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Montagne, J.

Publications and source records attributed to Montagne, J..

10 recordsLinked to original sources

Coordinated development of the male and female reproductive systems in the cestode Hymenolepis microstoma

BackgroundCestodes have complex hermaphroditic reproductive systems that produce massive numbers of eggs. This reproductive output is made possible by the continuous production of serially repeated sets of reproductive systems (proglottids). However, their reproductive development remains poorly understood. ResultsWe characterized reproductive development in the model cestode Hymenolepis microstoma by analyzing markers of cell proliferation, meiosis, and differentiation along the series of proglottids. Reproductive development begins with the formation of a central genital primordium, from which the reproductive ducts and gonads differentiate. Development is proterandrous, and testicular development is prolonged. In contrast, female reproductive development occurs over a short interval and is characterized by the coordinated differentiation of the ovary and vitelline gland. Entry of oocytes into meiosis is almost synchronous, and paralleled by cell proliferation in the vitelline gland. Subsequent growth of arrested oocytes and differentiation of vitelline cells occur in parallel. Insemination coincides with the onset of ovarian meiosis, indicating a close temporal coordination between male and female reproductive development. Finally, we show that gametogenesis and insemination proceed in adult worms maintained in vitro. ConclusionsOur findings show the coordination of reproductive development in a self-fertile hermaphrodite, and provide an experimental system for studying reproductive development in cestodes.

Developmental Biology↗

A chromosome-level assembly and functional genomic resources for the model annelid Capitella teleta

BackgroundThe polychaete Capitella teleta is a commonly used annelid for studies in evolutionary developmental biology, comparative genomics, conservation, and ecotoxicology. Over a decade ago, it was the first polychaete to have its genome sequenced and assembled, contributing to pioneering studies that transformed our understanding of animal genomes and their evolution. However, this early resource is now outdated compared to current genome sequencing standards, limiting the use of modern functional genomic tools that could further our understanding of numerous biological processes. ResultsWe combine long-read and short-read sequencing with Hi-C chromatin conformation capture data to assemble the chromosome-level nuclear and mitochondrial genomes of the laboratory strain of C. teleta. This reference assembly more accurately reflects the expected genome size for this polychaete ([~]243.6 Mb) and contains a highly complete, evolutionarily conserved gene repertoire. Notably, the nuclear and mitochondrial genomes are heavily rearranged, indicating a decoupling between gene family repertoire and chromosomal evolution. The analyses of multi-omic datasets available for C. teleta, including developmental time courses of bulk and single-cell RNA-seq, ATAC-seq, and EM-seq, using the new reference assembly, resulted in a significant quality improvement, allowing us to identify new cell-type-specific gene markers and gain additional insights of biological relevance. Finally, we generated a publicly available genome browser that ensures all these resources are easily findable, accessible, interoperable, and reusable. ConclusionsOur study provides state-of-the-art genomic resources for the polychaete model C. teleta, addressing a pressing community need that will open new research opportunities in animal and genome evolution.

genomics↗

Drosophila Sociality Influences Immune Peptide Load and Apoptosis-induced Tumor Suppression independently of the Antitumor Peptide Defensin

Patients social environment might influence cancer outcome. This potentially happens in Drosophila, as we previously reported that the social context influences the growth of intestinal tumors. To uncover the underlying social-induced physiological mechanisms, we performed RNA-seq of isogenized beheaded tumorous Drosophila. Importantly, expression of several immune peptides varied according to the social-induced tumor growth effect. Furthermore, ectopic expression in tumors of the apoptotic-inhibitor p35 suppressed the social-induced effect. Next, we challenged the immune peptide Defensin, previously reported to suppress imaginal disc tumor growth through a cell-death/JNK-dependent network. Nonetheless, the social-induced tumor suppression was maintained upon Defensin overexpression or JNKK-knockdown in tumors, and in defensin mutants. Surprisingly, tumor growth was reduced in the latter, indicating that Defensin sustains the growth of these intestinal tumors. In summary, our study indicates that the social context affects the immune response and that a given immune peptide may have opposite effects depending on tumor type.

cancer biology↗

Immune landscape of tertiary lymphoid structures in hepatocellular carcinoma (HCC) treated with neoadjuvant immune checkpoint blockade

Neoadjuvant immunotherapy is thought to produce long-term remissions through induction of antitumor immune responses before removal of the primary tumor. Tertiary lymphoid structures (TLS), germinal center-like structures that can arise within tumors, may contribute to the establishment of immunological memory in this setting, but understanding of their role remains limited. Here, we investigated the contribution of TLS to antitumor immunity in hepatocellular carcinoma (HCC) treated with neoadjuvant immunotherapy. We found that neoadjuvant immunotherapy induced the formation of TLS, which were associated with superior pathologic response, improved relapse free survival, and expansion of the intratumoral T and B cell repertoire. While TLS in viable tumor displayed a highly active mature morphology, in areas of tumor regression we identified an involuted TLS morphology, which was characterized by dispersion of the B cell follicle and persistence of a T cell zone enriched for ongoing antigen presentation and T cell-mature dendritic cell interactions. Involuted TLS showed increased expression of T cell memory markers and expansion of CD8+ cytotoxic and tissue resident memory clonotypes. Collectively, these data reveal the circumstances of TLS dissolution and suggest a functional role for late-stage TLS as sites of T cell memory formation after elimination of viable tumor. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=128 SRC="FIGDIR/small/562104v2_fig0.gif" ALT="Figure 0"> View larger version (30K): org.highwire.dtl.DTLVardef@859572org.highwire.dtl.DTLVardef@169ed2org.highwire.dtl.DTLVardef@11510f0org.highwire.dtl.DTLVardef@b39a4_HPS_FORMAT_FIGEXP M_FIG C_FIG O_LSTHighlightsC_LSTO_LIIn patients with hepatocellular carcinoma (HCC), tertiary lymphoid structures (TLS) are induced by neoadjuvant immunotherapy and are associated with favorable clinical outcomes. C_LIO_LITLS within the same tumor demonstrate extensive sharing of expanded granzyme K and granzyme B-expressing CD8+ T effector memory clonotypes, but the B cell repertoires of individual TLS are almost wholly distinct, consistent with independent germinal center reactions. C_LIO_LIWithin areas of viable tumor, mature TLS are characterized by high expression of CD21 and CD23, BCL6+ germinal center B cells, and close interactions between DCLAMP+ mature dendritic cells and CXCR5-CXCR3+ CD4 T peripheral helper cells within a T cell zone adjacent to the B cell follicle. C_LIO_LIWithin areas of tumor regression, an involuted TLS morphology is identified that is notable for dissolution of the B cell germinal center, retention of the T cell zone, and increased T cell memory. C_LI

immunology↗

Segmentation in tapeworms as a modified form of flatworm posterior regeneration involving Wnt and Hedgehog signalling

Tapeworms are parasitic flatworms that lack key features conventionally used to define the head-tail axis in free-living organisms, resulting in long standing questions regarding the true orientation of their main body axis. As adults, most tapeworms also exhibit a segmented body which has been considered an adaptation unique to the group. Anteroposterior (AP) patterning in free-living flatworms is controlled by {beta}-catenin-dependent Wnt signalling and positional control genes are expressed by their musculature in highly regionalised domains. Here we investigate the expression of Wnt and Hedgehog components during the strobilar phase of the tapeworm life cycle during which larval tissues are lost and replaced through the continuous production of new tissues. Results reveal previously unidentified centres of signalling associated with their neuromuscular system and show that segments are marked by secondary, AP axes in agreement with the polarity of the primary body axis. Neuromuscular expression and communication between Wnt and Hedgehog signalling is consistent with embryonic and regenerative growth in planarians and is a common mechanism for establishing AP-polarised boundaries in a diverse range of segmented animals. Taken together our results suggest that segmentation in tapeworms represents a modified form of posterior regeneration, a common feature among flatworms.

developmental biology↗

Stem cell proliferation and differentiation during larval metamorphosis of the model tapeworm Hymenolepis microstoma

Tapeworm larvae cause important diseases in humans and domestic animals. During infection, the first larval stage undergoes a metamorphosis where tissues are formed de novo from a population of stem cells called germinative cells. This process is difficult to study for human pathogens, as these larvae are infectious and difficult to maintain in the laboratory. In this work, we analyzed cell proliferation and differentiation during larval metamorphosis in the model tapeworm Hymenolepis microstoma, by in vivo labelling of proliferating cells with the thymidine analogue 5-ethynyl-2'-deoxyuridine (EdU), tracing their differentiation with a suite of specific molecular markers for different cell types. Proliferating cells are very abundant and fast-cycling during early metamorphosis: the total number of cells duplicates every ten hours, and the length of G2 is only 75 minutes. New tegumental, muscle and nerve cells differentiate from this pool of proliferating germinative cells, and these processes are very fast, as differentiation markers for neurons and muscle cells appear within 24 hours after exiting the cell cycle, and fusion of new cells to the tegumental syncytium can be detected after only 4 hours. Tegumental and muscle cells appear from early stages of metamorphosis (24 to 48 hours post-infection); in contrast, most markers for differentiating neurons appear later, and the detection of synapsin and neuropeptides correlates with scolex retraction. Finally, we identified populations of proliferating cells that express conserved genes associated with neuronal progenitors and precursors, suggesting the existence of tissue-specific lineages among germinative cells. These results provide for the first time a comprehensive view of the development of new tissues during tapeworm larval metamorphosis, providing a framework for similar studies in human and veterinary pathogens.

developmental biology↗

A Fatty Acid Anabolic Pathway in Specialized-Cells Remotely Controls Oocyte Activation in Drosophila

Pheromone-mediated partner recognition is crucial for maintenance of animal species. Here, we discover a metabolic link between pheromone and gamete physiology. In female genital tract, oocyte maturation is arrested at a specific meiotic-phase. Release of this arrest, called oocyte-activation, is triggered by a species-dependent signal. We show in Drosophila melanogaster that oenocytes, which produce the fatty acids (FAs) used as precursors of cuticular hydrocarbons (CHCs), including pheromones, are also essential for oocyte activation. We identified a set of FA-anabolic enzymes required within oenocytes for the synthesis of a particular FA that is not a CHC precursor but controls oocyte activation. Our study thus reveals that two tightly linked FA-anabolic pathways act in parallel, one to produce sexual pheromones, the other to initiate embryonic development. Given that pheromone-deficient Drosophila melanogaster females are highly attractive for males irrespective of their species, this oenocyte function might have evolved to prevent hybrid development.

physiology↗

Differential Metabolic Sensitivity of mTORC1- and mTORC2-Dependent Overgrowth

The glycolytic/lipogenic axis promotes the synthesis of energetic molecules and building blocks necessary to support cell growth, although the absolute requirement of this metabolic axis must be deeply investigated. Here, we used Drosophila genetics and focus on the mTOR signaling network that controls cell growth and homeostasis. mTOR is present in two distinct complexes, mTORC1 and mTORC2. The former directly responds to amino acids and energetic levels, whereas the latter is required to sustain the signaling response downstream of insulin-like-peptide (Ilp) stimulation. Either signaling branch can be independently modulated in most Drosophila tissues. We confirm this independency in the fat tissue. We show that ubiquitous over-activation of mTORC1 or Ilp signaling affects carbohydrate and lipid metabolism, supporting the use of Drosophila as a powerful model to study the link between growth and metabolism. We show that cell-autonomous restriction of glycolysis or lipogenesis in fat cells impedes overgrowth dependent on Ilp-but not mTORC1-signaling. Additionally, ubiquitous deficiency of lipogenesis (FASN mutants) results in a drop in mTORC1 but not Ilp signaling, whereas, at the cell-autonomous level, lipogenesis deficiency affects none of these signals in fat cells. These findings thus, reveal differential metabolic sensitivity of mTORC1- and Ilp-dependent overgrowth. Furthermore, they suggest that local metabolic defects may elicit compensatory pathways between neighboring cells, whereas enzyme knockdown in the whole organism results in animal death. Importantly, our study weakens the use of single inhibitors to fight mTOR-related diseases and strengthens the use of drug combination and selective tissue-targeting.

cell biology↗

Ultra-Efficient Short Read Sequencing of Immune Receptor Repertoires

AbstractT cell receptor (TCR) repertoire sequencing is increasingly employed to characterize adaptive immune responses. However, current TCR sequencing methodologies are complex and expensive, limiting the scale of feasible studies. Here we present Framework Region 3 AmplifiKation sequencing (FR3AK-seq), a simplified multiplex PCR-based approach for the ultra-efficient analysis of TCR complementarity determining region 3 (CDR3) repertoires. By using minimal primer sets targeting a conserved region adjacent to CDR3, undistorted amplicons are analyzed via short read, single-end sequencing. We find that FR3AK-seq is sensitive and quantitative, performing comparably to two industry standards. FR3AK-seq was utilized to quickly and inexpensively characterize the T cell infiltrates of muscle biopsies obtained from 145 patients with idiopathic inflammatory myopathies and controls. A cluster of related TCRs was identified in samples from patients with sporadic inclusion body myositis, suggesting the presence of a shared antigen-driven response. The ease and minimal cost of FR3AK-seq removes critical barriers to routine, large-scale TCR CDR3 repertoire analyses.

immunology↗

The Relationship Between Cancer Progression And Social Environment In Drosophila

The influence of oncogenic phenomena on the ecology and evolution of animal species is fast becoming an important research topic. Similar to host-pathogen interactions, cancer negatively affects host fitness, which should lead to the selection of host control mechanisms, including behavioral traits that best minimize the proliferation of malignant cells. Social behavior is one such trait, which is suggested to influence cancer progression. While the ecological benefits of sociality in gregarious species are widely acknowledged, only limited data are available on the role of the social environment on cancer progression. Here, we exposed adult Drosophila, with colorectal-like tumors, to different social environments. First, we show that cancerous flies kept in complete isolation exhibit increased tumor progression. Yet, more surprisingly, we find that cancerous flies, kept in groups with other noncancerous individuals, also develop tumors at a faster rate compared to those kept with other cancerous conspecifics, suggesting a strong impact of social group composition on cancer growth. Finally, we show that flies can discriminate between individuals at different stages of tumor growth and selectively choose their social environment accordingly. Control flies actively avoid flies with cancer but only at the later stages of tumor development, whereas cancerous flies display strong social interactions with cancerous flies in the early stages of tumor growth. Our study demonstrates the reciprocal links between cancer and social interactions, as well as highlighting how sociality impacts health and fitness in animals and its potential implications for disease ecology and ecosystem dynamics.

animal behavior and cognition↗