- Email: [email protected]
Role of neural EGF-like like protein 2 (NELL2) in neuronal differentiation of P19 cells
P.P. Kale, V. Addepalli / Int. J. Devl Neuroscience 30 (2012) 640–671
669
A chemical–genetic study of EphB signaling identifies kinasedependent developmental processes
a powerful tool kit to advance understanding of primate-specific cognitive circuit development.
M. Soskis 1,∗ , H. Ho 1 , B. Ataman 1 , C. Zhang 2 , K. Shokat 2 , M. Greenberg 1
doi:10.1016/j.ijdevneu.2012.03.331
1 2
Harvard Medical School, USA UCSF, USA
E-mail address: [email protected] (M. Soskis). EphB receptor tyrosine kinases regulate cell–cell contacts in a variety of processes ranging from axon guidance and topographic mapping to neuronal migration and synapse formation. EphBs bind to a group of ligands, ephrin-Bs, which span the plasma membrane, thus allowing for bidirectional signaling between cells. This complexity combined with the pleiotropic functions of EphBs throughout development has obscured the exact nature of endogenous EphB signaling. To refine our understanding of how EphBs signal, we used a chemical–genetic approach to reversibly inhibit EphB receptors in vitro and in vivo. By mutating a residue in the receptor’s ATP-binding pocket, its kinase activity is rendered sensitive to reversible inhibition by allele-specific PP1 derivatives. This approach blocks the kinase activity of EphBs while leaving their scaffolding and reverse signaling capabilities intact and can circumvent the developmental compensation that may occur after genetic lesions. Using this approach we generated knockin mice with mutations in EphB1, EphB2, and EphB3. We demonstrate that EphB kinase activity is required for several distinct processes in nervous system development. Further studies with these allelespecific kinase mice may be especially valuable in exploring the nature of EphB signaling in the adult brain. doi:10.1016/j.ijdevneu.2012.03.329
Comparative gene expression in developing cognitive circuits in common marmoset brain Hiromi Mashiko ∗ , Aya C. Yoshida, Satomi S. Kikuchi, Kimie Niimi, Eiki Takahashi, Jun Aruga, Hideyuki Okano, Tomomi Shimogori Lab for Molecular Mechanisms of Thalamus Development, RIKEN BSI, Japan E-mail address: [email protected] (H. Mashiko). The developmental mechanisms by which the cerebral cortex increased in size and complexity during primate evolution are essentially unknown. To understand the molecular mechanisms of how animals evolved higher brain functions such as cognition, self-awareness, and language ability, precise gene expression profiles are required. In particular, studies of gene expression during early developmental stages are important to reveal homologous and analogous structures. Here, we report gene expression analysis in the neonatal common marmoset (Callithrix jacchus) brain. Using in situ hybridization, we focused especially on areas known to control cognition and their connective areas, including the prefrontal cortex, pulvinar nuclei in the thalamus, hippocampus, primary and secondary visual cortices, and dorsal lateral geniculate nucleus in the thalamus. To directly compare gene expression, we selected marker genes whose expression patterns and functions have been studied in developing mouse brain. We found that many, but not all, cortical markers tested showed similar patterns between mouse and marmoset prefrontal cortex, supporting the idea that regional identity is conserved at a molecular level across mammalian super orders. However, some genes showed very different expression patterns in marmoset brain, providing insight into the combination of conserved and novel genes that resulted in the evolution of new areas and functions. Our results establish
Norepinephrine stimulates a distinct precursor population in the adult hippocampus Dhanisha Jhaveri a,∗ , Estella Newcombe a , Virginia Nink a , Geoffrey Osborne a , Vidita Vaidya b , Perry Bartlett a a
The Queensland Brain Institute, The University of Queensland, Brisbane, Queensland 4072, Australia b Department of Biological Sciences, Tata Institute of Fundamental Research, Mumbai 400005, India
E-mail address: [email protected] (D. Jhaveri). New neurons are continuously generated from a resident population of stem and precursor cells in the adult hippocampus. Accumulating evidence points to a direct role of newborn neurons in the functional hippocampal circuitry that leads to enhancement of mood, cognition and memory. The current thinking is that there is a homogenous pool of precursors in the hippocampus that generates these new neurons, all of which contribute equally to the hippocampal functions. Recent studies from our laboratory have shown that the majority of the precursor cells are latent and can be activated by either high levels of K+ (mimicking neural excitation) or by the monoamine neurotransmitter norepinephrine. Using selective pharmacological blockers, we have shown that 3 adrenergic receptors mediate this norepinephrine-dependent activation. Interestingly, we have found that norepinephrine activates a separate precursor population than KCl and that 3 adrenergic receptor antagonist fails to block KCl-mediated activation of hippocampal precursors. Together, these findings suggest that distinct precursor populations exist in the adult hippocampus, which are regulated by different molecular mechanisms. We are currently phenotyping the different precursor populations using a panel (∼87) of monoclonal antibodies to CD antigens. We believe that this knowledge will significantly enhance our understanding of heterogeneity amongst precursor populations in the adult hippocampus and guide the next generation of therapeutic strategies involving the use of adult-born neurons for the treatment of neuropsychiatric disorders. doi:10.1016/j.ijdevneu.2012.03.332
Role of neural EGF-like like protein 2 (NELL2) in neuronal differentiation of P19 cells B.J. Lee a,∗ , D.H. Kim a , S.G. Kang b a b
University of Ulsan, South Korea Inje University, South Korea
E-mail address: [email protected] (B.J. Lee). NELL2 was first identified as a mammalian homolog of chick NEL (neural EGF-like) protein. It was almost exclusively expressed in neurons of rat brain and has been suggested to play a role in neuroprotection and neural differentiation. However, no clear evidence has yet been available for detailed function of NELL2. In this study we tried to identify NELL2 function during neural differentiation of mouse embryonic carcinoma P19 cells. Endogenous expression of NELL2 in P19 cells increased in parallel with neural differentiation process induced by retinoic acid (RA). We found that rat NELL2 promoter contains RA response element (RARE) and that treatment of RA increased NELL2 promoter activity. Transfection of P19 cells with NELL2 expression vectors induced a dramatic increase in cell aggregation, which in turn, resulting in an increase of neurosphere
670
P.P. Kale, V. Addepalli / Int. J. Devl Neuroscience 30 (2012) 640–671
formation and neural differentiation without RA treatment. Moreover, NELL2 increased N-cadherin expression in the P19 cells. These data suggest that NELL2 plays a critical role in regulation of neuronal differentiation via regulating N-cadherin expression and cell aggregation. doi:10.1016/j.ijdevneu.2012.03.333
Three-dimensional (3D) modeling of the embryonic shark brain: A basic anatomical tool for studying the development of neuronal systems in a basal vertebrate I. Carrera a,∗ , J. Moss b , D. Davidson b , I. Rodríguez-Moldes a a
Department of Cell Biology and Ecology, University of Santiago de Compostela, CIBUS Bldg, 15706 Santiago de Compostela, Spain b MRC Human Genetics Unit, Institute of Genetics and Molecular Medicine, Western General Hospital, Edinburgh EH4 2XU, UK E-mail address: [email protected] (I. Carrera). The 3D representation of the developing brain provides a detailed framework to study the organization of neuronal systems from the embryo to the young adult. To gain insight into the origin and developmental distribution of the complex morphological structures in the gnathostome brain, we have performed 3D representations of the developing brain in the dogfish Scyliorhinus canicula and mapped the distribution pattern of several neuronal systems identified by their neurochemical contents. The dogfish S. canicula is emerging as an important model organism for comparative studies of brain organization and development because of its phylogenetic position as a basal vertebrate. Using complementary techniques such as immunohistochemistry and optical projection tomography (OPT) microscopy, we have created a 3D imaging of the most representative embryonic stages and their neuroanatomical early expression pattern based on transverse and sagittal sections guided by schematic drawings. Here we show a key stage in the development of the brain, the embryonic stage 28, timepoint in which the expression of neuroactive substances is beginning or already started to express in the main neuronal systems. The events occurring at this particular stage lead to the initiation of a new interaction pattern between neuroactive systems, related to the establishment of the anteroposterior and dorsoventral subdivisions of the brain, thus allowing data meaning for regionalization mapping in vertebrates’ comparative studies. With the 3D representation of different embryonic stages we intend to create the 4D Developing Elasmobranch Brain Atlas, which will be a key resource for understanding the organization of neuronal systems containing modulatory neuroactive substances in the elasmobranch brain, and a reference in the accurate embryonic staging in future evo-devo studies. Supported by: Spanish Dirección General de InvestigaciónFEDER (BFU2010-15816), Xunta de Galicia (10PXIB200051PR) and European Community-ASSEMBLE (grant agreement no. 227799). doi:10.1016/j.ijdevneu.2012.03.334
Mitigated seizure susceptibility in mice lacking CD137 S.G. Kang a,∗ , H.M. Lee b , C.H. Yun b , B.J. Lee b a
Inje University, South Korea b University of Ulsan, South Korea
E-mail address: [email protected] (S.G. Kang). CD137 is a member of tumor necrosis factor/nerve growth factor receptor superfamily. Interaction of CD137 with its ligand (CD137L) affects apoptosis, proliferation, and differentiation of immune cells. Expression of CD137 is not restricted to immune organs, but
detected in a wide variety of tissues such as brain, kidney, lung and heart. However, its role in brain is largely unknown. Recent study reported that CD137 deficiency in the mast cells results in reduced Ca2+ response on stimulation by antigen. Prolonged increase of intracellular free Ca2+ can cause hyper-excitation of neurons, which may result in seizure and neural cell death. Thus we tried to identify role of CD137 in generation of seizure and hippocampal neuronal cell death which induced by kainic acid (KA). Knock-out mice lacking with CD137 showed a decrease in seizure susceptibility and an increased survival rate in response to KA. KA-induced hippocampal neurodegeneration and astrocyte activation were significantly less severe in the CD137 KO mice than in wild mice. Moreover, CD137 KO mice revealed decrease in cfos and cyclooxygenase-2 expression induced by KA. Treatment of agonistic antibody against CD137 increased Ca2+ influx in the primary cultured hippocampal cells. These data suggest that CD137 plays a role in the KA-induced seizure and neurodegeneration via regulating Ca2+ influx. doi:10.1016/j.ijdevneu.2012.03.335
Behavioural dysfunction and delayed neuronal death associated with cholinergic hypofunction during global cerebral ischemia and the effect of rivastigmine, galantamine and choline A. Katyal ∗ , D. Kumaran, M. Udayabanu Dr. B.R. Ambedkar Center for Biomedical Research, University of Delhi, North Campus, New Delhi, India E-mail address: anju [email protected] (A. Katyal). Global cerebral ischemia associated with cardiac arrest engenders selective neuronal death in subiculum, CA1 and CA4 regions of hippocampus. The clinical and experimental evidences reveal that the cognitive dysfunction and neuronal death are the key features of global cerebral ischemia. Further compelling evidences point at direct correlation between increased hippocampal acetylcholine level and learning performance in spatial memory task. We hypothesized that stimulation of cholinergic system may be a potential therapeutic strategy to mitigate the GCI associated neuronal death and memory impairment. In the present study, an effort was made to evaluate the effect of rivastigmine, galantamine and choline on global cerebral ischemia induced memory impairment, cholinergic hypofunction, neuronal inflammation and delayed neuronal death in a mouse model of global cerebral ischemia. BCCAo followed by reperfusion induced spatial and associative memory impairment which was ameliorated with the cholinergic agonists. Cholinergic agonists mitigated the loss of muscarinic and nicotinic receptor expressions. Further in vehicle treated ischemic group choline acetyltransferase and acetylcholinesterase activities were decreased, PARP-1, NADPH oxidase and myeloperoxidase activities were markedly increased in hippocampal homogentae. The histopathological, TUNEL and immunoflurosence studies revealed that GCI/R induced delayed neuronal death, DNA fragmentation as well as increased the high mobility group box-1 protein expression in hippocampal CA1 region. However, choline, rivastigmine and galantamine treatment markedly attenuated the cholinergic hypofunction and neuronal death/inflammation during ischemia. Molecular mechanism of barrel cortex development controlled by thalamocortical axon innervation Asuka Matsui ∗ , Masaharu Ogawa, Aya C. Yoshida, Tomomi Shimogori Molecular Mechanisms of Thalamus Development, BSI RIKEN, Japan E-mail address: asuka [email protected] (A. Matsui).
669
A chemical–genetic study of EphB signaling identifies kinasedependent developmental processes
a powerful tool kit to advance understanding of primate-specific cognitive circuit development.
M. Soskis 1,∗ , H. Ho 1 , B. Ataman 1 , C. Zhang 2 , K. Shokat 2 , M. Greenberg 1
doi:10.1016/j.ijdevneu.2012.03.331
1 2
Harvard Medical School, USA UCSF, USA
E-mail address: [email protected] (M. Soskis). EphB receptor tyrosine kinases regulate cell–cell contacts in a variety of processes ranging from axon guidance and topographic mapping to neuronal migration and synapse formation. EphBs bind to a group of ligands, ephrin-Bs, which span the plasma membrane, thus allowing for bidirectional signaling between cells. This complexity combined with the pleiotropic functions of EphBs throughout development has obscured the exact nature of endogenous EphB signaling. To refine our understanding of how EphBs signal, we used a chemical–genetic approach to reversibly inhibit EphB receptors in vitro and in vivo. By mutating a residue in the receptor’s ATP-binding pocket, its kinase activity is rendered sensitive to reversible inhibition by allele-specific PP1 derivatives. This approach blocks the kinase activity of EphBs while leaving their scaffolding and reverse signaling capabilities intact and can circumvent the developmental compensation that may occur after genetic lesions. Using this approach we generated knockin mice with mutations in EphB1, EphB2, and EphB3. We demonstrate that EphB kinase activity is required for several distinct processes in nervous system development. Further studies with these allelespecific kinase mice may be especially valuable in exploring the nature of EphB signaling in the adult brain. doi:10.1016/j.ijdevneu.2012.03.329
Comparative gene expression in developing cognitive circuits in common marmoset brain Hiromi Mashiko ∗ , Aya C. Yoshida, Satomi S. Kikuchi, Kimie Niimi, Eiki Takahashi, Jun Aruga, Hideyuki Okano, Tomomi Shimogori Lab for Molecular Mechanisms of Thalamus Development, RIKEN BSI, Japan E-mail address: [email protected] (H. Mashiko). The developmental mechanisms by which the cerebral cortex increased in size and complexity during primate evolution are essentially unknown. To understand the molecular mechanisms of how animals evolved higher brain functions such as cognition, self-awareness, and language ability, precise gene expression profiles are required. In particular, studies of gene expression during early developmental stages are important to reveal homologous and analogous structures. Here, we report gene expression analysis in the neonatal common marmoset (Callithrix jacchus) brain. Using in situ hybridization, we focused especially on areas known to control cognition and their connective areas, including the prefrontal cortex, pulvinar nuclei in the thalamus, hippocampus, primary and secondary visual cortices, and dorsal lateral geniculate nucleus in the thalamus. To directly compare gene expression, we selected marker genes whose expression patterns and functions have been studied in developing mouse brain. We found that many, but not all, cortical markers tested showed similar patterns between mouse and marmoset prefrontal cortex, supporting the idea that regional identity is conserved at a molecular level across mammalian super orders. However, some genes showed very different expression patterns in marmoset brain, providing insight into the combination of conserved and novel genes that resulted in the evolution of new areas and functions. Our results establish
Norepinephrine stimulates a distinct precursor population in the adult hippocampus Dhanisha Jhaveri a,∗ , Estella Newcombe a , Virginia Nink a , Geoffrey Osborne a , Vidita Vaidya b , Perry Bartlett a a
The Queensland Brain Institute, The University of Queensland, Brisbane, Queensland 4072, Australia b Department of Biological Sciences, Tata Institute of Fundamental Research, Mumbai 400005, India
E-mail address: [email protected] (D. Jhaveri). New neurons are continuously generated from a resident population of stem and precursor cells in the adult hippocampus. Accumulating evidence points to a direct role of newborn neurons in the functional hippocampal circuitry that leads to enhancement of mood, cognition and memory. The current thinking is that there is a homogenous pool of precursors in the hippocampus that generates these new neurons, all of which contribute equally to the hippocampal functions. Recent studies from our laboratory have shown that the majority of the precursor cells are latent and can be activated by either high levels of K+ (mimicking neural excitation) or by the monoamine neurotransmitter norepinephrine. Using selective pharmacological blockers, we have shown that 3 adrenergic receptors mediate this norepinephrine-dependent activation. Interestingly, we have found that norepinephrine activates a separate precursor population than KCl and that 3 adrenergic receptor antagonist fails to block KCl-mediated activation of hippocampal precursors. Together, these findings suggest that distinct precursor populations exist in the adult hippocampus, which are regulated by different molecular mechanisms. We are currently phenotyping the different precursor populations using a panel (∼87) of monoclonal antibodies to CD antigens. We believe that this knowledge will significantly enhance our understanding of heterogeneity amongst precursor populations in the adult hippocampus and guide the next generation of therapeutic strategies involving the use of adult-born neurons for the treatment of neuropsychiatric disorders. doi:10.1016/j.ijdevneu.2012.03.332
Role of neural EGF-like like protein 2 (NELL2) in neuronal differentiation of P19 cells B.J. Lee a,∗ , D.H. Kim a , S.G. Kang b a b
University of Ulsan, South Korea Inje University, South Korea
E-mail address: [email protected] (B.J. Lee). NELL2 was first identified as a mammalian homolog of chick NEL (neural EGF-like) protein. It was almost exclusively expressed in neurons of rat brain and has been suggested to play a role in neuroprotection and neural differentiation. However, no clear evidence has yet been available for detailed function of NELL2. In this study we tried to identify NELL2 function during neural differentiation of mouse embryonic carcinoma P19 cells. Endogenous expression of NELL2 in P19 cells increased in parallel with neural differentiation process induced by retinoic acid (RA). We found that rat NELL2 promoter contains RA response element (RARE) and that treatment of RA increased NELL2 promoter activity. Transfection of P19 cells with NELL2 expression vectors induced a dramatic increase in cell aggregation, which in turn, resulting in an increase of neurosphere
670
P.P. Kale, V. Addepalli / Int. J. Devl Neuroscience 30 (2012) 640–671
formation and neural differentiation without RA treatment. Moreover, NELL2 increased N-cadherin expression in the P19 cells. These data suggest that NELL2 plays a critical role in regulation of neuronal differentiation via regulating N-cadherin expression and cell aggregation. doi:10.1016/j.ijdevneu.2012.03.333
Three-dimensional (3D) modeling of the embryonic shark brain: A basic anatomical tool for studying the development of neuronal systems in a basal vertebrate I. Carrera a,∗ , J. Moss b , D. Davidson b , I. Rodríguez-Moldes a a
Department of Cell Biology and Ecology, University of Santiago de Compostela, CIBUS Bldg, 15706 Santiago de Compostela, Spain b MRC Human Genetics Unit, Institute of Genetics and Molecular Medicine, Western General Hospital, Edinburgh EH4 2XU, UK E-mail address: [email protected] (I. Carrera). The 3D representation of the developing brain provides a detailed framework to study the organization of neuronal systems from the embryo to the young adult. To gain insight into the origin and developmental distribution of the complex morphological structures in the gnathostome brain, we have performed 3D representations of the developing brain in the dogfish Scyliorhinus canicula and mapped the distribution pattern of several neuronal systems identified by their neurochemical contents. The dogfish S. canicula is emerging as an important model organism for comparative studies of brain organization and development because of its phylogenetic position as a basal vertebrate. Using complementary techniques such as immunohistochemistry and optical projection tomography (OPT) microscopy, we have created a 3D imaging of the most representative embryonic stages and their neuroanatomical early expression pattern based on transverse and sagittal sections guided by schematic drawings. Here we show a key stage in the development of the brain, the embryonic stage 28, timepoint in which the expression of neuroactive substances is beginning or already started to express in the main neuronal systems. The events occurring at this particular stage lead to the initiation of a new interaction pattern between neuroactive systems, related to the establishment of the anteroposterior and dorsoventral subdivisions of the brain, thus allowing data meaning for regionalization mapping in vertebrates’ comparative studies. With the 3D representation of different embryonic stages we intend to create the 4D Developing Elasmobranch Brain Atlas, which will be a key resource for understanding the organization of neuronal systems containing modulatory neuroactive substances in the elasmobranch brain, and a reference in the accurate embryonic staging in future evo-devo studies. Supported by: Spanish Dirección General de InvestigaciónFEDER (BFU2010-15816), Xunta de Galicia (10PXIB200051PR) and European Community-ASSEMBLE (grant agreement no. 227799). doi:10.1016/j.ijdevneu.2012.03.334
Mitigated seizure susceptibility in mice lacking CD137 S.G. Kang a,∗ , H.M. Lee b , C.H. Yun b , B.J. Lee b a
Inje University, South Korea b University of Ulsan, South Korea
E-mail address: [email protected] (S.G. Kang). CD137 is a member of tumor necrosis factor/nerve growth factor receptor superfamily. Interaction of CD137 with its ligand (CD137L) affects apoptosis, proliferation, and differentiation of immune cells. Expression of CD137 is not restricted to immune organs, but
detected in a wide variety of tissues such as brain, kidney, lung and heart. However, its role in brain is largely unknown. Recent study reported that CD137 deficiency in the mast cells results in reduced Ca2+ response on stimulation by antigen. Prolonged increase of intracellular free Ca2+ can cause hyper-excitation of neurons, which may result in seizure and neural cell death. Thus we tried to identify role of CD137 in generation of seizure and hippocampal neuronal cell death which induced by kainic acid (KA). Knock-out mice lacking with CD137 showed a decrease in seizure susceptibility and an increased survival rate in response to KA. KA-induced hippocampal neurodegeneration and astrocyte activation were significantly less severe in the CD137 KO mice than in wild mice. Moreover, CD137 KO mice revealed decrease in cfos and cyclooxygenase-2 expression induced by KA. Treatment of agonistic antibody against CD137 increased Ca2+ influx in the primary cultured hippocampal cells. These data suggest that CD137 plays a role in the KA-induced seizure and neurodegeneration via regulating Ca2+ influx. doi:10.1016/j.ijdevneu.2012.03.335
Behavioural dysfunction and delayed neuronal death associated with cholinergic hypofunction during global cerebral ischemia and the effect of rivastigmine, galantamine and choline A. Katyal ∗ , D. Kumaran, M. Udayabanu Dr. B.R. Ambedkar Center for Biomedical Research, University of Delhi, North Campus, New Delhi, India E-mail address: anju [email protected] (A. Katyal). Global cerebral ischemia associated with cardiac arrest engenders selective neuronal death in subiculum, CA1 and CA4 regions of hippocampus. The clinical and experimental evidences reveal that the cognitive dysfunction and neuronal death are the key features of global cerebral ischemia. Further compelling evidences point at direct correlation between increased hippocampal acetylcholine level and learning performance in spatial memory task. We hypothesized that stimulation of cholinergic system may be a potential therapeutic strategy to mitigate the GCI associated neuronal death and memory impairment. In the present study, an effort was made to evaluate the effect of rivastigmine, galantamine and choline on global cerebral ischemia induced memory impairment, cholinergic hypofunction, neuronal inflammation and delayed neuronal death in a mouse model of global cerebral ischemia. BCCAo followed by reperfusion induced spatial and associative memory impairment which was ameliorated with the cholinergic agonists. Cholinergic agonists mitigated the loss of muscarinic and nicotinic receptor expressions. Further in vehicle treated ischemic group choline acetyltransferase and acetylcholinesterase activities were decreased, PARP-1, NADPH oxidase and myeloperoxidase activities were markedly increased in hippocampal homogentae. The histopathological, TUNEL and immunoflurosence studies revealed that GCI/R induced delayed neuronal death, DNA fragmentation as well as increased the high mobility group box-1 protein expression in hippocampal CA1 region. However, choline, rivastigmine and galantamine treatment markedly attenuated the cholinergic hypofunction and neuronal death/inflammation during ischemia. Molecular mechanism of barrel cortex development controlled by thalamocortical axon innervation Asuka Matsui ∗ , Masaharu Ogawa, Aya C. Yoshida, Tomomi Shimogori Molecular Mechanisms of Thalamus Development, BSI RIKEN, Japan E-mail address: asuka [email protected] (A. Matsui).