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Poole, R.

Publications and source records attributed to Poole, R..

3 recordsLinked to original sources

The replicative helicase CMG is required for the divergence of cell fates during asymmetric cell division in vivo

The mechanisms that enable differential gene expression in daughter cells produced by asymmetric cell divisions are not well understood. We discovered that the eukaryotic replicative helicase CMG (Cdc45-MCM-GINS) is required for this process in C. elegans. During C. elegans development, some dividing cells give rise to a daughter that survives and a daughter that dies. We found that PSF-2 GINS2, a component of C. elegans CMG, is necessary for the transcriptional burst of the pro-apoptotic gene egl-1 BH3-only, which occurs in the daughter that dies immediately following mother cell division. We present evidence that this requirement is independent of the function of CMG in DNA unwinding. We propose that the recently described histone chaperone activity of CMG causes epigenetic changes at the egl-1 locus during replication in mother cells, and that these changes are required for the increase in egl-1 transcription in the daughter that dies. We also find that PSF-2 is required for the divergence of other cell fates during C. elegans development, suggesting that this function is not restricted to the regulation of egl-1 expression. Our work uncovers a new and unexpected role of CMG in cell fate and a novel intrinsic mechanism for gene expression plasticity in the context of asymmetric cell division.

cell biology↗

Invasive slipper limpets Crepidula fornicata act like a sink, rather than source, of Vibrio spp.

A large knowledge gap exists regarding the disease profile and pathologic condition of the invasive, non-native, slipper limpet Crepidula fornicata. To help address this, we performed a yearlong health survey across two sites in South Wales UK - subtidal Swansea Bay and intertidal Milford Haven. In total, 1,800 limpets were screened systematically for haemolymph bacterial burdens using both general and vibrio-selective growth media (TSA +2% NaCl and TCBS, respectively), haemolymph (blood) inspection using microscopy, a PCR-based assay targeting Vibrio spp., and multi-tissue histology. Over 99% of haemolymph samples contained cultivable bacterial colony forming units, and 83% of limpets tested positive for the presence of vibrios via PCR (confirmed via Sanger sequencing). Vibrio presence did not vary greatly across sites, yet a strong temporal (seasonal) effect was observed - significantly higher bacterial loads during the summer. Binomial logistic regression models revealed larger (older) limpets were more likely to harbour vibrios, and the growth of bacteria on TCBS was a key predictor for PCR-based vibrio detection. Histological assessment of >340 animals revealed little evidence of inflammation, sepsis, or immune reactivity despite the gross bacterial numbers. We contend that slipper limpets are not susceptible to bacteriosis at either site surveyed, or do not to harbour vibrios known to be pathogenic to humans. The lack of susceptibility to local pathogenic bacteria may explain, in part, the invasion success of C. fornicata across this region.

microbiology↗

In-situ non-lethal rapid test to accurately detect the presence of the nematode parasite, Anguillicoloides crassus, in European eel, Anguilla anguilla.

Anguillicoloides crassus is an invasive nematode parasite of the European eel, Anguilla anguilla, and one of the primary drivers of eel population collapse. The presence of the parasite has been shown to impact many features of eel physiology and life history. Early detection of the parasite is vital to limit the spread of A. crassus. However, until recently, accurate diagnosis of infection could only be achieved via terminal dissection. To support A. anguilla fisheries management in the context of A. crassus we developed a rapid non-lethal and non-invasive environmental DNA method to detect the presence of the parasite in the swim bladder. Screening of 131 wild eels was undertaken between 2017 and 2019 Ireland and UK to validate the procedure. DNA extractions and PCR were conducted using both a Qiagen Stool kit at Glasgow University and in situ using Whatman qualitative filter paper No. 1 and a miniPCR DNA Discovery System. Primers were specifically designed from the cytochrome oxidase mtDNA gene region. In situ extraction and amplification takes approx. 3h for up to 16 individuals with higher specificity and sensitivity compare to the laboratory Qiagen kit extraction. The local diagnostic procedure demonstrated Positive Predictive Values at 96% and Negative Predictive Values at 87%. Our method will be a powerful tool in the hands of fisheries managers to help protect this iconic but critically endangered species. It will allow a non-invasive monitoring of the A. crassus dispersion across the European waters.

ecology↗