An introductory biology lab that uses enzyme histochemistry to teach students about skeletal muscle fiber types

2004 ◽  
Vol 28 (1) ◽  
pp. 23-28 ◽  
Author(s):  
Lauren J. Sweeney ◽  
Peter D. Brodfuehrer ◽  
Beth L. Raughley
Keyword(s):  
Skeletal Muscle ◽  
Enzyme Histochemistry ◽  
Fiber Type ◽  
Scientific Discovery ◽  
Muscle Fiber Types ◽  
Fiber Types ◽  
Introductory Biology ◽  
Laboratory Experiences ◽  
Biology Laboratory ◽  
Biology Lab

One important goal of introductory biology laboratory experiences is to engage students directly in all steps in the process of scientific discovery. Even when laboratory experiences are built on principles discussed in the classroom, students often do not adequately apply this background to interpretation of results they obtain in lab. This disconnect has been described at the level of medical education ( 4 ), so it should not be surprising that educators have struggled with this same phenomenon at the undergraduate level. We describe a new introductory biology lab that challenges students to make these connections. The lab utilizes enzyme histochemistry and morphological observations to draw conclusions about the composition of functionally different types of muscle fibers present in skeletal muscle. We report that students were not only successful at making these observations on a specific skeletal muscle, the gastrocnemius of the frog Rana pipiens, but that they were able to connect their results to the principles of fiber type differences that exist in skeletal muscles in all vertebrates.

Characteristics of Skeletal Muscle Fiber Types in the Miniature Pig and the Effect of Training

1973 ◽  
Vol 51 (11) ◽  
pp. 825-831 ◽  
Author(s):  
R. H. Fitts ◽  
F. J. Nagle ◽  
R. G. Cassens
Keyword(s):  
Skeletal Muscle ◽  
Enzyme Histochemistry ◽  
Fiber Type ◽  
Skeletal Muscle Fiber ◽  
Muscle Fiber Types ◽  
Fiber Types ◽  
Miniature Pig ◽  
Fast Red ◽  
Fiber Type Distribution ◽  
White Fiber

The fiber types present in miniature pig skeletal muscle were determined with enzyme histochemical techniques. Three distinct fiber types were found: a fast white fiber, a fast intermediate fiber, and a slow red fiber. The fiber types found in miniature pig (large mammal) skeletal muscle were different from those in rat (rodent) skeletal muscle where the fiber types are classified as fast white, slow intermediate, and fast red. The fiber type distribution in miniature pig skeletal muscle was not altered by either an endurance or sprint running program, despite physiologically measurable training effects. It is concluded that enzyme histochemistry is a good qualitative tool for assessing the fiber types present in a muscle but lacks the sensitivity to measure or quantitate changes due to training.

Blood flow to different skeletal muscle fiber types during contraction

1983 ◽  
Vol 245 (2) ◽  
pp. H265-H275 ◽  
Author(s):  
B. G. Mackie ◽  
R. L. Terjung
Keyword(s):  
Skeletal Muscle ◽  
Blood Flow ◽  
Muscle Fiber ◽  
Fiber Type ◽  
Oxidative Capacity ◽  
Skeletal Muscle Fiber ◽  
Muscle Fiber Types ◽  
Fiber Types ◽  
Blood Flows ◽  
Fast Twitch

Blood flow to fast-twitch red (FTR), fast-twitch white (FTW), and slow-twitch red (STR) muscle fiber sections of the gastrocnemius-plantaris-soleus muscle group was determined using 15 +/- 3-microns microspheres during in situ stimulation in pentobarbital-anesthetized rats. Steady-state blood flows were assessed during the 10th min of contraction using twitch (0.1, 0.5, 1, 3, and 5 Hz) and tetanic (7.5, 15, 30, 60, and 120/min) stimulation conditions. In addition, an earlier blood flow determination was begun at 3 min (twitch series) or at 30 s (tetanic series) of stimulation. Blood flow was highest in the FTR (220-240 ml X min-1 X 100 g-1), intermediate in the STR (140), and lowest in the FTW (70-80) section during tetanic contraction conditions estimated to coincide with the peak aerobic function of each fiber type. These blood flows are fairly proportional to the differences in oxidative capacity among fiber types. Further, their absolute values are similar to those predicted from the relationship between blood flow and oxidative capacity found by others for dog and cat muscles. During low-frequency contraction conditions, initial blood flow to the FTR and STR sections were excessively high and not dependent on contraction frequency. However, blood flows subsequently decreased to values in keeping with the relative energy demands. In contrast, FTW muscle did not exhibit this time-dependent relative hyperemia. Thus, besides the obvious quantitative differences between skeletal muscle fiber types, there are qualitative differences in blood flow response during contractions. Our findings establish that, based on fiber type composition, a heterogeneity in blood flow distribution can occur within a whole muscle during contraction.

Human Skeletal Muscle Fiber Types: Delineation, Development, and Distribution

1997 ◽  
Vol 22 (4) ◽  
pp. 307-327 ◽  
Author(s):  
Robert S. Staron
Keyword(s):  
Skeletal Muscle ◽  
Muscle Fiber ◽  
Human Skeletal Muscle ◽  
Fiber Type ◽  
Skeletal Muscle Fiber ◽  
Muscle Fiber Types ◽  
Fiber Types ◽  
Key Words Aging ◽  
Fiber Number ◽  
Human Limb

This brief review attempts to summarize a number of studies on the delineation, development, and distribution of human skeletal muscle fiber types. A total of seven fiber types can be identified in human limb and trunk musculature based on the pH stability/ability of myofibrillar adenosine triphosphatase (mATPase). For most human muscles, mATPase-based fiber types correlate with the myosin heavy chain (MHC) content. Thus, each histochemically identified fiber has a specific MHC profile. Although this categorization is useful, it must be realized that muscle fibers are highly adaptable and that innumerable fiber type transients exist. Also, some muscles contain specific MHC isoforms and/or combinations that do not permit routine mATPase-based fiber typing. Although the major populations of fast and slow are, for the most part, established shortly after birth, subtle alterations take place throughout life. These changes appear to relate to alterations in activity and/or hormonal levels, and perhaps later in life, total fiber number. Because large variations in fiber type distribution can be found within a muscle and between individuals, interpretation of data gathered from human muscle is often difficult. Key words: aging, myosin heavy chains, myogenesis, myofibrillar adenosine triphosphate

scholarly journals Calcineurin Is Necessary for the Maintenance but Not Embryonic Development of Slow Muscle Fibers

2005 ◽  
Vol 25 (15) ◽  
pp. 6629-6638 ◽  
Author(s):  
Misook Oh ◽  
Igor I. Rybkin ◽  
Victoria Copeland ◽  
Michael P. Czubryt ◽  
John M. Shelton ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Fiber Type ◽  
Oxidative Capacity ◽  
Muscle Fiber Types ◽  
Fiber Types ◽  
Specific Expression ◽  
Interacting Protein ◽  
Type Composition ◽  
Slow Fiber ◽  
Fast Twitch

ABSTRACT Skeletal muscles are a mosaic of slow and fast twitch myofibers. During embryogenesis, patterns of fiber type composition are initiated that change postnatally to meet physiological demand. To examine the role of the protein phosphatase calcineurin in the initiation and maintenance of muscle fiber types, we used a “Flox-ON” approach to obtain muscle-specific overexpression of the modulatory calcineurin-interacting protein 1 (MCIP1/DSCR1), an inhibitor of calcineurin. Myo-Cre transgenic mice with early skeletal muscle-specific expression of Cre recombinase were used to activate the Flox-MCIP1 transgene. Contractile components unique to type 1 slow fibers were absent from skeletal muscle of adult Myo-Cre/Flox-MCIP1 mice, whereas oxidative capacity, myoglobin content, and mitochondrial abundance were unaltered. The soleus muscles of Myo-Cre/Flox-MCIP1 mice fatigued more rapidly than the wild type as a consequence of the replacement of the slow myosin heavy chain MyHC-1 with a fast isoform, MyHC-2A. MyHC-1 expression in Myo-Cre/Flox-MCIP1 embryos and early neonates was normal. These results demonstrate that developmental patterning of slow fibers is independent of calcineurin, while the maintenance of the slow-fiber phenotype in the adult requires calcineurin activity.

Functionally distinct elements are required for expression of the AMPD1 gene in myocytes

1993 ◽  
Vol 13 (9) ◽  
pp. 5854-5860
Author(s):  
T Morisaki ◽  
E W Holmes
Keyword(s):  
Skeletal Muscle ◽  
Fiber Type ◽  
Transcriptional Start Site ◽  
Protein S ◽  
Muscle Fiber Types ◽  
Binding Motif ◽  
Fiber Types ◽  
Specific Expression ◽  
Tissue Specific ◽  
Ampd1 Gene

AMP deaminase (AMPD) is an enzyme found in all eukaryotic cells. Tissue-specific and stage-specific isoforms of this enzyme are found in vertebrates, and expression of these different isoforms is determined by selective expression of the multiple genes. The AMPD1 gene is expressed predominantly in skeletal muscle, in which transcript abundance is controlled by stage-specific and fiber type-specific signals. This enzyme activity is presumed to be important in skeletal muscle because a metabolic myopathy develops in individuals with an inherited deficiency of AMPD1. In the present study, cis- and trans-acting factors that control expression of AMPD1 have been identified. Two cis-acting elements located within 100 nucleotides of the transcriptional start site are required for muscle-specific expression of AMPD1. One element (-100 to -79) behaves like a tissue-specific enhancer, and it interacts with protein(s) found predominantly in nuclei of myoblasts and myotubes. This element is similar in sequence to an MEF2 binding motif, and it contains an A/T core that is essential for enhancer activity and binding of a nuclear protein(s). The second element (-60 to -40) has properties of a stage-specific promoter in that it is essential for muscle-specific expression of the AMPD1 promoter, does not confer muscle-specific expression on a heterologous promoter construct, and interacts with a protein(s) restricted to nuclei of differentiated myotubes. Interaction between these functionally distinct elements may be required for regulating the expression of AMPD1 during myocyte differentiation and in different muscle fiber types.

Strength, power, fiber types, and mRNA expression in trained men and women with different ACTN3 R577X genotypes

2009 ◽  
Vol 106 (3) ◽  
pp. 959-965 ◽  
Author(s):  
Barbara Norman ◽  
Mona Esbjörnsson ◽  
Håkan Rundqvist ◽  
Ted Österlund ◽  
Ferdinand von Walden ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Muscle Fiber ◽  
Fiber Type ◽  
Muscle Fiber Types ◽  
Vastus Lateralis Muscle ◽  
Fiber Types ◽  
Mrna Levels ◽  
Men And Women ◽  
Fiber Type Composition ◽  
Type Composition

α-Actinins are structural proteins of the Z-line. Human skeletal muscle expresses two α-actinin isoforms, α-actinin-2 and α-actinin-3, encoded by their respective genes ACTN2 and ACTN3. ACTN2 is expressed in all muscle fiber types, while only type II fibers, and particularly the type IIb fibers, express ACTN3. ACTN3 (R577X) polymorphism results in loss of α-actinin-3 and has been suggested to influence skeletal muscle function. The X allele is less common in elite sprint and power athletes than in the general population and has been suggested to be detrimental for performance requiring high power. The present study investigated the association of ACTN3 genotype with muscle power during 30-s Wingate cycling in 120 moderately to well-trained men and women and with knee extensor strength and fatigability in a subset of 21 men performing isokinetic exercise. Muscle biopsies were obtained from the vastus lateralis muscle to determine fiber-type composition and ACTN2 and ACTN3 mRNA levels. Peak and mean power and the torque-velocity relationship and fatigability output showed no difference across ACTN3 genotypes. Thus this study suggests that R577X polymorphism in ACTN3 is not associated with differences in power output, fatigability, or force-velocity characteristics in moderately trained individuals. However, repeated exercise bouts prompted an increase in peak torque in RR but not in XX genotypes, suggesting that ACTN3 genotype may modulate responsiveness to training. Our data further suggest that α-actinins do not play a significant role in determining muscle fiber-type composition. Finally, we show that ACTN2 expression is affected by the content of α-actinin-3, which implies that α-actinin-2 may compensate for the lack of α-actinin-3 and hence counteract the phenotypic consequences of the deficiency.

scholarly journals LncRNA-FKBP1C regulates muscle fiber type switching by affecting the stability of MYH1B

2021 ◽  
Vol 7 (1) ◽  
Author(s):  
Jia-ao Yu ◽  
Zhijun Wang ◽  
Xin Yang ◽  
Manting Ma ◽  
Zhenhui Li ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Protein Stability ◽  
Muscle Fiber ◽  
Fiber Type ◽  
Muscle Fibers ◽  
Fast Muscle ◽  
Muscle Fiber Types ◽  
Fiber Types ◽  
Regulatory Processes ◽  
Muscle Genes

AbstractLong non-coding RNAs (lncRNAs) are well-known to participate in a variety of important regulatory processes in myogenesis. In our previous RNA-seq study (accession number GSE58755), we found that lncRNA-FKBP1C was differentially expressed between White Recessive Rock (WRR) and Xinghua (XH) chicken. Here, we have further demonstrated that lncRNA-FKBP1C interacted directly with MYH1B by biotinylated RNA pull-down assay and RNA immunoprecipitation (RIP). Protein stability and degradation experiments identified that lncRNA-FKBP1C enhanced the protein stability of MYH1B. Overexpression of lncRNA-FKBP1C inhibited myoblasts proliferation, promoted myoblasts differentiation, and participated in the formation of skeletal muscle fibers. LncRNA-FKBP1C could downregulate the fast muscle genes and upregulate slow muscle genes. Conversely, its interference promoted cell proliferation, repressed cell differentiation, and drove the transformation of slow-twitch muscle fibers to fast-twitch muscle fibers. Similar results were observed after knockdown of the MYH1B gene, but the difference was that the MYH1B gene had no effects on fast muscle fibers. In short, these data demonstrate that lncRNA-FKBP1C could bound with MYH1B and enhance its protein stability, thus affecting proliferation, differentiation of myoblasts and conversion of skeletal muscle fiber types.

Purine salvage to adenine nucleotides in different skeletal muscle fiber types

2001 ◽  
Vol 91 (1) ◽  
pp. 231-238 ◽  
Author(s):  
Jeffrey J. Brault ◽  
Ronald L. Terjung
Keyword(s):  
Skeletal Muscle ◽  
De Novo ◽  
Adenine Nucleotide ◽  
Fiber Type ◽  
High Rate ◽  
Muscle Fiber Types ◽  
Fiber Types ◽  
De Novo Synthesis ◽  
Purine Salvage ◽  
Fast Twitch

Rates of purine salvage of adenine and hypoxanthine into the adenine nucleotide (AdN) pool of the different skeletal muscle phenotype sections of the rat were measured using an isolated perfused hindlimb preparation. Tissue adenine and hypoxanthine concentrations and specific activities were controlled over a broad range of purine concentrations, ranging from 3 to 100 times normal, by employing an isolated rat hindlimb preparation perfused at a high flow rate. Incorporation of [3H]adenine or [3H]hypoxanthine into the AdN pool was not meaningfully influenced by tissue purine concentration over the range evaluated (∼0.10–1.6 μmol/g). Purine salvage rates were greater ( P < 0.05) for adenine than for hypoxanthine (35–55 and 20–30 nmol · h−1 · g−1, respectively) and moderately different ( P < 0.05) among fiber types. The low-oxidative fast-twitch white muscle section exhibited relatively low rates of purine salvage that were ∼65% of rates in the high-oxidative fast-twitch red section of the gastrocnemius. The soleus muscle, characterized by slow-twitch red fibers, exhibited a high rate of adenine salvage but a low rate of hypoxanthine salvage. Addition of ribose to the perfusion medium increased salvage of adenine (up to 3- to 6-fold, P < 0.001) and hypoxanthine (up to 6- to 8-fold, P < 0.001), depending on fiber type, over a range of concentrations up to 10 mM. This is consistent with tissue 5-phosphoribosyl-1-pyrophosphate being rate limiting for purine salvage. Purine salvage is favored over de novo synthesis, inasmuch as delivery of adenine to the muscle decreased ( P < 0.005) de novo synthesis of AdN. Providing ribose did not alter this preference of purine salvage pathway over de novo synthesis of AdN. In the absence of ribose supplementation, purine salvage rates are relatively low, especially compared with the AdN pool size in skeletal muscle.

scholarly journals Function and Fiber-Type Specific Distribution of Hsp60 and αB-Crystallin in Skeletal Muscles: Role of Physical Exercise

Biology ◽  
2021 ◽  
Vol 10 (2) ◽  
pp. 77
Author(s):  
Daniela D’Amico ◽  
Roberto Fiore ◽  
Daniela Caporossi ◽  
Valentina Di Felice ◽  
Francesco Cappello ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Fiber Type ◽  
Mechanical Strain ◽  
Oxidative Capacity ◽  
Mitochondrial Activity ◽  
Muscle Fiber Types ◽  
Fiber Types ◽  
Specific Distribution ◽  
Αb Crystallin ◽  
The Impact

Skeletal muscle is a plastic and complex tissue, rich in proteins that are subject to continuous rearrangements. Skeletal muscle homeostasis can be affected by different types of stresses, including physical activity, a physiological stressor able to stimulate a robust increase in different heat shock proteins (HSPs). The modulation of these proteins appears to be fundamental in facilitating the cellular remodeling processes related to the phenomenon of training adaptations such as hypertrophy, increased oxidative capacity, and mitochondrial activity. Among the HSPs, a special attention needs to be devoted to Hsp60 and αB-crystallin (CRYAB), proteins constitutively expressed in the skeletal muscle, where their specific features could be highly relevant in understanding the impact of different volumes of training regimes on myofiber types and in explaining the complex picture of exercise-induced mechanical strain and damaging conditions on fiber population. This knowledge could lead to a better personalization of training protocols with an optimal non-harmful workload in populations of individuals with different needs and healthy status. Here, we introduce for the first time to the reader these peculiar HSPs from the perspective of exercise response, highlighting the control of their expression, biological function, and specific distribution within skeletal muscle fiber-types.

scholarly journals Functionally distinct elements are required for expression of the AMPD1 gene in myocytes.

1993 ◽  
Vol 13 (9) ◽  
pp. 5854-5860 ◽  
Author(s):  
T Morisaki ◽  
E W Holmes
Keyword(s):  
Skeletal Muscle ◽  
Fiber Type ◽  
Transcriptional Start Site ◽  
Protein S ◽  
Muscle Fiber Types ◽  
Binding Motif ◽  
Fiber Types ◽  
Specific Expression ◽  
Tissue Specific ◽  
Ampd1 Gene

AMP deaminase (AMPD) is an enzyme found in all eukaryotic cells. Tissue-specific and stage-specific isoforms of this enzyme are found in vertebrates, and expression of these different isoforms is determined by selective expression of the multiple genes. The AMPD1 gene is expressed predominantly in skeletal muscle, in which transcript abundance is controlled by stage-specific and fiber type-specific signals. This enzyme activity is presumed to be important in skeletal muscle because a metabolic myopathy develops in individuals with an inherited deficiency of AMPD1. In the present study, cis- and trans-acting factors that control expression of AMPD1 have been identified. Two cis-acting elements located within 100 nucleotides of the transcriptional start site are required for muscle-specific expression of AMPD1. One element (-100 to -79) behaves like a tissue-specific enhancer, and it interacts with protein(s) found predominantly in nuclei of myoblasts and myotubes. This element is similar in sequence to an MEF2 binding motif, and it contains an A/T core that is essential for enhancer activity and binding of a nuclear protein(s). The second element (-60 to -40) has properties of a stage-specific promoter in that it is essential for muscle-specific expression of the AMPD1 promoter, does not confer muscle-specific expression on a heterologous promoter construct, and interacts with a protein(s) restricted to nuclei of differentiated myotubes. Interaction between these functionally distinct elements may be required for regulating the expression of AMPD1 during myocyte differentiation and in different muscle fiber types.

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