PT  - JOURNAL ARTICLE
AU  - Richer, Petra
AU  - Speese, Sean D.
AU  - Logan, Mary A.
TI  - FLARIM v2.0, an improved method to quantify transcript-ribosome interactions <em>in vivo</em> in the adult <em>Drosophila</em> brain
AID  - 10.1101/2021.08.13.456301
DP  - 2021 Jan 01
TA  - bioRxiv
PG  - 2021.08.13.456301
4099  - http://biorxiv.org/content/early/2021/08/14/2021.08.13.456301.short
4100  - http://biorxiv.org/content/early/2021/08/14/2021.08.13.456301.full
AB  - Neural injury triggers striking immune reactions from glial cells, including significant transcriptional and morphological changes, but it is unclear how these events are coordinated to mount an effective immune response. Here, we present a new variant of the Fluorescence assay to detect ribosome interactions with mRNA (FLARIM), which we term FLARIM v2.0, to visualize single immune gene transcripts and association with ribosomes in glia responding to neurodegeneration. Specifically, using an in vivo axotomy assay in Drosophila, we show that matrix metalloproteinase-1 (Mmp-1) mRNAs and associated ribosomes are detected in distal processes of reactive glia where they are actively engulfing degenerating axonal material, suggesting that local translation is an important component of the glial immune response to axotomy. This work also validates our enhanced FLARIM assay as a promising tool to investigate mechanisms of mRNA transport and translation in a wide range of in vitro and in vivo paradigms.Competing Interest StatementThe authors have declared no competing interest.