Enrica Bianchi | The Wellcome Trust Sanger Institute (original) (raw)

Papers by Enrica Bianchi

Research paper thumbnail of Divergent evolution of vitamin B9 binding underlies Juno-mediated adhesion of mammalian gametes

Current biology : CB, Jan 8, 2016

The interaction between egg and sperm is the first necessary step of fertilization in all sexuall... more The interaction between egg and sperm is the first necessary step of fertilization in all sexually reproducing organisms. A decade-long search for a protein pair mediating this event in mammals culminated in the identification of the glycosylphosphatidylinositol (GPI)-anchored glycoprotein Juno as the egg plasma membrane receptor of sperm Izumo1 [1,2]. The Juno-Izumo1 interaction was shown to be essential for fertilization since mice lacking either gene exhibit sex-specific sterility, making these proteins promising non-hormonal contraceptive targets [1,3]. No structural information is available on how gamete membranes interact at fertilization, and it is unclear how Juno - which was previously named folate receptor (FR) 4, based on sequence similarity considerations - triggers membrane adhesion by binding Izumo1. Here, we report the crystal structure of Juno and find that the overall fold is similar to that of FRα and FRβ but with significant flexibility within the area that corres...

Research paper thumbnail of Traumatic events and life-style in ancient Italian populations

Collegium antropologicum

Traumatic lesions are commonly found in archaeological skeletal samples and provide useful inform... more Traumatic lesions are commonly found in archaeological skeletal samples and provide useful information about various behavioral and cultural aspects of the populations. Our aim was to evaluate the relationship between the distribution and types of skeletal traumatic lesions and the different life-styles of past populations. We examined three necropolises in central Italy. Pozzilli (VI-IV century BC) and Quadrella (I-IV century AD) are from the same geographical area (Molise) but belong to different periods; Novilara (IX-VI century BC) is located in Marche but belongs to the Iron Age like Pozzilli. The lesions observed at Pozzilli seem not to be accidental, whereas the traumas observed at Quadrella can be attributed to occasional, unintentional events. Cranial injuries observed at Novilara strengthen the hypothesis that the population was composed, at least in part, of warriors. Our results suggest the presence of a relationship between skeletal traumatic lesions and life-styles of p...

Research paper thumbnail of The challenges involved in elucidating the molecular basis of sperm-egg recognition in mammals and approaches to overcome them

Cell and Tissue Research

Sexual reproduction is used by many different organisms to create a new generation of genetically... more Sexual reproduction is used by many different organisms to create a new generation of genetically distinct progeny. Cells originating from separate sexes or mating types segregate their genetic material into haploid gametes which must then recognize and fuse with each other in a process known as fertilization to form a diploid zygote. Despite the central importance of fertilization, we know remarkably little about the molecular mechanisms that are involved in how gametes recognize each other, particularly in mammals, although the proteins that are displayed on their surfaces are almost certainly involved. This paucity of knowledge is largely due to both the unique biological properties of mammalian gametes (sperm and egg) which make them experimentally difficult to manipulate, and the technical challenges of identifying interactions between membrane-embedded cell surface receptor proteins. In this review, we will discuss our current knowledge of animal gamete recognition, highlighti...

Research paper thumbnail of Cross-species fertilization: the hamster egg receptor, Juno, binds the human sperm ligand, Izumo1

Philosophical transactions of the Royal Society of London. Series B, Biological sciences, Jan 5, 2015

Fertilization is the culminating event in sexual reproduction and requires the recognition and fu... more Fertilization is the culminating event in sexual reproduction and requires the recognition and fusion of the haploid sperm and egg to form a new diploid organism. Specificity in these recognition events is one reason why sperm and eggs from different species are not normally compatible. One notable exception is the unusual ability of zona-free eggs from the Syrian golden hamster (Mesocricetus auratus) to recognize and fuse with human sperm, a phenomenon that has been exploited to assess sperm quality in assisted fertility treatments. Following our recent finding that the interaction between the sperm and egg recognition receptors Izumo1 and Juno is essential for fertilization, we now demonstrate concordance between the ability of Izumo1 and Juno from different species to interact, and the ability of their isolated gametes to cross-fertilize each other in vitro. In particular, we show that Juno from the golden hamster can directly interact with human Izumo1. These data suggest that t...

Research paper thumbnail of Izumo meets Juno: preventing polyspermy in fertilization

Cell cycle (Georgetown, Tex.), 2014

Research paper thumbnail of Isolation and Analyses of Enriched Populations of Male Mouse Germ Cells by Sedimentation Velocity: The Centrifugal Elutriation

Methods in Molecular Biology, 2009

Research paper thumbnail of 1800 MHz radiofrequency (mobile phones, different Global System for Mobile communication modulations) does not affect apoptosis and heat shock protein 70 level in peripheral blood mononuclear cells from young and old donors

International Journal of Radiation Biology, 2004

Research paper thumbnail of FLASH is essential during early embryogenesis and cooperates with p73 to regulate histone gene transcription

Oncogene, 2011

Replication-dependent histone gene expression is a fundamental process occurring in S-phase under... more Replication-dependent histone gene expression is a fundamental process occurring in S-phase under the control of the cyclin-E/CDK2 complex. This process is regulated by a number of proteins, including Flice-Associated Huge Protein (FLASH) (CASP8AP2), concentrated in specific nuclear organelles known as HLBs. FLASH regulates both histone gene transcription and mRNA maturation, and its downregulation in vitro results in the depletion of the histone pull and cell-cycle arrest in S-phase. Here we show that the transcription factor p73 binds to FLASH and is part of the complex that regulates histone gene transcription. Moreover, we created a novel gene trap to disrupt FLASH in mice, and we show that homozygous deletion of FLASH results in early embryonic lethality, owing to arrest of FLASH(-/-) embryos at the morula stage. These results indicate that FLASH is an essential, non-redundant regulator of histone transcription and cell cycle during embryogenesis.

Research paper thumbnail of Phosphorylation of EIF4E by MNK Supports Proliferation of Pancreatic Cancer Cells

Research paper thumbnail of Post-transcriptional control of Gene expression in mouse early embryo development: A view from the tip of the Iceberg

Research paper thumbnail of Sam68 regulates translation of target mRNAs in male germ cells, necessary for mouse spermatogenesis

The Journal of Cell Biology, 2009

Research paper thumbnail of In Vitro Exposure of Human Lymphocytes to 900 MHz CW and GSM Modulated Radiofrequency: Studies of Proliferation, Apoptosis and Mitochondrial Membrane Potential

Radiation Research, 2004

The aim of this study was to investigate the nonthermal effects of radiofrequency (RF) fields on ... more The aim of this study was to investigate the nonthermal effects of radiofrequency (RF) fields on human immune cells exposed to a Global System for Mobile Communication (GSM) signal generated by a commercial cellular phone and by a sinusoidal non-modulated signal. To assess whether mobile phone RF-field exposure affects human immune cell functions, peripheral blood mononuclear cells (PBMCs) from healthy donors were exposed in vitro to a 900 MHz GSM or continuous-wave (CW) RF field 1 h/day for 3 days in a transverse electromagnetic mode (TEM) cell system (70-76 mW/kg average specific absorption rate, SAR). The cells were cultured for 48 or 72 h, and the following end points were studied: (1) mitogen-induced proliferation; (2) cell cycle progression; (3) spontaneous and 2-deoxy-D-ribose (dRib)-induced apoptosis; (4) mitochondrial membrane potential modifications during spontaneous and dRib-induced-apoptosis. Data obtained from cells exposed to a GSM-modulated RF field showed a slight decrease in cell proliferation when PBMCs were stimulated with the lowest mitogen concentration and a slight increase in the number of cells with altered distribution of phosphatidylserine across the membrane. On the other hand, cell cycle phases, mitochondrial membrane potential and susceptibility to apoptosis were found to be unaffected by the RF field. When cells were exposed to a CW RF field, no significant modifications were observed in comparison with sham-exposed cells for all the end points investigated.

Research paper thumbnail of Juno is the egg Izumo receptor and is essential for mammalian fertilization

Research paper thumbnail of Expression of stemness markers in mouse parthenogenetic-diploid blastocysts is influenced by slight variation of activation protocol adopted

In Vitro Cellular & Developmental Biology - Animal, 2010

The importance of obtaining stem cells through alternative methods has increased progressively in... more The importance of obtaining stem cells through alternative methods has increased progressively in the recent years due to the potential role that embryonic stem (ES) cells play in the field of regenerative medicine. In this regard, generation of parthenogenetic blastocysts allows the production of ethic-free ES cells without the need to manipulate normal embryos. Our work was aimed at clarifying whether variations in the method adopted to generate diploid parthenogenetic blastocysts could determine differences in the quality of blastocysts produced. In vitro development of mouse oocytes activated with three protocols, using Sr2+ and cytochalasin for different time, was compared with that of in vivo fertilized embryos. We have evaluated the efficiency of blastocyst formation and analysed the expression pattern of the stemness markers OCT4, CDX2, and NANOG. Our results indicate that the yield of diploid parthenogenotes and the segregation of the stemness marker OCT4 in the developing blastocyst are influenced by the parthenogenetic protocol adopted. Particularly, even if all methods tested allowed the production of blastocysts in vitro, the correct segregation of OCT4 occurred only in blastocysts developed from oocytes concomitantly treated for 4 h with Sr2+ and cytochalasin D. Our results indicate that the protocol employed to develop parthenogenetic blastocysts in vitro affects the quality of cells in the inner cell mass.

Research paper thumbnail of Ablation of the Sam68 gene impairs female fertility and gonadotropin-dependent follicle development

Human Molecular Genetics, 2010

Sam68 is a multifunctional RNA-binding protein highly expressed in the gonads, whose ablation cau... more Sam68 is a multifunctional RNA-binding protein highly expressed in the gonads, whose ablation causes male infertility. Herein, we have investigated Sam68 expression in the adult ovary and its function in female fertility. Immunohistochemistry showed that Sam68 was localized in the nucleus of oocytes and follicular cells at all stages of folliculogenesis. Sam68(-/-) females were severely subfertile, and they showed a delay in the age of first pregnancy, increased breeding time for successful pregnancy and yielded smaller litters. Morphological analyses indicated a significant reduction in the number of secondary and pre-antral follicles in the ovary. These defects were associated with alteration of oestrous cycles and a reduced number of ovulated oocytes, which were only partially restored by the administration of exogenous gonadotropins. Crosslinking/immunoprecipitation experiments showed that Sam68 directly binds the mRNAs for the follicle-stimulating hormone (FSH) and the luteinizing hormone receptors (Fshr and Lhcgr), which were downregulated in ovaries of adult knockout females. Stimulation of immature females with FSH-like pregnant mare serum gonadotropin (PMSG), or of follicular cells with the FSH second messenger analogue 8Br-cAMP, caused the upregulation of Sam68. The increase in Sam68 levels paralleled that of the Fshr and Lhcgr mRNAs in the pre-ovulatory follicle and was required to allow accumulation of these transcripts in follicular cells. These studies identify a new crucial function for Sam68 in the regulation of female fertility and indicate that this protein is required to insure proper expression of the gonadotropin receptor transcripts in pre-ovulatory follicles in adult ovary.

Research paper thumbnail of Dynamic expression of the RNA-binding protein Sam68 during mouse pre-implantation development

Gene Expression Patterns, 2008

Research paper thumbnail of Controllo traduzionale dell'espressione genica all'inizio dell'embriogenesi

Research paper thumbnail of Divergent evolution of vitamin B9 binding underlies Juno-mediated adhesion of mammalian gametes

Current biology : CB, Jan 8, 2016

The interaction between egg and sperm is the first necessary step of fertilization in all sexuall... more The interaction between egg and sperm is the first necessary step of fertilization in all sexually reproducing organisms. A decade-long search for a protein pair mediating this event in mammals culminated in the identification of the glycosylphosphatidylinositol (GPI)-anchored glycoprotein Juno as the egg plasma membrane receptor of sperm Izumo1 [1,2]. The Juno-Izumo1 interaction was shown to be essential for fertilization since mice lacking either gene exhibit sex-specific sterility, making these proteins promising non-hormonal contraceptive targets [1,3]. No structural information is available on how gamete membranes interact at fertilization, and it is unclear how Juno - which was previously named folate receptor (FR) 4, based on sequence similarity considerations - triggers membrane adhesion by binding Izumo1. Here, we report the crystal structure of Juno and find that the overall fold is similar to that of FRα and FRβ but with significant flexibility within the area that corres...

Research paper thumbnail of Traumatic events and life-style in ancient Italian populations

Collegium antropologicum

Traumatic lesions are commonly found in archaeological skeletal samples and provide useful inform... more Traumatic lesions are commonly found in archaeological skeletal samples and provide useful information about various behavioral and cultural aspects of the populations. Our aim was to evaluate the relationship between the distribution and types of skeletal traumatic lesions and the different life-styles of past populations. We examined three necropolises in central Italy. Pozzilli (VI-IV century BC) and Quadrella (I-IV century AD) are from the same geographical area (Molise) but belong to different periods; Novilara (IX-VI century BC) is located in Marche but belongs to the Iron Age like Pozzilli. The lesions observed at Pozzilli seem not to be accidental, whereas the traumas observed at Quadrella can be attributed to occasional, unintentional events. Cranial injuries observed at Novilara strengthen the hypothesis that the population was composed, at least in part, of warriors. Our results suggest the presence of a relationship between skeletal traumatic lesions and life-styles of p...

Research paper thumbnail of The challenges involved in elucidating the molecular basis of sperm-egg recognition in mammals and approaches to overcome them

Cell and Tissue Research

Sexual reproduction is used by many different organisms to create a new generation of genetically... more Sexual reproduction is used by many different organisms to create a new generation of genetically distinct progeny. Cells originating from separate sexes or mating types segregate their genetic material into haploid gametes which must then recognize and fuse with each other in a process known as fertilization to form a diploid zygote. Despite the central importance of fertilization, we know remarkably little about the molecular mechanisms that are involved in how gametes recognize each other, particularly in mammals, although the proteins that are displayed on their surfaces are almost certainly involved. This paucity of knowledge is largely due to both the unique biological properties of mammalian gametes (sperm and egg) which make them experimentally difficult to manipulate, and the technical challenges of identifying interactions between membrane-embedded cell surface receptor proteins. In this review, we will discuss our current knowledge of animal gamete recognition, highlighti...

Research paper thumbnail of Cross-species fertilization: the hamster egg receptor, Juno, binds the human sperm ligand, Izumo1

Philosophical transactions of the Royal Society of London. Series B, Biological sciences, Jan 5, 2015

Fertilization is the culminating event in sexual reproduction and requires the recognition and fu... more Fertilization is the culminating event in sexual reproduction and requires the recognition and fusion of the haploid sperm and egg to form a new diploid organism. Specificity in these recognition events is one reason why sperm and eggs from different species are not normally compatible. One notable exception is the unusual ability of zona-free eggs from the Syrian golden hamster (Mesocricetus auratus) to recognize and fuse with human sperm, a phenomenon that has been exploited to assess sperm quality in assisted fertility treatments. Following our recent finding that the interaction between the sperm and egg recognition receptors Izumo1 and Juno is essential for fertilization, we now demonstrate concordance between the ability of Izumo1 and Juno from different species to interact, and the ability of their isolated gametes to cross-fertilize each other in vitro. In particular, we show that Juno from the golden hamster can directly interact with human Izumo1. These data suggest that t...

Research paper thumbnail of Izumo meets Juno: preventing polyspermy in fertilization

Cell cycle (Georgetown, Tex.), 2014

Research paper thumbnail of Isolation and Analyses of Enriched Populations of Male Mouse Germ Cells by Sedimentation Velocity: The Centrifugal Elutriation

Methods in Molecular Biology, 2009

Research paper thumbnail of 1800 MHz radiofrequency (mobile phones, different Global System for Mobile communication modulations) does not affect apoptosis and heat shock protein 70 level in peripheral blood mononuclear cells from young and old donors

International Journal of Radiation Biology, 2004

Research paper thumbnail of FLASH is essential during early embryogenesis and cooperates with p73 to regulate histone gene transcription

Oncogene, 2011

Replication-dependent histone gene expression is a fundamental process occurring in S-phase under... more Replication-dependent histone gene expression is a fundamental process occurring in S-phase under the control of the cyclin-E/CDK2 complex. This process is regulated by a number of proteins, including Flice-Associated Huge Protein (FLASH) (CASP8AP2), concentrated in specific nuclear organelles known as HLBs. FLASH regulates both histone gene transcription and mRNA maturation, and its downregulation in vitro results in the depletion of the histone pull and cell-cycle arrest in S-phase. Here we show that the transcription factor p73 binds to FLASH and is part of the complex that regulates histone gene transcription. Moreover, we created a novel gene trap to disrupt FLASH in mice, and we show that homozygous deletion of FLASH results in early embryonic lethality, owing to arrest of FLASH(-/-) embryos at the morula stage. These results indicate that FLASH is an essential, non-redundant regulator of histone transcription and cell cycle during embryogenesis.

Research paper thumbnail of Phosphorylation of EIF4E by MNK Supports Proliferation of Pancreatic Cancer Cells

Research paper thumbnail of Post-transcriptional control of Gene expression in mouse early embryo development: A view from the tip of the Iceberg

Research paper thumbnail of Sam68 regulates translation of target mRNAs in male germ cells, necessary for mouse spermatogenesis

The Journal of Cell Biology, 2009

Research paper thumbnail of In Vitro Exposure of Human Lymphocytes to 900 MHz CW and GSM Modulated Radiofrequency: Studies of Proliferation, Apoptosis and Mitochondrial Membrane Potential

Radiation Research, 2004

The aim of this study was to investigate the nonthermal effects of radiofrequency (RF) fields on ... more The aim of this study was to investigate the nonthermal effects of radiofrequency (RF) fields on human immune cells exposed to a Global System for Mobile Communication (GSM) signal generated by a commercial cellular phone and by a sinusoidal non-modulated signal. To assess whether mobile phone RF-field exposure affects human immune cell functions, peripheral blood mononuclear cells (PBMCs) from healthy donors were exposed in vitro to a 900 MHz GSM or continuous-wave (CW) RF field 1 h/day for 3 days in a transverse electromagnetic mode (TEM) cell system (70-76 mW/kg average specific absorption rate, SAR). The cells were cultured for 48 or 72 h, and the following end points were studied: (1) mitogen-induced proliferation; (2) cell cycle progression; (3) spontaneous and 2-deoxy-D-ribose (dRib)-induced apoptosis; (4) mitochondrial membrane potential modifications during spontaneous and dRib-induced-apoptosis. Data obtained from cells exposed to a GSM-modulated RF field showed a slight decrease in cell proliferation when PBMCs were stimulated with the lowest mitogen concentration and a slight increase in the number of cells with altered distribution of phosphatidylserine across the membrane. On the other hand, cell cycle phases, mitochondrial membrane potential and susceptibility to apoptosis were found to be unaffected by the RF field. When cells were exposed to a CW RF field, no significant modifications were observed in comparison with sham-exposed cells for all the end points investigated.

Research paper thumbnail of Juno is the egg Izumo receptor and is essential for mammalian fertilization

Research paper thumbnail of Expression of stemness markers in mouse parthenogenetic-diploid blastocysts is influenced by slight variation of activation protocol adopted

In Vitro Cellular & Developmental Biology - Animal, 2010

The importance of obtaining stem cells through alternative methods has increased progressively in... more The importance of obtaining stem cells through alternative methods has increased progressively in the recent years due to the potential role that embryonic stem (ES) cells play in the field of regenerative medicine. In this regard, generation of parthenogenetic blastocysts allows the production of ethic-free ES cells without the need to manipulate normal embryos. Our work was aimed at clarifying whether variations in the method adopted to generate diploid parthenogenetic blastocysts could determine differences in the quality of blastocysts produced. In vitro development of mouse oocytes activated with three protocols, using Sr2+ and cytochalasin for different time, was compared with that of in vivo fertilized embryos. We have evaluated the efficiency of blastocyst formation and analysed the expression pattern of the stemness markers OCT4, CDX2, and NANOG. Our results indicate that the yield of diploid parthenogenotes and the segregation of the stemness marker OCT4 in the developing blastocyst are influenced by the parthenogenetic protocol adopted. Particularly, even if all methods tested allowed the production of blastocysts in vitro, the correct segregation of OCT4 occurred only in blastocysts developed from oocytes concomitantly treated for 4 h with Sr2+ and cytochalasin D. Our results indicate that the protocol employed to develop parthenogenetic blastocysts in vitro affects the quality of cells in the inner cell mass.

Research paper thumbnail of Ablation of the Sam68 gene impairs female fertility and gonadotropin-dependent follicle development

Human Molecular Genetics, 2010

Sam68 is a multifunctional RNA-binding protein highly expressed in the gonads, whose ablation cau... more Sam68 is a multifunctional RNA-binding protein highly expressed in the gonads, whose ablation causes male infertility. Herein, we have investigated Sam68 expression in the adult ovary and its function in female fertility. Immunohistochemistry showed that Sam68 was localized in the nucleus of oocytes and follicular cells at all stages of folliculogenesis. Sam68(-/-) females were severely subfertile, and they showed a delay in the age of first pregnancy, increased breeding time for successful pregnancy and yielded smaller litters. Morphological analyses indicated a significant reduction in the number of secondary and pre-antral follicles in the ovary. These defects were associated with alteration of oestrous cycles and a reduced number of ovulated oocytes, which were only partially restored by the administration of exogenous gonadotropins. Crosslinking/immunoprecipitation experiments showed that Sam68 directly binds the mRNAs for the follicle-stimulating hormone (FSH) and the luteinizing hormone receptors (Fshr and Lhcgr), which were downregulated in ovaries of adult knockout females. Stimulation of immature females with FSH-like pregnant mare serum gonadotropin (PMSG), or of follicular cells with the FSH second messenger analogue 8Br-cAMP, caused the upregulation of Sam68. The increase in Sam68 levels paralleled that of the Fshr and Lhcgr mRNAs in the pre-ovulatory follicle and was required to allow accumulation of these transcripts in follicular cells. These studies identify a new crucial function for Sam68 in the regulation of female fertility and indicate that this protein is required to insure proper expression of the gonadotropin receptor transcripts in pre-ovulatory follicles in adult ovary.

Research paper thumbnail of Dynamic expression of the RNA-binding protein Sam68 during mouse pre-implantation development

Gene Expression Patterns, 2008

Research paper thumbnail of Controllo traduzionale dell'espressione genica all'inizio dell'embriogenesi