RESUMO
BACKGROUND: Copy number variants (CNVs) associated with late-onset medical conditions are rare but important secondary findings in chromosomal microarray analysis (CMA) performed during pregnancy. Here, we critically review the cases at two tertiary centres to assess the criteria which guide the disclosure of such findings and develop a disclosure decision tool (DDT) aimed at facilitating disclosure decision. Parental decisions on receiving CNVs associated with risks for late-onset conditions were also recorded. METHODS: Prenatal CMAs in Hadassah and Shaare Zedek Medical Centers from November 2013 to October 2021 were reviewed for CNVs associated with late-onset conditions. The DDT proposed uses a five-parameter scoring system, which considers the severity, median age of onset, penetrance, understanding of genotype-phenotype correlation and actionability of the finding. RESULTS: Out of 16 238 prenatal CMAs, 16 (0.1%) harboured CNVs associated with late-onset conditions, 15 of which were disclosed. Outcome information was available on 13 of the 16 pregnancies, all of which continued to delivery. CONCLUSIONS: Our suggested DDT will help clinicians to quantitatively weigh the variables associated with CNVs of this type and arrive at a well thought out clinical decision regarding disclosure. Although the prevalence of late-onset conditions as a major finding in the prenatal setup is low, it is expected to rise with the increasing use of non-invasive CMA testing and whole exome and genome sequencing.
Assuntos
Aberrações Cromossômicas , Diagnóstico Pré-Natal , Feminino , Humanos , Gravidez , Revelação , Variações do Número de Cópias de DNA/genética , Análise em Microsséries , Resultado da GravidezRESUMO
Complex chromosomal rearrangements (CCRs), a class of structural variants (SVs) involving more than two chromosome breaks, were classically thought to be extremely rare. As advanced technologies become more available, it has become apparent that CCRs are more common than formerly thought, and are a substantial cause of genetic disorders. We attempted a novel approach for solving the mechanism of challenging CCRs, which involve repetitive sequences, by precisely identifying sequence-level changes and their order. Chromosomal microarray (CMA) and FISH analyses were used for interpretation of SVs detected by whole exome sequencing (WES). Breakpoint junctions were analyzed by Nanopore sequencing, a novel long-read whole genome sequencing tool. A large deletion identified by WES, encompassing the FOXF1 enhancer, was the cause of alveolar capillary dysplasia and respiratory insufficiency, resulting in perinatal death. CMA analysis of the newborn's mother revealed two duplications encompassing the deleted region in the proband, raising our hypothesis that the deletion resulted from the mother's CCR. Breakpoint junctions of complex SVs were determined at the nucleotide level using Nanopore long-read sequencing. According to sequencing results of breakpoint junctions, the CCR in the newborn was considered the consequence of at least one double-strand break during meiosis, and reassembly of DNA fragments by intra-chromosomal homologous recombination. Our comprehensive approach, combining cytogenetics and long-read sequencing, enabled delineation of the exact breakpoints in a challenging CCR, and proposal of a mechanism in which it arises. We suggest applying our integrative approach combining technologies for deciphering future challenging CCRs, enabling risk assessment in families.
Assuntos
Aberrações Cromossômicas , Genoma , Cromossomos , Análise Citogenética , Feminino , Genômica , Humanos , GravidezRESUMO
PURPOSE: To review cases of couples presented to our PGT-unit with copy number variants (CNVs) classified as variants of uncertain significance (VUS) in order to better understand their needs. METHODS: Retrospective cohort study conducted in a tertiary medical-center, 2014-2019. We reviewed files of all couples applying for genetic counseling with CNVs classified as VUS. The main outcomes measured: number of VUS findings and their description, PGT-M procedures planned and performed, IVF cycles, clinical pregnancy, and live birth rates (LBR). VUS were classified according to the American-College of Medical-Genetics and Genomics classification at time of first consultation, and updated-December 2018. RESULTS: Twenty-four couples presented with a total of 30 VUS. Twelve couples (50%) had isolated VUS and 12 (50%) had VUS diagnosed in addition to a pathogenic mutation. Initially, nine findings (30%) were defined as VUS; eight (27%) as likely benign (b-VUS); and 13 (43%) as likely pathogenic (p-VUS). PGT-M was recommended for 17/30 CNVs (56.6%), 12 (70%) of which, isolated VUS. No couple had other indications for IVF. To date, nine couples performed PGT-M for isolated VUS; LBR per-couple-55.5%. Five couples performed PGT-M for both pathogenic findings and VUS, LBR-80%. After reviewing VUS classifications, 30% remained unchanged, 20% were more severely defined, and 50% less severely defined. CONCLUSION: The genomic era enables detection of VUS whose definition is subject to change as additional information becomes available. The uncertainty of variants' clinical significance and changes in VUS definition over time complicates genetic counseling. Revised guidelines for VUS interpretation and reevaluation of patient counseling before each pregnancy must be practiced when counseling them regarding the justification of PGT-M for their diagnosed VUS.
Assuntos
Transtornos Cromossômicos/diagnóstico , Variações do Número de Cópias de DNA , Fertilização in vitro/métodos , Testes Genéticos/métodos , Diagnóstico Pré-Implantação/métodos , Adulto , Transtornos Cromossômicos/genética , Transferência Embrionária , Feminino , Humanos , Masculino , Gravidez , Resultado da Gravidez , Taxa de Gravidez , Estudos Retrospectivos , Adulto JovemRESUMO
Social spatial cognition refers to the interaction between self, place, and partners, with emphasis on the impact of the social environment on spatial behavior and on how individual spatial representations converge to form collective spatial behavior - i.e., common places and routes. Recent studies suggest that in addition to their mental representation (cognitive map) of the physical environment, humans and other animals also have a social cognitive map. We suggest that while social spatial cognition relies on knowledge of both the physical and the social environments, it is the latter hat predominates. This dominance is illustrated here in the modulation of spatial behavior according to dynamic social interactions, ranging from group formation to an attenuation of drug-induced stereotypy through the mere presence of a normal subject. Consequently we suggest that the numerous studies on the biobehavioral controlling mechanisms of spatial behavior (i.e. - the hippocampal formation, animal models for mental disorders) should also consider the social environment rather than solely focusing on the spatial behavior of lone animals.
Assuntos
Cognição , Comportamento Espacial , Animais , Meio Ambiente , Hipocampo , HumanosRESUMO
Lens abnormalities are a major cause of reduced vision and blindness. One mechanism that can lead to reduced lens transparency, i.e. cataract, is abnormal behavior of lens epithelial cells (LECs), the precursors of the transparent lens fiber cells. Here we describe a zebrafish mutation causing the embryonic lens epithelium to generate cellular masses comprising partially differentiated lens fiber cells. We identify the mutant gene as plod3, which encodes for Lysyl hydroxylase 3 (Lh3), an enzyme essential for modification of collagens, including Collagen IV, a main component of the lens capsule. We show that plod3-deficient lenses have abnormal lens epithelium from an early developmental stage, as well as abnormal lens capsules. Subsequently, upregulation of TGFß signaling takes place, which drives the formation of lens epithelial cellular masses. We identify a similar phenotype in Collagen IVα5-deficient embryos, suggesting a key role for the defective lens capsule in the pathogenesis. We propose that plod3 and col4a5 mutant zebrafish can serve as useful models for better understanding the biology of LECs during embryonic development and in formation of lens epithelium-derived cataract.
Assuntos
Glicosiltransferases/genética , Cápsula do Cristalino/embriologia , Cápsula do Cristalino/metabolismo , Pró-Colágeno-Lisina 2-Oxoglutarato 5-Dioxigenase/genética , Proteínas de Peixe-Zebra/genética , Actinas/genética , Actinas/metabolismo , Animais , Catarata/genética , Diferenciação Celular/fisiologia , Desenvolvimento Embrionário , Células Epiteliais/patologia , Epitélio/patologia , Glicosiltransferases/metabolismo , Cristalino/embriologia , Fenótipo , Pró-Colágeno-Lisina 2-Oxoglutarato 5-Dioxigenase/metabolismo , Peixe-Zebra , Proteínas de Peixe-Zebra/metabolismoRESUMO
In the present study, the dynamic process of group formation in eight unfamiliar rats was followed in order to reveal how the group becomes oriented together in time and space, in light of the complexity that accompanies grouping. The focus was on who, where, and when joined together. We found that rats preferred to be in companionship over remaining alone, with all the rats gradually shifting to share the same location as a resting place. Group formation can be viewed as a tri-phasic process, with some rats gradually becoming more social than others, and thus playing a key role in group formation. Starting with seemingly independent traveling, the rats gradually converged to share the same location as a terminal (home base) for roundtrips in the arena. Because such a terminal is considered as the organizer of an individual's spatial behavior, the shared home-base location may be viewed as the organizer of spatial behavior of the entire group. Despite huddling together, the rats continued to travel alone or in duos throughout the 3 h of testing. We suggest that resting together and traveling alone or in duos enabled the maintenance of communal relationship while reducing the complexity involved in traveling in relatively large groups. Taken together, the present results demonstrate the dynamic process during which unfamiliar rats shift from independent to group spatial behavior.
Assuntos
Comportamento Exploratório , Comportamento Social , Comportamento Espacial , Animais , RatosRESUMO
The eye primordium arises as a lateral outgrowth of the forebrain, with a transient fissure on the inferior side of the optic cup providing an entry point for developing blood vessels. Incomplete closure of the inferior ocular fissure results in coloboma, a disease characterized by gaps in the inferior eye and recognized as a significant cause of pediatric blindness. Here, we identify eight patients with defects in tissues of the superior eye, a congenital disorder that we term superior coloboma. The embryonic origin of superior coloboma could not be explained by conventional models of eye development, leading us to reanalyze morphogenesis of the dorsal eye. Our studies revealed the presence of the superior ocular sulcus (SOS), a transient division of the dorsal eye conserved across fish, chick, and mouse. Exome sequencing of superior coloboma patients identified rare variants in a Bone Morphogenetic Protein (Bmp) receptor (BMPR1A) and T-box transcription factor (TBX2). Consistent with this, we find sulcus closure defects in zebrafish lacking Bmp signaling or Tbx2b. In addition, loss of dorsal ocular Bmp is rescued by concomitant suppression of the ventral-specific Hedgehog pathway, arguing that sulcus closure is dependent on dorsal-ventral eye patterning cues. The superior ocular sulcus acts as a conduit for blood vessels, with altered sulcus closure resulting in inappropriate connections between the hyaloid and superficial vascular systems. Together, our findings explain the existence of superior coloboma, a congenital ocular anomaly resulting from aberrant morphogenesis of a developmental structure.
Assuntos
Receptores de Proteínas Morfogenéticas Ósseas Tipo I/genética , Coloboma/embriologia , Coloboma/genética , Citocromo P-450 CYP1B1/genética , Olho/embriologia , Adulto , Animais , Animais Geneticamente Modificados , Receptores de Proteínas Morfogenéticas Ósseas Tipo I/metabolismo , Embrião de Galinha , Embrião não Mamífero , Fator 6 de Diferenciação de Crescimento/genética , Fator 6 de Diferenciação de Crescimento/metabolismo , Humanos , Lactente , Camundongos , Proteínas com Domínio T/genética , Proteínas com Domínio T/metabolismo , Peixe-Zebra/embriologia , Peixe-Zebra/genética , Proteínas de Peixe-Zebra/genética , Proteínas de Peixe-Zebra/metabolismoRESUMO
The superficial ocular vasculature of the embryonic zebrafish develops in a highly stereotypic manner and hence provides a convenient model for studying molecular mechanisms that regulate vascular patterning. We have used transgenic zebrafish embryos in which all endothelial cells express enhanced Green Fluorescent Protein and small molecule inhibitors to examine the contribution of two signaling pathways, vascular endothelial growth factor (VEGF) and Hedgehog (Hh) pathways, to the development of the superficial system. We find that most, but not all vessels of the superficial system depend on VEGF signaling for their growth. Hh signaling appears to limit superficial vessel growth over the dorsal eye and is required to promote superficial vessel growth over the ventral eye. These effects of Hh signaling are indirect. Our initial analyses of factors that regulate growth and patterning of superficial ocular vessels suggest that early patterning events in the embryo during organogenesis stages could influence vascular patterning later on. By studying development of specific vascular systems it should be possible to identify new roles for signaling pathways in regulating vascular development.
Assuntos
Olho/embriologia , Regulação da Expressão Gênica no Desenvolvimento , Proteínas Hedgehog/metabolismo , Cristalino/embriologia , Vasos Retinianos/embriologia , Fator A de Crescimento do Endotélio Vascular/metabolismo , Peixe-Zebra/embriologia , Animais , Animais Geneticamente Modificados , Padronização Corporal , Olho/irrigação sanguínea , Proteínas Hedgehog/genética , Cristalino/irrigação sanguínea , Ligantes , Neovascularização Fisiológica/fisiologia , Organogênese , Fenótipo , Transdução de Sinais , Peixe-Zebra/genética , Proteínas de Peixe-Zebra/genéticaRESUMO
Limited resources result in competition among social animals. Nevertheless, social animals also have innate preferences for cooperative behavior. In the present study, 12 dyads of food-deprived rats were tested in four successive trials, and then re-tested as eight triads of food-deprived rats that were unfamiliar to each other. We found that the food-deprived dyads or triads of rats did not compete for the food available to them at regular spatially-marked locations that they had previously learnt. Rather, these rats traveled together to collect the baits. One rat, or two rats in some triads, lead (ran ahead) to collect most of the baits, but "leaders" differed across trials so that, on average, each rat ultimately collected similar amounts of baits. Regardless of which rat collected the baits, the rats traveled together with no substantial difference among them in terms of their total activity. We suggest that rats, which are a social species that has been found to display reciprocity, have evolved to travel and forage together and to share limited resources. Consequently, they displayed a sort of 'peace economy' that on average resulted in equal access to the baits across trials. For social animals, this type of dynamics is more relaxed, tolerant, and effective in the management of conflicts. Rather than competing for the limited available food, the food-deprived rats socialized and coexisted peacefully.
Assuntos
Comportamento Competitivo , Comportamento Cooperativo , Comportamento Alimentar , Privação de Alimentos , Animais , Masculino , Ratos , Ratos Sprague-DawleyRESUMO
Spatial organization is an extensively studied field, in which most of the research has been on how the physical environment is perceived and conceived. There is a consensus that physical attributes such as environment geometry and landmarks are key factors in shaping spatial cognition. Nevertheless, the numerous studies of spatial behavior have usually been carried out on individuals, thereby overlooking the possible impact of the social environment. In the present study, rats were exposed to an unfamiliar open-field, first alone and then in tetrads of unfamiliar individuals, in order to monitor and analyze when and how their individual spatial behavior converged to a group spatial behavior. We found that the unfamiliar rats spent most of their time in companionship, first with preferred partners and ultimately as a quartet. Specifically, group formation was dynamic and gradual, with the rats first forming duos, then trios, and ultimately a quartet. Trios and quartets mostly huddled in the same specific corner that became a shared home base, from which they took solo or duo roundtrips to the arena. The present study unveils how, by means of gradual interactions among self, place, and conspecifics, four unfamiliar rats organized together their social spatial behavior.
Assuntos
Comportamento Social , Comportamento Espacial , Animais , Cognição , Masculino , Ratos Sprague-DawleyRESUMO
BACKGROUND: Functions for the early embryonic vasculature in regulating development of central nervous system tissues, such as the retina, have been suggested by in vitro studies and by in vivo manipulations that caused additional ocular vessels to develop. Here, we use an avascular zebrafish embryo, cloche-/- (clo-/-), to begin to identify necessary developmental functions of the ocular vasculature in regulating development and patterning of the neural retina, in vivo. These studies are possible in zebrafish embryos, which do not yet rely upon the vasculature for tissue oxygenation. RESULTS: clo-/- embryos lacked early ocular vasculature and were microphthalmic, with reduced retinal cell proliferation and cell survival. Retinas of clo mutants were disorganized, with irregular synaptic layers, mispatterned expression domains of retinal transcription factors, morphologically abnormal Müller glia, reduced differentiation of specific retinal cell types, and sporadically distributed cone photoreceptors. Blockade of p53-mediated cell death did not completely rescue this phenotype and revealed ectopic cones in the inner nuclear layer. clo-/- embryos did not upregulate a molecular marker for hypoxia. CONCLUSIONS: The disorganized retinal phenotype of clo-/- embryos is consistent with a neural and glial developmental patterning role for the early ocular vasculature that is independent of its eventual function in gas exchange.
Assuntos
Fatores de Transcrição Hélice-Alça-Hélice Básicos/genética , Mutação , Retina/anormalidades , Retina/embriologia , Proteínas de Peixe-Zebra/genética , Peixe-Zebra/genética , Animais , Animais Geneticamente Modificados , Fatores de Transcrição Hélice-Alça-Hélice Básicos/fisiologia , Morte Celular , Diferenciação Celular , Proliferação de Células , Sobrevivência Celular , Embrião não Mamífero/metabolismo , Regulação da Expressão Gênica no Desenvolvimento , Hipóxia , Microscopia Confocal , Neuroglia/fisiologia , Neurônios/fisiologia , Fenótipo , Epitélio Pigmentado da Retina/metabolismo , Células-Tronco , Proteínas de Peixe-Zebra/fisiologiaRESUMO
BACKGROUND: The developing eye receives blood supply from two vascular systems, the intraocular hyaloid system and the superficial choroidal vessels. In zebrafish, a highly stereotypic and simple set of vessels develops on the surface of the eye prior to development of choroidal vessels. The origins and formation of this so-called superficial system have not been described. RESULTS: We have analyzed the development of superficial vessels by time-lapse imaging and identified their origins by photoconversion experiments in kdrl:Kaede transgenic embryos. We show that the entire superficial system is derived from a venous origin, and surprisingly, we find that the hyaloid system has, in addition to its previously described arterial origin, a venous origin for specific vessels. Despite arising solely from a vein, one of the vessels in the superficial system, the nasal radial vessel (NRV), appears to acquire an arterial identity while growing over the nasal aspect of the eye and this happens in a blood flow-independent manner. CONCLUSIONS: Our results provide a thorough analysis of the early development and origins of zebrafish ocular vessels and establish the superficial vasculature as a model for studying vascular patterning in the context of the developing eye.
Assuntos
Vasos Sanguíneos/embriologia , Olho/irrigação sanguínea , Peixe-Zebra/fisiologia , Animais , Animais Geneticamente ModificadosRESUMO
When animals explore an unfamiliar environment, they gather information that enables them to form a cognitive representation of that environment and to use it subsequently in traveling there. In the present study, rats were tested in a large arena as singles, then in dyads, and finally, again as singles, in order to examine the effect of the social environment on exploration. Traveling in dyads facilitated exploration compared to the behavior of the same rats when they explored alone. Specifically, each rat in a dyad traveled a greater distance with higher velocity and took wider turns compared to its lone traveling. Moreover, rats in dyads spent a long time together, shared a home base, and when traveling in the same direction, one rat was leading the other. In addition to exploring the same locations, leaders explored more "private" locations, not visited by the other rat. Features of the dyad behavior were carried over to the behavior of the same rats when tested as individuals, after the dyad trial. Compared to singles, dyads represent the first step toward grouping, and it is suggested that the conspicuous change between the behavior of a rat as single compared to its behavior when in a dyad should be greater than any further changes that may occur in spatial cognitive behavior of triads, quartets, or larger groups. In other words, while the present changes in spatial cognition observed in dyads represent a small step toward grouping, they are a giant leap for the individual.
Assuntos
Ratos/psicologia , Comportamento Social , Comportamento Espacial , Animais , Comportamento Exploratório , Masculino , Ratos Sprague-DawleyRESUMO
Ocular coloboma is a potentially blinding congenital eye malformation caused by failure of optic fissure closure during early embryogenesis. The optic fissure is a ventral groove that forms during optic cup morphogenesis, and through which hyaloid artery and vein enter and leave the developing eye, respectively. After hyaloid artery and vein formation, the optic fissure closes around them. The mechanisms underlying optic fissure closure are poorly understood, and whether and how this process is influenced by hyaloid vessel development is unknown. Here we show that a loss-of-function mutation in lmo2, a gene specifically required for hematopoiesis and vascular development, results in failure of optic fissure closure in zebrafish. Analysis of ocular blood vessels in lmo2 mutants reveals that some vessels are severely dilated, including the hyaloid vein. Remarkably, reducing vessel size leads to rescue of optic fissure phenotype. Our results reveal a new mechanism leading to coloboma, whereby malformed blood vessels interfere with eye morphogenesis.