BMC Biology最新文献

Background: Rapid morphological change is emerging as a consequence of climate change in many systems. It is intuitive to hypothesize that temporal morphological trends are driven by the same selective pressures that have established well-known ecogeographic patterns over spatial environmental gradients (e.g., Bergman's and Allen's rules). However, mechanistic understanding of contemporary morphological shifts is lacking.

Results: We combine morphological data and whole genome sequencing from a four-decade dataset in the migratory bird hermit thrush (Catharus guttatus) to test whether morphological shifts over time are accompanied by genetic change. Using genome-wide association, we identify alleles associated with body size, bill length, and wing length. Shifts in morphology and concordant shifts in morphology-associated alleles over time would support a genetic basis for the observed changes in morphology over recent decades, potentially an adaptive response to climate change. In our data, bill size decreases were paralleled by genetic shifts in bill size-associated alleles. On the other hand, alleles associated with body size showed no shift in frequency over time.

Conclusions: Together, our results show mixed support for evolutionary explanations of morphological response to climate change. Temporal shifts in alleles associated with bill size support the hypothesis that selection is driving temporal morphological trends. The lack of evidence for genetic shifts in body size alleles could be explained by a large role of plasticity or technical limitations associated with the likely polygenic architecture of body size, or both. Disentangling the mechanisms responsible for observed morphological response to changing environments will be vital for predicting future organismal and population responses to climate change.

Background: Alternative food sources are crucial for the survival and reproduction of moths during nectar scarcity. Noctuid moths make a better use of fermented food sources than moths from other families, while the underlying molecular and genetic basis remain unexplored. As the fermentation progresses, yeasts lysis and the accumulation of metabolic byproducts alter the composition and the volatile release of the sugary substrates. However, it is unclear whether and how this would affect the feeding preference of moths.

Results: Here, we identified eight compounds abundant in the dynamic volatile profiles of several sugary substrates during yeast fermentation. We showed that the cotton bollworm moths were attracted to the fermented sugary substrates while being repelled when the sugary substrates were over-fermented. The attraction and aversion were respectively mediated by isoamyl alcohol and octanoic acid. We deorphanized the olfactory receptors detecting these two compounds and found that they belonged to two distinct gene families and were functionally conserved across four noctuid subfamilies; HarmOR52 orthologues responded to the attractive isoamyl alcohol and HarmIR75q.1 orthologues responded to the aversive octanoic acid.

Conclusions: Our findings suggest that this functional conservation is an olfactory adaptation that has allowed noctuid moths to extend their diet to fermented food sources.

Background: Traditionally, transformed cell line monolayers have been the standard model for studying epithelial barrier and transport function. Recently, intestinal organoids were proposed as superior in recapitulating the intestine. Typically, 3D organoids are digested and seeded as monolayers on gelatinous matrix pre-coated surfaces for anchorage. As this coat could potentially act as a diffusion barrier, we aimed to generate robust human duodenum-derived organoid monolayers that do not need a gelatinous matrix for anchorage to improve upon existing models to study epithelial transport and barrier function.

Results: We characterized these monolayers phenotypically regarding polarization, tight junction formation and cellular composition, and functionally regarding uptake of nutrients, ion transport and cytokine-induced barrier dysfunction. The organoid monolayers recapitulated the duodenum phenotypically as well as functionally regarding glucose and short-chain fatty acid uptake. Tumour necrosis factor-alpha induced paracellular transport of 4-kDa Dextran and transcytosis of 44-kDa horseradish peroxidase. Notably, forskolin-stimulated chloride secretion was consistently lower when organoid monolayers were seeded on a layer of basement membrane extract (BME).

Conclusions: BME-free organoid monolayers represent an improved model for studying transcytotic, paracellular but especially transcellular transport. As BME is extracted from mice, our model furthers efforts to make organoid culture more animal-free.

Background: Intermuscular bones (IBs) are segmental intramembranous ossifications located within myosepta. They share similarities with tendon ossification, a form of heterotopic ossification (HO). The mechanisms underlying IB formation remain incompletely understood.

Results: In this study, we systematically analyzed transcriptome data across multiple tissues, species, time points, and resolutions in teleosts. First, we identified IB-specific expression genes using the tau index method. Through cross-species comparisons of the tendon development process, we discovered that candidate genes were primarily enriched in extracellular matrix organization, ossification, regulation of angiogenesis, and other related processes. We also revealed that some of these candidate genes are abnormally expressed in runx2b^-/- zebrafish, which lack IBs. To clarify the trajectory of cell differentiation during IB formation, we demonstrated that myoseptal stem cells differentiate into osteoblasts, fibroblasts, and tenocytes in runx2b^+/+ zebrafish. However, in runx2b^-/- zebrafish, the differentiation of myoseptal stem cell into osteoblast was inhibited, while differentiation into clec3bb + tenocyte and fibroblast was enhanced. Additionally, runx2b deficiency led to the upregulation of clec3bb expression in the clec3bb + tenocyte cluster. Notably, a compensatory effect was observed in cell differentiation and gene expression in runx2b^-/- zebrafish, suggesting that runx2b and the candidate genes, such as clec3bb, were involved in the gene network of IB development.

Conclusions: The results elucidate cell differentiation process during tendon ossification in teleosts and identify the key factor clec3bb involved in this process. These findings provide a foundation for understanding tendon ossification in teleosts and for further research on tendon ossification in mammals.

Background: Global climate change significantly impacts ecosystems, particularly through temperature fluctuations that affect insect physiology and behavior. As poikilotherms, insect pests such as the globally devastating diamondback moth (DBM), Plutella xylostella, are especially vulnerable to rising temperatures and extreme heat events, necessitating effective adaptive mechanisms.

Results: Here we demonstrate the roles of zinc finger proteins (ZFPs) in mediating thermal adaptability in DBM. We utilized a comprehensive approach involving cloning and bioinformatics analysis of three ZFPs, PxZNF568, PxZNF93, and PxZNF266, measurement of their expression levels in hot-evolved and control strains, and assessment of catalase enzymatic activity and total antioxidant capacity. We also employed CRISPR/Cas9 technology to create five stable homozygous knockout strains to elucidate ZFP functions in high-temperature tolerance. Survival rates under high-temperature stress and the critical thermal maxima (CTMax) of the knockout strains were significantly lower than the wild-type strain, and exhibited marked decreases in antioxidant capacity.

Conclusion: Findings reveal the importance of ZFPs in thermal adaptability of DBM, contributing critical insights for future pest management strategies in the context of a warming climate and laying the foundation for further exploration of ZFP functionality in agricultural pest control.

Background: Age-related kidney impairment, characterized by tubular epithelial cell senescence and renal fibrosis, poses a significant global public health threat. Although N6-methyladenosine (m6A) methylation is implicated in various pathological processes, its regulatory mechanism in kidney aging remains unclear.

Methods: An m6A-mRNA epitranscriptomic microarray was performed to identify genes with abnormal m6A modifications in aged human kidney tissues. Histological, immunohistochemical, and immunofluorescent staining, western blot, and RT-qPCR were employed to examine the biological functions of targeted genes and m6A methyltransferases both in vivo and in vitro. RNA immunoprecipitation, chromatin immunoprecipitation, ribosomal immunoprecipitation, and luciferase reporter assays were used to investigate the specific interactions between m6A methyltransferases, targeted genes, and their downstream signals.

Results: Significantly lower m6A modification levels were observed in aged human kidney tissues. GLIS1, identified as a "metabolic remodeling factor," showed significantly reduced protein levels with abnormal m6A modifications. The downregulation of GLIS1 induced cell senescence and renal fibrosis by shifting metabolic remodeling from fatty acid oxidation (FAO) to glycolysis. Additionally, the methylated GLIS1 mRNA was regulated by the abnormal expression of METTL3 and YTHDF1. Silencing METTL3/YTHDF1 weakened the translation of GLIS1 and disrupted the balance between FAO and glycolysis.

Conclusions: Our findings suggest that the m6A modification of GLIS1, activated by METTL3 and reduced in a YTHDF1-dependent manner, leads to kidney aging by regulating the metabolic shift from FAO to glycolysis. This mechanism provides a promising therapeutic target for kidney aging.

Background: Molecular interactions between proteins and their ligands are important for drug design. A pharmacophore consists of favorable molecular interactions in a protein binding site and can be utilized for virtual screening. Pharmacophores are easiest to identify from co-crystal structures of a bound protein-ligand complex. However, designing a pharmacophore in the absence of a ligand is a much harder task.

Results: In this work, we develop a deep learning method that can identify pharmacophores in the absence of a ligand. Specifically, we train a CNN model to identify potential favorable interactions in the binding site, and develop a deep geometric Q-learning algorithm that attempts to select an optimal subset of these interaction points to form a pharmacophore. With this algorithm, we show better prospective virtual screening performance, in terms of F1 scores, on the DUD-E dataset than random selection of ligand-identified features from co-crystal structures. We also conduct experiments on the LIT-PCBA dataset and show that it provides efficient solutions for identifying active molecules. Finally, we test our method by screening the COVID moonshot dataset and show that it would be effective in identifying prospective lead molecules even in the absence of fragment screening experiments.

Conclusions: PharmRL addresses the need for automated methods in pharmacophore design, particularly in cases where a cognate ligand is unavailable. Experimental results demonstrate that PharmRL generates functional pharmacophores. Additionally, we provide a Google Colab notebook to facilitate the use of this method.

Representative models of intestinal diseases are transforming our knowledge of the molecular mechanisms of disease, facilitating effective drug screening and avenues for personalised medicine. Despite the emergence of 3D in vitro intestinal organoid culture systems that replicate the genetic and functional characteristics of the epithelial tissue of origin, there are still challenges in reproducing the human physiological tissue environment in a format that enables functional readouts. Here, we describe the latest platforms engineered to investigate environmental tissue impacts, host-microbe interactions and enable drug discovery. This highlights the potential to revolutionise knowledge on the impact of intestinal infection and inflammation and enable personalised disease modelling and clinical translation.

Background: The order Rodentia is the largest group of mammals. Diversification of vocal communication has contributed to rodent radiation and allowed them to occupy diverse habitats and adopt different social systems. The mechanism by which efficient vocal sounds, which carry over surprisingly large distances, are generated is incompletely understood. Here we focused on the development and function of rhythmic mouth movements and laryngeal sound production. We studied spontaneously vocalizing California mice (Peromyscus californicus) through video and sound recordings. Mouth gape was estimated from video images and vocal characteristics were measured in synchronized sound recordings.

Results: California mice coordinated their mouth movements with laryngeal sound production but differently in two call types. In high-frequency whistles ("USV syllables"), mouth movements were present on postnatal day 1 but were reduced within the first 2 weeks of life. Mouth movements were prominently present during sustained vocalizations ("SV syllables"), and movements became more and more adjusted to syllable beginning and end. Maximum mouth gape was correlated with sound intensity and fundamental frequency of SV syllables. The effect on sound intensity was the strongest during postnatal development and most predictable when the mouth was closed by temporarily immobilizing the mandible in an elevated position.

Conclusions: This study demonstrates that rhythmic orofacial behavior not only plays a critical role in determining acoustic features of the vocal behavior of California mice but also shows remarkable adjustments during early development.

Background: The Antarctic krill Euphausia superba is a keystone species in the Southern Ocean ecosystem. This crustacean has an ancestral clock whose main components have been identified and characterized in the past few years. However, the second feedback loop, modulating clock gene expression through two transcription factors, VRI and PDP1, has yet to be described. The presence of this second regulatory mechanism is suggested by the identification of its negative component, vrille, at the transcriptional level.

Results: Here, we describe the second feedback loop of krill by identifying the positive component, pdp1, and functionally characterizing both pdp1 and vrille. Starting from the online transcriptome database KrillDB², we identified and cloned three putative pdp1 sequences which were subsequently analyzed for tissue expression and functional activity using luciferase assays, individually and in combination with two vrille isoforms. Among the pdp1 isoforms, Espdp1_3 displayed higher expression levels in relevant circadian districts than the other two. Furthermore, EsPDP1_3 and EsVRI_2 exhibited the expected positive and negative regulation of the V/P-box in our in vitro system. Finally, Espdp1_3 and Esvrille also showed rhythmic expression in light-dark cycles, supporting their involvement in the regulation of the main circadian clock of the Antarctic krill.

Conclusions: This study expands our knowledge about the molecular architecture of the Antarctic krill circadian clock by defining the components that take part in the modulation of clock expression, establishing a second feedback loop.