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1.
PeerJ ; 11: e15433, 2023.
Article in English | MEDLINE | ID: mdl-37214099

ABSTRACT

The miniMOS technique has been widely used in the C. elegans community to generate single copy insertions. A worm is considered as a potential insertion candidate if it is resistant to G418 antibiotics and does not express a co-injected fluorescence marker. If the expression of the extrachromosomal array is very low, it is possible for a worm to be mistakenly identified as a miniMOS candidate, as this low expression level can still confer resistance to G418 without producing a detectable fluorescence signal from the co-injection marker. This may increase the workload for identifying the insertion locus in the subsequent steps. In the present study, we modified the plasmid platform for miniMOS insertion by incorporating a myo-2 promoter-driven TagRFP or a ubiquitous H2B::GFP expression cassette into the targeting vector and introducing two loxP sites flanking the selection cassettes. Based on this new miniMOS tool kit, the removable fluorescence reporters can be used to visualize the single copy insertions, greatly reducing insertion locus identification efforts. In our experience, this new platform greatly facilitates the isolation of the miniMOS mutants.


Subject(s)
Caenorhabditis elegans , Genetic Engineering , Animals , Animals, Genetically Modified/genetics , Caenorhabditis elegans/genetics , Genetic Engineering/methods
2.
PLoS Genet ; 18(1): e1010029, 2022 01.
Article in English | MEDLINE | ID: mdl-35100258

ABSTRACT

Olfactory integration is important for survival in a natural habitat. However, how the nervous system processes signals of two odorants present simultaneously to generate a coherent behavioral response is poorly understood. Here, we characterize circuit basis for a form of olfactory integration in Caenorhabditis elegans. We find that the presence of a repulsive odorant, 2-nonanone, that signals threat strongly blocks the attraction of other odorants, such as isoamyl alcohol (IAA) or benzaldehyde, that signal food. Using a forward genetic screen, we found that genes known to regulate the structure and function of sensory neurons, osm-5 and osm-1, played a critical role in the integration process. Loss of these genes mildly reduces the response to the repellent 2-nonanone and disrupts the integration effect. Restoring the function of OSM-5 in either AWB or ASH, two sensory neurons known to mediate 2-nonanone-evoked avoidance, is sufficient to rescue. Sensory neurons AWB and downstream interneurons AVA, AIB, RIM that play critical roles in olfactory sensorimotor response are able to process signals generated by 2-nonanone or IAA or the mixture of the two odorants and contribute to the integration. Thus, our results identify redundant neural circuits that regulate the robust effect of a repulsive odorant to block responses to attractive odorants and uncover the neuronal and cellular basis for this complex olfactory task.


Subject(s)
Caenorhabditis elegans/physiology , Sensory Receptor Cells/physiology , Smell/physiology , Animals , Caenorhabditis elegans/genetics , Ketones/pharmacology , Mutation , Odorants , Pentanols/pharmacology , Smell/drug effects
3.
Article in Chinese | WPRIM (Western Pacific) | ID: wpr-329375

ABSTRACT

Introduced the "particles" item testing progress and some notes, using Coulter Multisizer, about medical devices and drugs wrapper, with an expectation of some help for the "particles" item tests in the solution in these area.


Subject(s)
Equipment Contamination , Equipment and Supplies , Materials Testing , Methods , Particle Size , Quality Control
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