C. elegans is an excellent model system for studying neuroscience using genetics because of its relatively simple nervous system, sequenced genome, and the availability of a large number of transgenic and mutant strains. Recently, microfluidic devices have been used for high-throughput genetic screens, replacing traditional methods of manually handling C. elegans. However, the orientation of nematodes within microfluidic devices is random and often not conducive to inspection, hindering visual analysis and overall throughput. In addition, while previous studies have utilized methods to bias head and tail orientation, none of the existing techniques allow for orientation along the dorso-ventral body axis. Here, we present the design of a simple and robust method for passively orienting worms into lateral body positions in microfluidic devices to facilitate inspection of morphological features with specific dorso-ventral alignments. Using this technique, we can position animals into lateral orientations with up to 84% efficiency, compared to 21% using existing methods. We isolated six mutants with neuronal development or neurodegenerative defects, showing that our technology can be used for on-chip analysis and high-throughput visual screens.
Spinal muscular atrophy (SMA) is a degenerative disorder that selectively deteriorates motor neurons due to a deficiency of survival motor neuron protein (SMN). The illness is the leading genetic cause of death in infants and is difficult to study in complex biological systems such as humans. A simpler model system, such as the nematode C. elegans, can be used to study potential mechanisms underlying this disease; C. elegans expresses the smn-1 gene, a homologue of SMN; powerful genetic tools in C. elegans research can be used to discover novel genes whose effect on SMN remains unknown or uncharacterized. Currently, conventional screening methods are time-consuming and laborious, as well as being subjective and mostly qualitative. To address these issues, we engineer an automated system capable of performing genetic suppressor screens on C. elegans using microfluidics in combination with custom image analysis software. We demonstrate the utility of this system by isolating 21 alleles that significantly suppress motor neuron degeneration at a screening rate of approximately 300 worms per hour. Many of these mutants also have improved motor function. These isolated alleles can potentially be further studied to understand mechanisms of protection against neurodegeneration. Our system is easily adaptable, providing a means to saturate screens not only implicated in the smn-1 pathway, but also for genes involved in other neurodegenerative phenotypes.
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