microPublication biology最新文献

As part of the Fly-CURE consortium, a mutant allele of Apoptosis inducing factor ( AIF ) was characterized using complementation mapping, genomic sequencing, and mosaic phenotypic analysis to investigate its role in cell growth control in Drosophila melanogaster . The AIF ^e04281 mutation dramatically reduced homozygous mutant clone size and caused morphological defects in genetically mosaic eyes. Sequencing confirmed a transposon insertion that truncates the AIF protein preceding conserved domains essential for mitochondrial function and apoptosis. The observed clone loss indicates a cell-autonomous requirement for AIF and supports the use of AIF ^e04281 as a loss-of-function background for genetic modifier screens on chromosome arm 2L.

Transgenic regulators and reporter controls such as GAL80 and RNA interference (RNAi) constructs targeting fluorescent proteins are invaluable tools in Drosophila research. During experiments designed to temporally regulate gene expression, we observed unexpected flight defects in aged flies with constitutive expression of temperature-sensitive GAL80 ^TS . Furthermore, ubiquitous expression of RNAi targeting EGFP or mCherry induced survival and motor impairments, even in the absence of corresponding fluorescent reporters. These findings suggest that genetic tools commonly assumed to be physiologically inert can have measurable effects on motor behaviour. Researchers employing these tools should therefore interpret behavioural data with caution and include appropriate controls.

Cyclic nucleotides (cAMP, cGMP) are critical second messengers that transduce diverse signaling events within cells, ranging from developmental processes and neuronal signaling to roles in cellular growth (Fajardo et al., 2014; Galande & Cote, 2025; Levy et al., 2011). Thus, the regulation of cyclic nucleotide levels is critical for the regulation of signal transduction pathways and cellular homeostasis.  We previously reported a role for cGMP in the negative regulation of bitter taste avoidance in C. elegans (Chaubey et al., 2023; Krzyzanowski et al., 2013; Krzyzanowski et al., 2016).  Phosphodiesterases (PDEs) are a class of enzymes that break down cyclic nucleotides, and we show here that multiple PDEs likely work together to modulate C. elegans bitter taste sensitivity.

Based on gene content similarity, Alatato and Audell are grouped to phage clusters FB and FR, respectively. Alatato has a 39,780bp genome and 71 genes of which 34 have a predicted function. Alatato is predicted to be temperate based on the identification of an immunity cassette with genes that encode a tyrosine integrase, helix-turn-helix DNA binding proteins and an excise. Audell has a 42,187bp genome and 67 genes of which 32 have a predicted function with five enzymes involved in thymine synthesis. Putative lysis cassettes with genes encoding endolysins and transmembrane proteins were identifiable in both phages.

Cross-species expression of proteins in mammalian cells and Xenopus oocytes remains a powerful strategy for functional studies. This can be facilitated by plasmids with dual promoters, enabling use in both systems without separate subcloning. We developed pXeDNA3.1-LacZ, a plasmid that supports expression in mammalian cells and oocytes. The construct combines a cytomegalovirus promoter for mammalian transcription with Xenopus β-globin untranslated regions to enhance oocyte translation, and incorporates XcmI-mediated TA cloning and blue/white selection. To validate it, we used human TRPV1 (HsTRPV1). The vector drove robust expression in HEK293G5A cells and Xenopus oocytes.pXeDNA3.1-LacZ streamlines assays and facilitates studies of membrane proteins.

GABA _A receptors are present in hindbrain and spinal cord networks, playing a pivotal role in regulating locomotion. In this study, we demonstrate that mutations in the gabra4 gene, which encodes the α4 subunit of GABA _A receptors, result in increased swim velocity of larval zebrafish. We also show that this gene is selectively expressed within spinal cord cerebrospinal fluid contacting neurons (CSF-cNs). Given the significance of these neurons in modulating locomotion, our findings support a model in which compromised α4 function leads to an increase in CSF-cN activity, causing a subtle, hyperactive swimming phenotype.

AmiCi24 is a novel siphoviral bacteriophage that was isolated from a soil sample collected in Sewell, NJ, USA using Arthrobacter globiformis B-2979 as the host. AmiCi24 has a genome consisting of 38,466 base pairs that encodes 68 predicted protein-coding genes. Based on gene content, AmiCi24 is assigned to actinobacteriophage cluster AS and subcluster of AS3 and is predicted to be temperate.

Septins are cytoskeletal proteins crucial for cell division and many other processes. In fission yeast, septins localize to the division site during septum formation. Recently, we conducted a study that elucidates the localizations and functionalities of epitope-tagged septins. However, some questions remain outstanding. Here we assessed the impacts of monomeric mEGFP and dimeric GFP(S65T) on the septin Spn4 tagged at either of its terminus. We found that septin levels were important for its function, Spn4-mEGFP localized normally to the division site, but GFP(S65T)-Spn4 formed elongated structures ectopically, further highlighting that dimeric tags are more disruptive to septin localizations.

Tepache is a traditional, homemade Mexican drink made by fermenting pineapple rinds. The natural probiotic bacteria in tepache are said to promote a healthy gut microbiome. This study assessed the microbial community in homemade tepache for diversity, survival in simulated gastric fluid, and antibiotic resistance. Simulated gastric passaging reduced total community numbers but the community density was not strongly impacted by exposure to tetracycline. Metagenomic analysis reveals a community dominated by Bacillus, Meyerozyma and Talaromyces. These results indicate that consuming home fermented beverages may provide helpful probiotic bacteria but could also expose the gut microbiome to antibiotic resistance genes.

Fluorescent proteins (FPs) are essential tools for live imaging, with brightness and photostability being key parameters when selecting an FP for experiments. Recently developed variants of monomeric StayGold (mSG) and mGold2 promise improvements in these properties, yet their performance can vary across model organisms and tissue contexts. To guide the selection of FPs for zebrafish imaging, we expressed membrane-targeted FP fusions in embryos and systematically compared their in vivo brightness and photostability against the widely used mNeonGreen (mNG). Among the mSG variants, mSG(A) displayed brightness comparable to mNG but with markedly increased photostability. In contrast, mSG(BJ) and mSG(E138D) showed reduced brightness, while retaining improved photostability. The mGold2 variants exhibited no improvement in photostability over mNG, but their increased brightness at 514 nm excitation allowed effective imaging at lower laser intensities, thereby extending usable imaging times. There were no significant differences detected between the two mGold2 variants. Overall, mSG(A) emerges as an optimal choice for long-term imaging, while mGold2 variants are advantageous when maximal brightness is required. These results provide practical benchmarks for FP selection in zebrafish embryonic imaging.