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Precise mapping of one classic and three novel GluRIIA mutants in Drosophila melanogaster
Journal article   Open access   Peer reviewed

Precise mapping of one classic and three novel GluRIIA mutants in Drosophila melanogaster

Bhagaban Mallik, Douglas J Brusich, Georgette Heyrman and C. Andrew Frank
microPublication biology, Vol.2023
01/01/2023
DOI: 10.17912/micropub.biology.000784
PMCID: PMC10276266
PMID: 37334199
url
https://doi.org/10.17912/micropub.biology.000784View
Published (Version of record) Open Access

Abstract

Mutation of the Drosophila melanogaster GluRIIA gene or pharmacological agents targeting it are commonly used to assess homeostatic synaptic function at the larval neuromuscular junction (NMJ). The commonly used mutation, GluRIIASP16, is a null allele created by a large and imprecise excision of a P-element which affects GluRIIA and multiple upstream genes. Here we mapped the exact bounds of the GluRIIASP16 allele, refined a multiplex PCR strategy for positive identification of GluRIIASP16 in homozygous or heterozygous backgrounds, and sequenced and characterized three new CRISPR-generated GluRIIA mutants. We found the three new GluRIIA alleles are apparent nulls that lack GluRIIA immunofluorescence signal at the 3rd instar larval NMJ and are predicted to cause premature truncations at the genetic level. Further, these new mutants have similar electrophysiological outcomes as GluRIIASP16, including reduced miniature excitatory postsynaptic potential (mEPSP) amplitude and frequency compared to controls, and they express robust homeostatic compensation as evidenced by normal excitatory postsynaptic potential (EPSP) amplitude and elevated quantal content. These findings and new tools extend the capacity of the D. melanogaster NMJ for assessment of synaptic function.
 Drosophila Genotype Data New Finding Phenotype Data

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