Isolation and characterization of novel 3,3'-iminodipropionitrile biodegrading Paracoccus communis, from an industrial wastewater treatment bioreactor.

Until now, bacteria able to degrade, 3,3'-iminodipropionitrile (IDPN), a neurotoxin that destroys vestibular hair cells, causing ototoxicity, culminating in irreversible movement disorders, had never been isolated. The aim of this study was to isolate a novel IDPN-biodegrading microorganism and characterize its metabolic pathway. Enrichment was performed by inoculating activated sludge from a wastewater treatment bioreactor that treated IDPN-contaminated wastewater in M9 salt medium, with IDPN as the sole carbon source. A bacterial strain with a spherical morphology that could grow at high concentrations was isolated on a solid medium. Growth of the isolated strain followed the Monod kinetic model. Based on the 16S rRNA gene, the isolate was Paracoccus communis. Whole-genome sequencing revealed that the isolated P. communis possessed the expected full metabolic pathway for IDPN biodegradation. Transcriptome analyses confirmed the overexpression of the gene encoding hydantoinase/oxoprolinase during the exponential growth phase under IDPN-fed conditions, suggesting that the enzyme involved in cleaving the imine bond of IDPN may promote IDPN biodegradation. Additionally, the newly discovered P. communis isolate seems to metabolize IDPN through cleavage of the imine bond in IDPN via nitrilase, nitrile hydratase, and amidase reactions. Overall, this study lays the foundation for the application of IDPN-metabolizing bacteria in the remediation of IDPN-contaminated environments.

When and why PM2.5 is high in Seoul, South Korea: Interpreting long-term (2015-2021) ground observations using machine learning and a chemical transport model.

Seoul has high PM2.5 concentrations and has not attained the national annual average standard so far. To understand the reasons, we analyzed long-term (2015-2021) hourly observations of aerosols (PM2.5, NO3-, NH4+, SO42-, OC, and EC) and gases (CO, NO2, and SO2) from Seoul and Baekryeong Island, a background site in the upwind region of Seoul. We applied the weather normalization method for meteorological conditions and a 3-dimensional chemical transport model, GEOS-Chem, to identify the effect of policy implementation and aerosol formation mechanisms. The monthly mean PM2.5 ranges between about 20 µg m-3 (warm season) and about 40 µg m-3 (cold season) at both sites, but the annual decreasing rates were larger at Seoul than at Baengnyeong (-0.7 µg m-3 a-1 vs. -1.8 µg m-3 a-1) demonstrating the effectiveness of the local air quality policies including the Special Act on Air Quality in the Seoul Metropolitan Area (SAAQ-SMA) and the seasonal control measures. The weather-normalized monthly mean data shows the highest PM2.5 concentration in March and the lowest concentration in August throughout the 7 years with NO3- accounting for about 40 % of the difference between the two months at both sites. Taking together with the GEOS-Chem model results, which reproduced the elevated NO3- in March, we concluded the elevated atmospheric oxidant level increases in HNO3 (which is not available from the observation) and the still low temperatures in March promote rapid production of NO3-. We used Ox (≡ O3 + NO2) from the observation and OH from the GEOS-Chem as a proxy for the atmospheric oxidant level which can be a source of uncertainty. Thus, direct observations of OH and HNO3 are needed to provide convincing evidence. This study shows that reducing HNO3 levels through atmospheric oxidant level control in the cold season can be effective in PM2.5 mitigation in Seoul.

- Invited Review - Hydrogen production and hydrogen utilization in the rumen: key to mitigating enteric methane production.

Molecular hydrogen (H2) and formate (HCOO-) are metabolic end products of many primary fermenters in the rumen ecosystem. Both play a vital role in fermentation where they are electron sinks for individual microbes in an anaerobic environment that lacks external electron acceptors. If H2 and/or formate accumulate within the rumen, the ability of primary fermenters to regenerate electron carriers may be inhibited and microbial metabolism and growth disrupted. Consequently, H2- and/or formate-consuming microbes such as methanogens and possibly homoacetogens play a key role in maintaining the metabolic efficiency of primary fermenters. There is increasing interest in identifying approaches to manipulate the rumen ecosystem for the benefit of the host and the environment. As H2 and formate are important mediators of interspecies interactions, an understanding of their production and utilization could be a significant starting point for the development of successful interventions aimed at redirecting electron flow and reducing methane emissions. We conclude by discussing in brief ruminant methane mitigation approaches as a model to help understand the fate of H2 and formate in the rumen ecosystem.

Genomic insight and physiological characterization of thermoacidophilic Alicyclobacillus isolated from Yellowstone National Park.

Introduction: Alicyclobacillus has been isolated from extreme environments such as hot springs, volcanoes, as well as pasteurized acidic beverages, because it can tolerate extreme temperatures and acidity. In our previous study, Alicyclobacillus was isolated during the enrichment of methane oxidizing bacteria from Yellowstone Hot Spring samples. Methods: Physiological characterization and genomic exploration of two new Alicyclobacillus isolates, AL01A and AL05G, are the main focus of this study to identify their potential relationships with a thermoacidophilic methanotroph (Methylacidiphilum) isolated from the same hot spring sediments. Results and discussion: In the present study, both Alicyclobacillus isolates showed optimal growth at pH 3.5 and 55°C, and contain ω-alicyclic fatty acids as a major lipid (ca. 60%) in the bacterial membrane. Genomic analysis of these strains revealed specific genes and pathways that the methanotroph genome does not have in the intermediary carbon metabolism pathway such as serC (phosphoserine aminotransferase), comA (phosphosulfolactate synthase), and DAK (glycerone kinase). Both Alicyclobacillus strains were also found to contain transporter systems for extracellular sulfate (ABC transporter), suggesting that they could play an important role in sulfur metabolism in this extreme environment. Genomic analysis of vitamin metabolism revealed Alicyclobacillus and Methylacidiphilum are able to complement each other's nutritional deficiencies, resulting in a mutually beneficial relationship, especially in vitamin B1(thiamin), B3 (niacin), and B7 (biotin) metabolism. These findings provide insights into the role of Alicyclobacillus isolates in geothermal environments and their unique metabolic adaptations to these environments.

Effects of dosing non-toxigenic Clostridia on the bacterial populations and immunological responses in the intestinal tract of lactating dairy cows.

Understanding the effects of dosing non-toxigenic Clostridia to cows is rare and has received little attention so far. In the present study, a total of eight lactating dairy cows were divided in two groups: control (n = 4) or Clostridia challenged (oral supplementation of five diverse strains of Paraclostridium bifermentans, n = 4). Bacterial communities were analyzed by qPCR and next-generation sequencing (NGS) in the buccal mucosa as well as digesta and mucosal samples of the gastrointestinal (GI) tract from rumen to rectum (10 compartments), as well as fecal samples. Transcriptomic analysis of barrier and immune-related gene expression was performed on rumen, jejunum, and liver samples. We observed increased microbial populations with the Clostridial challenge in the buccal tissues and the proximal GI tract (forestomach), correlating with Clostridial loads in the feed. Otherwise, there were no significant differences in microbial populations (p > 0.05) throughout the distal part of the GI tract. The NGS approach, however, revealed that the Clostridial challenge changed the relative abundance of gut and fecal microbiota. In particular, in the challenge group, no Bifidobacterium was observed in the mucosa-associated microbiota and abundance of Pseudomonadota increased in the feces. These results indicated potential adverse effects of Clostridia to cow health. In general, immune responses to the Clostridial challenge were weak. However, transcriptional analysis revealed the down-regulation of junction adhesion molecule encoding gene (-1.44 of log2 fold-change), which might impact intestinal permeability.

The introduction to air pollution on pregnancy outcome (APPO) study: a multicenter cohort study.

OBJECTIVE: The air pollution on pregnancy outcome (APPO) study is a prospective hospital-based cohort study designed to investigate the maternal and fetal effects of a particulate matter with an aerodynamic below 10 µm (PM10) and PM2.5 (below 2.5 µm) exposure. This study aims to analyze a relationship between particulate matter and adverse pregnancy outcomes and to find related biomarkers and develop management guidelines. METHODS: About 1,200 pregnant women are recruited for 3 years (from January 2021 to December 2023) from seven university hospitals to investigate the effects of particulate matter on pregnancy complications and adverse pregnancy outcomes. We collect biological samples by 5 mL of maternal venous blood and 15 mL of urine in each trimester of pregnancy, and 5 mL of umbilical cord blood and 2×2×2 cm of placental tissue are collected after delivery. In addition, by applying PM10 and PM2.5 concentration values and time-activity patterns from the time weighted average model, the individual predicted exposure of air pollution for the pregnant women are obtained. RESULTS: The average exposure of PM10 and PM2.5 of the participants in the entire period of pregnancy, was exceeded the World Health Organization air quality guidelines (an annual level, PM10 >15 µg/m3, PM2.5 >5 µg/m3). Moreover, it was revealed that the PM concentration was increasing toward the 3rd trimester of pregnancy. CONCLUSION: The APPO study will be able to identify the degree of exposure to air pollution in pregnant women and use it as basic data for estimating individual exposure to particulate matter. And the results of the APPO study will facilitate in the development of health management for pregnant women against air pollution.

Mitochondrial fragmentation and donut formation enhance mitochondrial secretion to promote osteogenesis.

Mitochondrial components have been abundantly detected in bone matrix, implying that they are somehow transported extracellularly to regulate osteogenesis. Here, we demonstrate that mitochondria and mitochondrial-derived vesicles (MDVs) are secreted from mature osteoblasts to promote differentiation of osteoprogenitors. We show that osteogenic induction stimulates mitochondrial fragmentation, donut formation, and secretion of mitochondria through CD38/cADPR signaling. Enhancing mitochondrial fission and donut formation through Opa1 knockdown or Fis1 overexpression increases mitochondrial secretion and accelerates osteogenesis. We also show that mitochondrial fusion promoter M1, which induces Opa1 expression, impedes osteogenesis, whereas osteoblast-specific Opa1 deletion increases bone mass. We further demonstrate that secreted mitochondria and MDVs enhance bone regeneration in vivo. Our findings suggest that mitochondrial morphology in mature osteoblasts is adapted for extracellular secretion, and secreted mitochondria and MDVs are critical promoters of osteogenesis.

Biofilm characteristics for providing resilient denitrification in a hydrogen-based membrane biofilm reactor.

In a hydrogen-based membrane biofilm reactor (H2-MBfR), the biofilm thickness is considered to be one of the most important factors for denitrification. Thick biofilms in MBfRs are known for low removal fluxes owing to their resistance to substrate transport. In this study, the H2-MBfR was operated under various loading rates of oxyanions, such as NO3-N, SO4-S, and ClO4- at an H2 flux of 1.06 e- eq/m2-d. The experiment was initiated with NO3-N, SO4-S, and ClO4- loadings of 0.464, 0.026, and 0.211 e- eq/m2-d, respectively, at 20 °C. Under the most stressful conditions, the loading rates increased simultaneously to 1.911, 0.869, and 0.108 e- eq/m2-d, respectively, at 10 °C. We observed improved performance in significantly thicker biofilms (approximately 2.7 cm) compared to previous studies using a denitrifying H2-MBfR for 120 days. Shock oxyanion loadings led to a decrease in total nitrogen (TN) removal by 20 to 30%, but TN removal returned to 100% within a few days. Similarly, complete denitrification was observed, even at 10 °C. The protective function and microbial diversity of the thick biofilm may allow stable denitrification despite stress-imposing conditions. In the microbial community analysis, heterotrophs were dominant and acetogens accounted for 11% of the biofilm. Metagenomic results showed a high abundance of functional genes involved in organic carbon metabolism and homoacetogenesis. Owing to the presence of organic compounds produced by acetogens and autotrophs, heterotrophic denitrification may occur simultaneously with autotrophic denitrification. As a result, the total removal flux of oxyanions (1.84 e- eq/m2-d) far exceeded the H2 flux (1.06 e- eq/m2-d). Thus, the large accumulation of biofilms could contribute to good resilience and enhanced removal fluxes.

Different contribution of exoelectrogens in methanogenesis via direct interspecies electron transfer (DIET) by the different substrate in continuous anaerobic bioreactor.

Direct interspecies electron transfer (DIET) is a syntrophic mechanism for electron transfer between exo- and endoelectrogens. Previous studies have demonstrated that methanogenesis performance was significantly improved via the DIET mechanism through conductive materials (CMs) under batch conditions with a single substrate, while that under continuous condition is still under investigation. To investigate how the DIET via CM on methanogenesis performance was changed in response to the different substrates (acetate versus glucose)-fed in continuous anaerobic bioreactors, continuous bioreactors were operated by cross-feeding with acetate and glucose. Acetate-fed conditions showed 0.40 day shorten lagtime, 1.88- and 1.22-folds higher methane production rate, and ultimate methane production than glucose-fed conditions, respectively. Burkholderiaceae- and Anaerolineaceae-related exo-electrogenic populations were enriched with low abundance of Geobacter species in batch reactors. Furthermore, influent substrates affected the distribution of the enriched populations. Taken together, the results suggested that different syntrophic associations contributed methane production by DIET in continuous bioreactors.

Current biotechnologies on depolymerization of polyethylene terephthalate (PET) and repolymerization of reclaimed monomers from PET for bio-upcycling: A critical review.

The production of polyethylene terephthalate (PET) has drastically increased in the past half-century, reaching 30 million tons every year. The accumulation of this recalcitrant waste now threatens diverse ecosystems. Despite efforts to recycle PET wastes, its rate of recycling remains limited, as the current PET downcycling is mostly unremunerative. To address this problem, PET bio-upcycling, which integrates microbial depolymerization of PET followed by repolymerization of PET-derived monomers into value-added products, has been suggested. This article critically reviews current understanding of microbial PET hydrolysis, the metabolic mechanisms involved in PET degradation, PET hydrolases, and their genetic improvement. Furthermore, this review includes the use of meta-omics approaches to search PET-degrading microbiomes, microbes, and putative hydrolases. The current development of biosynthetic technologies to convert PET-derived materials into value-added products is also comprehensively discussed. The integration of various depolymerization and repolymerization biotechnologies enhances the prospects of a circular economy using waste PET.

Magnetic Field Alignment and Optical Anisotropy of MoS2 Nanosheets Dispersed in a Liquid Crystal Polymer.

Molybdenum disulfide (MoS2) nanosheets exhibit anisotropic optical and electronic properties, stemming from their shape and electronic structure. Unveiling this anisotropy for study and usage in materials and devices requires the ability to control the orientation of dispersed nanosheets, but to date this has proved a challenging proposition. Here, we demonstrate magnetic field driven alignment of MoS2 nanosheets in a liquid crystal (LC) polymer and unveil the optical properties of the resulting anisotropic assembly. Nanosheet optical anisotropy is observed spectroscopically by Raman and direction-dependent photoluminescence (PL) measurements. Resulting data indicate significantly lower PL emission due to optical excitation with electric field oscillation out of plane, parallel to the MoS2 c-axis, than that associated with perpendicular excitation, with the dichroic ratio Iperp/Ipar = 3. The approach developed here provides a useful route to elucidate anisotropic optical properties of MoS2 nanosheets and to utilize such properties in new materials and devices.

Enrichment of Ca. Jettenia in sequencing batch reactors operated with low nitrogen loading rate and high influent nitrogen concentration.

To apply the anammox processes into the mainstream of domestic wastewater treatment plants, two laboratory-scale sequence batch reactors have been developed and used with two different activated sludges seeded in each sequence batch reactors with gradually increases influent total nitrogen concentrations under low nitrogen loading rates. During 320 days of operation, both sequence batch reactors showed high specific anammox activity (0.68 - 0.75 kgN kg-1VSS d-1) and a nitrogen removal efficiency of 97.50%. To monitor changes in microbial community dynamics during enrichment, high-throughput sequencing analysis was performed. Members taxonomically affiliated with Candidatus Jettenia were markedly enriched and predominant in both sequence batch reactors in response to the increasing influent total nitrogen concentrations. These results suggest that Candidatus Jettenia might be a prominent anammox genus under low nitrogen loading rate with high total nitrogen concentration conditions and could be suitably applied to the mainstream process of domestic wastewater treatment systems.

Genome Sequence of a Thermoacidophilic Methanotroph Belonging to the Verrucomicrobiota Phylum from Geothermal Hot Springs in Yellowstone National Park: A Metagenomic Assembly and Reconstruction.

Verrucomicrobiotal methanotrophs are thermoacidophilic methane oxidizers that have been isolated from volcanic and geothermal regions of the world. We used a metagenomic approach that entailed obtaining the whole genome sequence of a verrucomicrobiotal methanotroph from a microbial consortium enriched from samples obtained from Nymph Lake (89.9 °C, pH 2.73) in Yellowstone National Park in the USA. To identify and reconstruct the verrucomicrobiotal genome from Illumina NovaSeq 6000 sequencing data, we constructed a bioinformatic pipeline with various combinations of de novo assembly, alignment, and binning algorithms. Based on the marker gene (pmoA), we identified and assembled the Candidatus Methylacidiphilum sp. YNP IV genome (2.47 Mbp, 2392 ORF, and 41.26% GC content). In a comparison of average nucleotide identity between Ca. Methylacidiphilum sp. YNP IV and Ca. Methylacidiphilum fumariolicum SolV, its closest 16S rRNA gene sequence relative, is lower than 95%, suggesting that Ca. Methylacidiphilum sp. YNP IV can be regarded as a different species. The Ca. Methylacidiphilum sp. YNP IV genome assembly showed most of the key genes for methane metabolism, the CBB pathway for CO2 fixation, nitrogen fixation and assimilation, hydrogenases, and rare earth elements transporter, as well as defense mechanisms. The assembly and reconstruction of a thermoacidophilic methanotroph belonging to the Verrucomicrobiota phylum from a geothermal environment adds further evidence and knowledge concerning the diversity of biological methane oxidation and on the adaptation of this geochemically relevant reaction in extreme environments.

Fast Photoswitchable Order-Disorder Transitions in Liquid-Crystalline Block Co-oligomers.

Optically driven ordering transitions are rarely observed in macromolecular systems, often because of kinetic limitations. Here, we report a series of block co-oligomers (BCOs) that rapidly order and disorder at room temperature in response to optical illumination, and the absence thereof. The system is a triblock where rigid azobenzene (Azo) mesogens are attached to each end of a flexible siloxane chain. UV-induced trans-to-cis Azo isomerization, and vice versa in the absence of UV light, drive disordering and ordering of lamellar superstructures and smectic mesophases, as manifested by liquefaction and solidification of the material, respectively. The impacts of chemical structure on BCO self-assembly and photoswitching kinetics are explored by in situ microscopy and X-ray measurements for different mesogen end groups (NO2 or CN), and different carbon chain lengths (0C or 12C) between the siloxane and the mesogen. The presence of the 12C spacer leads to hierarchical ordering with smectic layers of mesogens existing alongside larger length-scale lamellae, versus only smectic ordering without the spacer. These hierarchically ordered BCOs display highly persistent lamellar sheets that contrast with the tortuous, low-persistence "fingerprint"-type structures seen in conventional block copolymers. The reordering kinetics upon removal of UV illumination are extremely rapid (<5 s). This fast response is due to the electron-withdrawing NO2 and CN, which facilitate cis-to-trans isomerization via thermal relaxation at room temperature without additional stimuli. This work elucidates structure-property relationships in photoswitching BCOs and advances the possibility of developing systems in which ordered nanostructures can be easily optically written and erased.

Current understanding and perspectives in anaerobic digestion based on genome-resolved metagenomic approaches.

Anaerobic digestion (AD) is a technique that can be used to treat high concentrations of various organic wastes using a consortium of functionally diverse microorganisms under anaerobic conditions. Methane gas, a beneficial by-product of the AD process, is a renewable energy source that can replace fossil fuels following purification. However, detailed functional roles and metabolic interactions between microbial populations involved in organic waste removal and methanogenesis are yet to be known. Recent metagenomic approaches based on advanced high-throughput sequencing techniques have enabled the exploration of holistic microbial taxonomy and functionality of complex microbial populations involved in the AD process. Gene-centric and genome-centric analyses based on metagenome-assembled genomes are a platform that can be used to study the composition of microbial communities and their roles during AD. This review looks at how these up-to-date metagenomic analyses can be applied to promote our understanding and improved the development of the AD process.

Stable cross-linked lyotropic gyroid mesophases from single-head/single-tail cross-linkable monomers.

A single-head/single-tail surfactant with a polymerizable group at each end is presented as a new simplified motif for intrinsically cross-linkable, gyroid-phase lyotropic mesogens. The resulting nanoporous polymer networks exhibit excellent structural stability in various solvents and are capable of molecular size discrimination.

Microbiome degrading linear alkylbenzene sulfonate in activated sludge.

As the most widely used anionic surfactant, linear alkylbenzene sulfonate (LAS) requires biological alkane degradation when it is treated using an activated sludge (AS) process in a wastewater treatment plant because of its structural carboxylic unavailability. As consumption of LAS is gradually increasing, LAS loading into the WWTP is accordingly increasing. However, fewer studies have examined the involvement of the AS microbial community in the LAS degradation. In this study, metagenomic approaches were used to define microbiomes involved in LAS degradation in AS, with a particular focus on ω-hydroxylation. The abundance and diversity of alkane-degrading genes were investigated, and these genes were integrated with reconstructed metagenome-assembled genomes (MAGs). Additionally, the association of functional genes and MAGs with respect to LAS degradation was investigated. The results showed that alkB and cytochrome P450 genes were only shared within specific MAGs. Unique sets of genes with diverse abundances were detected in each sample. The MAGs with the alkB and cytochrome P450 genes were strongly associated with the other MAGs and involved in positive commensal interactions. The findings provided significant insights into how the AS microbiomes, which have continuously treated anionic surfactants for decades, potentially metabolize LAS and interact with commensal bacteria.

Rapid Fabrication by Lyotropic Self-Assembly of Thin Nanofiltration Membranes with Uniform 1 Nanometer Pores.

Nanostructured materials with precisely defined and water-bicontinuous 1-nm-scale pores are highly sought after as advanced materials for next-generation nanofiltration membranes. While several self-assembled systems appear to satisfy this need, straightforward fabrication of such materials as submicron films with high-fidelity retention of their ordered nanostructure represents a nontrivial challenge. We report the development of a lyotropic liquid crystal mesophase that addresses the aforementioned issue. Films as thin as ∼200 nm are prepared on conventional support membranes using solution-based methods. Within these films, the system is composed of a hexagonally ordered array of ∼3 nm diameter cylinders of cross-linked polymer, embedded in an aqueous medium. The cylinders are uniformly oriented in the plane of the film, providing a transport-limiting dimension of ∼1 nm, associated with the space between the outer surfaces of nearest-neighbor cylinders. These membranes exhibit molecular weight cutoffs of ∼300 Da for organic solutes and are effective in rejecting dissolved salts, and in particular, divalent species, while exhibiting water permeabilities that rival or exceed current state-of-the-art commercial nanofiltration membranes. These materials have the ability to address a broad range of nanofiltration applications, while structure-property considerations suggest several avenues for potential performance improvements.

GDF11 promotes osteogenesis as opposed to MSTN, and follistatin, a MSTN/GDF11 inhibitor, increases muscle mass but weakens bone.

Growth and differentiation factor 11 (GDF11) and myostatin (MSTN) are closely related transforming growth factor ß (TGF-ß) family members, but their biological functions are quite distinct. While MSTN has been widely shown to inhibit muscle growth, GDF11 regulates skeletal patterning and organ development during embryogenesis. Postnatal functions of GDF11, however, remain less clear and controversial. Due to the perinatal lethality of Gdf11 null mice, previous studies used recombinant GDF11 protein to prove its postnatal function. However, recombinant GDF11 and MSTN proteins share nearly identical biochemical properties, and most GDF11-binding molecules have also been shown to bind MSTN, generating the possibility that the effects mediated by recombinant GDF11 protein actually reproduce the endogenous functions of MSTN. To clarify the endogenous functions of GDF11, here, we focus on genetic studies and show that Gdf11 null mice, despite significantly down-regulating Mstn expression, exhibit reduced bone mass through impaired osteoblast (OB) and chondrocyte (CH) maturations and increased osteoclastogenesis, while the opposite is observed in Mstn null mice that display enhanced bone mass. Mechanistically, Mstn deletion up-regulates Gdf11 expression, which activates bone morphogenetic protein (BMP) signaling pathway to enhance osteogenesis. Also, mice overexpressing follistatin (FST), a MSTN/GDF11 inhibitor, exhibit increased muscle mass accompanied by bone fractures, unlike Mstn null mice that display increased muscle mass without fractures, indicating that inhibition of GDF11 impairs bone strength. Together, our findings suggest that GDF11 promotes osteogenesis in contrast to MSTN, and these opposing roles of GDF11 and MSTN must be considered to avoid the detrimental effect of GDF11 inhibition when developing MSTN/GDF11 inhibitors for therapeutic purposes.

Growth differentiation factor 11 locally controls anterior-posterior patterning of the axial skeleton.

Growth and differentiation factor 11 (GDF11) is a transforming growth factor ß family member that has been identified as the central player of anterior-posterior (A-P) axial skeletal patterning. Mice homozygous for Gdf11 deletion exhibit severe anterior homeotic transformations of the vertebrae and craniofacial defects. During early embryogenesis, Gdf11 is expressed predominantly in the primitive streak and tail bud regions, where new mesodermal cells arise. On the basis of this expression pattern of Gdf11 and the phenotype of Gdf11 mutant mice, it has been suggested that GDF11 acts to specify positional identity along the A-P axis either by local changes in levels of signaling as development proceeds or by acting as a morphogen. To further investigate the mechanism of action of GDF11 in the vertebral specification, we used a Cdx2-Cre transgene to generate mosaic mice in which Gdf11 expression is removed in posterior regions including the tail bud, but not in anterior regions. The skeletal analysis revealed that these mosaic mice display patterning defects limited to posterior regions where Gdf11 expression is deficient, whereas displaying normal skeletal phenotype in anterior regions where Gdf11 is normally expressed. Specifically, the mosaic mice exhibited seven true ribs, a pattern observed in wild-type (wt) mice (vs. 10 true ribs in Gdf11-/- mice), in the anterior axis and nine lumbar vertebrae, a pattern observed in Gdf11 null mice (vs. six lumbar vertebrae in wt mice), in the posterior axis. Our findings suggest that GDF11, rather than globally acting as a morphogen secreted from the tail bud, locally regulates axial vertebral patterning.