RESUMO
The zebrafish is an important animal system for modeling human diseases. This includes kidney dysfunction as the embryonic kidney (pronephros) shares considerable molecular and morphological homology with the human nephron. A key clinical indicator of kidney disease is proteinuria, but a high-throughput readout of proteinuria in the zebrafish is currently lacking. To remedy this, we used the Tol2 transposon system to generate a transgenic zebrafish line that uses the fabp10a liver-specific promoter to over-express a nanoluciferase molecule fused with the D3 domain of Receptor-Associated Protein (a type of molecular chaperone) which we term NL-D3. Using a luminometer, we quantified proteinuria in NL-D3 zebrafish larvae by measuring the intensity of luminescence in the embryo medium. In the healthy state, NL-D3 is not excreted, but when embryos were treated with chemicals that affected either proximal tubular reabsorption (cisplatin, gentamicin) or glomerular filtration (angiotensin II, Hanks Balanced Salt Solution, Bovine Serum Albumin), NL-D3 is detected in fish medium. Similarly, depletion of several gene products associated with kidney disease (nphs1, nphs2, lrp2a, ocrl, col4a3, and col4a4) also induced NL-D3 proteinuria. Treating col4a4 depleted zebrafish larvae (a model of Alport syndrome) with captopril reduced proteinuria in this system. Thus, our findings validate the use of the NL-D3 transgenic zebrafish as a robust and quantifiable proteinuria reporter. Hence, given the feasibility of high-throughput assays in zebrafish, this novel reporter will permit screening for drugs that ameliorate proteinuria, thereby prioritizing candidates for further translational studies.
Assuntos
Nefrite Hereditária , Peixe-Zebra , Angiotensina II/metabolismo , Animais , Animais Geneticamente Modificados , Captopril/metabolismo , Cisplatino , Gentamicinas/metabolismo , Humanos , Glomérulos Renais/metabolismo , Nefrite Hereditária/genética , Síndrome Nefrótica , Proteinúria/tratamento farmacológico , Proteinúria/genética , Proteinúria/metabolismo , Soroalbumina Bovina/metabolismo , Peixe-Zebra/genéticaRESUMO
The 13th International Podocyte Conference was held in Manchester, UK, and online from July 28 to 30, 2021. Originally planned for 2020, this biannual meeting was postponed by a year because of the coronavirus disease 2019 (COVID-19) pandemic and proceeded as an innovative hybrid meeting. In addition to in-person attendance, online registration was offered, and this attracted 490 conference registrations in total. As a Podocyte Conference first, a day for early-career researchers was introduced. This premeeting included talks from graduate students and postdoctoral researchers. It gave early career researchers the opportunity to ask a panel, comprising academic leaders and journal editors, about career pathways and the future for podocyte research. The main meeting over 3 days included a keynote talk and 4 focused sessions each day incorporating invited talks, followed by selected abstract presentations, and an open panel discussion. The conference concluded with a Patient Day, which brought together patients, clinicians, researchers, and industry representatives. The Patient Day was an interactive and diverse day. As well as updates on improving diagnosis and potential new therapies, the Patient Day included a PodoArt competition, exercise and cooking classes with practical nutrition advice, and inspirational stories from patients and family members. This review summarizes the exciting science presented during the 13th International Podocyte Conference and demonstrates the resilience of researchers during a global pandemic.
Assuntos
COVID-19 , Podócitos , COVID-19/epidemiologia , Humanos , Pesquisa Translacional BiomédicaRESUMO
BACKGROUND: CD151 is a cell-surface molecule of the tetraspanin family. Its lateral interaction with laminin-binding integrin É3ß1 is important for podocyte adhesion to the glomerular basement membrane (GBM). Deletion of Cd151 in mice induces glomerular dysfunction, with proteinuria and associated focal glomerulosclerosis, disorganisation of GBM and tubular cystic dilation. Despite this, CD151 is not routinely screened for in patients with nephrotic-range proteinuria. We aimed to better understand the relevance of CD151 in human kidney disease. METHODS: Next-generation sequencing (NGS) was used to detect the variant in CD151. Electron and light microscopy were used to visualise the filtration barrier in the patient kidney biopsy, and immunoreactivity of patient red blood cells to anti-CD151/MER2 antibodies was performed. Further validation of the CD151 variant as disease-causing was performed in zebrafish using CRISPR-Cas9. RESULTS: We report a young child with nail dystrophy and persistent urinary tract infections who was incidentally found to have nephrotic-range proteinuria. Through targeted NGS, a novel, homozygous truncating variant was identified in CD151, a gene rarely reported in patients with nephrotic syndrome. Electron microscopy imaging of patient kidney tissue showed thickening of GBM and podocyte effacement. Immunofluorescence of patient kidney tissue demonstrated that CD151 was significantly reduced, and we did not detect immunoreactivity to CD151/MER2 on patient red blood cells. CRISPR-Cas9 depletion of cd151 in zebrafish caused proteinuria, which was rescued by injection of wild-type CD151 mRNA, but not CD151 mRNA containing the variant sequence. CONCLUSIONS: Our results indicate that a novel variant in CD151 is associated with nephrotic-range proteinuria and microscopic haematuria and provides further evidence for a role of CD151 in glomerular disease. Our work highlights a functional testing pipeline for future analysis of patient genetic variants. A higher resolution version of the Graphical abstract is available as Supplementary information.
Assuntos
Nefropatias , Podócitos , Animais , Criança , Humanos , Membrana Basal Glomerular/patologia , Integrina alfa3beta1 , Nefropatias/genética , Nefropatias/complicações , Laminina/genética , Podócitos/patologia , Proteinúria/etiologia , RNA Mensageiro , Tetraspanina 24/genética , Peixe-ZebraRESUMO
Not all encounters with pathogens are stochastic and insects can adjust their immune management in relation to cues associated with the likelihood of infection within a life cycle as well as across generations. In this study we show that female insects (bed bugs) up-regulate immune function in their copulatory organ in anticipation of mating by using feeding cues. Male bed bugs only mate with recently fed females and do so by traumatic insemination (TI). Consequently, there is a tight temporal correlation between female feeding and the likelihood of her being infected via TI. Females that received predictable access to food (and therefore predictable insemination and infection cycles) up-regulated induced immunity (generic antibacterial activity) in anticipation of feeding and mating. Females that received unpredictable (but the same mean periodicity) access to food did not. Females that anticipated mating-associated immune insult received measurable fitness benefits (survival and lifetime reproductive success) despite laying eggs at the same rate as females that were not able to predict these cycles. Given that mating is a time of increased likelihood of infection in many organisms, and is often associated with temporal cues such as courtship and/or feeding, we propose that anticipation of mating-associated infection in females may be more widespread than is currently evidenced.
Assuntos
Percevejos-de-Cama/fisiologia , Comportamento Alimentar/fisiologia , Imunidade/fisiologia , Inseminação/imunologia , Preferência de Acasalamento Animal/fisiologia , Animais , Percevejos-de-Cama/microbiologia , Sinais (Psicologia) , Feminino , Trato Gastrointestinal/imunologia , Trato Gastrointestinal/microbiologia , Masculino , Fatores SexuaisRESUMO
BACKGROUND: Lowe syndrome and Dent-2 disease are caused by mutations in the OCRL gene, which encodes for an inositol 5-phosphatase. The renal phenotype associated with OCRL mutations typically comprises a selective proximal tubulopathy, which can manifest as Fanconi syndrome in the most extreme cases. METHODS: Here, we report a 12-year-old male with nephrotic-range proteinuria and focal segmental glomerulosclerosis on renal biopsy. As a glomerular pathology was suspected, extensive investigation of tubular function was not performed. RESULTS: Surprisingly, whole exome sequencing identified a genetic variant in OCRL (c1467-2A>G) that introduced a novel splice mutation leading to skipping of exon 15. In situ hybridisation of adult human kidney tissue and zebrafish larvae showed OCRL expression in the glomerulus, supporting a role for OCRL in glomerular function. In cultured podocytes, we found that OCRL associated with the linker protein IPIP27A and CD2AP, a protein that is important for maintenance of the podocyte slit diaphragm. CONCLUSION: Taken together, this work suggests a previously under-appreciated role for OCRL in glomerular function and highlights the importance of investigating tubular function in patients with persistent proteinuria.
Assuntos
Glomerulosclerose Segmentar e Focal/genética , Glomérulos Renais/metabolismo , Síndrome Oculocerebrorrenal/genética , Animais , Criança , Canais de Cloreto , Glomerulosclerose Segmentar e Focal/complicações , Humanos , Masculino , Mutação , Síndrome Oculocerebrorrenal/complicações , Monoéster Fosfórico Hidrolases , Podócitos/metabolismo , Proteinúria/etiologia , Sequenciamento do Exoma , Peixe-ZebraRESUMO
The hepatocyte nuclear factor-1ß (Hnf1b) transcription factor is a key regulator of kidney tubule formation and is associated with a syndrome of renal cysts and early onset diabetes. To further our understanding of Hnf1b in the developing zebrafish kidney, we performed RNA sequencing analysis of proximal tubules from hnf1b-deficient larvae. This analysis revealed an enrichment of gene transcripts encoding transporters of the solute carrier (SLC) superfamily, including multiple members of slc2 and slc5 glucose transporters. An investigation of expression of slc2a1a, slc2a2, and slc5a2 as well as a poorly studied glucose/mannose transporter encoded by slc5a9 revealed that these genes undergo dynamic spatiotemporal changes during tubule formation and maturation. A comparative analysis of zebrafish SLC genes with those expressed in mouse proximal tubules showed a substantial overlap at the level of gene families, indicating a high degree of functional conservation between zebrafish and mammalian proximal tubules. Taken together, our findings are consistent with a role for Hnf1b as a critical determinant of proximal tubule transport function by acting upstream of a large number of SLC genes and validate the zebrafish as a physiologically relevant model of the mammalian proximal tubule.
Assuntos
Perfilação da Expressão Gênica , Fator 1-beta Nuclear de Hepatócito/genética , Fator 1-beta Nuclear de Hepatócito/fisiologia , Túbulos Renais Proximais/metabolismo , Proteínas de Peixe-Zebra/genética , Proteínas de Peixe-Zebra/fisiologia , Peixe-Zebra/genética , Animais , Animais Geneticamente Modificados , Proteínas de Transporte/genética , Proteínas de Transporte/metabolismo , Regulação da Expressão Gênica no Desenvolvimento , Larva , Camundongos , RNA/biossíntese , RNA/genética , Especificidade da EspécieRESUMO
During zebrafish embryogenesis the pronephric kidney arises from a small population of posterior mesoderm cells that then undergo expansion during early stages of renal organogenesis. While wnt8 is required for posterior mesoderm formation during gastrulation, it is also transiently expressed in the post-gastrula embryo in the intermediate mesoderm, the precursor to the pronephros and some blood/vascular lineages. Here, we show that knockdown of wnt8a, using a low dose of morpholino that does not disrupt early mesoderm patterning, reduces the number of kidney and blood cells. For the kidney, wnt8a deficiency decreases renal progenitor growth during early somitogenesis, as detected by EdU incorporation, but has no effect on apoptosis. The depletion of the renal progenitor pool in wnt8a knockdown embryos leads to cellular deficits in the pronephros at 24 hpf that are characterised by a shortened distal-most segment and stretched proximal tubule cells. A pulse of the canonical Wnt pathway agonist BIO during early somitogenesis is sufficient to rescue the size of the renal progenitor pool while longer treatment expands the number of kidney cells. Taken together, these observations indicate that Wnt8, in addition to its well-established role in posterior mesoderm patterning, also plays a later role as a factor that expands the renal progenitor pool prior to kidney morphogenesis.
Assuntos
Proteínas do Citoesqueleto/metabolismo , Embrião não Mamífero/citologia , Rim/citologia , Rim/embriologia , Células-Tronco/citologia , Proteínas Wnt/metabolismo , Proteínas de Peixe-Zebra/metabolismo , Peixe-Zebra/embriologia , Peixe-Zebra/metabolismo , Animais , Apoptose/efeitos dos fármacos , Células Sanguíneas/citologia , Células Sanguíneas/efeitos dos fármacos , Padronização Corporal/efeitos dos fármacos , Contagem de Células , Proliferação de Células/efeitos dos fármacos , Embrião não Mamífero/efeitos dos fármacos , Técnicas de Silenciamento de Genes , Indóis/farmacologia , Túbulos Renais/efeitos dos fármacos , Túbulos Renais/patologia , Mesoderma/efeitos dos fármacos , Mesoderma/embriologia , Mesoderma/metabolismo , Morfolinos/farmacologia , Oximas/farmacologia , Pronefro/citologia , Pronefro/embriologia , Células-Tronco/efeitos dos fármacos , Células-Tronco/metabolismoRESUMO
The nephron is the functional subunit of the vertebrate kidney and plays important osmoregulatory and excretory roles during embryonic development and in adulthood. Despite its central role in kidney function, surprisingly little is known about the molecular and cellular processes that control nephrogenesis. The zebrafish pronephric kidney, comprising two nephrons, provides a visually accessible and genetically tractable model system for a better understanding of nephron formation. Using this system, various developmental processes, including the commitment of mesoderm to a kidney fate, renal tubule proliferation, and migration, can be studied during nephrogenesis. Here, we discuss some of these processes in zebrafish with a focus on the pathways that influence renal tubule cell morphogenesis.
Assuntos
Túbulos Renais/embriologia , Morfogênese/fisiologia , Organogênese/fisiologia , Peixe-Zebra/embriologia , AnimaisRESUMO
Loss of kidney function underlies many renal diseases. Mammals can partly repair their nephrons (the functional units of the kidney), but cannot form new ones. By contrast, fish add nephrons throughout their lifespan and regenerate nephrons de novo after injury, providing a model for understanding how mammalian renal regeneration may be therapeutically activated. Here we trace the source of new nephrons in the adult zebrafish to small cellular aggregates containing nephron progenitors. Transplantation of single aggregates comprising 10-30 cells is sufficient to engraft adults and generate multiple nephrons. Serial transplantation experiments to test self-renewal revealed that nephron progenitors are long-lived and possess significant replicative potential, consistent with stem-cell activity. Transplantation of mixed nephron progenitors tagged with either green or red fluorescent proteins yielded some mosaic nephrons, indicating that multiple nephron progenitors contribute to a single nephron. Consistent with this, live imaging of nephron formation in transparent larvae showed that nephrogenic aggregates form by the coalescence of multiple cells and then differentiate into nephrons. Taken together, these data demonstrate that the zebrafish kidney probably contains self-renewing nephron stem/progenitor cells. The identification of these cells paves the way to isolating or engineering the equivalent cells in mammals and developing novel renal regenerative therapies.
Assuntos
Rim/citologia , Rim/crescimento & desenvolvimento , Néfrons/citologia , Regeneração/fisiologia , Células-Tronco/citologia , Peixe-Zebra/crescimento & desenvolvimento , Envelhecimento/fisiologia , Animais , Animais Geneticamente Modificados , Proliferação de Células , Rim/lesões , Rim/metabolismo , Larva , Modelos Animais , Néfrons/crescimento & desenvolvimento , Organogênese , Transplante de Células-TroncoRESUMO
The nephron is the functional unit that executes the homeostatic roles of the kidney in vertebrates. Critical to this function is the physical arrangement of the glomerular blood filter attached to a tubular epithelium that is subdivided into specialized proximal and distal segments. During embryogenesis, nephron progenitors undergo a mesenchymal-epithelial transition (MET) and adopt different segment-specific cell fates along the proximo-distal axis of the nephron. The molecular basis of how these segments arise remains largely unknown. Recent studies using the zebrafish have identified the Hnf1beta transcription factor (Hnf1b) as a major regulator of tubular segmentation. In Hnf1b-deficient zebrafish embryos, nephron progenitors fail to adopt the proximo-distal segmentation pattern of the nephron, yet still undergo MET. This observation suggests that the functional segmentation of renal tubular epithelial cells is independent of pathways that induce their epithelialization. Here we review this new role of Hnf1b for nephron segmentation during zebrafish and mouse kidney development.
Assuntos
Fator 1-beta Nuclear de Hepatócito/metabolismo , Néfrons/embriologia , Néfrons/metabolismo , Proteínas de Peixe-Zebra/metabolismo , Animais , Fator 1-beta Nuclear de Hepatócito/fisiologia , Humanos , Organogênese/fisiologia , Proteínas de Peixe-Zebra/fisiologiaRESUMO
Nephrons comprise a blood filter and an epithelial tubule that is subdivided into proximal and distal segments, but what directs this patterning during kidney organogenesis is not well understood. Using zebrafish, we found that the HNF1ß paralogues hnf1ba and hnf1bb, which encode homeodomain transcription factors, are essential for normal segmentation of nephrons. Embryos deficient in hnf1ba and hnf1bb did not express proximal and distal segment markers, yet still developed an epithelial tubule. Initiating hnf1ba/b expression required Pax2a and Pax8, but hnf1ba/b-deficient embryos did not exhibit the expected downregulation of pax2a and pax8 at later stages of development, suggesting complex regulatory loops involving these molecules. Embryos deficient in hnf1ba/b also did not express the irx3b transcription factor, which is responsible for differentiation of the first distal tubule segment. Reciprocally, embryos deficient in irx3b exhibited downregulation of hnf1ba/b transcripts in the distal early segment, suggesting a segment-specific regulatory circuit. Deficiency of hnf1ba/b also led to ectopic expansion of podocytes into the proximal tubule domain. Epistasis experiments showed that the formation of podocytes required wt1a, which encodes the Wilms' tumor suppressor-1 transcription factor, and rbpj, which encodes a mediator of canonical Notch signaling, downstream or parallel to hnf1ba/b. Taken together, these results suggest that Hnf1ß factors are essential for normal segmentation of nephrons during kidney organogenesis.
Assuntos
Fator 1-beta Nuclear de Hepatócito/metabolismo , Néfrons/embriologia , Proteínas de Peixe-Zebra/metabolismo , Animais , Caderinas/metabolismo , Regulação para Baixo , Embrião não Mamífero/metabolismo , Proteínas de Homeodomínio/metabolismo , Fator de Transcrição PAX2/metabolismo , Fator de Transcrição PAX8 , Fatores de Transcrição Box Pareados/metabolismo , Podócitos/fisiologia , Receptores Notch/metabolismo , Transdução de Sinais , Fatores de Transcrição/metabolismo , Tretinoína/metabolismo , Peixe-ZebraRESUMO
AIMS: Rare, deleterious genetic variants in FLT4 are associated with Tetralogy of Fallot (TOF), the most common cyanotic congenital heart disease (CHD). Distinct genetic variants in FLT4 are also an established cause of Milroy disease, the most prevalent form of primary hereditary lymphoedema. Phenotypic features of these two conditions are non-overlapping, implying pleiotropic cellular mechanisms during development. METHODS AND RESULTS: Here, we show that FLT4 variants identified in TOF patients, when expressed in primary human endothelial cells, cause aggregation of FLT4 protein in the perinuclear endoplasmic reticulum, activating proteostatic and metabolic signalling, whereas lymphoedema-associated FLT4 variants and wildtype FLT4 do not. FLT4 TOF variants display characteristic gene expression profiles in key developmental signalling pathways, revealing a role for FLT4 in cardiogenesis distinct from its role in lymphatic development. Inhibition of proteostatic signalling abrogates these effects, identifying potential avenues for therapeutic intervention. Depletion of flt4 in zebrafish caused cardiac phenotypes of reduced heart size and altered heart looping. These phenotypes were rescued with coinjection of wildtype human FLT4 mRNA, but incompletely or not at all by mRNA harbouring FLT4 TOF variants. CONCLUSIONS: Taken together, we identify a pathogenic mechanism for FLT4 variants predisposing to TOF that is distinct from the known dominant negative mechanism of Milroy-causative variants. FLT4 variants give rise to conditions of the two circulatory subdivisions of the vascular system via distinct developmental pleiotropic molecular mechanisms. TRANSLATIONAL PERSPECTIVE: Proteostatic dysfunction, if confirmed as a mechanism of CHD pathogenesis for other predisposing genes, may identify pathways to therapeutic interventions. Distinguishing mechanistically how variants in FLT4 give rise to CHD may have potential to individualise genetic counselling in affected families.
RESUMO
Modern bed bugs are resistant to multiple insecticide classes, particularly the pyrethroids. The efficacy of pyrethroid-impregnated mattress liners marketed for bed bug management has been variable. This study evaluated the efficacy of a permethrin-impregnated mattress liner, ActiveGuard, against 24 bed bug strains, consisting of both Cimex hemipterus (F.) and Cimex lectularius L. A 'mat assay', employing an allethrin-impregnated mat, was used to establish the pyrethroid resistance profile of all strains. Three experiments were conducted to evaluate the effect of ActiveGuard exposure on bed bug knockdown: 1) exposing the bed bugs continuously on the liner for up to 24 d, 2) holding the bed bugs on the liner for either 4 or 6 h, and 3) placing a noninsecticide treated fabric above the liner with the bed bugs held continuously on top. Our results indicated that all modern strains (collected within the last 15 years during the current resurgence) were pyrethroid-resistant, although the magnitude of resistance was highly variable between strains. In the continuous exposure study, an incomplete knockdown was recorded for most modern bed bug strains, with some having no knockdown even up to 7 d of constant exposure. In the 4 or 6 h exposure study, the level of knockdown was reduced even further, and very few bed bugs were knocked down in the double fabric study. The results of this study indicate that pyrethroid-impregnated mattress liners are not likely to be effective in the management of most modern bed bug infestations involving either C. hemipterus or C. lectularius.
Assuntos
Percevejos-de-Cama , Inseticidas , Piretrinas , Animais , Piretrinas/farmacologia , Inseticidas/farmacologia , Permetrina , Aletrinas/farmacologiaRESUMO
Previous studies have highlighted a role for the Notch signalling pathway during pronephrogenesis in the amphibian Xenopus laevis, and in nephron development in the mammalian metanephros, yet a mechanism for this function remains elusive. Here, we further the understanding of how Notch signalling patterns the early X. laevis pronephros anlagen, a function that might be conserved in mammalian nephron segmentation. Our results indicate that early phase pronephric Notch signalling patterns the medio-lateral axis of the dorso-anterior pronephros anlagen, permitting the glomus and tubules to develop in isolation. We show that this novel function acts through the Notch effector gene hrt1 by upregulating expression of wnt4. Wnt-4 then patterns the proximal pronephric anlagen to establish the specific compartments that span the medio-lateral axis. We also identified pronephric expression of lunatic fringe and radical fringe that is temporally and spatially appropriate for a role in regulating Notch signalling in the dorso-anterior region of the pronephros anlagen. On the basis of these results, along with data from previous publications, we propose a mechanism by which the Notch signalling pathway regulates a Wnt-4 function that patterns the proximal pronephric anlagen.
Assuntos
Néfrons/embriologia , Receptores Notch/metabolismo , Proteínas Wnt/metabolismo , Xenopus laevis/embriologia , Animais , Embrião não Mamífero/metabolismo , Regulação da Expressão Gênica no Desenvolvimento , Glicosiltransferases/metabolismo , Néfrons/metabolismo , Fatores de Transcrição/metabolismo , Regulação para Cima , Proteína Wnt4 , Proteínas de Xenopus/metabolismoRESUMO
Increased female reproductive rates usually result in accelerated senescence. This correlation provides a link between the evolutionary conflict of the sexes and aging when ejaculate components elevate female reproductive rates at the cost of future reproduction. It is not clear whether this female cost is manifest as shorter lifespan or an earlier onset or a steeper rate of reproductive senescence. It also is unclear whether beneficial ejaculates release females from reproductive trade-offs and, if so, which senescence parameters are affected. We examined these issues in the bedbug, Cimex lectularius, a long-lived insect that shows reduced female lifespan as well as female reproductive senescence at the male-determined mating frequency. We demonstrate experimentally that, independently of the mating frequency, females receiving more ejaculate show increased reproductive rates and enter reproductive senescence later than females receiving less ejaculate. The rate of reproductive senescence did not differ between treatments, and reproductive rates did not predict mortality. The ejaculate effects were consistent in inter- and intra-population crosses, suggesting they have not evolved recently and are not caused by inbreeding. Our results suggest that ejaculate components compensate for the costs of elevated female reproductive rates in bedbugs by delaying the onset of reproductive senescence. Ejaculate components that are beneficial to polyandrous females could have arisen because male traits that protect the ejaculate have positive pleiotropic effects and/or because female counteradaptations to antagonistic male traits exceed the neutralization of those traits. That males influence female reproductive senescence has important consequences for trade-offs between reproduction and longevity and for studies of somatic senescence.
Assuntos
Fertilidade , Insetos/fisiologia , Reprodução/fisiologia , Sêmen , Comportamento Sexual Animal , Animais , Feminino , MasculinoRESUMO
Basement membranes (BMs) are ubiquitous extracellular matrices whose composition remains elusive, limiting our understanding of BM regulation and function. By developing a bioinformatic and in vivo discovery pipeline, we define a network of 222 human proteins and their animal orthologs localized to BMs. Network analysis and screening in C. elegans and zebrafish uncovered BM regulators, including ADAMTS, ROBO, and TGFß. More than 100 BM network genes associate with human phenotypes, and by screening 63,039 genomes from families with rare disorders, we found loss-of-function variants in LAMA5, MPZL2, and MATN2 and show that they regulate BM composition and function. This cross-disciplinary study establishes the immense complexity of BMs and their impact on in human health.
Assuntos
Caenorhabditis elegans , Peixe-Zebra , Animais , Membrana Basal/metabolismo , Caenorhabditis elegans/genética , Caenorhabditis elegans/metabolismo , Matriz Extracelular/genética , Matriz Extracelular/metabolismo , Proteínas da Matriz Extracelular/metabolismo , Humanos , Peixe-Zebra/genéticaRESUMO
The glomerular basement membrane (GBM) is a key component of the glomerular capillary wall and is essential for kidney filtration. The major components of the GBM include laminins, type IV collagen, nidogens and heparan sulfate proteoglycans. In addition, the GBM harbours a number of other structural and regulatory components and provides a reservoir for growth factors. New technologies have improved our ability to study the composition and assembly of basement membranes. We now know that the GBM is a complex macromolecular structure that undergoes key transitions during glomerular development. Defects in GBM components are associated with a range of hereditary human diseases such as Alport syndrome, which is caused by defects in the genes COL4A3, COL4A4 and COL4A5, and Pierson syndrome, which is caused by variants in LAMB2. In addition, the GBM is affected by acquired autoimmune disorders and metabolic diseases such as diabetes mellitus. Current treatments for diseases associated with GBM involvement aim to reduce intraglomerular pressure and to treat the underlying cause where possible. As our understanding about the maintenance and turnover of the GBM improves, therapies to replace GBM components or to stimulate GBM repair could translate into new therapies for patients with GBM-associated disease.
Assuntos
Doença Antimembrana Basal Glomerular/etiologia , Nefropatias Diabéticas/etiologia , Membrana Basal Glomerular/patologia , Membrana Basal Glomerular/fisiopatologia , Síndromes Miastênicas Congênitas/etiologia , Nefrite Hereditária/etiologia , Síndrome Nefrótica/etiologia , Distúrbios Pupilares/etiologia , HumanosRESUMO
The rapid evolution of ejaculate components is considered to be largely driven by sexual selection. Less attention has been paid to the fact that sperm and microorganisms frequently meet; we consequently predict selection for substances that protect a male's ejaculate. We report, for the first time, bacteriolytic activity (lysozyme-like immune activity [LLA]) in the ejaculate of an animal, the common bedbug Cimex lectularius. We also show that in almost half the males LLA in the seminal fluid exceeded LLA in the hemolymph. We detected no antimicrobial peptide activity in seminal fluid. Because lysozymes degrade only bacteria, our results suggest that sperm-microbe interactions are probably important in the evolution of ejaculate components and thereby provide a route for natural selection to account for some of the diversity of seminal components.
Assuntos
Anti-Infecciosos/metabolismo , Percevejos-de-Cama/imunologia , Muramidase/metabolismo , Sêmen/imunologia , Animais , Anti-Infecciosos/farmacologia , Percevejos-de-Cama/enzimologia , Percevejos-de-Cama/microbiologia , Hemolinfa/enzimologia , Masculino , Testes de Sensibilidade Microbiana , Micrococcus luteus/efeitos dos fármacos , Muramidase/farmacologia , Seleção Genética , Sêmen/enzimologia , Sêmen/microbiologiaRESUMO
BACKGROUND: Bed bugs (Heteroptera: Cimicidae) are a group of blood-feeding ectoparasites. They mainly specialize on bats and birds, but a few species are important human pests. They exhibit several unique adaptations for their parasitic lifestyle. Among those, bed bug aggregations represent a striking example of a sub-social structure. However, their benefits for the bed bugs as well as their potential for bed bug control are largely unexplored. Young nymphs are known to disperse from the aggregations much less than older ones or adults. We therefore found possible that the aggregation age structure is connected with success in finding host and tested the effect of presence of adults on nymphal feeding success. RESULTS: We tested the effect of presence of adults on feeding success of first-instar nymphs using an artificial feeding system. We found that presence of fed adults causes larger proportion of nymphs to feed. CONCLUSIONS: Based on our data, fed bed bugs seem to trigger the young nymphs to actively forage. Since the first instar is much less viable than later stages, our finding points to an adaptive behavior that economizes on foraging energy cost. In the context of bed bug control, knowledge on such behavior emphasizes the prevention of fed bed bugs from returning to harborages. Bed bug traps may thus be used not just as means of bed bug monitoring, but also as means of control.