenbolone acetate and estradiol implant on steady-state IGF-I mRNA concentrations in the liver of wethers and in the longissimus muscle of steers. J. Anim. Sci. (in press).

White, M. E., M. R. Hathaway, A. J. Lepine, and W. R. Dayton. 1997. Characterization and identification of a novel 37 kDa insulin-like growth factor binding portein in canine serum. Submitted: General and Comp. Endocrinology. (in review).

Nebraska

Zeece, M. G., Q. Chu, T. L. Woods, S. J. Jones, and W. J. Reville. 1996. Determination of 3-methylhistidine by capillary electrophoresis. J. of Capillary Electrophor. 1:55-59.

Chu, Q., B. T. Evans, and M. G. Zeece. 1997. Quantitative separation of 4-hydroxyproline from bovine skeletal muscle collagen by micellar electrokinetic capillary electrophoresis. 1997. J. of Chromatogr. B.692:293-301.

Chu, Q., M. O’Dwyer, and M. G. Zeece. 1997. Capillary electrophoretic analysis of cathepsin D action on hemoglobin. J. of Capillary Electrophor. (in press).

Woods, T.L., Smith, C.W.,. Zeece, M.G., and Jones, S.J. 1997. Conditions for the culture of bovine embryonic myogenic cells. Tiss. Cell. 29:207-215

New York

Byrem, T.M., D.H. Beermann and T.F. Robinson. The beta-agonist cimaterol directly enhances chronic protein accretion in skeletal muscle. J. Anim. Sci. (in press) 1997.

Knaus, W.F., D.H. Beermann, T.F. Robinson, D.G. Fox and K.D. Finnerty. Effects of a dietary mixture of meat and bone meal, feather meal, blood meal and fish meal on nitrogen utilization in finishing Holstein steers. J. Anim. Sci. (in press) 1998.

Moloney, A.P. D.H. Beermann, D. Gerrard, T.F. Robinson and K.D. Finnerty. Temporal changes in skeletal muscle IGF-I mRNa abundance and nitrogen metabolism responses to abomasal casein infusion in steers. J. Anim. Sci. (in press). 1998.

Ohio

Velleman, S.G., W. Bacon, R. Whitmoyer, and S.J. Hosso. 1997. Changes in distribution of glycosaminoglycans during the progression of cholesterol induced atherosclerosis in Japanese quail. Atherosclerosis (accepted).

Velleman, S.G. and C.S. Coy. 1997. Transforming growth factor ß gene expression in avian low score normal pectoral muscle. Poultry Sci. (accepted).

Velleman, S.G., D.C. McFarland, Z. Li, N.H. Ferrin, R. Whitmoyer, and J.E. Dennis. 1997. Alterations in sarcomere structure, collagen organization, mitochondrial activity, and protein metabolism in the avian Low Score Normal muscle weakness. Development Growth and Differentiation (in press).

Velleman, S.G. and C.S. Coy. 1997. Decorin and Collagen Type I gene expression in avian low score normal pectoral muscle. Poultry Sci. 76:878-881.

Velleman, S.G., R.A. Patterson, and K.E. Nestor. 1997. Identification of decorin and chondroitin sulfate proteoglycans in turkey skeletal muscle. Poultry Sci. 76:506-510.

Velleman, S.G., J.R. Racela, C. Faustman, S.D. Zimmerman, and R.J. McCormick. 1996. Partial characterization of ovine skeletal muscle proteoglycans and fibrillar collagens. Connective Tissue Res. 34:175-190.

Li, Z., S.G. Velleman, D.C. McFarland, Pesall, K.K. Gilkerson, N.H. Ferrin, and Y. Yun. 1997. Characterization of satellite cells derived from chickens with the Low Score Normal (LSN) muscle weakness. Cytobios. (submitted).

South Dakota

Ernst, C. W., D. C. McFarland, and M. E. White. 1996. Expression of Insulin-like Growth Factor II (IGF-II), IGF Binding Protein-2 and Myogenin During Differentiation of Myogenic Satellite Cells Derived From the Turkey. Differentiation. 61:25-33.

Ye, W. V., D. C. McFarland, K. K. Gilkerson, and J. E. Pesall. 1996. The Role of Platelet-derived Growth Factor in Turkey Skeletal Muscle Development. Cytobios. 88:53-62.

Yun, Y., D. C. McFarland, J. E. Pesall, K. K. Gilkerson, L. S. VanderWal, and N. H. Ferrin. 1997. Variation in Response to Growth Factor Stimuli in Satellite Cell Populations. Comparative Biochemistry and Physiology. 117A(4):463-470.

McFarland, D.C., K. K. Gilkerson, J. E. Pesall, R. H. Wellenreiter, N. H. Ferrin, W. V. Ye, Y. Yun, and L.S. Vander Wal. 1997. Comparison of Growth Factor Receptors and Metabolic Characteristics of Satellite Cells Derived from the Biceps Femoris and Pectoralis Major Muscles of the Turkey. General and Comparative Endocrinology. 105:114-120.

Velleman, S. G., D. C. McFarland, Z. Li, N. H. Ferrin, R. Whitmeyer, and J. E. Dennis. Alterations in Sarcomere Structure, Collagen Organization, Mitochondrial Activity and Protein Metabolism in the Low Score Normal Muscle Weakness. Development, Growth and Differentiation. (In Press)  

Velleman, S. G., D. C. McFarland, and C. S. Coy. Skeletal Muscle Satellite Cells: Identification of a Heparin Sulfate Proteoglycan. Basic and Applied Myology. (In Press).

McFarland, D. C. Nutritional and Developmental Roles of Insulin-like Growth Factors Among Species: A Brief History and Introduction. Journal of Nutrition. (In Press).

Li, Z., S.G. Velleman, D. C. McFarland, J. E. Pesall, K. K. Gilkrson, N. H. Ferrin and Y. Yun. Characterization of Satellite Cells Derived from Chickens with the Low Score Normal (LSN) Muscle Weakness. (Submitted).

McFarland, D. C., K. K. Gilkerson, J. E. Pesall, N. H. Ferrin, and R. H. Wellenreiter. In Vitro Characteristics of Myogenic Satellite Cells Derived from the Pectoralis Major and Biceps Femoris Muscles of the Chicken. (Submitted)

USDA - US-MARC, Clay Center

Barkhouse, K. L., L. D. Van Vleck, L. V. Cundiff, M. Koohmaraie, D. D. Lunstra, and J. D. Crouse. 1996. Prediction of breeding values for tenderness of market animals from measurements of bulls. J. Anim. Sci. 74:2612-2621.

Doumit, M. E., S. M. Lonergan, J. R. Arbona, J. Killefer, and M. Koohmaraie. 1996. Development of an enzyme-linked immunosorbent assay (ELISA) for quantification of skeletal muscle calpastatin. J. Anim. Sci. 74:2679-2686.

Koohmaraie, M., M. E. Doumit, and T. L. Wheeler. 1996. Meat toughening does not occur when rigor shortening is prevented. J. Anim. Sci. 74:2935-2942.

Dorsa, W. J., G. R. Siragusa, C. N. Cutter, E. D. Berry, and M. Koohmaraie. 1997. Efficacy of using a sponge sampling method to recover low levels of Escherichia coli 0157:H7, Salmonella typhimurium, and aerobic bacteria from beef carcass surface tissue. Food Microbiol. 14:63-69.

Boleman, S. J., S. L. Boleman, R. K. Miller, H. R. Cross, T. L. Wheeler, M. Koohmaraie, S. D. Shackelford, M. F. Miller, R. L. West, D. D. Johnson, and J. W. Savell. 1997. consuemr evaluation of been of known categories of tenderness. J. Anim. Sci. 75:1521-1524.

Harris, J. J., J. W. Savell, and M. Koohmaraie. 1997. Live animal performance, carcass traits, and meat palatability of calf- and yearling-fed cloned steers. J. Anim. Sci. 75:986-992.

Shackelford, S. D., T. L. Wheeler, and M. Koohmaraie. 1997. Effect of the callipyge phenotype and cooking method on tenderness of several major lamb muscles. J. Anim. Sci. 75:2100-2105.

Shackelford, S. D., T. L. Wheeler, and M. Koohmaraie. 1997. Repeatability of tenderness measurements in beef round muscles. J. Anim. Sci. 75:2411-2416.

Shackelford, S. D., T. L. Wheeler,a and M. Koohmaraie. 1997. Tenderness classification of beef. I. Evaluation of beef longissimus shear force at 1 or 2 days postmortem as a predictor of aged beef tenderness. J. Anim. Sci. 75:2417-2422.

Wheeler, T. L., S. D. Shackelford, L. P. Johnson, M. F. Miller, R. K. Miller, and M. Koohmaraie. 1997. A comparison of Warner-Bratzler shear force assessment within and among institutions. J. Anim. Sci. 75:2423-2432.

Siragusa, G. R., C. N. Cutter, W. J. Dorsa, and M.Koohmaraie. 1997. Microbial load monitoring of mean animal and poultry carcasses using a rapid filtration microbial ATP bioluminescence assay. In: P.E. Stanley, R. Smither, W. J. Simpson (Eds.), A Practical Guide to Industrial Uses of ATP-Luminescence in Rapid Microbiology. Cara Technology Ltd., Lingfield, UK. (Book Chapter).

Washington

Hossner, K. L., R. H. McCusker, Jr. and M. V. Dodson. 1997. Insulin-like growth factors and their binding proteins in domestic animals. Animal Science 64(1):1-15. [listed last year as ‘in press’].

Stewart, N. T., M. Foss, U. Carraro, M. Cantini, K. Byrne, J. L. Vierck, Y. Chen, E. Greene and M. V. Dodson. 1997. Muscle regeneration is modulated by satellite cell--macrophage interactions at the site of muscle injury: Prospective clinical applications. J. Equine Veterinary Science 17(4):172-177, 217-219. [listed last year as ‘in press’].

Hossner, K. L., R, Yemm, J. Vierck and M. V. Dodson. 1997. Insulin-like growth factor (IGF)-I and -II and IGFBP secretion by ovine satellite cell strains grown alone or in co-culture with 3T3-L1 preadipocytes. In Vitro: Cellular and Developmental Biology. 33(10):[in press].

Dodson, M. V., J. L. Vierck, K. L. Hossner, K. Byrne and J. P. McNamara. 1997. The development and utility of a defined muscle and fat co-culture system. Tissue and Cell 29(5):[in press].

Krabbenhoft, E. A., K. Shultz, B. A. O’Reilly, Y. Chen, N. T. Stewart and M. V. Dodson. 1997. A simplified method of analysis of cell--conditioned medium for IGF activity. Methods in Cell Science 19(3):[in press].

Byrne, K. M., W. C. Davis, M. Holmes, A. L. Brassfield and T. C. McGuire. 1997. Cytokine RNA expression in an equine CD4+ subset differentiated by expression of a novel 46 kDa surface protein. Veterinary Immunology and Immunopathology 58:[in press].

Byrne, K. M., X. Cheng, J. L. Vierck, S. Erickson, E. A. Greene, S. K. Duckett and M. V. Dodson. Use of a 96-well plate reader to evaluate proliferation of equine satellite cells in vitro: Application for proliferation studies. Methods in Cell Science [submitted].

Krabbenhoft, E. A., J. L. Vierck, K. L. Hossner and M. V. Dodson. Use of computer enhancement technology to detect the presence of free IGF-I: Application for use with dot-blot techniques. Biochem. Biophys. Res. Comm. [submitted]

Wisconsin

Trombitas, K., M. L. Greaser, and G. H. Pollack. 1997. Interaction between titin and thin filaments in intact cardiac muscle. J. Muscle Res. Cell Motil. 18:1-7.

Fassel, T. A. and M. L. Greaser. 1997. Uranyl acetate as a primary fixative for skeletal muscle. Micros. Res. Tech. 37:600-601.

Swartz, D. R., R. L. Moss, and M. L. Greaser. 1997. Characteristics of TnC binding to the myofibrillar thin filament: extraction of TnC is not random along the length of the thin filament. Biophys. J. 293-305.

Pietrzak, M., M. L. Greaser, and A. A. Sosnicki. 1997. Effect of rapid rigor mortis processes on protein functionality in Pectoralis major muscle of domestic turkeys. J. Animal Sci. 75:2106-2116.

Huang, X. P., Y. Pi, A. J. Lokuta, M. L. Greaser, and J. W. Walker. 1997. Arachidonic acid stimulates protein kinase C- redistribution in heart cells. J. Cell Sci. 110:1625-1634.

Mozdziak, P. E., E. Schultz, and R. G. Cassens, 1997. Myonuclear accretion is a major determinant of avian skeletal muscle growth. Am. J. Physiol. 272: C565-C571.

Published Abstracts

Alabama

Mulvaney, D. R., D. B. Anderson, R. C. Smith, J. D. Muegge, J. L.Wallace, J. L. Roth, A. J. Wuethrich, D. Clarke, and L. P. Muegge. Intra-uterine environment and embryonic/fetal development: consequences of perturbation. J. Anim. Sci. 75 (Suppl. 2): Review presented at 1997 annual ASAS.

Indiana

Berg, E. P., A. L. Grant, J. C. Forrest, and D. E. Gerrard. 1996. Electrophoretic separation of porcine myosin heavy chain isoforms. J. Anim. Sci. 74(Suppl. 1):139.

Depreux, F. F. S., C. S. Okamura and D. E. Gerrard. 1996. Development of monoclonal antibodies that recognize type IIa and IIb myosin heavy chain isoforms in porcine longissimus dorsi muscle. J. Anim. Sci. 74(Suppl. 1):151.

Depreux, F. F. S., C. S. Okamura and D. E. Gerrard. 1996. Evaluation of muscle fiber type in the longissimus dorsi muscle of halothane-positive pigs. J. Anim. Sci. 74(Suppl. 1):46.

Leininger, J. W., C. S. Okamura and D. E. Gerrard. 1996. Developmental expression of the type I and II IGF receptor in bovine muscle. J. Anim. Sci. 74(Suppl. 1):46.

Leininger, J. W., F. F. S. Depreux and D. E. Gerrard. 1996. Developmental expression of myogenin in fetal bovine and porcine muscle. J. Anim. Sci. 74(Suppl. 1):140.

Reecy, J. M., C. A. Bidwell, and A. L. Grant. 1996. Different DNA elements control porcine alpha-actin transcription in cultured myotubes and skeletal muscle. J. Anim. Sci. 74(Suppl. 1):139.

Blanton, J. R., A. L. Grant, D. C. McFarland, J. P. Robinson, and C. A. Bidwell. 1997. Isolation of porcine myoblasts by flow cytometry. J. Anim. Sci. 75(Suppl. 1):165.

Jacobi, S. K., D. E. Gerrard, C. A. Bidwell and A. L. Grant. 1997. Direct injection of DNA into skeletal muscle of pigs for delivery of recombinant protein. J. Anim. Sci. 75(Suppl. 1):165.

Iowa

Sernett, S. W., R. M. Bellin, T. W. Huiatt, and R. M. Robson. 1997. Molecular characterization and interaction studies of synemin, an intermediate filament protein. J. Anim. Sci. 75(Suppl. 1): 55.

Sernett, S. W., R. M. Bellin, T. W. Huiatt, and R. M. Robson. 1997. Molecular properties of synemin and its interactions with specific proteins in the muscle cell cytoskeleton. Proc. Recip. Meat Conf. 50 (in press).

Bellin, R.M., S.W. Sernett, T. W. Huiatt, and R. M. Robson. 1997. Molecular interactions of specific domains of the intermediate filament protein synemin. Mol. Biol. Cell (in press).

Carlsson, L., M. G. Price, R. Robson, J. Breckler, G. Wiche, Z. Li, D. Paulin, and L.-E. Thornell. 1997. Expression of IF-related proteins in desmin knock-out mice. Mol. Biol. Cell (in press).

Michigan

Izevbigie, E. B. and W. G. Bergen. 1996. Properties of beta receptors and response to beta agonists in C2C12 cells. J. Anim. Sci. 74(Suppl. 1):139.

Izevbigie, E. B. and W. G. Bergen. 1996. Ontogeny of beta-adrenergic receptors in cultured porcine satellite cells. J. Anim. Sci. (Suppl. 1):139.

Nebraska

Klaasmeyer, J.G., Smith, C.W., Grant, D., Woods, T.L., and Jones, S.J. 1997. Changes in ribosomal recruitment during translation in differentiating C2C12 myogenic cells. J. Anim. Sci. 75:160 (Suppl. 1)

Smith, C.W., Klaasmeyer, J. G., Woods, T.L. and Jones, S.J. 1997. Insulin, FGF, IGFI, IGFII, and PDGF alter the recruitment of ribosomes to polysomes for protein synthesis in C2C12 myogenic cells. J. Anim. Sci. 75:160 (Suppl. 1)

Motherway, M. M., W. J. Reville, A. M. O’Donovon, S. Sullivan, and M. G. Zeece. 1996. The assessment of myofibrillar breakdown during conditioning of meat. Irish Journal of Agricultural and Food Research 35:2.

Chu, Q., M. O’Dwyer, and M. G. Zeece. 1997. Analysis of cathepsin D action on hemoglobin by micellar electrokinetic capillary electrophoresis. Presented at HPCE-97. Anaheim, CA. Jan. 25-30.

Chu, Q., W. J. Reville, and M. G. Zeece. 1997. Use of Capillary electrophoretic analysis to study multicatalytic proteinase. Present at 3rd Annual Porteasome workshop. Clermont-Ferrand France, March 18-20.

New York

Knaus, W., D.H. Beermann, T.F. Robinson, and D.G. Fox. Use of the Cornell Net Carbohydrate and Protein System (CNCPS) model to optimize amino acid availability in finishing Holstein steers with a mixture of dietary animal protein sources. J. Anim. Sci. 75 (Suppl.1):131. 1997.

Robinson, T.F., D.H. Beermann, T.C. Perry, D.J. Ketchen and D.G. Fox. Effects of feeding a diet balanced using the 1996 Beef NRC to meet amino acid requirements in feedlot steers. J. Anim. Sci. 75 (Suppl.1):260. 1997.

Robinson, T.F., A.P. Moloney, D.H. Beermann, K.D. Finnerty and D.G. Fox. Nitrogen retention in Holstein steers offered isoenergetic rations containing increasing amounts of a blend of protein sources of low ruminal degradability. Proc. Nutrition Society 56:182A. 1997.

Ohio

Velleman, S. G. 1997. Alterations in sarcomere structure and collagen organization in the avian low score normal muscle weakness. Poultry Sci. 76 (suppl.1):72.

Liu, X. and Velleman, S. G. 1997. Isolation and partial characterization of proteoglycan during turkey embryonic skeletal muscle development. Poultry Sci. 76 (suppl. 1):72.

South Dakota

McFarland, D., K. Gilkerson, J. Pesall, N. Ferrin, and R. Wellenreiter. 1997. In Vitro Characteristics of Myogenic Satellite Cells Derived from the Pectoralis Major and Biceps Femoris Muscles of the Broiler Chicken. FASEB Journal 11(3):A419.

Blanton, Jr., J. R., A. L. Grant, D. C. McFarland, J. P. Robinson and C. A. Bidwell. 1997. Isolation of Porcine Myoblasts by Flow Cytometry. J. Animal Science 75:165, (suppl 1).

McFarland, D. C., J. E. Pesall, K. K. Gilkerson, and L. S. Vander Wal. 1997. Can Cell Culture Techniques Be Used in Genetic Selection of Poultry? Poultry Science 76:22, (suppl 1).

USDA - US-MARC, Clay Center

Casas, E., J. W. Keele, S. D. Shackelford, T. S. Sonstegard, T. P. Smith, M. Koohmaraie, S. M. Kappes, and R. T. Stone. 1997. Association of the double-muscling locus on bovine chromosome two (BTA2) with carcass traits. J. Anim. Sci. 75 (Suppl. 1):149.

Shackelford, S. D., T. L. Wheeler,a and M. Koohmaraie. 1997. Tenderness classification of beef. J. Anim. Sci. 75 (Suppl. 1):176.

Lorenzen, C. L., M. L. Fiorotto, H. C. Freetly, S. D. Shackelford, T. L. Wheeler, J. W. Savell, and M. Koohmaraie. 1997. Muscle protein synthesis and protein degradation play a role in callipyge-induced muscle hypertrophy. J. Anim. Sci. 75(Suppl. 1):177.

Echternkamp, S. E., K. E. Gregory, and M. Koohmaraie. 1997. Comparison of growth and carcass traits between freemartin and co-twin males. J. Anim. Sci. 75(Suppl. 1):180.

Wisconsin

Mozdziak, P. E., M. L. Greaser, and E. Schultz, 1997. Myogenin and MyoD expression are more closely related to myosin heavy chain isoform composition than muscle mass. J. Animal Sci. 75 (Suppl. 1):121.

Berri, E., J. Wolff, and M. L. Greaser. 1997. Characterization of the titin PEVK fragment from different species. Biophys. J. 72:A189.

Trombitas, K., M. L. Greaser, and G.H. Pollack. 1997. Diffusion of antibodies into mechanically skinned muscle fibers. Biophys. J. 72:A276.

Diffee, G. M., J. R. Patel, F. C. Reinach, X. P. Huang, M. L. Greaser, and R. L. Moss. 1997. Interaction between phosphorylation and Ca++ binding to myosin regulatory light chain from smooth and striated muscle. Biophys. J. 72:A331.

Non-Refereed Publications

Proceedings, Book Chapters etc.

Iowa

Bellin, R. M., S. W. Sernett, and R. M. Robson. 1997. Synemin. In T. Kreis and R. Vale, eds., Guidebook to the Cytoskeletal and Motor Proteins, Second Edition, Oxford Univ. Press (in press).

Robson, R. M., E. Huff-Lonergan, F. C. Parrish, Jr., C.-Y. Ho, M. H. Stromer, T. W. Huiatt, R. M. Bellin, and S. W. Sernett. 1997. Postmortem changes in the myofibrillar and other cytoskeletal proteins in muscle. Proc. Recip. Meat Conf. 50 (in press).

Minnesota

Dayton, W. R., M. R. Hathaway, and B. J. Johnson. 1997. Effects of a combined trenbolone acetate and estradiol implant on carcass composition and biological parameters of feedlot cattle. Proccedings "Impact of Implants on Performance and Carcass Value of Beef Cattle". Oklahoma State University (in press).

New York

Beermann, D.H., T.F. Robinson, O.H. Cheng and K.D. Finnerty. Enhancing amino acid availability and protein accretion in growing ruminants. Proceedings 49th Annual Reciprocal Meat Conference, pp. 39-44. 1996.

Beermann, D.H. Hormonal aspects of muscle growth. Proceedings Satellite Symposium on beef production with special respect to beef quality. 48th Annual Meeting of the European Association for Animal Production. pp. 1-14. 1997.

Beermann, D.H., T.F. Robinson, W.F. Knaus and D.G. Fox. Formulation of protein supplements to provide ideal amounts of absorbed amino acids in growing cattle. Proceedings of the Cornell Nutrition Conference pp.172-180. 1997.

Bell, A.W., D.E. Bauman, D.H. Beermann and R.J. Harrell. Nutrition, development and efficacy of growth modifiers in livestock species. J. Nutr. Suppl. 1 (in press) 1997.

Cheng, O.H and D.H. Beermann. A summary and guide to analytical methods for protein, RNA and DNA determinations in skeletal muscle and other tissues. Proceedings 50th Annual Reciprocal Meats Conference. (in press)1997.

Theses and Dissertation

Alabama

Kelly R. L. 1997. Developmental expressionof MRF genes in bovine muscle and cultures of fetal skeletal muscle. M. S. Thesis, Auburn University, AL.

Minnesota

Yang, F. 1997. Identification of Insulin-like growth factor binding proteins produced by cultured embryonic muscle cells and the effecects of insulin-like growth factor-I on binding protein production. M. S. Thesis.

Johnson, B. J. 1997. Characterizationof an IGFBP-free medium for cultured porcine myogenic cells and the effect of growth factor addition to this medium on steady-state IGFBP mRNA levels in differentiating porcine myogenic cells. Ph.D. Dissertation.

Nebraska

Woods, T.L. 1997. Bovine embryonic myogenic cells: Conditions for culture and response of protein turnover and ribosome populations with either insulin of dexamethasone. Ph.D. Dissertation

APPROVED:

______________________________________________             ____________________
Dr. Everett Bandman, Chair of Technical Committee, 1996-97          Date

______________________________________________            _____________________
Dr. Colin Scanes, Administrative Advisor                                          Date