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Karl J Kramer - One of the best experts on this subject based on the ideXlab platform.
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Development and ultrastructure of the rigid dorsal and flexible ventral Cuticles of the elytron of the red flour beetle, Tribolium castaneum.
Insect biochemistry and molecular biology, 2017Co-Authors: Mi Young Noh, Subbaratnam Muthukrishnan, Karl J Kramer, Yasuyuki ArakaneAbstract:Insect exoskeletons are composed of the Cuticle, a biomaterial primarily formed from the linear and relatively rigid polysaccharide, chitin, and structural proteins. This extracellular material serves both as a skin and skeleton, protecting insects from environmental stresses and mechanical damage. Despite its rather limited compositional palette, Cuticles in different anatomical regions or developmental stages exhibit remarkably diverse physicochemical and mechanical properties because of differences in chemical composition, molecular interactions and morphological architecture of the various layers and sublayers throughout the Cuticle including the envelope, epiCuticle and proCuticle (exoCuticle and endoCuticle). Even though the ultrastructure of the arthropod Cuticle has been studied rather extensively, its temporal developmental pattern, in particular, the synchronous development of the functional layers in different Cuticles during a molt, is not well understood. The beetle elytron, which is a highly modified and sclerotized forewing, offers excellent advantages for such a study because it can be easily isolated at precise time points during development. In this study, we describe the morphogenesis of the dorsal and ventral Cuticles of the elytron of the red flour beetle, Tribolium castaneum, during the period from the 0 d-old pupa to the 9 d-old adult. The deposition of exoCuticle and mesoCuticle is substantially different in the two Cuticles. The dorsal Cuticle is four-fold thicker than the ventral. Unlike the ventral Cuticle, the dorsal contains a thicker exoCuticle consisting of a large number of horizontal laminae and vertical pore canals with pore canal fibers and rib-like veins and bristles as well as a mesoCuticle, lying right above the enodCuticle. The degree of sclerotization appears to be much greater in the dorsal Cuticle. All of these differences result in a relatively thick and tanned rigid dorsal Cuticle and a much thinner and less pigmented membrane-like ventral Cuticle.
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cuticular protein with a low complexity sequence becomes cross linked during insect Cuticle sclerotization and is required for the adult molt
Scientific Reports, 2015Co-Authors: Neal T Dittmer, Karl J Kramer, Subbaratnam Muthukrishnan, Yasuyuki ArakaneAbstract:In the insect Cuticle, structural proteins (CPs) and the polysaccharide chitin are the major components. It has been hypothesized that CPs are cross-linked to other CPs and possibly to chitin by quinones or quinone methides produced by the laccase2-mediated oxidation of N-acylcatechols. In this study we investigated functions of TcCP30, the third most abundant CP in protein extracts of elytra (wing covers) from Tribolium castaneum adults. The mature TcCP30 protein has a low complexity and highly polar amino acid sequence. TcCP30 is localized with chitin in horizontal laminae and vertically oriented columnar structures in rigid Cuticles, but not in soft and membranous Cuticles. Immunoblot analysis revealed that TcCP30 undergoes laccase2-mediated cross-linking during Cuticle maturation in vivo, a process confirmed in vitro using recombinant rTcCP30. We identified TcCPR27 and TcCPR18, the two most abundant proteins in the elytra, as putative cross-linking partners of TcCP30. RNAi for the TcCP30 gene had no effect on larval and pupal growth and development. However, during adult eclosion, ~70% of the adults were unable to shed their exuvium and died. These results support the hypothesis that TcCP30 plays an integral role as a cross-linked structural protein in the formation of lightweight rigid Cuticle of the beetle.
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Tribolium castaneum RR-1 cuticular protein TcCPR4 is required for formation of pore canals in rigid Cuticle.
PLoS genetics, 2015Co-Authors: Mi Young Noh, Subbaratnam Muthukrishnan, Karl J Kramer, Yasuyuki ArakaneAbstract:Insect Cuticle is composed mainly of structural proteins and the polysaccharide chitin. The CPR family is the largest family of Cuticle proteins (CPs), which can be further divided into three subgroups based on the presence of one of the three presumptive chitin-binding sequence motifs denoted as Rebers-Riddiford (R&R) consensus sequence motifs RR-1, RR-2 and RR-3. The TcCPR27 protein containing the RR-2 motif is one of the most abundant CPs present both in the horizontal laminae and in vertical pore canals in the proCuticle of rigid Cuticle found in the elytron of the red flour beetle, Tribolium castaneum. Depletion of TcCPR27 by RNA interference (RNAi) causes both unorganized laminae and pore canals, resulting in malformation and weakening of the elytron. In this study, we investigated the function(s) of another CP, TcCPR4, which contains the RR-1 motif and is easily extractable from elytra after RNAi to deplete the level of TcCPR27. Transcript levels of the TcCPR4 gene are dramatically increased in 3 d-old pupae when adult Cuticle synthesis begins. Immunohistochemical studies revealed that TcCPR4 protein is present in the rigid Cuticles of the dorsal elytron, ventral abdomen and leg but not in the flexible Cuticles of the hindwing and dorsal abdomen of adult T. castaneum. Immunogold labeling and transmission electron microscopic analyses revealed that TcCPR4 is predominantly localized in pore canals and regions around the apical plasma membrane protrusions into the proCuticle of rigid adult Cuticles. RNAi for TcCPR4 resulted in an abnormal shape of the pore canals with amorphous pore canal fibers (PCFs) in their lumen. These results support the hypothesis that TcCPR4 is required for achieving proper morphology of the vertical pore canals and PCFs that contribute to the assembly of a Cuticle that is both lightweight and rigid.
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proteomic and transcriptomic analyses of rigid and membranous Cuticles and epidermis from the elytra and hindwings of the red flour beetle tribolium castaneum
Journal of Proteome Research, 2012Co-Authors: Neal T Dittmer, Karl J Kramer, Richard W. Beeman, Yasuaki Hiromasa, John M. Tomich, Michael R. KanostAbstract:The insect Cuticle is a composite biomaterial made up primarily of chitin and proteins. The physical properties of the Cuticle can vary greatly from hard and rigid to soft and flexible. Understanding how different Cuticle types are assembled can aid in the development of novel biomimetic materials for use in medicine and technology. Toward this goal, we have taken a combined proteomics and transcriptomics approach with the red flour beetle, Tribolium castaneum, to examine the protein and gene expression profiles of the elytra and hindwings, appendages that contain rigid and soft Cuticles, respectively. Two-dimensional gel electrophoresis analysis revealed distinct differences in the protein profiles between elytra and hindwings, with four highly abundant proteins dominating the elytral Cuticle extract. MALDI/TOF mass spectrometry identified 19 proteins homologous to known or hypothesized cuticular proteins (CPs), including a novel low complexity protein enriched in charged residues. Microarray analysis id...
Yasuyuki Arakane - One of the best experts on this subject based on the ideXlab platform.
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Development and ultrastructure of the rigid dorsal and flexible ventral Cuticles of the elytron of the red flour beetle, Tribolium castaneum.
Insect biochemistry and molecular biology, 2017Co-Authors: Mi Young Noh, Subbaratnam Muthukrishnan, Karl J Kramer, Yasuyuki ArakaneAbstract:Insect exoskeletons are composed of the Cuticle, a biomaterial primarily formed from the linear and relatively rigid polysaccharide, chitin, and structural proteins. This extracellular material serves both as a skin and skeleton, protecting insects from environmental stresses and mechanical damage. Despite its rather limited compositional palette, Cuticles in different anatomical regions or developmental stages exhibit remarkably diverse physicochemical and mechanical properties because of differences in chemical composition, molecular interactions and morphological architecture of the various layers and sublayers throughout the Cuticle including the envelope, epiCuticle and proCuticle (exoCuticle and endoCuticle). Even though the ultrastructure of the arthropod Cuticle has been studied rather extensively, its temporal developmental pattern, in particular, the synchronous development of the functional layers in different Cuticles during a molt, is not well understood. The beetle elytron, which is a highly modified and sclerotized forewing, offers excellent advantages for such a study because it can be easily isolated at precise time points during development. In this study, we describe the morphogenesis of the dorsal and ventral Cuticles of the elytron of the red flour beetle, Tribolium castaneum, during the period from the 0 d-old pupa to the 9 d-old adult. The deposition of exoCuticle and mesoCuticle is substantially different in the two Cuticles. The dorsal Cuticle is four-fold thicker than the ventral. Unlike the ventral Cuticle, the dorsal contains a thicker exoCuticle consisting of a large number of horizontal laminae and vertical pore canals with pore canal fibers and rib-like veins and bristles as well as a mesoCuticle, lying right above the enodCuticle. The degree of sclerotization appears to be much greater in the dorsal Cuticle. All of these differences result in a relatively thick and tanned rigid dorsal Cuticle and a much thinner and less pigmented membrane-like ventral Cuticle.
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cuticular protein with a low complexity sequence becomes cross linked during insect Cuticle sclerotization and is required for the adult molt
Scientific Reports, 2015Co-Authors: Neal T Dittmer, Karl J Kramer, Subbaratnam Muthukrishnan, Yasuyuki ArakaneAbstract:In the insect Cuticle, structural proteins (CPs) and the polysaccharide chitin are the major components. It has been hypothesized that CPs are cross-linked to other CPs and possibly to chitin by quinones or quinone methides produced by the laccase2-mediated oxidation of N-acylcatechols. In this study we investigated functions of TcCP30, the third most abundant CP in protein extracts of elytra (wing covers) from Tribolium castaneum adults. The mature TcCP30 protein has a low complexity and highly polar amino acid sequence. TcCP30 is localized with chitin in horizontal laminae and vertically oriented columnar structures in rigid Cuticles, but not in soft and membranous Cuticles. Immunoblot analysis revealed that TcCP30 undergoes laccase2-mediated cross-linking during Cuticle maturation in vivo, a process confirmed in vitro using recombinant rTcCP30. We identified TcCPR27 and TcCPR18, the two most abundant proteins in the elytra, as putative cross-linking partners of TcCP30. RNAi for the TcCP30 gene had no effect on larval and pupal growth and development. However, during adult eclosion, ~70% of the adults were unable to shed their exuvium and died. These results support the hypothesis that TcCP30 plays an integral role as a cross-linked structural protein in the formation of lightweight rigid Cuticle of the beetle.
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Tribolium castaneum RR-1 cuticular protein TcCPR4 is required for formation of pore canals in rigid Cuticle.
PLoS genetics, 2015Co-Authors: Mi Young Noh, Subbaratnam Muthukrishnan, Karl J Kramer, Yasuyuki ArakaneAbstract:Insect Cuticle is composed mainly of structural proteins and the polysaccharide chitin. The CPR family is the largest family of Cuticle proteins (CPs), which can be further divided into three subgroups based on the presence of one of the three presumptive chitin-binding sequence motifs denoted as Rebers-Riddiford (R&R) consensus sequence motifs RR-1, RR-2 and RR-3. The TcCPR27 protein containing the RR-2 motif is one of the most abundant CPs present both in the horizontal laminae and in vertical pore canals in the proCuticle of rigid Cuticle found in the elytron of the red flour beetle, Tribolium castaneum. Depletion of TcCPR27 by RNA interference (RNAi) causes both unorganized laminae and pore canals, resulting in malformation and weakening of the elytron. In this study, we investigated the function(s) of another CP, TcCPR4, which contains the RR-1 motif and is easily extractable from elytra after RNAi to deplete the level of TcCPR27. Transcript levels of the TcCPR4 gene are dramatically increased in 3 d-old pupae when adult Cuticle synthesis begins. Immunohistochemical studies revealed that TcCPR4 protein is present in the rigid Cuticles of the dorsal elytron, ventral abdomen and leg but not in the flexible Cuticles of the hindwing and dorsal abdomen of adult T. castaneum. Immunogold labeling and transmission electron microscopic analyses revealed that TcCPR4 is predominantly localized in pore canals and regions around the apical plasma membrane protrusions into the proCuticle of rigid adult Cuticles. RNAi for TcCPR4 resulted in an abnormal shape of the pore canals with amorphous pore canal fibers (PCFs) in their lumen. These results support the hypothesis that TcCPR4 is required for achieving proper morphology of the vertical pore canals and PCFs that contribute to the assembly of a Cuticle that is both lightweight and rigid.
Neal T Dittmer - One of the best experts on this subject based on the ideXlab platform.
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cuticular protein with a low complexity sequence becomes cross linked during insect Cuticle sclerotization and is required for the adult molt
Scientific Reports, 2015Co-Authors: Neal T Dittmer, Karl J Kramer, Subbaratnam Muthukrishnan, Yasuyuki ArakaneAbstract:In the insect Cuticle, structural proteins (CPs) and the polysaccharide chitin are the major components. It has been hypothesized that CPs are cross-linked to other CPs and possibly to chitin by quinones or quinone methides produced by the laccase2-mediated oxidation of N-acylcatechols. In this study we investigated functions of TcCP30, the third most abundant CP in protein extracts of elytra (wing covers) from Tribolium castaneum adults. The mature TcCP30 protein has a low complexity and highly polar amino acid sequence. TcCP30 is localized with chitin in horizontal laminae and vertically oriented columnar structures in rigid Cuticles, but not in soft and membranous Cuticles. Immunoblot analysis revealed that TcCP30 undergoes laccase2-mediated cross-linking during Cuticle maturation in vivo, a process confirmed in vitro using recombinant rTcCP30. We identified TcCPR27 and TcCPR18, the two most abundant proteins in the elytra, as putative cross-linking partners of TcCP30. RNAi for the TcCP30 gene had no effect on larval and pupal growth and development. However, during adult eclosion, ~70% of the adults were unable to shed their exuvium and died. These results support the hypothesis that TcCP30 plays an integral role as a cross-linked structural protein in the formation of lightweight rigid Cuticle of the beetle.
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proteomic and transcriptomic analyses of rigid and membranous Cuticles and epidermis from the elytra and hindwings of the red flour beetle tribolium castaneum
Journal of Proteome Research, 2012Co-Authors: Neal T Dittmer, Karl J Kramer, Richard W. Beeman, Yasuaki Hiromasa, John M. Tomich, Michael R. KanostAbstract:The insect Cuticle is a composite biomaterial made up primarily of chitin and proteins. The physical properties of the Cuticle can vary greatly from hard and rigid to soft and flexible. Understanding how different Cuticle types are assembled can aid in the development of novel biomimetic materials for use in medicine and technology. Toward this goal, we have taken a combined proteomics and transcriptomics approach with the red flour beetle, Tribolium castaneum, to examine the protein and gene expression profiles of the elytra and hindwings, appendages that contain rigid and soft Cuticles, respectively. Two-dimensional gel electrophoresis analysis revealed distinct differences in the protein profiles between elytra and hindwings, with four highly abundant proteins dominating the elytral Cuticle extract. MALDI/TOF mass spectrometry identified 19 proteins homologous to known or hypothesized cuticular proteins (CPs), including a novel low complexity protein enriched in charged residues. Microarray analysis id...
Gaetan Guignard - One of the best experts on this subject based on the ideXlab platform.
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thirty three years 1986 2019 of fossil plant Cuticle studies using transmission electron microscopy a review
Review of Palaeobotany and Palynology, 2019Co-Authors: Gaetan GuignardAbstract:Abstract Although the Cuticle is a very thin layer of a few micrometers, it is one of the main remains found among fossil plants. Apparently very fragile, ultrathin Cuticle sections of few dozen nanometers thick are studied by using TEM studies and statistical measurements. The ultrastructure details yielded are of significance not only for taxonomy, but also for paleoenvironment and evolution of fossil plant. After an historical introduction and a review of the methods to obtain and observe the ultrathin sections, although data are still not enough numerous, detailed information and interesting facts of Cuticle ultrastructure emerge from the studied plant groups. This paper gives a review of the main results of transmission electron microscopy -TEM- of fossil plant Cuticles since the pioneering study 33 years ago.
Mi Young Noh - One of the best experts on this subject based on the ideXlab platform.
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Development and ultrastructure of the rigid dorsal and flexible ventral Cuticles of the elytron of the red flour beetle, Tribolium castaneum.
Insect biochemistry and molecular biology, 2017Co-Authors: Mi Young Noh, Subbaratnam Muthukrishnan, Karl J Kramer, Yasuyuki ArakaneAbstract:Insect exoskeletons are composed of the Cuticle, a biomaterial primarily formed from the linear and relatively rigid polysaccharide, chitin, and structural proteins. This extracellular material serves both as a skin and skeleton, protecting insects from environmental stresses and mechanical damage. Despite its rather limited compositional palette, Cuticles in different anatomical regions or developmental stages exhibit remarkably diverse physicochemical and mechanical properties because of differences in chemical composition, molecular interactions and morphological architecture of the various layers and sublayers throughout the Cuticle including the envelope, epiCuticle and proCuticle (exoCuticle and endoCuticle). Even though the ultrastructure of the arthropod Cuticle has been studied rather extensively, its temporal developmental pattern, in particular, the synchronous development of the functional layers in different Cuticles during a molt, is not well understood. The beetle elytron, which is a highly modified and sclerotized forewing, offers excellent advantages for such a study because it can be easily isolated at precise time points during development. In this study, we describe the morphogenesis of the dorsal and ventral Cuticles of the elytron of the red flour beetle, Tribolium castaneum, during the period from the 0 d-old pupa to the 9 d-old adult. The deposition of exoCuticle and mesoCuticle is substantially different in the two Cuticles. The dorsal Cuticle is four-fold thicker than the ventral. Unlike the ventral Cuticle, the dorsal contains a thicker exoCuticle consisting of a large number of horizontal laminae and vertical pore canals with pore canal fibers and rib-like veins and bristles as well as a mesoCuticle, lying right above the enodCuticle. The degree of sclerotization appears to be much greater in the dorsal Cuticle. All of these differences result in a relatively thick and tanned rigid dorsal Cuticle and a much thinner and less pigmented membrane-like ventral Cuticle.
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Tribolium castaneum RR-1 cuticular protein TcCPR4 is required for formation of pore canals in rigid Cuticle.
PLoS genetics, 2015Co-Authors: Mi Young Noh, Subbaratnam Muthukrishnan, Karl J Kramer, Yasuyuki ArakaneAbstract:Insect Cuticle is composed mainly of structural proteins and the polysaccharide chitin. The CPR family is the largest family of Cuticle proteins (CPs), which can be further divided into three subgroups based on the presence of one of the three presumptive chitin-binding sequence motifs denoted as Rebers-Riddiford (R&R) consensus sequence motifs RR-1, RR-2 and RR-3. The TcCPR27 protein containing the RR-2 motif is one of the most abundant CPs present both in the horizontal laminae and in vertical pore canals in the proCuticle of rigid Cuticle found in the elytron of the red flour beetle, Tribolium castaneum. Depletion of TcCPR27 by RNA interference (RNAi) causes both unorganized laminae and pore canals, resulting in malformation and weakening of the elytron. In this study, we investigated the function(s) of another CP, TcCPR4, which contains the RR-1 motif and is easily extractable from elytra after RNAi to deplete the level of TcCPR27. Transcript levels of the TcCPR4 gene are dramatically increased in 3 d-old pupae when adult Cuticle synthesis begins. Immunohistochemical studies revealed that TcCPR4 protein is present in the rigid Cuticles of the dorsal elytron, ventral abdomen and leg but not in the flexible Cuticles of the hindwing and dorsal abdomen of adult T. castaneum. Immunogold labeling and transmission electron microscopic analyses revealed that TcCPR4 is predominantly localized in pore canals and regions around the apical plasma membrane protrusions into the proCuticle of rigid adult Cuticles. RNAi for TcCPR4 resulted in an abnormal shape of the pore canals with amorphous pore canal fibers (PCFs) in their lumen. These results support the hypothesis that TcCPR4 is required for achieving proper morphology of the vertical pore canals and PCFs that contribute to the assembly of a Cuticle that is both lightweight and rigid.
