Marcello Ceci | University of Tuscia (Università degli Studi della Tuscia, Viterbo) (original) (raw)
Papers by Marcello Ceci
Cellular and Molecular Life Sciences
Dystrophin (dys) mutations predispose Duchenne muscular disease (DMD) patients to brain and retin... more Dystrophin (dys) mutations predispose Duchenne muscular disease (DMD) patients to brain and retinal complications. Although different dys variants, including long dys products, are expressed in the retina, their function is largely unknown. We investigated the putative role of full-length dystrophin in the homeostasis of neuro-retina and its impact on synapsis stabilization and cell fate. Retinas of mdx mice, the most used DMD model which does not express the 427-KDa dys protein (Dp427), showed overlapped cell death and impaired autophagy. Apoptotic neurons in the outer plexiform/inner nuclear layer and the ganglion cell layer had an impaired autophagy with accumulated autophagosomes. The autophagy dysfunction localized at photoreceptor axonal terminals and bipolar, amacrine, and ganglion cells. The absence of Dp427 does not cause a severe phenotype but alters the neuronal architecture, compromising mainly the pre-synaptic photoreceptor terminals and their post-synaptic sites. The a...
Fishes
Antimicrobial peptides (AMPS) are ancestral components in the evolution of immunity from protozoa... more Antimicrobial peptides (AMPS) are ancestral components in the evolution of immunity from protozoans to metazoans. Their expression can be constitutive or inducible by infectious challenge. Although characterized in detail in their structure and activity, the temporal and spatial expression of AMPS during vertebrate embryogenesis is still poorly understood. In the present study, we identified selected AMPs in zebrafish, and characterized their expression during early development, and upon experimental immune challenge in adult animals, with the goal of establishing this genetically-tractable model system for further AMP studies. By mining available genomic databases, zebrafish AMP sequences homologous to AMPs from other vertebrates were selected for further study. These included parasin I and its enzyme cathepsin D, β-defensin (DB1), liver-expressed antimicrobial peptide 2 (LEAP2), bactericidal permeability-increasing protein (BPI), and chromogranin-A and-B (CgA and CgB). Specific primers were designed for RT-PCR amplification of each AMP gene of interest and amplicons between 242 bp and 504 bp were obtained from RNA extracted from adult zebrafish. Sequencing of the amplicons and alignment of their deduced amino acid sequences with those from AMPs from other vertebrate species confirmed their identity. The temporal expression of AMPs was investigated by RT-PCR analysis in fertilized oocytes, embryos, and adult individuals. Parasin I and chatepsin D transcripts were detectable immediately after fertilization, while the transcripts for CgA and CgB became evident starting at 48 h post fertilization. Mature transcripts of LEAP2 and DB1 were detectable only in the adult zebrafish, while BPI transcripts were detectable starting from the 12th day post fertilization. To explore the possible upregulation of AMP expression by infectious challenge, experiments were carried out in adult zebrafish by intraperitoneal injection of a cocktail of lipopolysaccharide and lipoteichoic acid. Except for CgA and CgB, amplicons corresponding to all tested AMPs showed stronger signals in the experimental animals as compared to the unchallenged controls. This study provided information on the early expression of AMPs in zebrafish from ontogeny to adulthood and their inducibility by microbials. This information could be useful to actuate new prophylactic strategies as an alternative to the use of antibiotics in culture.
Experimental Cell Research
Cell death discovery, 2018
Zebrafish could be an interesting translational model to understand and improve the post-infarcti... more Zebrafish could be an interesting translational model to understand and improve the post-infarction trial and possible regeneration in humans. The adult zebrafish is able to regenerate efficiently after resecting nearly 20% of the ventricular apex. This process requires the concert activation of the epicardium and endocardium, as well as trans-differentiation of pre-existing cardiomyocytes that together replace the lost tissue. The molecular mechanisms involved in this activation process are not completely clarified. In this work, in order to investigate if the downregulation of these miRNAs (miRs) are linked with the activation of epicardium, the expressions of miR-133a, b and miR-1 during regeneration were analysed. qPCR analyses in whole-heart, or from distinct dissected epicardial cells comparing to regenerative clot (containing cardiomyocytes, fibroblasts and endocardial cells) by a laser-micro-dissector, have indicated that already at 24 h there is a downregulation of miRs: (1...
Human Molecular Genetics, 2017
TDP-43 is a well known RNA binding protein involved in the pathogenesis of Amyotrophic Lateral Sc... more TDP-43 is a well known RNA binding protein involved in the pathogenesis of Amyotrophic Lateral Sclerosis (ALS) and Frontotemporal Lobar Dementia (FTLD). In physiological conditions, TDP-43 mainly localizes in the nucleus and shuttles, at least in neurons, to the cytoplasm to form TDP-43 RNA granules. In the nucleus, TDP-43 participates to the expression and splicing of RNAs, while in the cytoplasm its functions range from transport to translation of specific mRNAs. However, if loss or gain of these TDP-43 functions are affected in ALS/FTLD pathogenesis is not clear. Here, we report that TDP-43 localizes on ribosomes not only in primary neurons but also in SH-SY5Y human neuroblastoma cells. We find that binding of TDP-43 to the translational machinery is mediated by an interaction with a specific ribosomal protein, RACK1, and that an increase in cytoplasmic TDP-43 represses global protein synthesis, an effect which is rescued by overexpression of RACK1. Ribosomal loss of RACK1, which excludes TDP-43 from the translational machinery, remarkably reduces formation of TDP-43 cytoplasmic inclusions in neuroblastoma cells. Finally, we corroborate the interaction between TDP-43 and RACK1 on polyribosomes of neuroblastoma cells with mis-localization of RACK1 on TDP-43 positive cytoplasmic inclusions in motor neurons of ALS patients. In conclusions, results from this study suggest that TDP-43 represents a translational repressor not only for specific mRNAs but for overall translation and that its binding to polyribosomes through RACK1 may promote, under conditions inducing ALS pathogenesis, the formation of cytoplasmic inclusions.
Circulation, Oct 28, 2008
PLoS ONE, 2012
In neurons, specific mRNAs are transported in a translationally repressed manner along dendrites ... more In neurons, specific mRNAs are transported in a translationally repressed manner along dendrites or axons by transport ribonucleic-protein complexes called RNA granules. ZBP1 is one RNA binding protein present in transport RNPs, where it transports and represses the translation of cotransported mRNAs, including b-actin mRNA. The release of b-actin mRNA from ZBP1 and its subsequent translation depends on the phosphorylation of ZBP1 by Src kinase, but little is known about how this process is regulated. Here we demonstrate that the ribosomal-associated protein RACK1, another substrate of Src, binds the b-actin mRNA/ZBP1 complex on ribosomes and contributes to the release of b-actin mRNA from ZBP1 and to its translation. We identify the Src binding and phosphorylation site Y246 on RACK1 as the critical site for the binding to the bactin mRNA/ZBP1 complex. Based on these results we propose RACK1 as a ribosomal scaffold protein for specific mRNA-RBP complexes to tightly regulate the translation of specific mRNAs.
During adulthood, the neurotrophin Nerve Growth Factor (NGF) sensitizes nociceptors, thereby incr... more During adulthood, the neurotrophin Nerve Growth Factor (NGF) sensitizes nociceptors, thereby increasing the response to noxious stimuli. The relationship between NGF and pain is supported by genetic evidence: mutations in the NGF TrkA receptor in patients affected by an hereditary rare disease (Hereditary Sensory and Autonomic Neuropathy type IV, HSAN IV) determine a congenital form of severe pain insensitivity, with mental retardation, while a mutation in NGFB gene, leading to the aminoacid substitution R100W in mature NGF, determines a similar loss of pain perception, without overt cognitive neurological defects (HSAN V). The R100W mutation provokes a reduced processing of proNGF to mature NGF in cultured cells and a higher percentage of neurotrophin secreted is in the proNGF form. Moreover, using Surface Plasmon Resonance we showed that the R100W mutation does not affect NGF binding to TrkA, while it abolishes NGF binding to p75NTR receptors. However, it remains to be clarified whether the major impact of the mutation is on the biological function of proNGF or of mature NGF and to what extent the effects of the R100W mutation on the HSAN V clinical phenotype are developmental, or whether they reflect an impaired effectiveness of NGF to regulate and mediate nociceptive transmission in adult sensory neurons. Here we show that the R100 mutation selectively alters some of the signaling pathways activated downstream of TrkA NGF receptors. NGFR100 mutants maintain identical neurotrophic and neuroprotective properties in a variety of cell assays, while displaying a significantly reduced pain-inducing activity in vivo (n = 8-10 mice/group). We also show that proNGF has a significantly reduced nociceptive activity, with respect to NGF. Both sets of results jointly contribute to elucidating the mechanisms underlying the clinical HSAN V manifestations, and to clarifying which receptors and intracellular signaling cascades participate in the pain sensitizing action of NGF.
PLoS ONE, 2012
Nerve Growth Factor (NGF) is being considered as a therapeutic candidate for Alzheimer's disease ... more Nerve Growth Factor (NGF) is being considered as a therapeutic candidate for Alzheimer's disease (AD) treatment but the clinical application is hindered by its potent pro-nociceptive activity. Thus, to reduce systemic exposure that would induce pain, in recent clinical studies NGF was administered through an invasive intracerebral gene-therapy approach. Our group demonstrated the feasibility of a non-invasive intranasal delivery of NGF in a mouse model of neurodegeneration. NGF therapeutic window could be further increased if its nociceptive effects could be avoided altogether. In this study we exploit forms of NGF, mutated at residue R100, inspired by the human genetic disease HSAN V (Hereditary Sensory Autonomic Neuropathy Type V), which would allow increasing the dose of NGF without triggering pain. We show that ''painless'' hNGF displays full neurotrophic and anti-amyloidogenic activities in neuronal cultures, and a reduced nociceptive activity in vivo. When administered intranasally to APPxPS1 mice (n = 8), hNGFP61S/R100E prevents the progress of neurodegeneration and of behavioral deficits. These results demonstrate the in vivo neuroprotective and antiamyloidogenic properties of hNGFR100 mutants and provide a rational basis for the development of ''painless'' hNGF variants as a new generation of therapeutics for neurodegenerative diseases.
Toxicology in Vitro, 2001
Herbicides are chemical compounds widely used in agriculture. As their intensive application is b... more Herbicides are chemical compounds widely used in agriculture. As their intensive application is becoming a cause of environmental pollution, detailed and more sophisticated investigations are needed to understand better their consequences at the biological level. After herbicides are dispersed in the ®elds, they establish chemical interactions with both target and non-target plants. In both cases, herbicides can interact with the plant reproductive apparatus; consequently they could play a role during the fertilisation process in higher plants. Using an antibody to the a-tubulin subunit in immuno¯uorescence and immunoelectron microscopy techniques, we investigated the distribution of microtubules in Nicotiana tabacum pollen tubes grown under in vitro conditions in the presence of ®ve dierent herbicides selected among those used frequently in central Italy. Herbicides have a speci®c eect on the microtubular apparatus of both pollen tube and generative cell. In addition to other tests and assays, these results suggest that the microtubule cytoskeleton of pollen tubes can be used as a bioindicator for studying the toxicity eects induced by herbicides.
STEM CELLS, 2014
Adult neurogenesis is a multistep process regulated by several extrinsic factors, including neuro... more Adult neurogenesis is a multistep process regulated by several extrinsic factors, including neurotrophins. Among them, little is known about the role of nerve growth factor (NGF) in the neurogenic niches of the mouse. Here we analyzed the biology of adult neural stem cells (NSCs) from the subventricular zone (SVZ) of AD11 anti-NGF transgenic mice, in which the expression of the recombinant antibody aD11 leads to a chronic postnatal neutralization of endogenous NGF. We showed that AD11-NSCs proliferate 10-fold less, with respect to their control counterparts, and display a significant impairment in their ability to differentiate into b-tubulin positive neurons. We found a considerable reduction in the number of SVZ progenitors and neuroblasts also in vivo, which correlates with a lower number of newborn neurons in the olfactory bulbs of AD11 mice and a severe deficit in the ability of these mice to discriminate between different odors. We also demonstrated that, in AD11 mice, the morphology of both SVZ-resident and neurosphere-derived astrocytes is significantly altered. We were able to reproduce the AD11 phenotype in vitro, by acutely treating wild type NSCs with the anti-NGF antibody, further demonstrating that both the proliferation and the differentiation defects are due to the NGF deprivation. Consistently, the proliferative impairment of AD11 progenitors, as well as the atrophic morphology of AD11 astrocytes, can be partly rescued in vitro and in vivo by exogenous NGF addition. Altogether, our results demonstrate a causal link between NGF signaling and proper proliferation and differentiation of neural stem cells from the SVZ.
PLoS ONE, 2012
ABSTRACT [This corrects the article on p. e37555 in vol. 7.].
Medicine & Science in Sports & Exercise, 2007
Medicine & Science in Sports & Exercise, 2006
Page 1. Slide S e SS iONS Official Journal of the American College of Sports Medicine Vol. 38 No.... more Page 1. Slide S e SS iONS Official Journal of the American College of Sports Medicine Vol. 38 No. 5 Supplement S #583-760WEDNESDAY, MAY 31 | #761-918THURSDAY, JUNE 1 | #919-1067FRIDAY, JUNE 2 | #1068-1124 - SATURDAY, JUNE 3 ...
Journal of Molecular and Cellular Cardiology, 2004
Cardiomyocytes (CMCs) adapt to physiological or pathological stimuli by undergoing molecular chan... more Cardiomyocytes (CMCs) adapt to physiological or pathological stimuli by undergoing molecular changes which differentiate according to the specificity of the stimulus and eventually generate a phenotype with peculiar molecular characteristics. Here, we review the literature on the molecular mechanisms activated in the CMC during physiologic adaptation to stress, as opposed to maladaptation. The critical role of the IGF-1 receptor/PI3K/Akt signaling pathway during this process is described, including effector targets regulating inotropism and cell size.
Journal of Molecular and Cellular Cardiology, 2006
whilst undifferentiated and differentiate into CM. We examined the capacity of HESC-CM to survive... more whilst undifferentiated and differentiate into CM. We examined the capacity of HESC-CM to survive in the mouse heart. Methods: HESC, genetically marked by GFP (green fluorescent protein), were co-cultured with visceral endoderm-like cells, which induce differentiation into CM. Spontaneously beating areas, originating from HESC, were dissected and dissociated into small clumps containing 15% CM. In each SCID mouse 2-10 5 cells were injected intramyocardially. Between 2 and 4 weeks, HESC-CM were traced in cryosections by GFP-epifluorescence and characterized by immunofluorescent staining with humanand cardiomyocyte specific antibodies. Results: GFP-expressing cells were found both in scarred myocardium and aligned with host cardiomyocytes. Human origin of these cells was confirmed by staining for human nuclei and human mitochondria. 85% of the grafted human cells expressed a-actinin and had a sarcomeric structure. While mouse CM expressed only myosin light chain subtype MLC-2V, transplanted GFP cells expressed exclusively MLC-2A protein and not MLC-2V, indicating the absence of fusion with host CM. Cell survival was 0.2-1.4% of the total amount of transplanted cells and 2.0-12% of CM. Discussion: HESC-CM are able to survive in the mouse heart for at least 4 weeks after intramyocardial transplantation independently of fusion with host CM. The proportion of aactinin positive HESC-derived cells increases from 15% pre-to 85% post-transplantation, suggesting selective survival of HESC-CM in the heart, and/or differentiation of primed HESC into CM after transplantation.
Cellular and Molecular Life Sciences
Dystrophin (dys) mutations predispose Duchenne muscular disease (DMD) patients to brain and retin... more Dystrophin (dys) mutations predispose Duchenne muscular disease (DMD) patients to brain and retinal complications. Although different dys variants, including long dys products, are expressed in the retina, their function is largely unknown. We investigated the putative role of full-length dystrophin in the homeostasis of neuro-retina and its impact on synapsis stabilization and cell fate. Retinas of mdx mice, the most used DMD model which does not express the 427-KDa dys protein (Dp427), showed overlapped cell death and impaired autophagy. Apoptotic neurons in the outer plexiform/inner nuclear layer and the ganglion cell layer had an impaired autophagy with accumulated autophagosomes. The autophagy dysfunction localized at photoreceptor axonal terminals and bipolar, amacrine, and ganglion cells. The absence of Dp427 does not cause a severe phenotype but alters the neuronal architecture, compromising mainly the pre-synaptic photoreceptor terminals and their post-synaptic sites. The a...
Fishes
Antimicrobial peptides (AMPS) are ancestral components in the evolution of immunity from protozoa... more Antimicrobial peptides (AMPS) are ancestral components in the evolution of immunity from protozoans to metazoans. Their expression can be constitutive or inducible by infectious challenge. Although characterized in detail in their structure and activity, the temporal and spatial expression of AMPS during vertebrate embryogenesis is still poorly understood. In the present study, we identified selected AMPs in zebrafish, and characterized their expression during early development, and upon experimental immune challenge in adult animals, with the goal of establishing this genetically-tractable model system for further AMP studies. By mining available genomic databases, zebrafish AMP sequences homologous to AMPs from other vertebrates were selected for further study. These included parasin I and its enzyme cathepsin D, β-defensin (DB1), liver-expressed antimicrobial peptide 2 (LEAP2), bactericidal permeability-increasing protein (BPI), and chromogranin-A and-B (CgA and CgB). Specific primers were designed for RT-PCR amplification of each AMP gene of interest and amplicons between 242 bp and 504 bp were obtained from RNA extracted from adult zebrafish. Sequencing of the amplicons and alignment of their deduced amino acid sequences with those from AMPs from other vertebrate species confirmed their identity. The temporal expression of AMPs was investigated by RT-PCR analysis in fertilized oocytes, embryos, and adult individuals. Parasin I and chatepsin D transcripts were detectable immediately after fertilization, while the transcripts for CgA and CgB became evident starting at 48 h post fertilization. Mature transcripts of LEAP2 and DB1 were detectable only in the adult zebrafish, while BPI transcripts were detectable starting from the 12th day post fertilization. To explore the possible upregulation of AMP expression by infectious challenge, experiments were carried out in adult zebrafish by intraperitoneal injection of a cocktail of lipopolysaccharide and lipoteichoic acid. Except for CgA and CgB, amplicons corresponding to all tested AMPs showed stronger signals in the experimental animals as compared to the unchallenged controls. This study provided information on the early expression of AMPs in zebrafish from ontogeny to adulthood and their inducibility by microbials. This information could be useful to actuate new prophylactic strategies as an alternative to the use of antibiotics in culture.
Experimental Cell Research
Cell death discovery, 2018
Zebrafish could be an interesting translational model to understand and improve the post-infarcti... more Zebrafish could be an interesting translational model to understand and improve the post-infarction trial and possible regeneration in humans. The adult zebrafish is able to regenerate efficiently after resecting nearly 20% of the ventricular apex. This process requires the concert activation of the epicardium and endocardium, as well as trans-differentiation of pre-existing cardiomyocytes that together replace the lost tissue. The molecular mechanisms involved in this activation process are not completely clarified. In this work, in order to investigate if the downregulation of these miRNAs (miRs) are linked with the activation of epicardium, the expressions of miR-133a, b and miR-1 during regeneration were analysed. qPCR analyses in whole-heart, or from distinct dissected epicardial cells comparing to regenerative clot (containing cardiomyocytes, fibroblasts and endocardial cells) by a laser-micro-dissector, have indicated that already at 24 h there is a downregulation of miRs: (1...
Human Molecular Genetics, 2017
TDP-43 is a well known RNA binding protein involved in the pathogenesis of Amyotrophic Lateral Sc... more TDP-43 is a well known RNA binding protein involved in the pathogenesis of Amyotrophic Lateral Sclerosis (ALS) and Frontotemporal Lobar Dementia (FTLD). In physiological conditions, TDP-43 mainly localizes in the nucleus and shuttles, at least in neurons, to the cytoplasm to form TDP-43 RNA granules. In the nucleus, TDP-43 participates to the expression and splicing of RNAs, while in the cytoplasm its functions range from transport to translation of specific mRNAs. However, if loss or gain of these TDP-43 functions are affected in ALS/FTLD pathogenesis is not clear. Here, we report that TDP-43 localizes on ribosomes not only in primary neurons but also in SH-SY5Y human neuroblastoma cells. We find that binding of TDP-43 to the translational machinery is mediated by an interaction with a specific ribosomal protein, RACK1, and that an increase in cytoplasmic TDP-43 represses global protein synthesis, an effect which is rescued by overexpression of RACK1. Ribosomal loss of RACK1, which excludes TDP-43 from the translational machinery, remarkably reduces formation of TDP-43 cytoplasmic inclusions in neuroblastoma cells. Finally, we corroborate the interaction between TDP-43 and RACK1 on polyribosomes of neuroblastoma cells with mis-localization of RACK1 on TDP-43 positive cytoplasmic inclusions in motor neurons of ALS patients. In conclusions, results from this study suggest that TDP-43 represents a translational repressor not only for specific mRNAs but for overall translation and that its binding to polyribosomes through RACK1 may promote, under conditions inducing ALS pathogenesis, the formation of cytoplasmic inclusions.
Circulation, Oct 28, 2008
PLoS ONE, 2012
In neurons, specific mRNAs are transported in a translationally repressed manner along dendrites ... more In neurons, specific mRNAs are transported in a translationally repressed manner along dendrites or axons by transport ribonucleic-protein complexes called RNA granules. ZBP1 is one RNA binding protein present in transport RNPs, where it transports and represses the translation of cotransported mRNAs, including b-actin mRNA. The release of b-actin mRNA from ZBP1 and its subsequent translation depends on the phosphorylation of ZBP1 by Src kinase, but little is known about how this process is regulated. Here we demonstrate that the ribosomal-associated protein RACK1, another substrate of Src, binds the b-actin mRNA/ZBP1 complex on ribosomes and contributes to the release of b-actin mRNA from ZBP1 and to its translation. We identify the Src binding and phosphorylation site Y246 on RACK1 as the critical site for the binding to the bactin mRNA/ZBP1 complex. Based on these results we propose RACK1 as a ribosomal scaffold protein for specific mRNA-RBP complexes to tightly regulate the translation of specific mRNAs.
During adulthood, the neurotrophin Nerve Growth Factor (NGF) sensitizes nociceptors, thereby incr... more During adulthood, the neurotrophin Nerve Growth Factor (NGF) sensitizes nociceptors, thereby increasing the response to noxious stimuli. The relationship between NGF and pain is supported by genetic evidence: mutations in the NGF TrkA receptor in patients affected by an hereditary rare disease (Hereditary Sensory and Autonomic Neuropathy type IV, HSAN IV) determine a congenital form of severe pain insensitivity, with mental retardation, while a mutation in NGFB gene, leading to the aminoacid substitution R100W in mature NGF, determines a similar loss of pain perception, without overt cognitive neurological defects (HSAN V). The R100W mutation provokes a reduced processing of proNGF to mature NGF in cultured cells and a higher percentage of neurotrophin secreted is in the proNGF form. Moreover, using Surface Plasmon Resonance we showed that the R100W mutation does not affect NGF binding to TrkA, while it abolishes NGF binding to p75NTR receptors. However, it remains to be clarified whether the major impact of the mutation is on the biological function of proNGF or of mature NGF and to what extent the effects of the R100W mutation on the HSAN V clinical phenotype are developmental, or whether they reflect an impaired effectiveness of NGF to regulate and mediate nociceptive transmission in adult sensory neurons. Here we show that the R100 mutation selectively alters some of the signaling pathways activated downstream of TrkA NGF receptors. NGFR100 mutants maintain identical neurotrophic and neuroprotective properties in a variety of cell assays, while displaying a significantly reduced pain-inducing activity in vivo (n = 8-10 mice/group). We also show that proNGF has a significantly reduced nociceptive activity, with respect to NGF. Both sets of results jointly contribute to elucidating the mechanisms underlying the clinical HSAN V manifestations, and to clarifying which receptors and intracellular signaling cascades participate in the pain sensitizing action of NGF.
PLoS ONE, 2012
Nerve Growth Factor (NGF) is being considered as a therapeutic candidate for Alzheimer's disease ... more Nerve Growth Factor (NGF) is being considered as a therapeutic candidate for Alzheimer's disease (AD) treatment but the clinical application is hindered by its potent pro-nociceptive activity. Thus, to reduce systemic exposure that would induce pain, in recent clinical studies NGF was administered through an invasive intracerebral gene-therapy approach. Our group demonstrated the feasibility of a non-invasive intranasal delivery of NGF in a mouse model of neurodegeneration. NGF therapeutic window could be further increased if its nociceptive effects could be avoided altogether. In this study we exploit forms of NGF, mutated at residue R100, inspired by the human genetic disease HSAN V (Hereditary Sensory Autonomic Neuropathy Type V), which would allow increasing the dose of NGF without triggering pain. We show that ''painless'' hNGF displays full neurotrophic and anti-amyloidogenic activities in neuronal cultures, and a reduced nociceptive activity in vivo. When administered intranasally to APPxPS1 mice (n = 8), hNGFP61S/R100E prevents the progress of neurodegeneration and of behavioral deficits. These results demonstrate the in vivo neuroprotective and antiamyloidogenic properties of hNGFR100 mutants and provide a rational basis for the development of ''painless'' hNGF variants as a new generation of therapeutics for neurodegenerative diseases.
Toxicology in Vitro, 2001
Herbicides are chemical compounds widely used in agriculture. As their intensive application is b... more Herbicides are chemical compounds widely used in agriculture. As their intensive application is becoming a cause of environmental pollution, detailed and more sophisticated investigations are needed to understand better their consequences at the biological level. After herbicides are dispersed in the ®elds, they establish chemical interactions with both target and non-target plants. In both cases, herbicides can interact with the plant reproductive apparatus; consequently they could play a role during the fertilisation process in higher plants. Using an antibody to the a-tubulin subunit in immuno¯uorescence and immunoelectron microscopy techniques, we investigated the distribution of microtubules in Nicotiana tabacum pollen tubes grown under in vitro conditions in the presence of ®ve dierent herbicides selected among those used frequently in central Italy. Herbicides have a speci®c eect on the microtubular apparatus of both pollen tube and generative cell. In addition to other tests and assays, these results suggest that the microtubule cytoskeleton of pollen tubes can be used as a bioindicator for studying the toxicity eects induced by herbicides.
STEM CELLS, 2014
Adult neurogenesis is a multistep process regulated by several extrinsic factors, including neuro... more Adult neurogenesis is a multistep process regulated by several extrinsic factors, including neurotrophins. Among them, little is known about the role of nerve growth factor (NGF) in the neurogenic niches of the mouse. Here we analyzed the biology of adult neural stem cells (NSCs) from the subventricular zone (SVZ) of AD11 anti-NGF transgenic mice, in which the expression of the recombinant antibody aD11 leads to a chronic postnatal neutralization of endogenous NGF. We showed that AD11-NSCs proliferate 10-fold less, with respect to their control counterparts, and display a significant impairment in their ability to differentiate into b-tubulin positive neurons. We found a considerable reduction in the number of SVZ progenitors and neuroblasts also in vivo, which correlates with a lower number of newborn neurons in the olfactory bulbs of AD11 mice and a severe deficit in the ability of these mice to discriminate between different odors. We also demonstrated that, in AD11 mice, the morphology of both SVZ-resident and neurosphere-derived astrocytes is significantly altered. We were able to reproduce the AD11 phenotype in vitro, by acutely treating wild type NSCs with the anti-NGF antibody, further demonstrating that both the proliferation and the differentiation defects are due to the NGF deprivation. Consistently, the proliferative impairment of AD11 progenitors, as well as the atrophic morphology of AD11 astrocytes, can be partly rescued in vitro and in vivo by exogenous NGF addition. Altogether, our results demonstrate a causal link between NGF signaling and proper proliferation and differentiation of neural stem cells from the SVZ.
PLoS ONE, 2012
ABSTRACT [This corrects the article on p. e37555 in vol. 7.].
Medicine & Science in Sports & Exercise, 2007
Medicine & Science in Sports & Exercise, 2006
Page 1. Slide S e SS iONS Official Journal of the American College of Sports Medicine Vol. 38 No.... more Page 1. Slide S e SS iONS Official Journal of the American College of Sports Medicine Vol. 38 No. 5 Supplement S #583-760WEDNESDAY, MAY 31 | #761-918THURSDAY, JUNE 1 | #919-1067FRIDAY, JUNE 2 | #1068-1124 - SATURDAY, JUNE 3 ...
Journal of Molecular and Cellular Cardiology, 2004
Cardiomyocytes (CMCs) adapt to physiological or pathological stimuli by undergoing molecular chan... more Cardiomyocytes (CMCs) adapt to physiological or pathological stimuli by undergoing molecular changes which differentiate according to the specificity of the stimulus and eventually generate a phenotype with peculiar molecular characteristics. Here, we review the literature on the molecular mechanisms activated in the CMC during physiologic adaptation to stress, as opposed to maladaptation. The critical role of the IGF-1 receptor/PI3K/Akt signaling pathway during this process is described, including effector targets regulating inotropism and cell size.
Journal of Molecular and Cellular Cardiology, 2006
whilst undifferentiated and differentiate into CM. We examined the capacity of HESC-CM to survive... more whilst undifferentiated and differentiate into CM. We examined the capacity of HESC-CM to survive in the mouse heart. Methods: HESC, genetically marked by GFP (green fluorescent protein), were co-cultured with visceral endoderm-like cells, which induce differentiation into CM. Spontaneously beating areas, originating from HESC, were dissected and dissociated into small clumps containing 15% CM. In each SCID mouse 2-10 5 cells were injected intramyocardially. Between 2 and 4 weeks, HESC-CM were traced in cryosections by GFP-epifluorescence and characterized by immunofluorescent staining with humanand cardiomyocyte specific antibodies. Results: GFP-expressing cells were found both in scarred myocardium and aligned with host cardiomyocytes. Human origin of these cells was confirmed by staining for human nuclei and human mitochondria. 85% of the grafted human cells expressed a-actinin and had a sarcomeric structure. While mouse CM expressed only myosin light chain subtype MLC-2V, transplanted GFP cells expressed exclusively MLC-2A protein and not MLC-2V, indicating the absence of fusion with host CM. Cell survival was 0.2-1.4% of the total amount of transplanted cells and 2.0-12% of CM. Discussion: HESC-CM are able to survive in the mouse heart for at least 4 weeks after intramyocardial transplantation independently of fusion with host CM. The proportion of aactinin positive HESC-derived cells increases from 15% pre-to 85% post-transplantation, suggesting selective survival of HESC-CM in the heart, and/or differentiation of primed HESC into CM after transplantation.