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  • 1
    Online Resource
    Online Resource
    Pore properties and pharmacological features of the P2X receptor channel in airway ciliated cells
    Wiley ; 2006
    In:  The Journal of Physiology Vol. 571, No. 3 ( 2006-03), p. 503-517
    Details
    In: The Journal of Physiology, Wiley, Vol. 571, No. 3 ( 2006-03), p. 503-517
    Abstract: Airway ciliated cells express an ATP‐gated P2X receptor channel of unknown subunit composition (P2X cilia ) which is modulated by Na + and by long exposures to ATP. P2X cilia was investigated by recording currents from freshly dissociated rabbit airway ciliated cells with the patch‐clamp technique in the whole‐cell configuration. During the initial continuous exposure to extracellular ATP, P2X cilia currents gradually increase in magnitude (priming), yet the permeability to N ‐methyl‐ d ‐glucamine (NMDG) does not change, indicating that priming does not arise from a progressive change in pore diameter. Na + , which readily permeates P2X cilia receptor channels, was found to inhibit the channel extracellular to the electric field. The rank order of permeability to various monovalent cations is: Li + , Na + , K + , Rb + , Cs + , NMDG + and TEA + , with a relative permeability of 1.35, 1.0, 0.99, 0.91, 0.79, 0.19 and 0.10, respectively. The rank order for the alkali cations follows an Eisenman series XI for a high‐strength field site. Ca 2+ has been estimated to be 7‐fold more permeant than Na + . The rise in [Ca 2+ ] i in ciliated cells, induced by the activation of P2X cilia , is largely inhibited by either Brilliant Blue G or KN‐62, indicating that P2X 7 may be a part of P2X cilia . P2X cilia is augmented by Zn 2+ and by ivermectin, and P2X 4 receptor protein is detected by immunolabelling at the basal half of the cilia, strongly suggesting that P2X 4 is a component of P2X cilia receptor channels. Taken together, these results suggest that P2X cilia is either assembled from P2X 4 and P2X 7 subunits, or formed from modified P2X 4 subunits.
    Type of Medium: Online Resource
    ISSN: 0022-3751 , 1469-7793
    URL: Article
    DOI: 10.1113/jphysiol.2005.103408
    RVK:
    WA 15000
    Language: English
    Publisher: Wiley
    Publication Date: 2006
    detail.hit.zdb_id: 1475290-6
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  • 2
    Online Resource
    Online Resource
    The identification of the phosphorylated 150/160-kDa proteins of sarcoplasmic reticulum, their kinase and their association with the ryanodine receptor
    Elsevier BV ; 1996
    In:  Biochimica et Biophysica Acta (BBA) - Biomembranes Vol. 1283, No. 1 ( 1996-08), p. 89-100
    Details
    In: Biochimica et Biophysica Acta (BBA) - Biomembranes, Elsevier BV, Vol. 1283, No. 1 ( 1996-08), p. 89-100
    Type of Medium: Online Resource
    ISSN: 0005-2736
    URL: Article
    DOI: 10.1016/0005-2736(96)00079-X
    RVK:
    WA 15000
    Language: English
    Publisher: Elsevier BV
    Publication Date: 1996
    detail.hit.zdb_id: 2209384-9
    SSG: 12
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  • 3
    Online Resource
    Online Resource
    Insulin and gender: An insulin-like gene expressed exclusively in the androgenic gland of the male crayfish
    Elsevier BV ; 2007
    In:  General and Comparative Endocrinology Vol. 150, No. 2 ( 2007-1), p. 326-336
    Details
    In: General and Comparative Endocrinology, Elsevier BV, Vol. 150, No. 2 ( 2007-1), p. 326-336
    Type of Medium: Online Resource
    ISSN: 0016-6480
    URL: Article
    DOI: 10.1016/j.ygcen.2006.09.006
    Language: English
    Publisher: Elsevier BV
    Publication Date: 2007
    detail.hit.zdb_id: 1467679-5
    SSG: 12
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  • 4
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    The eyestalk–androgenic gland–testis endocrine axis in the crayfish Cherax quadricarinatus
    Elsevier BV ; 2002
    In:  General and Comparative Endocrinology Vol. 127, No. 2 ( 2002-6), p. 147-156
    Details
    In: General and Comparative Endocrinology, Elsevier BV, Vol. 127, No. 2 ( 2002-6), p. 147-156
    Type of Medium: Online Resource
    ISSN: 0016-6480
    URL: Article
    DOI: 10.1016/S0016-6480(02)00031-X
    Language: English
    Publisher: Elsevier BV
    Publication Date: 2002
    detail.hit.zdb_id: 1467679-5
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  • 5
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    Temporal Silencing of an Androgenic Gland-Specific Insulin-Like Gene Affecting Phenotypical Gender Differences and Spermatogenesis
    The Endocrine Society ; 2009
    In:  Endocrinology Vol. 150, No. 3 ( 2009-03-01), p. 1278-1286
    Details
    In: Endocrinology, The Endocrine Society, Vol. 150, No. 3 ( 2009-03-01), p. 1278-1286
    Abstract: Androgenic glands (AGs) of the freshwater prawn Macrobrachium rosenbergii were subjected to endocrine manipulation, causing them to hypertrophy. Transcripts from these glands were used in the construction of an AG cDNA subtractive library. Screening of the library revealed an AG-specific gene, termed the M. rosenbergii insulin-like AG (Mr-IAG) gene. The cDNA of this gene was then cloned and fully sequenced. The cysteine backbone of the predicted mature Mr-IAG peptide (B and A chains) showed high similarity to that of other crustacean AG-specific insulin-like peptides. In vivo silencing of the gene, by injecting the prawns with Mr-IAG double-stranded RNA, temporarily prevented the regeneration of male secondary sexual characteristics, accompanied by a lag in molt and a reduction in growth parameters, which are typically higher in males of the species. In terms of reproductive parameters, silencing of Mr-IAG led to the arrest of testicular spermatogenesis and of spermatophore development in the terminal ampullae of the sperm duct, accompanied by hypertrophy and hyperplasia of the AGs. This study constitutes the first report of the silencing of a gene expressed specifically in the AG, which caused a transient adverse effect on male phenotypical gender differences and spermatogenesis. Temporal silencing of a newly identified insulin-like gene from prawn androgenic gland inhibits primary spermatogenesis, male secondary sex characteristics, and growth.
    Type of Medium: Online Resource
    ISSN: 0013-7227 , 1945-7170
    URL: Article
    DOI: 10.1210/en.2008-0906
    Language: English
    Publisher: The Endocrine Society
    Publication Date: 2009
    detail.hit.zdb_id: 2011695-0
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  • 6
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    Online Resource
    Image_2.jpeg
    Frontiers Media SA ; 2023
    In:  Frontiers in Marine Science Vol. 9 ( 2023-1-9)
    Details
    In: Frontiers in Marine Science, Frontiers Media SA, Vol. 9 ( 2023-1-9)
    Abstract: Crustacean cell line immortalization has gained a great deal of attention in recent decades for both scientific and applied reasons. Our goal in this study was to advance the state of art towards establishing an immortalized cell line by improving the proliferation rates of primary cells isolated from embryos of the giant freshwater prawn Macrobrachium rosenbergii by using a lentivirus expressing the Ras oncogene. The choice of Ras derived from its involvement in various cellular pathways, such as cell growth, differentiation, and survival, and its use as a tool for in-vitro immortalization, e.g., a specific mutated Ras ( Ras V12 ) was used to generate an arthropod cell line. Complementarily, in-silico screening of M. rosenbergii transcriptomic libraries for Ras expression indicated that Ras is already expressed at very early stages of embryo development. In the current study, we transduced primary M. rosenbergii embryonic cells with a lentivirus expressing Ras V12 by using the white spot syndrome virus ( WSSV IE1 ) promoter. Expression and sequencing (as followed by sequencing cDNA, confocal microscopy and FACS analysis) of the mutated Ras in the transduced cells confirmed that the lentivirus was successfully integrated into the genome. The lenti -MrRas transduction rate was 23% in the total primary cell population and more than 80% in a sub-population of cells with high granularity. Proliferation of lenti- MrRas transfected cells was enhanced to almost 1200% of the seeding density by the end of our experiment (18 days), which was double that of the control. We were thus successful in enhancing the longevity of embryonic primary cell cultures by ectopic expression of the mutated Ras protein, but the improvement was not sufficient for immortalization.
    Type of Medium: Online Resource
    ISSN: 2296-7745
    URL: Article
    URL: Article
    URL: Article
    URL: Article
    DOI: 10.3389/fmars.2022.1100971
    DOI: 10.3389/fmars.2022.1100971.s001
    DOI: 10.3389/fmars.2022.1100971.s002
    DOI: 10.3389/fmars.2022.1100971.s003
    Language: Unknown
    Publisher: Frontiers Media SA
    Publication Date: 2023
    detail.hit.zdb_id: 2757748-X
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  • 7
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    Online Resource
    In Situ Crosslinking of Highly Porous Chitosan Scaffolds for Bone Regeneration: Production Parameters and In Vitro Characterization
    Wiley ; 2017
    In:  Macromolecular Materials and Engineering Vol. 302, No. 10 ( 2017-10)
    Details
    In: Macromolecular Materials and Engineering, Wiley, Vol. 302, No. 10 ( 2017-10)
    Abstract: Various methods of chitosan scaffold production are reported in the literature so far. Here, in situ crosslinking with glutaraldehyde is reported for the first time. It combines pore formation and chitosan crosslinking in a single step. This combination allows incorporation of fragile molecules into 3D porous chitosan scaffolds produced by simple and gentle lyophilization. In this study, parameters of in situ crosslinking of porous chitosan scaffold formation as well as their effect on degradation and bioactivity of the scaffolds are examined. The scaffolds are characterized in the context of their prospective application as bone substitute material. The addition of calcium phosphate phases (hydroxyapatite, brushite) to the macroporous chitosan scaffolds allows manipulation of the bioactivity that is investigated by incubation in simulated body fluid (SBF). The bioactivity is significantly influenced by the modus of changing the fluid (static, daily‐, and twice‐a‐week change). Scaffolds are morphologically characterized by means of scanning electron microscopy, and the mechanical stability is tested after incubation in SBF and phosphate‐buffered saline.
    Type of Medium: Online Resource
    ISSN: 1438-7492 , 1439-2054
    URL: Issue
    URL: Article
    DOI: 10.1002/mame.v302.10
    DOI: 10.1002/mame.201700147
    Language: English
    Publisher: Wiley
    Publication Date: 2017
    detail.hit.zdb_id: 2004372-7
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  • 8
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    Crayfish hemocyanin on chitin bone substitute scaffolds promotes the proliferation and osteogenic differentiation of human mesenchymal stem cells
    Wiley ; 2020
    In:  Journal of Biomedical Materials Research Part A Vol. 108, No. 3 ( 2020-03), p. 694-708
    Details
    In: Journal of Biomedical Materials Research Part A, Wiley, Vol. 108, No. 3 ( 2020-03), p. 694-708
    Abstract: Crustacean chitin–hemocyanin–calcium mineral complexes were designed as bone biomimetics, with emphasis on their ability to bind or release calcium ions. Chitin scaffolds were prepared by dissolving chitin flakes in LiCl/dimethylacetamide, followed by gel formation and freeze‐drying. Some of these scaffolds were modified by incorporation of CaCO 3 . In some of the chitin–CaCO 3 scaffolds, macroporosity was introduced by HCl treatment. Hemocyanin from the crayfish Cherax quadricarinatus was used to further modify the chitin scaffolds by dip coating. Cytocompatibility, cellular adherence and proliferation of human mesenchymal stem cells (hMSCs) were evaluated in terms of cell number as reflected in lactate dehydrogenase activity. The chitin, chitin–CaCO 3 , and porous chitin–CaCO 3 scaffolds were all found to facilitate cell attachment. Hemocyanin dip‐coating of these scaffolds led to increased initial cell adhesion, enhanced proliferation, and osteogenic differentiation. Since the hemocyanin loading of the scaffolds was impaired by sterilization by gamma‐irradiation (as required for biomedical applications), the hemocyanin loading was performed on previously sterilized scaffolds. All scaffolds facilitated osteogenic differentiation of osteoblasts, with the highest cell ALP‐activity being found on hemocyanin‐modified porous chitin–CaCO 3 scaffolds. Thus, chitin–hemocyanin scaffolds enhanced the initial stages of bone cell development and could serve as promising biomaterials for bone regeneration.
    Type of Medium: Online Resource
    ISSN: 1549-3296 , 1552-4965
    URL: Issue
    URL: Article
    DOI: 10.1002/jbm.a.v108.3
    DOI: 10.1002/jbm.a.36849
    Language: English
    Publisher: Wiley
    Publication Date: 2020
    detail.hit.zdb_id: 1477192-5
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  • 9
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    Online Resource
    Intracellular Ca2+ Regulates the Phosphorylation and the Dephosphorylation of Ciliary Proteins Via the NO Pathway
    Rockefeller University Press ; 2004
    In:  The Journal of General Physiology Vol. 124, No. 5 ( 2004-11-01), p. 527-540
    Details
    In: The Journal of General Physiology, Rockefeller University Press, Vol. 124, No. 5 ( 2004-11-01), p. 527-540
    Abstract: The phosphorylation profile of ciliary proteins under basal conditions and after stimulation by extracellular ATP was investigated in intact tissue and in isolated cilia from porcine airway epithelium using anti-phosphoserine and anti-phosphothreonine specific antibodies. In intact tissue, several polypeptides were serine phosphorylated in the absence of any treatment (control conditions). After stimulation by extracellular ATP, changes in the phosphorylation pattern were detected on seven ciliary polypeptides. Serine phosphorylation was enhanced for three polypeptides (27, 37, and 44 kD), while serine phosphorylation was reduced for four polypeptides (35, 69, 100, and 130 kD). Raising intracellular Ca2+ with ionomycin induced identical changes in the protein phosphorylation profile. Inhibition of the NO pathway by inhibiting either NO syntase (NOS), guanylyl cyclase (GC), or cGMP-dependent protein kinase (PKG) abolished the changes in phosphorylation induced by ATP. The presence of PKG within the axoneme was demonstrated using a specific antibody. In addition, in isolated permeabilized cilia, submicromolar concentrations of cGMP induced protein phosphorylation. Taken together, these results suggest that the axoneme is an integral part of the intracellular NO pathway. The surprising observation that ciliary activation is accompanied by sustained dephosphorylation of ciliary proteins via NO pathway was not detected in isolated cilia, suggesting that the protein phosphatases were either lost or deactivated during the isolation procedure. This work reveals that any pharmacological manipulation that abolished phosphorylation and dephosphorylation also abolished the enhancement of ciliary beating. Thus, part or all of the phosphorylated polypeptides are likely directly involved in axonemal regulation of ciliary beating.
    Type of Medium: Online Resource
    ISSN: 1540-7748 , 0022-1295
    URL: Article
    DOI: 10.1085/jgp.200409153
    Language: English
    Publisher: Rockefeller University Press
    Publication Date: 2004
    detail.hit.zdb_id: 1477246-2
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  • 10
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    Hemocyanin with phenoloxidase activity in the chitin matrix of the crayfish gastrolith
    The Company of Biologists ; 2013
    In:  Journal of Experimental Biology
    Details
    In: Journal of Experimental Biology, The Company of Biologists
    Abstract: Gastroliths are transient extracellular calcium deposits formed by the crayfish Cherax quadricarinatus von Martens on both sides of the stomach wall during pre-molt. Gastroliths are made of a rigid chitinous organic matrix, constructed as sclerotized chitin-protein microfibrils within which calcium carbonate is deposited. Although gastroliths share many characteristics with the exoskeleton, they are simpler in structure and relatively homogenous in composition, making them an excellent cuticle-like model for the study of cuticular proteins. In searching for molt-related proteins involved in gastrolith formation, two integrated approaches were employed, namely the isolation and mass spectrometric analysis of proteins from the gastrolith matrix, and 454-sequencing of mRNAs from both the gastrolith-forming and sub-cuticular epithelia. SDS-PAGE separation of gastrolith proteins revealed a set of bands at apparent molecular weights of 75-85 kDa, of which peptide sequencing following mass spectrometry matched the deduced amino acid sequences of seven hemocyanin transcripts. This assignment was then examined by immunoblot analysis using anti-hemocyanin antibodies, also used to determine the spatial distribution of the proteins in situ. Apart from contributing to oxygen transport, crustacean hemocyanins were previously suggested as being involved in several aspects of the molt cycle, including hardening of the new post-molt exoskeleton via phenoloxidation. The phenoloxidase activity of gastrolith hemocyanins was demonstrated. It was also noted that hemocyanin transcript expression during pre-molt was specific to the hepatopancreas. Our results thus reflect a set of functionally versatile proteins, expressed in a remote metabolic tissue and dispersed via the hemolymph to perform different roles in various organs and structures.
    Type of Medium: Online Resource
    ISSN: 1477-9145 , 0022-0949
    URL: Article
    DOI: 10.1242/jeb.080945
    Language: English
    Publisher: The Company of Biologists
    Publication Date: 2013
    detail.hit.zdb_id: 1482461-9
    SSG: 12
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