scholarly journals Ontogeny of a synaptophysin-mediated GABA transmission mechanism from the ciliary band-associated strand to the ciliary band during the development of the sea urchin Hemicentrotus pulcherrimus

2018 ◽  
Author(s):  
Hideki Katow ◽  
Hiromi Yoshida ◽  
Tomoko Katow ◽  
Masato Kiyomoto
Keyword(s):  
Sea Urchin ◽  
Single Copy ◽  
Single Band ◽  
Swimming Activity ◽  
Transmission Mechanism ◽  
Ciliary Band ◽  
Gaba A ◽  
Spatiotemporal Expression ◽  
Acid Protein ◽  
Gaba Transmission

Swimming activity of the sea urchin larva depends on ciliary beating primarily in the circumoral ciliary band (CB) and is regulated by several neurotransmitters, such as 5HT, dopamine and -aminobutyric acid (GABA). Accordingly, the larval swimming activity is severely inhibited by 3-mercaptopropionic acid [a glutamate decarboxylase (GAD) inhibitor]. Although GABA is detected in the CB, GAD is absent. GAD is expressed in the spatially segregated nearby ciliary band-associated strand (CBAS). Thus, it is assumed that GABA transmission extends from the CBAS to the CB. Here, we examined the synaptic transmission mechanism by focusing on the spatiotemporal expression pattern of synaptophysin (Syp), a synaptic vesicle glycoprotein. The sea urchin has a single copy of the Syp gene, which encodes a 266-amino acid protein with possibly 4 transmembrane domains. We generated an anti-Syp antibody (Ab). Immunoblotting (IB) detected Ab binding to a single band at approximately 38 kDa. Whole-mount immunohistochemistry (WMIHC) detected high intensity Ab binding to the CBAS. Syp was initially detected at the mesenchyme blastula stage (mBL) as a single band by IB. Accordingly, WMIHC detected Syp in the cytoplasm of small patches of several ectodermal cells at the mBL stage. Syp has also been detected in the cytoplasm of blastocoelar cells from the prism stage to the 2-arm pluteus stage (2aPL). By the 4aPL stage, Syp was expressed in the CBAS and was moderately expressed among the blastocoelar cells. Distinctive co-localization of GABA and Syp was not detected until the 2aPL stage. Beginning at the 4aPL stage, GABA was detected in the CB and Syp-positive puncta in the CBAS. In the CB, GABA was co-localized with the GABA-A receptor (GABAAR). Thus, the GABA signal may be transmitted from GAD in the CBAS through a Syp-mediated system to the CB and then, in the CB, to the basal body of the cilia through GABAAR.

scholarly journals Ontogeny of a synaptophysin-mediated GABA transmission mechanism from the ciliary band-associated strand to the ciliary band during the development of the sea urchin Hemicentrotus pulcherrimus

2018 ◽  
Author(s):  
Hideki Katow ◽  
Hiromi Yoshida ◽  
Tomoko Katow ◽  
Masato Kiyomoto
Keyword(s):  
Sea Urchin ◽  
Single Copy ◽  
Single Band ◽  
Swimming Activity ◽  
Transmission Mechanism ◽  
Ciliary Band ◽  
Gaba A ◽  
Spatiotemporal Expression ◽  
Acid Protein ◽  
Gaba Transmission

Swimming activity of the sea urchin larva depends on ciliary beating primarily in the circumoral ciliary band (CB) and is regulated by several neurotransmitters, such as 5HT, dopamine and -aminobutyric acid (GABA). Accordingly, the larval swimming activity is severely inhibited by 3-mercaptopropionic acid [a glutamate decarboxylase (GAD) inhibitor]. Although GABA is detected in the CB, GAD is absent. GAD is expressed in the spatially segregated nearby ciliary band-associated strand (CBAS). Thus, it is assumed that GABA transmission extends from the CBAS to the CB. Here, we examined the synaptic transmission mechanism by focusing on the spatiotemporal expression pattern of synaptophysin (Syp), a synaptic vesicle glycoprotein. The sea urchin has a single copy of the Syp gene, which encodes a 266-amino acid protein with possibly 4 transmembrane domains. We generated an anti-Syp antibody (Ab). Immunoblotting (IB) detected Ab binding to a single band at approximately 38 kDa. Whole-mount immunohistochemistry (WMIHC) detected high intensity Ab binding to the CBAS. Syp was initially detected at the mesenchyme blastula stage (mBL) as a single band by IB. Accordingly, WMIHC detected Syp in the cytoplasm of small patches of several ectodermal cells at the mBL stage. Syp has also been detected in the cytoplasm of blastocoelar cells from the prism stage to the 2-arm pluteus stage (2aPL). By the 4aPL stage, Syp was expressed in the CBAS and was moderately expressed among the blastocoelar cells. Distinctive co-localization of GABA and Syp was not detected until the 2aPL stage. Beginning at the 4aPL stage, GABA was detected in the CB and Syp-positive puncta in the CBAS. In the CB, GABA was co-localized with the GABA-A receptor (GABAAR). Thus, the GABA signal may be transmitted from GAD in the CBAS through a Syp-mediated system to the CB and then, in the CB, to the basal body of the cilia through GABAAR.

scholarly journals The gene encoding the transcription factor SCIP has features of an expressed retroposon.

1991 ◽  
Vol 11 (9) ◽  
pp. 4642-4650 ◽  
Author(s):  
R Kuhn ◽  
E S Monuki ◽  
G Lemke
Keyword(s):  
Transcription Factor ◽  
Schwann Cells ◽  
Cpg Island ◽  
Single Copy ◽  
Gene Encoding ◽  
Intronless Gene ◽  
Gene Comparison ◽  
Acid Protein ◽  
Central Nervous Systems ◽  
Glial Progenitor Cells

SCIP is a POU domain transcription factor expressed by glial progenitor cells in the peripheral and central nervous systems (dividing Schwann cells and O-2A cells, respectively), where it appears to act as a repressor of myelin-specific genes. We have isolated genomic clones encoding the rat SCIP gene. Comparison of the structure of these clones with genomic Southern blots and SCIP cDNAs demonstrates that SCIP is encoded in a single-copy, intronless gene that has the general features of an expressed retroposon. This gene contributes to an extended CpG island. It is transcribed to produce a 3.1-kb mRNA that encodes a 451-amino-acid protein with a predicted molecular mass of 45 kDa. Immunopurified SCIP antibodies specifically recognize a nuclear protein of this size in cultured proliferating Schwann cells, and gel shift analyses demonstrate that this protein is the predominant octamer-binding protein in these cells.

The gene encoding the transcription factor SCIP has features of an expressed retroposon

1991 ◽  
Vol 11 (9) ◽  
pp. 4642-4650
Author(s):  
R Kuhn ◽  
E S Monuki ◽  
G Lemke
Keyword(s):  
Transcription Factor ◽  
Schwann Cells ◽  
Cpg Island ◽  
Single Copy ◽  
Gene Encoding ◽  
Intronless Gene ◽  
Gene Comparison ◽  
Acid Protein ◽  
Central Nervous Systems ◽  
Glial Progenitor Cells

SCIP is a POU domain transcription factor expressed by glial progenitor cells in the peripheral and central nervous systems (dividing Schwann cells and O-2A cells, respectively), where it appears to act as a repressor of myelin-specific genes. We have isolated genomic clones encoding the rat SCIP gene. Comparison of the structure of these clones with genomic Southern blots and SCIP cDNAs demonstrates that SCIP is encoded in a single-copy, intronless gene that has the general features of an expressed retroposon. This gene contributes to an extended CpG island. It is transcribed to produce a 3.1-kb mRNA that encodes a 451-amino-acid protein with a predicted molecular mass of 45 kDa. Immunopurified SCIP antibodies specifically recognize a nuclear protein of this size in cultured proliferating Schwann cells, and gel shift analyses demonstrate that this protein is the predominant octamer-binding protein in these cells.

Characterization and Cloning of Cutinase from Ascochyta rabiei

1997 ◽  
Vol 52 (3-4) ◽  
pp. 197-208 ◽  
Author(s):  
Raimund Tenhaken ◽  
Martin Arnemann ◽  
Gabriele Köhler ◽  
Wolfgang Barz
Keyword(s):  
Ascochyta Blight ◽  
Apparent Molecular Weight ◽  
Single Copy ◽  
Cosmid Library ◽  
Ascochyta Rabiei ◽  
Serine Esterase ◽  
Acid Protein ◽  
Strong Homology ◽  
Hydroxylated Fatty Acids ◽  
Organophosphorous Compounds

Abstract Ascochyta rabiei, the causal agent of Ascochyta blight on chickpea plants, secretes a cutinase in the culture filtrate when it is induced by cutin or hydroxylated fatty acids. This cutinase is the main esterase in the culture fluids. The enzyme was purified to homogeneity by three successive chromatographic steps. It showed an apparent molecular weight of 22 kD in SDS-PAGE and cleaved ester bonds of 3H-labelled cutin or p-nitrophenylbutyrate with maximal activities around pH 8. As a serine esterase, cutinase is strongly inhibited by organophosphorous compounds and the most effective inhibitor 2,3,5-trichloropyridine-6-(0-methyl-O-n-butyl)-phosphateester (MAT 9564) shows a K1 value of 0.8 nᴍ. The cutinase gene was cloned from a genomic cosmid library by screening with two oligonucleotides directed against cutinase consensus peptides. The gene was subcloned to a 1.7 Kb Sa1l/Hindlll-insert and sequenced. The cutinase gene codes for a 223 amino acid protein with strong homology to other fungal cutinase sequences. The purified cutinase is encoded by a single copy gene.

scholarly journals Characterization of two developmentally regulated sea urchin U2 small nuclear RNA promoters: a common required TATA sequence and independent proximal and distal elements.

1992 ◽  
Vol 12 (2) ◽  
pp. 650-660 ◽  
Author(s):  
B Stefanovic ◽  
W F Marzluff
Keyword(s):  
Sea Urchin ◽  
Core Promoter ◽  
Sequence Similarity ◽  
Single Copy ◽  
Upstream Sequence ◽  
Small Nuclear Rna ◽  
Developmentally Regulated ◽  
Sequence Element ◽  
Nuclear Rna ◽  
Tata Sequence

The promoters of two U2 small nuclear RNA genes isolated from the sea urchin Lytechinus variegatus were mapped by microinjection of genes into sea urchin zygotes. One gene, LvU2E, is expressed only in oocytes and embryos and is found in a tandemly repeated gene set, while the other gene, LvU2L, is a single-copy gene and is expressed in embryos and somatic cells. The promoters each contain a TATA sequence at -25 which is required for expression, a proximal sequence element (PSE) centered at -55 required for expression, a sequence at -100 which couples the core promoter (PSE plus TATA box) to the upstream element, and an upstream sequence which stimulates expression fourfold. The PSE together with the TATA sequence is sufficient to determine the transcription start site. There is no sequence similarity between the -100 and PSE sequences of the two genes. The -100 sequences can be interchanged between the two genes. The LvU2E PSE functions in the context of the LvU2L gene, but the LvU2L PSE functions poorly in the context of the LvU2E gene.

scholarly journals DNA methylation pattern changes during development of a sea urchin

1992 ◽  
Vol 283 (3) ◽  
pp. 751-753 ◽  
Author(s):  
J Fronk ◽  
G A Tank ◽  
J P Langmore
Keyword(s):  
Sea Urchin ◽  
Cytosine Methylation ◽  
Single Copy ◽  
Methylation Pattern ◽  
Developmentally Regulated ◽  
Partial Methylation ◽  
Methylation Of Dna ◽  
Cytoplasmic Actin ◽  
The Family ◽  
Endonuclease Digestion

Cytosine methylation of developmentally regulated genes of the sea urchin Strongylocentrotus purpuratus was studied by using restriction-endonuclease digestion and Southern blotting. The single-copy bindin gene, the family of five cytoplasmic actin genes and the 400-fold-repeated set of five early histone genes were mostly unmethylated, but some sites exhibited partial methylation that varied throughout development. This shows that in echinoderms the methylation of DNA is not confined to the non-transcribed portion of the genome, as previously believed [Bird, Tagart & Smith (1979) Cell 17, 889-901], and may play a role in transcriptional regulation.

scholarly journals SpCoel1: a sea urchin profilin gene expressed specifically in coelomocytes in response to injury.

1992 ◽  
Vol 3 (4) ◽  
pp. 403-414 ◽  
Author(s):  
L C Smith ◽  
R J Britten ◽  
E H Davidson
Keyword(s):  
Sea Urchin ◽  
Sequence Similarity ◽  
Single Copy ◽  
Untranslated Regions ◽  
Amino Acid Sequence Similarity ◽  
Signal Transduction System ◽  
Reading Frame ◽  
Purple Sea Urchin ◽  
Copy Gene ◽  
External Injury

SpCoel1 is a single copy gene that is specifically expressed in most of the coelomocytes of the adult purple sea urchin, Strongylocentrotus purpuratus. The 4-kb transcript from this gene has a relatively short (426 nucleotide) open reading frame (ORF) with long 3' and 5' untranslated regions. The ORF encodes a protein that has strong amino acid sequence similarity to profilins from yeast to mammals. Transcript titrations of SpCoel1 show significant increases per coelomocyte in animals that have been physiologically challenged. Increases in transcript levels are of similar magnitudes between animals receiving different treatments, such as injuries from needle punctures or from injections of foreign cells. The evidence presented here implies a molecular mechanism by which this lower deuterostome defense system responds to external insult, viz that an external "injury signal" activates a signal transduction system, which in turn mediates the alterations in cytoskeletal state that are required for coelomocyte activation.

Transcription of similar sets of rare maternal RNAs and rare nuclear RNAs in sea urchin blastulae and adult coelomocytes

Development ◽  
1985 ◽  
Vol 85 (1) ◽  
pp. 131-149
Author(s):  
Kenneth C. Kleene ◽  
Tom Humphreys
Keyword(s):  
Sea Urchin ◽  
Sea Urchins ◽  
Single Copy ◽  
Cell Populations ◽  
Maternal Rna ◽  
Class A ◽  
Nuclear Rna ◽  
Rare Class ◽  
Copy Dna ◽  
Single Copy Dna

We studied the sequences transcribed in the rare class of hnRNA and the rare maternal RNA set in blastula embryos and a tissue of adult sea urchins, coelomocytes. About 26 % of labelled single-copy DNA formed hybrids which bound to hydroxyapatite after three cycles of hybridization with nuclear RNA from blastulae and coelomocytes. This corresponds to transcription of about 50 % of the single-copy genome by both cell populations. To compare the rare hnRNA sequences synthesized by blastulae and coelomocytes directly, labelled single-copy DNA was hybridized with blastula nuclear RNA to high RNA C0t, fractionated into sequences complementary and non-complementary to blastula nuclear RNA by chromotography on hydroxyapatite, and then each fraction was rehybridized with nuclear RNA from blastulae and coelomocytes. About 62 % of the labelled DNA complementary to blastula nuclear RNA and about 1·5 % of the labelled DNA non-complementary to blastula nuclear RNA hybridized with nuclear RNA from both cell populations. Thus, coelomocytes and blastula embryos transcribe essentially the same single-copy sequences in the rare hnRN A class. A probe for the rare maternal RNA set was isolated by hybridizing single-copy DNA with total egg RNA to high RNA C0t. 65–67 % of this probe hybridized with whole-cell RNA from eggs, blastulae, plutei and coelomocytes demonstrating that essentially all rare maternal RNAs are present, and presumably transcribed, in blastulae, plutei and coelomocytes.

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