Biology subjects

Miccoli, A.

Publications and source records attributed to Miccoli, A..

3 recordsLinked to original sources

bioRxiv2025

Mugil cephalus Low-Density Lipoprotein Receptors: protein family characterization and identification of vitellogenin receptors

Vitellogenin receptors (VtgRs) are pivotal to the successful reproductive event in oviparous species by mediating the uptake of vitellogenin (Vtg) into developing oocytes, ensuring proper yolk formation and embryonic development. In this study, we characterized the low-density lipoprotein receptor (LDLR) family in flathead mullet Mugil cephalus, to which VtgRs belong, and placed a focus on identifying and analyzing vitellogenin receptors. Using a comprehensive approach that combined LDLR orthology inference across 13 species, protein domain analysis and 3D structure prediction, synteny evaluation, functional validation through RNA-seq-derived transcript models and phylogenetic analyses, we identified 87 LDLR family members and characterized two putative vitellogenin receptors belonging to the Lr8/VLDLR and Lrp13/LRX+1 subfamilies. These receptors exhibited conserved domain architectures, and the syntenic arrangements of the genes shared with orthologs in other teleost species as well as the ovary-specific expression profiles were consistent with the functional roles of VtgRs in mediating vitellogenin uptake during oocyte development. Phylogenetic analyses confirmed their evolutionary conservation, while structural modeling revealed key features related to ligand binding and functional specialization. The characterization of vitellogenin receptors in the flathead mullet not only advances the understanding of the reproductive biology in an aquaculture-relevant species but also lays the groundwork for future biotechnological interventions aimed at enhancing reproductive management and genetic improvement. Continued exploration of these avenues promises to improve the sustainability and economic viability of aquaculture practices, making it a fertile area for both basic research and applied biotechnology.

physiology↗

bioRxiv2024

Thrombospondins 1 and 4 undergo coordinated transport to multicore cytotoxic granules to regulate SMAP biogenesis and function in CTL-mediatedcytotoxicity

Supramolecular Attack Particles (SMAPs) are particulate entities, characterized by a cytotoxic core enriched in granzymes and perforin surrounded by a glycoproteic shell, released by CTLs and NK cells. Prior proteomic analysis identified thrombospondin-1 (TSP-1) and thrombospondin-4 (TSP-4) as putative components of SMAPs. While TSP-1 has been validated as a component of the SMAP shell and shown to contribute significantly to CTL-mediated killing, the expression and function of TSP-4 in CTLs, and its interplay with TSP-1 in SMAP biogenesis and function, has not been investigated as yet. Here we demonstrate that TSP-4 and TSP-1 have a complementary expression profile during in vitro human CD8+ T cell differentiation to CTLs and sequentially localize to lytic granules (LG), with TSP-4 being required for TSP-1 association with LGs. Correlative light microscopy identified the TSP-enriched LGs as the SMAP-containing multicore granules. We show by STED microscopy a heterogeneity among TSP-enriched LGs, the most abundant population being positive for both TSP-4 and TSP-1. We also show that TSP-1 and TSP-4 are co-released in association with SMAPs at immune synapses formed on planar supported lipid bilayers, as assessed by dSTORM imaging. Finally, we provide evidence that TSP-4 is required for CTL- and SMAP-mediated cell killing. Of note, we found that chronic lymphocytic leukemia (CLL) cell supernatants, which suppress CTL mediated killing, also suppress expression of TSP-4 as well as of cytolytic effectors and impair SMAP biogenesis. These results identify TSP-4 as a key player in SMAP structure and activity and suggest that SMAPs may be a new target for immune suppression by CLL.

immunology↗

bioRxiv2021

Microlitter pollution in the marine environment and preliminary evidences of in vitro cytotoxic effects on two Mediterranean commercial fish species

Marine litter is composed mainly of plastics and is recognized as a serious threats to marine ecosystems. Ecotoxicological approaches have started elucidating the potential severity of microplastics (MPs) in controlled laboratory studies with pristine materials but no information exist on marine environmental microlitter as a whole. Here, we characterized the litter in the coastal Northern Tyrrhenian sea and in the stomach of two fish species of socio-economic importance, and exposed primary cell cultures of mucosal and lymphoid organs to marine microlitter for evaluating possible cytotoxic effects. An average of 0.30 {+/-} 0.02 microlitter items m-3 was found in water samples. FT-IR analysis revealed that plastic particles, namely HDPE, polyamide and polypropylene were present in 100% and 83.3% of Merluccius merluccius and Mullus barbatus analyzed, which overall ingested 14.67 {+/-} 4.10 and 5.50 {+/-} 1.97 items/individual, respectively. Moreover, microlitter was confirmed as a vector of microorganisms. Lastly, the apical end-point of viability was found to be significantly reduced in splenic cells exposed in vitro to two microlitter conditions. Considering the role of the spleen in the mounting of adaptive immune responses, our results warrant more in-depth investigations for clarifying the actual susceptibility of these two species to anthropogenic microlitter. HighlightsO_LI0.30 {+/-} 0.02 microlitter items m-3 were found at the surface of coastal Northern Tyrrhenian sea C_LIO_LI14.67 {+/-} 4.10 and 5.50 {+/-} 1.97 items/individual were retrieved from the stomach of hakes and mullets C_LIO_LIThe ingested microlitter contained plastic items C_LIO_LIMicrolitter was validated as a carrier of bacteria, fungi and flagellates C_LIO_LISplenic cells exposed to two microlitter conditions for 72 hours suffered cytotoxicity C_LI

zoology↗