The ADAMTS (a disintegrin and metalloproteinase with thrombospondin type-1 motifs) metalloprotease family comprises 19 members, and they play critical roles in development, angiogenesis and coagulation. These enzymes are...The ADAMTS (a disintegrin and metalloproteinase with thrombospondin type-1 motifs) metalloprotease family comprises 19 members, and they play critical roles in development, angiogenesis and coagulation. These enzymes are also pivotal in the turnover of extracellular matrix proteins in various tissues, and their dysregulation has been linked to diseases such as arthritis, atherosclerosis, cancer and inflammation. These secreted zinc metalloproteases are composed of multiple domains and are localized to the extracellular matrix and present in circulation. The metalloprotease member 7 of this family or ADAMTS7 was cloned in 2004. However, in 2011, it gained significant attention when its gene locus was associated with an increased risk of atherosclerosis and coronary artery disease. Despite its strong association with human diseases, the biological functions, substrate targets and processing of ADAMTS7 remain poorly understood, limiting our comprehension of its roles in both health and disease. This review highlights the structural organization of ADAMTS7, its expression and regulation, known substrates, and its involvement in various pathophysiological processes. Additionally, perspectives on future research directions and priorities are presented. ADAMTS7 remains a compelling target for drug and biomarker discovery, though much about this metalloprotease remains to be uncovered.
The ability to learn and form memory is critical for survival, yet even genetically similar individuals can vary considerably in their cognitive performance. Using the pond snail Lymnaea stagnalis, we investigated how in...The ability to learn and form memory is critical for survival, yet even genetically similar individuals can vary considerably in their cognitive performance. Using the pond snail Lymnaea stagnalis, we investigated how individual sensitivity to stress influences configural learning-a higher-order form of associative learning in which the simultaneous exposure to two contrasting stimuli, such as a predatory odour and an appetitive taste, results in the appetitive stimulus becoming associated with risk and evoking anti-predator behaviours. We used freshly collected, predator-naive snails from Margo Lake, Canada. While group-level data suggested the Margo strain failed to learn, individual-level analysis revealed that some snails successfully formed configural memories, while others did not. We hypothesized that this divergence reflects differences in individual (predator-related) stress responsiveness, which may modulate the engagement of memory-related molecular pathways. To test this, we measured expression levels of selected genes in the central ring ganglia. Snails that formed configural memories showed significantly higher expression of stress-responsive genes, components of the serotonin pathway and markers of neuroplasticity, along with increased endocannabinoid turnover. These findings suggest that individual variation in stress reactivity can drive adaptive differences in cognitive performance, offering new insights into the molecular and behavioural mechanisms underlying learning and memory.
Calcium (Ca²+) release from intracellular stores, Ca²+ entry across the plasma membrane and their coordination via store-operated Ca²+ entry (SOCE) are critical for receptor-activated Ca²+ oscillations. However, the prec...Calcium (Ca²+) release from intracellular stores, Ca²+ entry across the plasma membrane and their coordination via store-operated Ca²+ entry (SOCE) are critical for receptor-activated Ca²+ oscillations. However, the precise mechanism of Ca²+ oscillations and whether their control loop resides at the plasma membrane or intracellularly remains unresolved. By examining the dynamics of stromal interaction molecule 1 (STIM1), an endoplasmic reticulum (ER)-localized Ca²+ sensor that activates the Orai1 channel on the plasma membrane for SOCE, in mast cells, we found that a significant proportion of cells exhibited STIM1 oscillations with the same periodicity as Ca²+ oscillations. These cortical oscillations, shared with ER-plasma membrane (ER-PM) contact site proteins, were only detectable using total internal reflection fluorescence microscopy. Notably, STIM1 oscillations could occur independently of Ca²+ oscillations. Simultaneous imaging of cytoplasmic Ca²+ and ER Ca²+ with CEPIA1er revealed that receptor activation does not deplete ER Ca²+, whereas receptor activation without extracellular Ca²+ influx induces cyclic ER Ca²+ depletion. However, under such non-physiological conditions, cyclic ER Ca²+ oscillations lead to sustained STIM1 recruitment, indicating that oscillatory Ca²+ release is neither necessary nor sufficient for STIM1 oscillations. Using optogenetic tools to manipulate ER-PM contact site dynamics, we found that persistent ER-PM contact sites reduced the amplitude of Ca²+ oscillations without alteration of oscillation frequency. Together, these findings suggest an active cortical mechanism governs the rapid dissociation of ER-PM contact sites, thereby controlling amplitude of oscillatory Ca²+ dynamics during receptor-induced Ca²+ oscillations.
Cellular myelocytomatosis oncogene (MYC) transcription factors are encoded by a family of genes that include the prototype member MYC, MYCN and MYCL, and most human cancers display expression alterations of MYC genes. MY...Cellular myelocytomatosis oncogene (MYC) transcription factors are encoded by a family of genes that include the prototype member MYC, MYCN and MYCL, and most human cancers display expression alterations of MYC genes. MYC is regulated at multiple levels, and its stability and activity are modulated by protein phosphorylation. Although there is a reasonable knowledge of the kinases required for MYC modifications, the counteracting phosphatases have been understudied. Here, we have investigated the role of the chromatin-associated protein phosphatase 1 (PP1) regulatory subunit CDCA2, also known as Repo-Man, in the regulation of MYC proteins in cancer cells. Using RNA interference and degron-mediated degradation of CDCA2, we have demonstrated that the PP1 subunit is required for cMYC and MYCN stabilization and viability of triple-negative breast cancer, neuroblastoma and colon cancer cells. Proximity ligation assays indicate that both cMYC and MYCN are in close proximity to CDCA2 in vivo. Furthermore, we have shown that CDC2A is a bona fide MYC target gene in cancer cells, revealing a reciprocal regulatory loop that could be exploited for therapeutic purposes.
Chemosensory systems play an essential role in insect survival and reproduction, mediating behaviours such as host location, mating and oviposition. The rice grasshopper Oxya intricata is an important rice pest in Asia,...Chemosensory systems play an essential role in insect survival and reproduction, mediating behaviours such as host location, mating and oviposition. The rice grasshopper Oxya intricata is an important rice pest in Asia, yet the molecular basis of its olfaction remains poorly understood. In this study, we sequenced antennal transcriptomes from adult males and females and systematically identified chemosensory-related genes. A total of 104 candidates were identified, including 14 odorant-binding proteins (OBPs), 12 chemosensory proteins (CSPs), 67 odorant receptors (ORs), nine ionotropic receptors (IRs) and two sensory neuron membrane proteins (SNMPs). Expression profiling by quantitative real-time PCR revealed broad antennal enrichment across families, with subsets showing antenna-specific or sex-biased patterns, suggesting differential chemosensory deployment between tissues and sexes. Several OBPs, CSPs and ORs were also elevated in maxillary palps or tarsi, raising the possibility that these genes may be associated with contact or near-field chemosensation. Collectively, these results provide the first comprehensive survey of chemosensory genes in O. intricata and establish a molecular resource for future functional analyses of odour coding. The identified gene set offers tractable targets for ligand screening and behavioural assays, and may inform future efforts to develop semiochemical-based monitoring and management strategies for this economically important pest.
Ca2+ imaging in the living tissues of experimental animals currently presents limitations due to motion artefacts, variable expression levels and the low basal fluorescence of available probes. These issues are even more...Ca2+ imaging in the living tissues of experimental animals currently presents limitations due to motion artefacts, variable expression levels and the low basal fluorescence of available probes. These issues are even more troublesome when performing Ca2+ imaging in contractile tissues and moving organelles. Despite the well-known advantages of ratiometric sensors, their application in experimental animals is still limited. This study addresses these challenges in the fruit fly Drosophila melanogaster, introducing the development and characterization of novel transgenic fly lines expressing advanced FRET-based Cameleon probes for cytosolic and mitochondrial Ca2+ imaging. A series of Ca2+ imaging experiments in selected tissues demonstrates their functionality. These newly developed tools guarantee precise and quantitative measurement of Ca2+ in various cellular contexts, which will contribute to advancing Ca2+ research in Drosophila.
The existence of sex differences in human pre-implantation development remains an open question, with previous attempts based on human observations yielding inconclusive results. In this study, we combined manual annotat...The existence of sex differences in human pre-implantation development remains an open question, with previous attempts based on human observations yielding inconclusive results. In this study, we combined manual annotation and deep learning analysis of a dataset comprising 515 time-lapse embryo movies to investigate whether birth sex influences early developmental dynamics. While manual assessment did not identify any developmental timing parameters that reliably distinguish male from female embryos, a deep learning model trained and tested on these videos achieves a statistically significant sex prediction accuracy of 61%. Importantly, our analyses identified the period after the eight-cell stage as critical for accurate prediction, indicating that subtle sex-related differences may begin to emerge around day 3 of human embryogenesis. Studying sex differences at this early stage may enhance our understanding of why some embryos fail to develop and why sex ratios can be skewed in the context of in vitro fertilization. More broadly, our findings open the possibility for an early, non-invasive detection tool that could assist in identifying and addressing sex-related embryonic developmental abnormalities.
SIRT6, a highly conserved member of the sirtuin family, plays a critical role in diverse cellular processes, including gene regulation, DNA damage response and maintaining nuclear lamina integrity. These functions are es...SIRT6, a highly conserved member of the sirtuin family, plays a critical role in diverse cellular processes, including gene regulation, DNA damage response and maintaining nuclear lamina integrity. These functions are essential in contexts such as differentiation, metabolic regulation, cancer development and ageing. Given the multifaceted influence of SIRT6 on cellular activities, there is an increasing interest in elucidating the regulatory mechanisms governing its enzymatic functions. SIRT6 exhibits two NAD+-dependent activities: deacetylation and ADP-ribosylation, with current research predominantly focusing on the former. However, the latter-its (ADP-ribosyl)transferase activity-remains underexplored, particularly concerning the specific amino acid targets it modifies and the (ADP-ribosyl)hydrolases that can reverse these modifications. In this study, we have utilized biochemical assays and proteomic techniques to investigate these aspects, revealing that SIRT6 transfers ADP-ribosyl moieties onto histidine and tyrosine residues. In addition, we reveal that the (ADP-ribosyl)hydrolase ARH3 has significant activity in erasing SIRT6-derived ADP-ribosylation in cells.
Choanoflagellates are heterotrophic holozoans that are classified into two groups based on their morphology: loricates, which possess silica-based extracellular structures, and craspedids, which do not. Loricate choanofl...Choanoflagellates are heterotrophic holozoans that are classified into two groups based on their morphology: loricates, which possess silica-based extracellular structures, and craspedids, which do not. Loricate choanoflagellate taxonomy has historically been based on selected aspects of their lorica morphology and on their division mode, either tectiform or nudiform. Here, we characterize two new loricate strains, BEAP0094 and BEAP0360, that display unexpected morphological features when compared to their closest genetic relatives. The ribosomal gene sequence of BEAP0094 closely matched Pseudostephanoeca paucicostata, but its lorica lacked the characteristic anterior ring found in all Stephanoeca species. Instead, its lorica resembled the Acanthocorbis genus, raising the possibility of the existence of either multiple lorica morphologies in closely related species or multiple species sharing the same ribosomal 18S gene. In contrast, BEAP0360 presented a morphological match to Stephanoeca cauliculata, but its sequence did not, suggesting that different species could share the same lorica architecture. BEAP0360, here described as Cepoeca plumata (n. gen. n. sp.), possesses a key phylogenetic placement, potentially as the earliest-branching nudiform loricate. Our findings are inconsistent with a strict classification based on lorica morphology and support the usage of genetic data as primary criterion for genus-level taxonomic assignment.
Sensory processing deficits are common in neurodevelopmental disorders (NDDs); however, we lack a full understanding of the circuits impacted. The superior colliculus (SC) is a sensorimotor region that directs complex be...Sensory processing deficits are common in neurodevelopmental disorders (NDDs); however, we lack a full understanding of the circuits impacted. The superior colliculus (SC) is a sensorimotor region that directs complex behaviours, which recent work suggests is adversely impacted in NDDs. However, our understanding of cellular diversity in the SC lags in comparison to other regions, limiting our ability to parse circuit changes in NDDs. A goal of neuroscience has been to elucidate the diversity of neurons in the brain. Analysis of action potential shape in extracellular recordings has revealed subpopulations in several regions, allowing for insights into subtype-specific function in the intact brain. Here, we utilized semi-automated clustering methods to classify neurons in the mouse SC based on features of extracellularly recorded waveforms to identify five putative cell types. Secondary analysis of firing statistics and visual tuning properties supported cluster segregation. Interestingly, the proportions of units assigned to each cluster differed in a mouse model of fragile X syndrome (Fmr1-/y). Furthermore, we observed changes in waveform properties and firing statistics between genotypes in a subtype-specific manner. Taken together, these data add to our understanding of neuronal diversity in the SC and alterations of visual circuit organization and function in NDDs.
Anti-apoptotic B-cell lymphoma 2 (BCL-2) controls inositol 1,4,5-trisphosphate receptor (IP3R)-mediated Ca²+ signalling. As cells typically express all three IP3R isoforms in variable abundances that assemble in hetero-t...Anti-apoptotic B-cell lymphoma 2 (BCL-2) controls inositol 1,4,5-trisphosphate receptor (IP3R)-mediated Ca²+ signalling. As cells typically express all three IP3R isoforms in variable abundances that assemble in hetero-tetrameric channels, the specific effects of BCL-2 on each isoform remain unclear. Here, we employed a reductionist approach using HEK293 cells triple-IP3R knockout reconstituted with a single IP3R isoform to elucidate the impact of BCL-2 on Ca2+ signalling by homo-tetrameric IP3R channels. Co-immunoprecipitation experiments demonstrated that BCL-2 interacts with each IP3R isoform. Live-cell Ca²+ imaging revealed that BCL-2 overexpression suppresses Ca²+ signals evoked by any of the three IP3R isoforms. Moreover, BCL-2 overexpression impaired mitochondrial Ca²+ uptake following IP3R-mediated Ca²+ release, irrespective of the IP3R isoform present. To investigate the effects on single IP3R-channel activity, we performed Ca²+-puff analysis using TIRF microscopy in response to UV-flash photolysis of caged IP3. BCL-2 overexpression reduced the number of Ca²+ puffs across all IP3R isoforms without affecting the amplitude or duration of individual puffs. Thus, BCL-2 acts as a universal inhibitor of all three IP3R isoforms, highlighting its critical role in fine-tuning intracellular Ca2+ dynamics to promote cell survival and its potential as a therapeutic target in cancer via its role in Ca2+ signalling.
The recycling of urea nitrogenous by urease presents a valuable approach for optimizing nitrogen utilization in organisms with limited nitrogen resources. In silkworms, haemolymph urea concentration correlates with ureas...The recycling of urea nitrogenous by urease presents a valuable approach for optimizing nitrogen utilization in organisms with limited nitrogen resources. In silkworms, haemolymph urea concentration correlates with urease activity, and the urease is derived from their exclusive food source, mulberry leaves. Although mulberry-derived urease has been shown to cross the silkworm midgut barrier, the mechanism of its translocation into the haemolymph remains unknown. In the present study, we investigated the transport mechanism of mulberry urease into the silkworm haemolymph using Western blotting and immunoelectron microscopy. Our results revealed that exosomes and autophagy-related factors, which are regulated by 20-hydroxyecdysone (20E), mediate the stage-specific transport of mulberry urease from the midgut to the haemolymph in silkworm larvae. Urease deficiency impaired silkworm growth and development, significantly reducing cocoon shell weight and egg production. These findings highlight the profound influence of host plants on insect phenotypic traits and reveal a novel cross-kingdom molecular transport mechanism. Our results not only deepen the understanding of plant-insect interactions but also offer practical insights for sericulture improvement, particularly in formulating artificial diets to enhance silk production.
Heat shock protein 90s (HSP90s) are molecular chaperones often produced by cells in response to hostile conditions. Here, we characterized the importance of HSP90 to growth/development and survival of the blood fluke Sch...Heat shock protein 90s (HSP90s) are molecular chaperones often produced by cells in response to hostile conditions. Here, we characterized the importance of HSP90 to growth/development and survival of the blood fluke Schistosoma mansoni, focusing on life stages that parasitize the human host. Western blotting revealed striking HSP90 upregulation in developing schistosomula in vitro, and pharmacological assays revealed that the HSP90 inhibitors gedunin, 17-N-allylamino-17-demethoxygeldanamycin (17-AAG) and 5-(2-amino-4-chloro-7-((4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-methylpent-4-yn-2-ol (EC-144) profoundly attenuated the viability/development of skin schistosomula to lung and liver stages. In vitro liver stage, ex vivo adult male/female worms and eggs were also killed. Strikingly, 17-AAG and EC-144 blocked stem cell proliferation in the skin, lung and liver schistosomula, and the testicular lobes and ovaries of adult worms. Small interfering RNA-mediated knockdown of the cytoplasmic HSP90 alpha isoform 2 (Smp_072330) also attenuated stem cell proliferation and restricted schistosomulum growth, supporting the importance of this isoform. 17-AAG and EC-144 attenuated the phosphorylation of schistosomula Akt/protein kinase B, extracellular signal-regulated kinase and p38 mitogen-activated protein kinase, suggesting HSP90 regulates core 'system-based' signalling pathways in the parasite; glucose uptake was also reduced. Collectively, these novel findings highlight the importance of HSP90 to intra-mammalian schistosome life stages and identify HSP90 as a possible drug target to control human schistosomiasis.
Transmembrane proteins (TMEMs) constitute a large family of proteins that span biological membranes and are distributed across various cellular organelles, playing key roles in maintaining cellular homeostasis. Increasin...Transmembrane proteins (TMEMs) constitute a large family of proteins that span biological membranes and are distributed across various cellular organelles, playing key roles in maintaining cellular homeostasis. Increasing evidence has revealed that dysregulation of TMEMs is closely associated with cancer development and progression. Therefore, a deeper understanding of the relationship between TMEMs and cancer is essential. Different TMEMs can function either as oncogenes or tumour suppressors, depending on the context. In this review, we explore the involvement of TMEMs in cancer, categorizing them into three groups based on their roles: oncogenic, tumour-suppressive or dual-function (both oncogenic and tumour-suppressive). We summarize the roles of various TMEMs in different cancer types, highlighting both well-characterized proteins and those identified through database screening, even if their exact molecular mechanisms remain unclear. Where possible, we include known signalling pathways associated with these TMEMs. This review highlights the critical roles of the TMEM protein family and encourages further research into their mechanisms, prognostic value and potential as targets for cancer therapy.
Exploring the evolution of gene networks associated with metabolic/energetic homeostasis can yield key insights into the adaptive landscapes governing the physiology of extant lineages. Here, we investigate a key hormona...Exploring the evolution of gene networks associated with metabolic/energetic homeostasis can yield key insights into the adaptive landscapes governing the physiology of extant lineages. Here, we investigate a key hormonal module of energy metabolism in reptiles. Ghrelin (GHRL), also known as the 'hunger hormone', is a multifunctional gastric peptide, involved in appetite, food intake and body weight regulation. We examined the genomes of 112 species comprising members of the Squamata, Testudines, Crocodilia and Rhynchocephalia and provided ample evidence that GHRL was independently lost in snakes (32 species), chameleons (four species) and toadhead agamas (two species). In accordance, the enzyme responsible for ghrelin acylation and essential for its activity, MBOAT4 (membrane bound O-acyltransferase domain containing 4), is also eroded in these lineages. We suggest that the loss of this hormonal signalling system parallels critical modifications in energy metabolism, such as lower energy expenditure during rest, possibly linked with their unique ability to undergo large periods of fasting.
Necroptosis is a form of regulated cell death (RCD) that evolved as a defence against pathogenic infection. Unlike caspase-dependent RCD, necroptosis, in its canonical form, is driven by receptor-interacting protein kina...Necroptosis is a form of regulated cell death (RCD) that evolved as a defence against pathogenic infection. Unlike caspase-dependent RCD, necroptosis, in its canonical form, is driven by receptor-interacting protein kinase 1 and 3 (RIPK1 and RIPK3) signalling, culminating in the activation of the pseudokinase mixed lineage kinase domain-like protein (MLKL). Central to this process is the interaction between MLKL and its upstream regulator, RIPK3, forming a functional module called the necrosome that governs the spatiotemporal execution of cell death. Despite progress in our understanding of necroptotic signalling, key open questions remain. The structural organization of MLKL influences its interaction with RIPK3, yet the precise features of their binding surfaces and their regulation are not fully resolved. Additionally, the high-order supramolecular assembly of the necrosome and its transition between different states remain poorly understood, particularly regarding how RIPK3 and MLKL configurations impact necrosome activity and stability. In this review, we summarize current knowledge on the evolution, structure and regulation of the RIPK3-MLKL axis and discuss models of their activation in light of recent discoveries.
Fucosyltransferase 8 (FUT8), a glycosyltransferase responsible for core fucosylation, is overexpressed in numerous cancers and promotes many malignant processes such as cell proliferation, invasion and migration. Transfo...Fucosyltransferase 8 (FUT8), a glycosyltransferase responsible for core fucosylation, is overexpressed in numerous cancers and promotes many malignant processes such as cell proliferation, invasion and migration. Transforming growth factor-β (TGF-β) stimulation promotes epithelial-mesenchymal transition (EMT), a pivotal process indicating the invasion and metastasis of glioblastoma (GBM). However, the mechanism underlying the impact of FUT8 on the TGF-β signalling pathway in GBM progression remains largely unexplored. Our data revealed that FUT8 was highly expressed in patients with GBM and was associated with poor outcomes. FUT8 knockdown inhibited TGF-β-induced EMT, whereas FUT8 overexpression promoted TGF-β-induced EMT in vitro and in vivo. Mechanistic investigations revealed that FUT8 expression increased during TGF-β stimulation. In addition, the core fucosylation of TGF-β receptor complexes decreased after FUT8 knockdown. Moreover, the expression of E2F4, a transcription factor upregulated upon TGF-β stimulation, was shown to directly regulate the expression of FUT8 via a TGF-β-induced non-Smad signalling pathway. Our results elucidated a new mechanism facilitated by E2F4-FUT8-mediated receptor core fucosylation that promotes TGF-β signalling and EMT, ultimately driving the invasion and metastasis of GBM cells.
Ageing and age-related diseases are the result of complex biological processes that progressively cause deterioration of cellular and tissue function. Among the key hallmarks of ageing are epigenetic alterations and geno...Ageing and age-related diseases are the result of complex biological processes that progressively cause deterioration of cellular and tissue function. Among the key hallmarks of ageing are epigenetic alterations and genomic instability, both of which are closely interconnected and significantly contribute to the ageing process. The epigenome, encompassing both DNA and histone modifications, regulates gene expression and maintains genomic integrity throughout life. With age, these regulatory systems become dysregulated, leading to genome-wide changes in chromatin structure, histone modifications and the reactivation of transposable elements (TEs). TEs, typically silenced in heterochromatic regions, become active in aged cells, contributing to genomic instability, mutagenesis, inflammation and metabolic disruption. Despite their significant implications, the role of TEs in the ageing process remains underexplored, and the interplay between epigenomic remodelling and TE activity remains poorly understood. In this review, we explore the molecular mechanisms underlying epigenetic alterations and TE reactivation during ageing, the impact of these changes on genomic stability and the potential therapeutic interventions targeting this interplay. By deciphering the role of epigenetic modifications and TE derepression in the ageing process, we aim to highlight novel avenues for anti-ageing and pro-longevity strategies.
The endoplasmic reticulum (ER) is an interconnected network of membrane-bound tubules and sheets stretching throughout the cytoplasm of all eukaryotic cells including plant cells. The ER is highly dynamic and undergoes c...The endoplasmic reticulum (ER) is an interconnected network of membrane-bound tubules and sheets stretching throughout the cytoplasm of all eukaryotic cells including plant cells. The ER is highly dynamic and undergoes constant remodelling. A properly formed ER is essential for cell growth, development and cellular responses to stresses. It is known that the dynamics of the cytoskeleton is linked to the formation and/or remodelling of a functional ER. Over the past 20 years, research has revealed that a set of ER localized ER-shaping proteins play crucial roles in building a functional ER. Recent research also indicates that maintaining a functional ER, in particular under stressful conditions, requires a proper turnover of the ER mediated by selective autophagy of the ER. In this review, we discuss the current understanding of functions of reticulons and atlastins, two classes of ER-shaping proteins in the formation of the ER in both animal and plant cells, with an emphasis on the plant system. We also discuss how the two classes of proteins may interplay to maintain a proper ER and how their actions may be regulated. Finally, we briefly mention how autophagy of the ER may be regulated during cell development and stress responses.