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Dean R Tolan - One of the best experts on this subject based on the ideXlab platform.
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struCture of human brain fruCtose 1 6 bis phosphate Aldolase linking isozyme struCture with funCtion
Protein Science, 2009Co-Authors: Tracy L Arakaki, Dean R Tolan, John A Pezza, Michelle A Cronin, Christopher E Hopkins, Danna B Zimmer, Karen N AllenAbstract:FruCtose-1,6-(bis)phosphate Aldolase is a ubiquitous enzyme that Catalyzes the reversible aldol Cleavage of fruCtose-1,6-(bis)phosphate and fruCtose 1-phosphate to dihydroxyaCetone phosphate and either glyCeral-dehyde-3-phosphate or glyCeraldehyde, respeCtively. Vertebrate Aldolases exist as three isozymes with different tissue distributions and kinetiCs: Aldolase A (musCle and red blood Cell), Aldolase B (liver, kidney, and small intestine), and Aldolase C (brain and neuronal tissue). The struCtures of human Aldolases A and B are known and herein we report the first struCture of the human Aldolase C, solved by X-ray Crystallography at 3.0 A resolution. StruCtural differenCes between the isozymes were expeCted to aCCount for isozyme-speCifiC aCtivity. However, the struCtures of isozymes A, B, and C are the same in their overall fold and aCtive site struCture. The subtle Changes observed in aCtive site residues Arg42, Lys146, and Arg303 are insuffiCient to Completely aCCount for the tissue-speCifiC isozymiC differenCes. Consequently, the struCtural analysis has been extended to the isozyme-speCifiC residues (ISRs), those residues Conserved among paralogs. A Complete analysis of the ISRs in the Context of this struCture demonstrates that in several Cases an amino aCid residue that is Conserved among Aldolase C orthologs prevents an interaCtion that oCCurs in paralogs. In addition, the struCture Confirms the Clustering of ISRs into disCrete patChes on the surfaCe and reveals the existenCe in Aldolase C of a patCh of eleCtronegative residues loCalized near the C terminus. Together, these struCtural Changes highlight the differenCes required for the tissue and kinetiC speCifiCity among Aldolase isozymes.
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IdentifiCation of Neuronal Isozyme SpeCifiC Residues by Comparison of Goldfish Aldolase C to Other Aldolases
Comparative Biochemistry and Physiology Part A: Physiology, 1997Co-Authors: Tanya Z Berardini, Mariola Drygas-williams, Gloria V. Callard, Dean R TolanAbstract:AbstraCt A 2061 bp CDNA enCoding a goldfish (Carassius auratus) Aldolase was isolated from a goldfish brain library. The deduCed 362 amino aCid sequenCe is more similar to vertebrate brain (Aldolase C) and musCle Aldolases (Aldolase A) than to the liver isozymes (Aldolase B). Northern blot analysis indiCates strong expression of the mRNA in brain but not in liver or musCle, whiCh indiCates that this is Aldolase C rather than Aldolase A. Analysis of all known vertebrate Aldolase amino aCid sequenCes reveals five residues; Leu-57, Arg-314, Thr-324, Glu-332, and Gly-350 that are present exClusively in Aldolase Cs. The goldfish Clone possesses all five residues. The residues are primarily loCated in the Carboxyl-terminal region of the enzyme and may play a role in determining the neuronal isozyme-speCifiC properties of the enzyme. Furthermore, the existenCe of an Aldolase C in a teleost fish has impliCations with respeCt to the timing of genome dupliCation events that are thought to have been CritiCal in vertebrate evolution.
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NonCoordinate Changes in the steady‐state mrna expressed from Aldolase A and Aldolase C genes during differentiation of ChiCken myoblasts
Journal of Cellular Biochemistry, 1995Co-Authors: Rachel L. Meighan-mantha, Dean R TolanAbstract:: In ChiCkens, as in all vertebrates, tissue-speCifiC expression of Aldolase isozymes A, B, and C is developmentally Coordinated. These developmental transitions in Aldolase expression have been studied most extensively by Charting enzyme aCtivity during normal and abnormal development of speCifiC vertebrate tissues. Indeed, Aldolase expression has been a key marker for normal differentiation and for retrodifferentiation during CarCinogenesis. Aldolase expression during ChiCken myoblast differentiation offers a model for investigating the regulatory meChanisms of these developmental transitions at the level of gene expression. For these studies, CDNAs enCoding the most isozyme-speCifiC regions of both ChiCken Aldolase A and C were Cloned. The ChiCken Aldolase A CDNA represents the first report of this sequenCe. Aldolase steady-state mRNA expression was measured during ChiCken myoblast differentiation in primary Cultures using RNase proteCtion assays with CRNA probes generated from these Aldolase CDNA Clones. Steady-state mRNA for Aldolase C, the predominant embryoniC Aldolase isozyme in ChiCkens, did not signifiCantly Change throughout myoblast differentiation. In Contrast, expression of steady-state mRNA for Aldolase A, the only Aldolase isozyme found in adult-skeletal musCle, was not deteCted until after myoblast fusion was approximately 50% Completed. Aldolase A expression gradually inCreased throughout myoblast differentiation until approximately 48 h after fusion was Completed when there was a dramatiC inCrease. These results are Contrasted with those of Turner et al. (1974) [Dev Biol 37:63-89] that showed a Coordinated switCh in isozyme aCtivities between the embryoniC Aldolase C and the musCle-speCifiC Aldolase A. This disCordant expression indiCates that the Aldolase A and C genes may employ different regulatory meChanisms during myoblast differentiation.
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nonCoordinate Changes in the steady state mrna expressed from Aldolase a and Aldolase C genes during differentiation of ChiCken myoblasts
Journal of Cellular Biochemistry, 1995Co-Authors: Rachel L Meighanmantha, Dean R TolanAbstract:In ChiCkens, as in all vertebrates, tissue-speCifiC expression of Aldolase isozymes A, B, and C is developmentally Coordinated. These developmental transitions in Aldolase expression have been studied most extensively by Charting enzyme aCtivity during normal and abnormal development of speCifiC vertebrate tissues. Indeed, Aldolase expression has been a key marker for normal differentiation and for retrodifferentiation during CarCinogenesis. Aldolase expression during ChiCken myoblast differentiation offers a model for investigating the regulatory meChanisms of these developmental transitions at the level of gene expression. For these studies, CDNAs enCoding the most isozyme-speCifiC regions of both ChiCken Aldolase A and C were Cloned. The ChiCken Aldolase A CDNA represents the first report of this sequenCe. Aldolase steady-state mRNA expression was measured during ChiCken myoblast differentiation in primary Cultures using RNase proteCtion assays with CRNA probes generated from these Aldolase CDNA Clones. Steady-state mRNA for Aldolase C, the predominant embryoniC Aldolase isozyme in ChiCkens, did not signifiCantly Change throughout myoblast differentiation. In Contrast, expression of steady-state mRNA for Aldolase A, the only Aldolase isozyme found in adult skeletal musCle, was not deteCted until after myoblast fusion was approximately 50% Completed. Aldolase A expression gradually inCreased throughout myoblast differentiation until approximately 48 h after fusion was Completed when there was a dramatiC inCrease. These results are Contrasted with those of Turner et al. (1974) [Dev Biol 37:63–89] that showed a Coordinated switCh in isozyme aCtivities between the embryoniC Aldolase C and the musCle-speCifiC Aldolase A. This disCordant expression indiCates that the Aldolase A and C genes may employ different regulatory meChanisms during myoblast differentiation.
Karl Herrup - One of the best experts on this subject based on the ideXlab platform.
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Purkinje Cell expression of the mouse Aldolase C gene in transgeniC miCe is direCted by an upstream regulatory element
Brain research. Molecular brain research, 2005Co-Authors: Rita R. Romito-digiacomo, Ernst U. Walther, Elizabeth A. Williams, Karl HerrupAbstract:We have sought to understand the regulation of the expression pattern of Aldolase C (Zebrin II) in Cerebellar Purkinje Cells. Normally, Aldolase C is expressed in a series of sagittal stripes of Purkinje Cells interrupted by stripes of little or no expression. GenomiC Aldolase C:LaCZ fusion genes with 1.8 kb of sequenCe 5' to the transCription start site drive CNS expression of LaCZ only in astroCytes and Cells of the pia mater. If the 5' portion of the transgene is extended to a full 5.0 kb, expression is reliably observed in Purkinje Cells, yet none of the astroCyte expression is lost. We broke the additional 3.0 kb into 1.0 kb fragments and tested eaCh for Purkinje Cell enhanCer aCtivity when appended to the original 1.8 kb ConstruCt. We show that the 886 bp region from nuCleotide -2796 to -3682 (relative to the start of transCription) Contains virtually all of the Purkinje Cell enhanCer aCtivity. However, neither the full 5.0 kb nor the 886 bp region direCted a striped expression pattern, as is seen for the endogenous gene. Taken together, our study loCalizes a Purkinje Cell enhanCer to a small 5' region of the Aldolase C gene and illustrates that the element(s) responsible for the normal anatomiCally Complex pattern of Aldolase C expression are separate from those Conferring Cell-type speCifiCity. The relationship of these findings to previous work in other laboratories is disCussed.
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GenomiC sequenCes of Aldolase C (Zebrin II) direCt laCZ expression exClusively in non-neuronal Cells of transgeniC miCe
Proceedings of the National Academy of Sciences of the United States of America, 1998Co-Authors: Ernst U. Walther, Richard Hawkes, Suzan Dziennis, Martin Dichgans, Stephen M. Maricich, Rita R. Romito, Fei Yang, Saul L. Zackson, Karl HerrupAbstract:Aldolase C is regarded as the brain-speCifiC form of fruCtose-1,6-bisphosphate Aldolase whereas Aldolase A is regarded as musCle-speCifiC. In situ hybridization of mouse Central nervous system using isozyme-speCifiC probes revealed that Aldolase A and C are expressed in Complementary Cell types. With the exCeption of Cerebellar Purkinje Cells, Aldolase A mRNA is found in neurons; Aldolase C message is deteCted in astroCytes, some Cells of the pia mater, and Purkinje Cells. We isolated Aldolase C genomiC Clones that span the entire protein Coding region from 1.5 kb 5′ to the transCription start site to 0.5 kb 3′ to the end of the last exon. The baCterial gene, laCZ, was inserted in two different loCations and the ConstruCts tested in transgeniC miCe. When the protein Coding sequenCes were replaCed with laCZ, three of five transgeniC lines expressed β-galaCtosidase only in Cells of the pia mater; one line also expressed in astroCyte-like Cells. When laCZ was inserted into the final exon (and all struCtural gene sequenCes were retained) transgene expression was observed in astroCytes in all regions of the Central nervous system as well as in pial Cells. Thus, with the exCeption of Purkinje Cell expression, the behavior of the full-length transgene mimiCs the endogenous Aldolase C gene. The results with the shorter transgene suggest that additional enhanCer elements exist within the intrageniC sequenCes. The absenCe of Purkinje Cell staining suggests that the Cis elements required for this expression must be loCated outside of the sequenCes used in this study.
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Aldolase C/zebrin II and the regionalization of the Cerebellum
Journal of Molecular Neuroscience, 1995Co-Authors: Richard Hawkes, Karl HerrupAbstract:The Cerebellum is Comprised of multiple bands of Cells, eaCh with CharaCteristiC afferent and efferent projeCtions, and patterns of gene expression. The most studied example of a striped pattern of expression is the antigen reCognized by monoClonal antibody antizebrin II. Zebrin II is expressed by subsets of Purkinje Cells that form an array of parasagittal bands that extend rostroCaudally throughout the Cerebellar Cortex, separated by similar bands of Purkinje Cells that do not express zebrin II. ReCent Cloning studies have revealed that the zebrin II antigen is the respiratory isoenzyme Aldolase C. This artiCle reviews the Cellular and moleCular Compartmentation of the Cerebellum together with the moleCular biology of the Aldolase C gene, and speCulates on possible reasons for a striped pattern of expression.
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The Cloning of zebrin II reveals its identity with Aldolase C.
Development (Cambridge England), 1994Co-Authors: Andrew H. Ahn, Richard Hawkes, Suzan Dziennis, Karl HerrupAbstract:The sagittal organization of the mammalian Cerebellum Can be observed at the anatomiCal, physiologiCal and bioChemiCal level. Previous sCreening of monoClonal antibodies produCed in our laboratory has identified two intraCellular antigens, zebrin I and II, that oCCur exClusively in adult Cerebellar Purkinje Cells. As their name suggests, the zebrin antibody staining of the Purkinje Cell population is not uniform. Rather, zebrin-positive Purkinje Cells are organized in stripes or bands that run from anterior to posterior aCross most of the Cerebellum; interposed between the zebrin-positive Cells are bands of Purkinje Cells that are zebrin-negative. Comparison of the position of the antigeniC bands with the anatomy of afferent projeCtions suggests that the bands are Congruent with the basiC developmental and funCtional ‘Compartments’ of the Cerebellum. We report the isolation of CDNA Clones of the 36 × 10(3) M(r) antigen, zebrin II, by sCreening of a mouse Cerebellum CDNA expression library. SequenCe analysis reveals a 98% identity between our Clone and the glyColytiC isozyme, Aldolase C. In order to more rigorously demonstrate the identity of the two proteins, we stained adult Cerebellum with an independent monoClonal antibody raised against Aldolase C. Anti-Aldolase staining oCCurs in a previously unreported pattern of sagittal bands of Purkinje Cells; the pattern is identiCal to that revealed by the zebrin II monoClonal. Further, in situ hybridization of antisense Aldolase C riboprobe shows that the aCCumulation of zebrin II/Aldolase C mRNA Corresponds to the pattern of the zebrin antigen in Purkinje Cells. Zebrin II/Aldolase C gene expression is thus regulated at the level of transCription (or mRNA stability). In light of previous work that has demonstrated the Cell-autonomous and developmentally regimented expression of zebrin II, further studies of the regulation of this gene may lead to insights about the determination of Cerebellar Compartmentation.
Richard Hawkes - One of the best experts on this subject based on the ideXlab platform.
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zebrin ii Aldolase C expression in the Cerebellum of the western diamondbaCk rattlesnake Crotalus atrox
PLOS ONE, 2015Co-Authors: Joel W Aspden, Carol L Armstrong, Richard Hawkes, Tobias Kohl, David J Graham, Andrew N Iwaniuk, Cristian Gutierrezibanez, Douglas R WylieAbstract:Aldolase C, also known as Zebrin II (ZII), is a glyColytiC enzyme that is expressed in Cerebellar Purkinje Cells of the vertebrate Cerebellum. In both mammals and birds, ZII is expressed heterogeneously, suCh that there are sagittal stripes of Purkinje Cells with high ZII expression (ZII+), alternating with stripes of Purkinje Cells with little or no expression (ZII-). The patterns of ZII+ and ZII- stripes in the Cerebellum of birds and mammals are strikingly similar, suggesting that it may have first evolved in the stem reptiles. In this study, we examined the expression of ZII in the Cerebellum of the western diamondbaCk rattlesnake (Crotalus atrox). In Contrast to birds and mammals, the Cerebellum of the rattlesnake is muCh smaller and simpler, Consisting of a small, unfoliated dome of Cells. A pattern of alternating ZII+ and ZII- sagittal stripes Cells was not observed: rather all Purkinje Cells were ZII+. This suggests that ZII stripes have either been lost in snakes or that they evolved Convergently in birds and mammals.
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Zebrin II / Aldolase C expression in the Cerebellum of the western diamondbaCk rattlesnake (Crotalus atrox).
PLOS ONE, 2015Co-Authors: Joel W Aspden, Carol L Armstrong, Cristián Gutiérrez-ibáñez, Richard Hawkes, Tobias Kohl, David J Graham, Andrew N Iwaniuk, Douglas R WylieAbstract:Aldolase C, also known as Zebrin II (ZII), is a glyColytiC enzyme that is expressed in Cerebellar Purkinje Cells of the vertebrate Cerebellum. In both mammals and birds, ZII is expressed heterogeneously, suCh that there are sagittal stripes of Purkinje Cells with high ZII expression (ZII+), alternating with stripes of Purkinje Cells with little or no expression (ZII-). The patterns of ZII+ and ZII- stripes in the Cerebellum of birds and mammals are strikingly similar, suggesting that it may have first evolved in the stem reptiles. In this study, we examined the expression of ZII in the Cerebellum of the western diamondbaCk rattlesnake (Crotalus atrox). In Contrast to birds and mammals, the Cerebellum of the rattlesnake is muCh smaller and simpler, Consisting of a small, unfoliated dome of Cells. A pattern of alternating ZII+ and ZII- sagittal stripes Cells was not observed: rather all Purkinje Cells were ZII+. This suggests that ZII stripes have either been lost in snakes or that they evolved Convergently in birds and mammals.
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Aldolase C/zebrin II expression in the neonatal rat forebrain reveals Cellular heterogeneity within the subventriCular zone and early astroCyte differentiation.
The Journal of neuroscience : the official journal of the Society for Neuroscience, 2001Co-Authors: Susan M. Staugaitis, Richard Hawkes, Marielba Zerlin, Joel M. Levine, James E. GoldmanAbstract:During late gestational and early postnatal development, proliferating Cells in the subventriCular zones of the lateral ventriCles (SVZ) migrate into the gray and white matter of the forebrain and differentiate into astroCytes and oligodendroCytes. BeCause the Cellular Composition and struCture of the neonatal SVZ is poorly understood, we performed a differential display PCR sCreen to identify genes preferentially expressed therein. One highly expressed gene enCoded Aldolase C. We used a speCifiC monoClonal antibody, Aldolase C/zebrin II (ALDC/ZII), in Combination with markers of glial lineage and proliferation, to CharaCterize the Cells that express this gene. In the neonatal SVZ, ALDC/ZII-positive Cells, whiCh are generally polygonal and display several proCesses, have a nonuniform spatial distribution. They do not express vimentin, GFAP, or NG2. A subset of ALDC/ZII-positive Cells inCorporates bromodeoxyuridine, but progenitors identified by β-galaCtosidase expression after infeCtion with reCombinant BAG virus do not show ALDC/ZII immunoreaCtivity. Outside of the SVZ, β-galaCtosidase-positive/ALDC/ZII-positive Cells have an astroCytiC phenotype, suggesting that immunoreaCtivity was aCquired after exit from the SVZ. These studies demonstrate that the neonatal SVZ is Composed of different populations of Cells that Can be CharaCterized by their antigeniC phenotype, their proliferative CapaCity, and their spatial distributions. Nonrandom distributions of different Cell types within the SVZ may permit the formation of miCroenvironments that stimulate the produCtion of Cells with speCifiC potentials at appropriate points in development. Analysis of ALDC/ZII expression by astroCyte lineage Cells in the neonatal Cerebral Cortex and white matter may reveal insights into the phenotype and behavior of undifferentiated astroCyte progenitors.
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GenomiC sequenCes of Aldolase C (Zebrin II) direCt laCZ expression exClusively in non-neuronal Cells of transgeniC miCe
Proceedings of the National Academy of Sciences of the United States of America, 1998Co-Authors: Ernst U. Walther, Richard Hawkes, Suzan Dziennis, Martin Dichgans, Stephen M. Maricich, Rita R. Romito, Fei Yang, Saul L. Zackson, Karl HerrupAbstract:Aldolase C is regarded as the brain-speCifiC form of fruCtose-1,6-bisphosphate Aldolase whereas Aldolase A is regarded as musCle-speCifiC. In situ hybridization of mouse Central nervous system using isozyme-speCifiC probes revealed that Aldolase A and C are expressed in Complementary Cell types. With the exCeption of Cerebellar Purkinje Cells, Aldolase A mRNA is found in neurons; Aldolase C message is deteCted in astroCytes, some Cells of the pia mater, and Purkinje Cells. We isolated Aldolase C genomiC Clones that span the entire protein Coding region from 1.5 kb 5′ to the transCription start site to 0.5 kb 3′ to the end of the last exon. The baCterial gene, laCZ, was inserted in two different loCations and the ConstruCts tested in transgeniC miCe. When the protein Coding sequenCes were replaCed with laCZ, three of five transgeniC lines expressed β-galaCtosidase only in Cells of the pia mater; one line also expressed in astroCyte-like Cells. When laCZ was inserted into the final exon (and all struCtural gene sequenCes were retained) transgene expression was observed in astroCytes in all regions of the Central nervous system as well as in pial Cells. Thus, with the exCeption of Purkinje Cell expression, the behavior of the full-length transgene mimiCs the endogenous Aldolase C gene. The results with the shorter transgene suggest that additional enhanCer elements exist within the intrageniC sequenCes. The absenCe of Purkinje Cell staining suggests that the Cis elements required for this expression must be loCated outside of the sequenCes used in this study.
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Aldolase C/zebrin II and the regionalization of the Cerebellum
Journal of Molecular Neuroscience, 1995Co-Authors: Richard Hawkes, Karl HerrupAbstract:The Cerebellum is Comprised of multiple bands of Cells, eaCh with CharaCteristiC afferent and efferent projeCtions, and patterns of gene expression. The most studied example of a striped pattern of expression is the antigen reCognized by monoClonal antibody antizebrin II. Zebrin II is expressed by subsets of Purkinje Cells that form an array of parasagittal bands that extend rostroCaudally throughout the Cerebellar Cortex, separated by similar bands of Purkinje Cells that do not express zebrin II. ReCent Cloning studies have revealed that the zebrin II antigen is the respiratory isoenzyme Aldolase C. This artiCle reviews the Cellular and moleCular Compartmentation of the Cerebellum together with the moleCular biology of the Aldolase C gene, and speCulates on possible reasons for a striped pattern of expression.
Izumi Sugihara - One of the best experts on this subject based on the ideXlab platform.
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The entire trajeCtories of single pontoCerebellar axons and their lobular and longitudinal terminal distribution patterns in multiple Aldolase C‐positive Compartments of the rat Cerebellar Cortex
The Journal of comparative neurology, 2019Co-Authors: Izumi Sugihara, Yoshikazu ShinodaAbstract:The mammalian Cerebellar Cortex is Compartmentalized, both anatomiCally and histoChemiCally, into multiple parasagittal bands. To CharaCterize the multiple zonal patterns of pontoCerebellar mossy fiber projeCtion, single neurons in the basilar pontine nuCleus (BPN) were labeled by injeCting biotinylated dextran amine into the BPN, and the entire axonal trajeCtory of single labeled neurons (n = 25) was reConstruCted in relation to Aldolase C Compartments of Purkinje Cells in rats. Single pontoCerebellar axons, after passing through the Contralateral middle Cerebellar pedunCle, ran transversely in the deep Cerebellar white matter toward and often aCross the midline, and on their ways, gave rise to 2-10 primary Collaterals at almost right angles in speCifiC lobules only Contralaterally or bilaterally with Contralateral predominanCe. EaCh primary Collateral further branChed in a parasagittal plane to form a strip-shaped longitudinal termination zone with rosette-type swellings Clustered in Aldolase C-positive Compartments in a single or multiple lobules, mainly in Compartment 4+//5+, 5+//6+, and 6+//7+. Axons arising from the Central, rostral, and lateral part of the BPN projeCted with multiple branChes, mainly to simple lobule, Crus II and paramedian lobule, to Crus I and dorsal parafloCCulus, and to ventral parafloCCulus and lobule IXC, respeCtively. The results showed the pontoCerebellar projeCtion is Closely related to lobular and Compartmental organization of the Cerebellum. A Comparison of single axon morphologies of different mossy fiber systems indiCates that the projeCtion pattern of single pontoCerebellar neurons with multiple Collaterals innervating different longitudinal Compartments arranged in a mediolateral direCtion represents a general feature of mossy fiber projeCtion.
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Single axonal morphology and termination to Cerebellar Aldolase C stripes CharaCterize distinCt spinoCerebellar projeCtion systems originating from the thoraCiC spinal Cord in the mouse.
The Journal of comparative neurology, 2017Co-Authors: Yuanjun Luo, Radhika Pooja Patel, Gideon Anokye Sarpong, Kazuma Sasamura, Izumi SugiharaAbstract:The spinoCerebellar projeCtion has an essential role in sensorimotor Coordination of limbs and the trunk. Multiple groups of spinoCerebellar projeCtions have been identified in retrograde labeling studies. In this study, we aimed at CharaCterizing projeCtion patterns of these groups using a Combination of anterograde labeling of the thoraCiC spinal Cord and Aldolase C immunostaining of longitudinal stripes of the Cerebellar Cortex in the mouse. We reConstruCted 22 single spinoCerebellar axons, wholly in the Cerebellum and brain stem and partly, in the spinal Cord. They were Classified into three groups, (a) non-Crossed axons of Clarke's Column neurons (NCC, 8 axons), (b) non-Crossed axons of marginal Clarke's Column neurons (NMCC, 7 axons), and (C) Crossed axons of neurons in the medial ventral horn (CMVH, 7 axons), based on previous retrograde labeling studies. While NCC axons projeCted mainly to multiple bilateral stripes in vermal lobules II-IV and VIII-IX, and the ipsilateral medial Cerebellar nuCleus, NMCC axons projeCted mainly to ipsilateral stripes in paravermal lobules II-V and Copula pyramidis, and the anterior interposed nuCleus. CMVH axons projeCted bilaterally to multiple stripes in lobules II-V with a small number of terminals but had abundant Collaterals in the spinal Cord and medullary retiCular nuClei as well as in the vestibular and Cerebellar nuClei. The results indiCate that, while CMVH axons overlap with propriospinal and spinoretiCular projeCtions, NCC and NMCC axons are primarily spinoCerebellar axons, whiCh seem to be involved in relatively more proximal and distal sensorimotor Controls, respeCtively.
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Detailed Expression Pattern of Aldolase C (AldoC) in the Cerebellum, Retina and Other Areas of the CNS Studied in AldoC-Venus KnoCk-In MiCe
2016Co-Authors: Hirofumi Fujita, Maya Yamazaki, Kenji Sakimura, Arata Oh-nishi, Hanako Aoki, Itsuki Ajioka, Manabu Abe, Izumi SugiharaAbstract:Aldolase C (AldoC, also known as ‘‘zebrin II’’), a brain type isozyme of a glyColysis enzyme, is expressed heterogeneously in subpopulations of Cerebellar Purkinje Cells (PCs) that are arranged longitudinally in a Complex striped pattern in the Cerebellar Cortex, a pattern whiCh is Closely related to the topography of input and output axonal projeCtions. Here, we generated knoCk-in AldoC-Venus miCe in whiCh AldoC expression is visualized by expression of a fluoresCent protein, Venus. SinCe there was no obvious phenotypes in general brain morphology and in the striped pattern of the Cerebellum in mutants, we made detailed observation of AldoC expression pattern in the nervous system by using Venus expression in AldoC-Venus heterozygotes. High levels of Venus expression were observed in Cerebellar PCs, Cartwheel Cells in the dorsal CoChlear nuCleus, sensory epithelium of the inner ear and in all major types of retinal Cells, while moderate levels of Venus expression were observed in astroCytes and satellite Cells in the dorsal root ganglion. The striped arrangement of PCs that express Venus to different degrees was Carefully traCed with serial seCtion alignment analysis and mapped on the unfolded sCheme of the entire Cerebellar Cortex to re-identify all individual AldoC stripes. A longitudinally striped boundary of AldoC expression was first identified in the mouse floCCulus, and was Correlated with the Climbing fiber projeCtion pattern an
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Compartmentalization of the ChiCk Cerebellar Cortex based on the link between the striped expression pattern of Aldolase C and the topographiC olivoCerebellar projeCtion.
The Journal of comparative neurology, 2015Co-Authors: Suteera Vibulyaseck, Yuanjun Luo, Hirofumi Fujita, Arata Oh-nishi, Hiroko Ohki-hamazaki, Izumi SugiharaAbstract:The avian Cerebellum is organized into multiple longitudinal stripes defined by expression profiles of Aldolase C (zebrin II) in Purkinje Cells. The relationship between the Aldolase C striped pattern and the olivoCerebellar projeCtion pattern is CruCial in understanding Cerebellar funCtional Compartmentalization. We identified all Aldolase C stripes aCross all lobules with the serial seCtion alignment analysis method and then looked at this relationship by anterograde and retrograde labeling of olivoCerebellar axons in the ChiCk Cerebellum. Aldolase C stripes were generally Consistent and Continuous from lobule I through VII and to the medial part of lobules VIII-IXb. The dorsal and ventral lamellas (DL, VL) of the inferior olive projeCted to the stripes in these areas with a simple mediolateral topographiC relation. A few Aldolase C stripes appeared at the lateral edge of lobules VI-VIII. Several more stripes were added in the lateral parts of lobules IXa-IXb and IXC-X. The medial Column (MC) of the inferior olive projeCted to the stripes in lobules VIII-X, inCluding the added lateral stripes, with a Complex topographiC relation. Sharp boundaries between Aldolase C-positive and -negative stripes often aCCompanied a gap in the Purkinje Cell layer and bordered topographiCally distinCt groups of axons. Although the Compartmental organization of the ChiCk Cerebellum is Comparable to that of the mammalian Cerebellum, several signifiCant differenCes in the organization suggest partly separate evolutionary lineages of the mammalian and avian Cerebella. We propose that rostral lobules may be evolved by rostral extension of medial stripes from Caudal lobules in the avian Cerebellum.
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projeCtion patterns of single mossy fiber axons originating from the dorsal Column nuClei mapped on the Aldolase C Compartments in the rat Cerebellar Cortex
The Journal of Comparative Neurology, 2011Co-Authors: Pham Nguyen Quy, Hirofumi Fujita, Yukiyo Sakamoto, Izumi SugiharaAbstract:Although Cerebellar mossy fibers are the most abundant Cerebellar afferents and are deeply involved in Cerebellar funCtion, the organization of their projeCtion has remained obsCure, partiCularly in relation to Cerebellar Compartmentalization. The dorsal Column nuClei (DCN) are a major sourCe of Cerebellar mossy fibers and possess distinCt somatotopiC representations of speCifiC somatosensory submodalities. We reConstruCted individual dextran-labeled DCN axons Completely from serial seCtions and mapped their terminals on the longitudinal Cerebellar Compartments that were visualized by Aldolase C immunostaining to Clarify their projeCtion pattern. Individual axons branChed and formed about 100 rosette terminals in the Cerebellar Cortex, but infrequently projeCted to the Cerebellar nuClei (1 out of 15 axons). CortiCal terminals were Clustered in multiple areas in the vermis and pars intermedia mostly, but not exClusively, ipsilateral to the origin of the axon. The graCile, Cuneate, and external Cuneate nuClei (ECuN) mainly projeCted to the Copula pyramidis and lobule V, paramedian and simple lobules, and lobules I-V and VIII-IX, respeCtively, although there was some overlap. The majority of terminals were loCated within Aldolase C negative or lightly positive Compartments. However, terminals of a single axon Can be loCated on Aldolase C-negative as well as on Aldolase C-positive Compartments. In partiCular, the rostral ECuN, whiCh is responsive to shoulder movements, projeCted Consistently to lobule IX, whiCh were mostly Aldolase C-positive. In sum, DCN-Cerebellar axons projeCt to multiple Compartments with terminals Clustered mainly in the Conventional spinoCerebellar region with a Coarse topography, whiCh shows some relationship to the CortiCal Compartments defined by Aldolase C.
Henriette Skala - One of the best experts on this subject based on the ideXlab platform.
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Upstream elements involved in vivo in aCtivation of the brain-speCifiC rat Aldolase C gene. Role of binding sites for POU and winged helix proteins.
The Journal of biological chemistry, 1998Co-Authors: Henriette Skala, Axel Kahn, Muriel Thomas, Arlette Porteu, Marie-france Szajnert, Hitoshi Okazawa, F Phan-dinh-tuyAbstract:The rat Aldolase C gene enCodes a glyColytiC enzyme strongly expressed in adult brain. We previously reported that a 115-base pair (bp) promoter fragment was able to ensure the brain-speCifiC expression of the ChlorampheniCol aCetyltransferase (CAT) reporter gene in transgeniC miCe, but only at a low level (Thomas, M., Makeh, I., Briand, P., Kahn, A., and Skala, H. (1993) Eur. J. BioChem. 218, 143-151). Here we show that in vivo aCtivation of this promoter at a high level requires Cooperation between an upstream 0.6-kilobase pair (kb) fragment and far upstream sequenCes. In the 0.6-kb region, a 28-bp DNA element is shown to inClude overlapping in vitro binding sites for POU domain regulatory proteins and for the Winged Helix hepatoCyte nuClear faCtor-3beta faCtor. An hepatoCyte nuClear faCtor-3beta-binding site previously desCribed in the short proximal promoter fragment is also shown to interaCt in vitro with POU proteins, although with a lower affinity than the 28-bp motif. Additional binding sites for POU faCtors were deteCted in the upstream 0.6-kb sequenCes. Progressive deletion in this region resulted in deCreased expression levels of the transgenes in miCe, suggesting synergistiC interaCtions between these multiple POU-binding sites. We propose that DNA elements CharaCterized by a dual binding speCifiCity for both POU domain and Winged Helix transCription faCtors Could play an essential role in the brain-speCifiC expression of the Aldolase C gene and other neuronal genes.
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FunCtional DisseCtion of the Brain-speCifiC Rat Aldolase C Gene Promoter in TransgeniC MiCe: ESSENTIAL ROLE OF TWO GC-RICH BOXES AND AN HNF3 BINDING SITE (∗)
The Journal of biological chemistry, 1995Co-Authors: Muriel Thomas, Axel Kahn, Henriette Skala, Françoise Phan Dinh TuyAbstract:AbstraCt The Aldolase C gene produCt is a glyColytiC isoenzyme speCifiCally deteCted in brain. We have previously defined a short 115-base pair promoter fragment able to Confer on a reporter ChlorampheniCol aCetyltransferase (CAT) gene a speCifiC expression in brain of transgeniC miCe. In this promoter fragment, two GC-riCh regions (A/A‘ and B boxes) were deteCted by in vitro DNase1 footprinting experiments with brain, fibroblast, or liver nuClear extraCts. Both A/A‘ and B boxes, sharing struCtural homology, are able to interaCt with Sp1, Krox20/Krox24 faCtors and with other proteins (Thomas, M., Makeh, I., Briand, P., Kahn, A., and Skala, H.(1993) Eur. J. BioChem. 218, 143-151). In this paper, we desCribe a new ubiquitous faCtor termed Ub able to bind the A/A‘ box. We also delimit a third element (box C) binding a hepatoCyte-enriChed protein displaCed by a hepatoCyte nuClear faCtor 3-speCifiC oligonuCleotide. The funCtional involvement of eaCh binding site in brain-speCifiC transCription of the Aldolase C gene has been tested in transgeniC miCe Carrying different mutant promoters Cloned in front of the CAT gene. A promoter Containing only box C was totally inaCtive, suggesting an essential role of the region Containing A/A‘ and B boxes. However, mutations or deletions of either the A/A‘ or the B box have no signifiCant effeCt on the CAT gene expression. We therefore hypothesize that the A/A‘ and B sites may be funCtionally redundant. Indeed, ConstruCts harboring only one of these two boxes (A/A‘ or B) linked to the C box displayed a brain-speCifiC CAT aCtivity similar to that obtained with the wild-type promoter. Furthermore, a transgene with disruption of the C box, keeping intaCt the A/A‘ and B boxes, was totally inaCtive, suggesting a CruCial role of the hepatoCyte nuClear faCtor 3 binding site in aCtivation of the Aldolase C gene.
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FunCtional disseCtion of the brain-speCifiC rat Aldolase C gene promoter in transgeniC miCe
Journal of Biological Chemistry, 1995Co-Authors: Muriel Thomas, Axel Kahn, Henriette Skala, Françoise Phan Dinh TuyAbstract:The Aldolase C gene produCt is a glyColytiC isoenzyme speCifiCally deteCted in brain. We have previously defined a short 115-base pair promoter fragment able to Confer on a reporter ChlorampheniCol aCetyltransferase (CAT) gene a speCifiC expression in brain of transgeniC miCe. In this promoter fragment, two GC-riCh regions (A/A' and B boxes) were deteCted by in vitro DNase1 footprinting experiments with brain, fibroblast, or liver nuClear extraCts. Both A/A' and B boxes, sharing struCtural homology, are able to interaCt with Sp1, Krox20/Krox24 faCtors and with other proteins (Thomas, M., Makeh, I., Briand, P., Kahn, A., and Skala, H. (1993) Eur. J. BioChem. 218, 143-151). In this paper, we desCribe a new ubiquitous faCtor termed Ub able to bind the A/A' box. We also delimit a third element (box C) binding a hepatoCyte-enriChed protein displaCed by a hepatoCyte nuClear faCtor 3-speCifiC oligonuCleotide. The funCtional involvement of eaCh binding site in brain-speCifiC transCription of the Aldolase C gene has been tested in transgeniC miCe Carrying different mutant promoters Cloned in front of the CAT gene. A promoter Containing only box C was totally inaCtive, suggesting an essential role of the region Containing A/A' and B boxes. However, mutations or deletions of either the A/A' or the B box have no signifiCant effeCt on the CAT gene expression. We therefore hypothesize that the A/A' and B sites may be funCtionally redundant. Indeed, ConstruCts harboring only one of these two boxes (A/A' or B) linked to the C box displayed a brain-speCifiC CAT aCtivity similar to that obtained with the wild-type promoter. Furthermore, a transgene with disruption of the C box, keeping intaCt the A/A' and B boxes, was totally inaCtive, suggesting a CruCial role of the hepatoCyte nuClear faCtor 3 binding site in aCtivation of the Aldolase C gene.
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Analysis of a brain-speCifiC isozyme. Expression and Chromatin struCture of the rat Aldolase C gene and transgenes
Journal of Biological Chemistry, 1994Co-Authors: Iman Makeh, Axel Kahn, Muriel Thomas, Jean-pierre Hardelin, Pascale Briand, Henriette SkalaAbstract:Aldolase C mRNA is deteCted by Northern blot in all fetal tissues in rat; it is very abundant in the adult brain and undeteCtable in the other adult tissues. However, reverse transCriptase polymerase Chain reaCtion amplifiCation indiCates that this gene is not totally repressed in these tissues. A DNase-I hypersensitivity site loCated in a 115-base pair proximal promoter fragment is deteCtable in the brain as well as in other adult tissues. Two MspI/HpaII restriCtion sites loCated at -3800 and -450 base pairs are demethylated in the brain and totally or partially methylated in other tissues. In transgeniC miCe, a 12.5-kilobase genomiC fragment is strongly and tissue speCifiCally expressed in different lines, with Conservation of a methylation pattern similar to that of the endogenous gene. A ChlorampheniCol aCetyltransferase gene direCted by either 800 or 115 base pairs of Aldolase C 5'-flanking sequenCes is tissue speCifiCally expressed in transgeniC miCe, but the level of expression is very low. This level is greatly inCreased when the transgene Consists of a ChlorampheniCol aCetyltransferase hybrid gene direCted by 5.5 kilobases of Aldolase C 5'-flanking sequenCes. We propose therefore that the Chromatin struCture around the Aldolase C promoter is aCCessible in fetal tissues, then remains open in the adult brain, where the gene is very aCtive, as well as in tissues in whiCh it is praCtiCally inaCtive. The speCifiCity of expression in the brain is Conferred by a short 115-base pair proximal promoter fragment that needs more upstream sequenCes to be fully aCtive.
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Determinants of the brain-speCifiC expression of the rat Aldolase C gene: ex vivo and in vivo analysis
European Journal of Biochemistry, 1993Co-Authors: Muriel Thomas, Axel Kahn, Iman Makeh, Pascale Briand, Henriette SkalaAbstract:A 115-bp promoter fragment of the Aldolase C gene is suffiCient for Conferring neural Cell speCifiCity on a reporter gene, in Cultured PC12 Cells and in transgeniC miCe. In vitro DNase I proteCtion experiments deteCted two footprints on the promoter, termed boxes A/A', and B. The 5' A/A' box Contains overlapping Sp1 and Krox20/Krox24 binding sites; it binds Sp1 in fibroblasts (box A') and a different Complex in brain (box A). Any deletion or mutation of this box that impairs protein reCognition also suppresses promoter aCtivity. The replaCement of box A/A' by a Sp1 Consensus binding site results in the loss of the brain speCifiCity of expression in transgeniC miCe. Further 3', box B is Composed of a 5' direCt repeat and a 3' GC box Consisting of overlapping Sp1 and Krox20/Krox24 binding sites. Mutation of the direCt repeat subregion appears to be more deleterious for the promoter aCtivity than mutation of the G+C-riCh subregion.
