Tag Archives: Histology

Brain and Muscles

Among mammals, primates exhibit a trend toward increasing encephalisation. Attempts to explain the development of this trend focus on the energetic and metabolic trade-offs required to increase brain mass. The most widely discussed are variants of the Expensive Tissue Hypothesis (ETH) which proposes for any increase in brain mass other metabolically expensive tissues must decrease in size. The brain is metabolically costly with primates having larger brain sizes than other mammals and devoting up to 20% more basal metabolic rate to brain maintenance. Brain maintenance relies on aerobic cellular respiration processes, thus requiring oxygen to efficiently function. In a resting-state, up to 90% of brain maintenance is sourced from aerobic respiration. The brain does not source oxygen directly but relies on aerobic cellular respiration, converting glucose into adenosine triphosphate (ATP) to produce energy. In humans, the brain consumes, on average, around 30% of total glucose allocation. Skeletal muscle is another expensive tissue type. Muscle consumes up to 30% of resting energy expenditure with nearly 100-fold increase during high activity. Mammals have nearly 50% of their total body mass accounted for by muscle mass while primates have only 35% of total body mass accounted for by muscle mass. Of primates, humans possess 50% less muscle mass than expected for body size. Skeletal muscle comprises a mixture of fibers known as Type I (slow-twitch for prolonged activity) and Type II (fast-twitch for short, sudden activity). Both fiber types require constant oxygen supply and glucose to convert to ATP via mitochondria. Although Type II fibers consume a higher net-amount of glucose than Type I, this is done for short periods of time. Type I fibers used for prolonged activities possess greater capillary density and more mitochondria than Type II, potentially allowing significantly more efficient conversion of glucose to ATP. This could suggest muscle mass is in direct competition with the brain through glucose requirement and that any increase in brain size could require a corresponding decrease in muscle mass as evidenced in primates, especially humans.

Muchlinski et al. (2018) examined the potential trade-off between muscle mass and brain size in non-human primates. Several skeletal muscles were dissected from primate cadavers and immunohistochemistry used to isolate muscle fiber types. Body mass strongly influenced endocranial volume and muscle mass in the primate species so variables were size adjusted. Results indicated an increase in endocranial volume was associated with a decrease in muscle mass. Type I muscle fibers were negatively correlated with endocranial volume but a positive correlation between Type II and endocranial volume was not statistically significant. In general, the primates sampled possessed more Type II than Type I muscle fibers. These results are encouraging but potential bias could be introduced from the small sample size and muscle selection with larger postural and locomotor muscles, erector spinae and scalenes, not examined as the minimum sample content for immunohistochemistry could not be dissected in very small primate species. The use of published literature for endocranial volumes and body mass may introduce additional issues. Despite this, the assumption that muscle may be in direct competition with the brain appears metabolically and energetically viable and a potential avenue for proper consideration in evolutionary primatology.

Alannah Pearson

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Brian Metscher

Following bachelor’s work in applied physics at Caltech and a first career as a research engineer at NASA/JPL, Brian Metscher completed his PhD in the then-new interdiscipline of evo-devo at the University of California, Irvine. He did postdoctoral research on the development and evolution of appendages and teeth at The Natural History Museum (London) and Penn State University, and then served five years as an Assistant Professor in southern Indiana. During the summers he carried out research at Yale University and came to the University of Vienna in 2006, to set up the imaging lab in the Department of Theoretical Biology, where he is now Senior Scientist. He helped to establish X-ray microtomography as an essential method for imaging ex vivo biological samples, especially embryos and invertebrates. His lab is developing new and refined sample preparation and imaging methods, with applications including molecular imaging and imaging of specific cells types. He coordinates a MicroCT Methods Forum. Here a brief interview …

What are the basic principles of these methods mixing histology and digital anatomy?

MicroCT provides 3D images of intact samples at resolutions that overlap with what is achieved by light microscopy of sectioned material. Contrast-enhanced X-ray images give only histomorphological information, so microCT images are a powerful complement to traditional histology, which takes advantage of a vast array of stains with different tissue specificities. MicroCT gives a 3D overview and context for more detailed section-based images from histology (and also electron microscope).

So, you stain specimens before microtomographic scan … what about these staining techniques?

The familiar X-ray images of bones or teeth inside the body are possible because the dense calcium-rich materials absorb a lot more X-ray energy than the soft tissues around them – skin, muscle, and internal organs, which are made up mostly of proteins and water. To make soft tissues clearly visible in X-ray images, it helps to add a contrast agent: this can be a suspension of an iodine- or barium-containing liquid injected or swallowed, as is common in clinical radiology examinations. In the case of non-living samples (ex vivo imaging, most of what I do), the sample can be stained with a substance that actually binds to the tissues and has a higher X-ray absorption. The contrast stains used most often are inorganic iodine, phosphotungstic acid, and (less frequently) osmium tetroxide. None of these is specific to any one tissue type, but they do allow the different tissues and structures to be distinguished clearly in the X-ray images.

What kind of expertise, career, and tools are necessary to work in this field?

As with any kind of biological imaging, it is necessary to have a good understanding both of the biological systems under study and of how the imaging systems actually work. So a strong background in microscopy, histology, and image acquisition and analysis is important. And one must always complement one’s own expertise with good working collaborations with partners in other fields.

What is, at present, the most intriguing current challenge?

We would really like to make microCT imaging more tissue- and molecule-specific. Thus I have collaborative projects to test new staining methods and calibrate their functions in microCT images with histological baselines. And my lab is working on refining the antibody imaging method we published a few years ago to make this a more robust and routine method for 3D imaging of gene expression and other molecular patterns in developmental, comparative, and medical-related research.


Ontogenetic changes in human crania

cranial-growth-postThe brain growth pattern in humans is distinctive among primates. In the past decades several hypotheses have been proposed to analyze cranial ontogenetic changes (i.e shape and size variations) in humans (e.g. Moss and Young, 1960; Lieberman et al., 2002; Bruner, 2004; Neubauer et al., 2009). The recent study conducted by García Gil et al. (2015) presents a preliminary approach to the histological variations of the vault bones in three individuals of different ages (child, adolescent and young adult). According to their results, it is possible to identify three different histological phases of cranial growth. In the child, vault bones are primarily composed of avascular lamellar bone (widely vascularized). In contrast, the adolescent bones show a larger extension of mineralized regions (highly remodeled areas) and low levels of vascularization, with a much reduced diploe. In the adult, the vault bone is highly vascularized and the diploe is largely expanded. The authors suggest that the sealing of the cranial bone surfaces helps to minimize the bone porosity while increases bone expansion (during childhood) and thickness (during youth). This “sealing process” could play a main role controlling head thermoregulation until the brain finishes its maturation. When confirmed on larger samples, these results can introduce new perspectives in functional craniology.

Gizéh Rangel de Lázaro