Using qRT-PCR to Assess Changes in Rat Muscle Tissue Under Microgravity
Start Date
August 2024
End Date
August 2024
Location
ALT 207
Abstract
A problem that astronauts face in space is muscle atrophy. While consistent exercise multiple times a week helps slow down muscle loss, it is not a perfect prevention method. A way to mitigate these effects is to supplement exercise with something else that physiologically maintains muscle mass. Sialic acid (SA), a sugar glycan attached to the outermost glycan chain on skeletal muscle cells, is a molecule that aids the development, regeneration, and contractility of skeletal muscle cells. This molecule is placed on the outside of muscle cells by proteins called sialyltransferases (STs) and taken off by sialidases. Preliminary data shows that decreased SA in muscle cells leads to muscle atrophy in other diseases, like GNE myopathy, and may play a role in atrophy associated with microgravity. Therefore, to simulate microgravity in a rat model, animal’s hind limbs were suspended for two weeks, while others were under normal conditions for comparison. Semiquantitative real-time polymerase chain reaction (qRT-PCR), a method used to detect and quantify RNA using a known internal standard gene to compare to a targeted gene, was used to quantify the gene expression of STs and sialidases in these rats. RNA was extracted from their gastrocnemius and soleus muscles and then converted to cDNA. The cDNA specific to STs and sialidases was quantified. Methodological data and preliminary development showed that while further developments are needed for the methodology of qRT-PCR, the results could lead to the possibility of therapies that help astronauts maintain their muscle mass in space.
Using qRT-PCR to Assess Changes in Rat Muscle Tissue Under Microgravity
ALT 207
A problem that astronauts face in space is muscle atrophy. While consistent exercise multiple times a week helps slow down muscle loss, it is not a perfect prevention method. A way to mitigate these effects is to supplement exercise with something else that physiologically maintains muscle mass. Sialic acid (SA), a sugar glycan attached to the outermost glycan chain on skeletal muscle cells, is a molecule that aids the development, regeneration, and contractility of skeletal muscle cells. This molecule is placed on the outside of muscle cells by proteins called sialyltransferases (STs) and taken off by sialidases. Preliminary data shows that decreased SA in muscle cells leads to muscle atrophy in other diseases, like GNE myopathy, and may play a role in atrophy associated with microgravity. Therefore, to simulate microgravity in a rat model, animal’s hind limbs were suspended for two weeks, while others were under normal conditions for comparison. Semiquantitative real-time polymerase chain reaction (qRT-PCR), a method used to detect and quantify RNA using a known internal standard gene to compare to a targeted gene, was used to quantify the gene expression of STs and sialidases in these rats. RNA was extracted from their gastrocnemius and soleus muscles and then converted to cDNA. The cDNA specific to STs and sialidases was quantified. Methodological data and preliminary development showed that while further developments are needed for the methodology of qRT-PCR, the results could lead to the possibility of therapies that help astronauts maintain their muscle mass in space.
