Molecular cloning and expression of a novel homeobox gene AHox1 of the ascidian, Halocynthia roretzi

Development ◽  
1991 ◽  
Vol 111 (3) ◽  
pp. 821-828
Author(s):  
H. Saiga ◽  
A. Mizokami ◽  
K.W. Makabe ◽  
N. Satoh ◽  
T. Mita
Keyword(s):  
Digestive Tract ◽  
Blot Hybridization ◽  
Amino Acid Level ◽  
Homeobox Gene ◽  
Cloning And Expression ◽  
Northern Blot Hybridization ◽  
Halocynthia Roretzi ◽  
Adult Tissues ◽  
Pharyngeal Epithelium

We have isolated a novel ascidian homeobox gene, designated AHox1, by screening the genomic DNA of Halocynthia roretzi with the Bombyx mori Antennapedia type homeobox as a probe. The AHox1 gene encodes a protein that consists of 741 amino acids. The homeobox of AHox1 is interrupted by 2 introns each of which is about 300 bp in length and it shows about 70% similarity at a deduced amino acid level to that of Drosophila H2.0. This suggests that AHox1 is one of the most diverged homeobox genes so far characterized. Northern blot hybridization with an AHox1 probe showed the presence of single transcripts approximately 2.8 kb in length in larvae, juveniles and some adult tissues. The expression of AHox1 is scarcely detected during the course of early development but it increases to a moderate level at the larval stage. After metamorphosis, the level of AHox1 expression increases as development proceeds. In situ hybridization to the juvenile 7 days after metamorphosis showed that the site of AHox1 expression is the epithelium of digestive tract. Among the adult tissues examined, digestive tract, digestive gland and coelomic cells were the major sites of the expression of AHox1. In gonad, body wall muscle and pharyngeal epithelium, the expression of AHox1 is relatively weak. These results suggest that AHox1 is primarily expressed in the tissues of endodermal origin and that the gene expression may be associated with differentiation of the endodermal tissues.

scholarly journals Molecular characterization and expression of the stratification-related cytokeratins 4 and 15.

1988 ◽  
Vol 106 (4) ◽  
pp. 1249-1261 ◽  
Author(s):  
R E Leube ◽  
B L Bader ◽  
F X Bosch ◽  
R Zimbelmann ◽  
T Achtstaetter ◽  
...  
Keyword(s):  
Basal Cell ◽  
Blot Hybridization ◽  
Northern Blot Hybridization ◽  
Type I ◽  
Specific Expression ◽  
Genes Encoding ◽  
Cell Layers ◽  
Hybridization In Situ ◽  
Stratified Epithelia

A number of human cytokeratins are expressed during the development of stratified epithelia from one-layered polar epithelia and continue to be expressed in several adult epithelial tissues. For studies of the regulation of the synthesis of stratification-related cytokeratins in internal tissues, we have prepared cDNA and genomic clones encoding cytokeratin 4, as a representative of the basic (type II) cytokeratin subfamily and cytokeratin 15, as representative of the acidic (type I) subfamily, and determined their nucleotide sequences. The specific expression of mRNAs encoding these two polypeptides in certain stratified tissues and cultured cell lines is demonstrated by Northern blot hybridization. Hybridization in situ with antisense riboprobes and/or synthetic oligonucleotides shows the presence of cytokeratin 15 mRNA in all layers of esophagus, whereas cytokeratin 4 mRNA tends to be suprabasally enriched, although to degrees varying in different regions. We conclude that the expression of the genes encoding these stratification-related cytokeratins starts already in the basal cell layer and does not depend on vertical differentiation and detachment from the basal lamina. Our results also show that simple epithelial and stratification-related cytokeratins can be coexpressed in basal cell layers of certain stratified epithelia such as esophagus. Implications of these findings for epithelial differentiation and the formation of squamous cell carcinomas are discussed.

scholarly journals Cloning and expression analysis of murine phospholipase D1

1997 ◽  
Vol 326 (3) ◽  
pp. 745-753 ◽  
Author(s):  
William C. COLLEY ◽  
Yelena M. ALTSHULLER ◽  
Christopher K. SUE-LING ◽  
Neal G. COPELAND ◽  
Debra J. GILBERT ◽  
...  
Keyword(s):  
Blot Hybridization ◽  
Cell Types ◽  
Adult Brain ◽  
Cloning And Expression ◽  
Complex Signal ◽  
Northern Blot Hybridization ◽  
Rnase Protection ◽  
Phospholipase D1 ◽  
In Situhybridization ◽  
Embryo Sections

Activation of phosphatidylcholine-specific phospholipase D (PLD) occurs as part of the complex signal-transduction cascade initiated by agonist stimulation of tyrosine kinase and G-protein-coupled receptors. A variety of mammalian PLD activities have been described, and cDNAs for two PLDs recently reported (human PLD1 and murine PLD2). We describe here the cloning and chromosomal localization of murine PLD1. Northern-blot hybridization and RNase protection analyses were used to examine the expression of murine PLD1 and PLD2 in a variety of cell lines and tissues. PLD1 and PLD2 were expressed in all RNA samples examined, although the absolute expression of each isoform varied, as well as the ratio of PLD1 to PLD2. Moreover, in situhybridization of adult brain and murine embryo sections revealed high levels of expression of individual PLDs in some cell types and no detectable expression in others. Thus the two PLDs probably carry out distinct roles in restricted subsets of cells rather than ubiquitous roles in all cells.

Post-transcriptional block in oxytocin gene expression within the seminiferous tubules of the bovine testis

1994 ◽  
Vol 140 (1) ◽  
pp. 63-72 ◽  
Author(s):  
H Ungefroren ◽  
M Davidoff ◽  
R Ivell
Keyword(s):  
Gene Expression ◽  
Leydig Cells ◽  
Blot Hybridization ◽  
Seminiferous Tubules ◽  
Transcriptional Level ◽  
Gene Transcript ◽  
Northern Blot Hybridization ◽  
Rnase Protection ◽  
Low Level

Abstract Northern blot hybridization showed that bovine and sheep testis, unlike testes from other mammals, contain moderate levels of an apparently normal oxytocin gene transcript. In situ hybridization localized this mRNA to within the seminiferous tubules, possibly in the Sertoli cells. Conflicting with this result, immunohistochemistry showed that both oxytocin and the syngeneic neurophysin I epitopes are both clearly restricted to the Leydig cells, being expressed here at a low level. Since illegitimate transcription from spurious start sites can lead to a lack of translation product, the integrity of the major ruminant testicular transcripts of the oxytocin gene was checked using differential hybridization, RNase protection and multiple polymerase chain reaction assays. All tests showed the transcripts to have a normal, translatable composition and to be transcribed from the conventional 5' initiation site. Therefore, the block in oxytocin gene expression within the tubules is probably due to a lesion at the post-transcriptional level. The low level peptide expression in the Leydig cells can probably be attributed to the presence of functional transcripts in these cells, which are below the level of significant detection for the in situ hybridization assay. Journal of Endocrinology (1994) 140, 63–72

Detection and characterization of latent HSV RNA by in Situ and northern blot hybridization in guinea pigs

Virology ◽  
1991 ◽  
Vol 181 (2) ◽  
pp. 793-797 ◽  
Author(s):  
Rae Lyn Burke ◽  
Karin Hartog ◽  
Kenneth D. Croen ◽  
Jeffrey M. Ostrove
Keyword(s):  
Northern Blot ◽  
Guinea Pigs ◽  
Blot Hybridization ◽  
Northern Blot Hybridization

A maternal homeobox gene, Bombyx caudal, forms both mRNA and protein concentration gradients spanning anteroposterior axis during gastrulation

Development ◽  
1994 ◽  
Vol 120 (2) ◽  
pp. 277-285 ◽  
Author(s):  
X. Xu ◽  
P.X. Xu ◽  
Y. Suzuki
Keyword(s):  
Protein Concentration ◽  
Blot Hybridization ◽  
Homeobox Gene ◽  
Peptide Sequence ◽  
Northern Blot Hybridization ◽  
Concentration Gradients ◽  
Anteroposterior Axis ◽  
Maternal Transcripts ◽  
Cad Protein ◽  
Blastoderm Stage

We have isolated a caudal (cad) homologue from a cDNA library of Bombyx mori embryos. The Bombyx cad cDNA encodes a protein of 244 amino acids. The homology between Drosophila and Bombyx homeodomains is 80%. Similar to Drosophila cad, there is no YPWM peptide sequence along the upstream of homeodomain. Northern blot hybridization with a Bombyx cad probe revealed the presence of single maternal transcript of 2.3 kb. A stronger signal of the transcripts was detected in unfertilized eggs and in eggs up to 36 hours after deposition. The transcripts decreased rapidly by 2 days and a weak signal was maintained until hatching. To analyse its spatial expression pattern, we have established a novel frozen sectioning method for in situ hybridization and immunohistochemistry experiments. The results showed that Bombyx cad transcripts accumulated first in the nurse cells and transferred into the oocyte at a defined time during oogenesis. The maternal transcripts of Bombyx cad formed a concentration gradient spanning the anteroposterior axis during the gastrulation stage and were restricted to the anal pad, the most posterior domain, after 2 days of embryogenesis; the Drosophila cad mRNA revealed the corresponding expression profile during the syncytial blastoderm stage. The Bombyx cad protein was not detected in the ovary and the first 9 hours of eggs, but was first detected evenly during cellular blastoderm stage. During gastrulation, Bombyx cad protein concentration gradients shifted along the anteroposterior axis coinciding with the shifting of the mRNA concentration gradients.(ABSTRACT TRUNCATED AT 250 WORDS)

scholarly journals Extended expression of MaKN1 contributes to the leaf morphology in aquatic forms of Myriophyllum aquaticum

Botany ◽  
2015 ◽  
Vol 93 (9) ◽  
pp. 611-621
Author(s):  
M.D. Shafiullah ◽  
Christian R. Lacroix
Keyword(s):  
Apical Meristem ◽  
Leaf Morphology ◽  
Expression Patterns ◽  
Homeobox Gene ◽  
Leaf Primordia ◽  
Leaf Primordium ◽  
Myriophyllum Aquaticum ◽  
Knox Gene ◽  
Leaf Morphogenesis

Myriophyllum aquaticum (Vell.) Verdc. is heterophyllous in nature with highly dissected simple leaves consisting of several lobes. KNOX (KNOTTED1-LIKE HOMEOBOX) genes are believed to have played an important role in the evolution of leaf diversity. Up-regulation of KNOX during leaf primordium initiation can lead to leaf dissection in plants with simple leaves and, if overexpressed, can produce ectopic meristems on leaves. A previous study on KNOX gene expression in the aerial form of this species showed that this gene is expressed in the shoot apical meristem (SAM), as well as in leaf primordia P0 to P8. Based on these results, it was hypothesized that the prolonged expression of the MaKN1 (Myriophyllum aquaticum Knotted1-like homeobox) gene beyond P8, might play an important role in the generation of more lobes, longer lobes, and hydathode formation in the aquatic leaves of M. aquaticum. The technique of in situ hybridization was carried out using a previously sequenced 300 bp fragment of MaKN1 to determine the expression patterns of this gene in the shoot of aquatic forms of the plant. Expression patterns of MaKN1 revealed that the SAM and leaf primordia of aquatic forms of M. aquaticum at levels P0 (youngest) to P4 were distributed throughout these structures. The level of expression of this MaKN1 gene progressively became more localized to lobes in older leaf primordia (levels P5 to P12). Previous studies of aerial forms of this plant showed MaKN1 expression until P8. Our results with aquatic forms show that the highly dissected leaf morphology in aquatic forms was the result of the prolonged expression of MaKN1 beyond P8. This resulted in the formation of elongated and slightly more numerous lobes, and hydathodes in aquatic forms. These findings support the view that KNOX genes are important developmental regulators of leaf morphogenesis and have played an important role in the evolution of leaf forms in the plant kingdom.

scholarly journals Location of the initial cleavage sites in mouse pre-rRNA.

1983 ◽  
Vol 3 (8) ◽  
pp. 1501-1510 ◽  
Author(s):  
L H Bowman ◽  
W E Goldman ◽  
G I Goldberg ◽  
M B Hebert ◽  
D Schlessinger
Keyword(s):  
Blot Hybridization ◽  
Northern Blot Hybridization ◽  
28S Rrna ◽  
Cleavage Sites ◽  
S1 Nuclease ◽  
5.8S Rrna ◽  
Initial Cleavage ◽  
S1 Nuclease Mapping ◽  
Mapping Techniques ◽  
Nuclease Mapping

The locations of three cleavages that can occur in mouse 45S pre-rRNA were determined by Northern blot hybridization and S1 nuclease mapping techniques. These experiments indicate that an initial cleavage of 45S pre-rRNA can directly generate the mature 5' terminus of 18S rRNA. Initial cleavage of 45S pre-rRNA can also generate the mature 5' terminus of 5.8S rRNA, but in this case cleavage can occur at two different locations, one at the known 5' terminus of 5.8S rRNA and another 6 or 7 nucleotides upstream. This pattern of cleavage results in the formation of cytoplasmic 5.8S rRNA with heterogeneous 5' termini. Further, our results indicate that one pathway for the formation of the mature 5' terminus of 28S rRNA involves initial cleavages within spacer sequences followed by cleavages which generate the mature 5' terminus of 28S rRNA. Comparison of these different patterns of cleavage for mouse pre-rRNA with that for Escherichia coli pre-rRNA implies that there are fundamental differences in the two processing mechanisms. Further, several possible cleavage signals have been identified by comparing the cleavage sites with the primary and secondary structure of mouse rRNA (see W. E. Goldman, G. Goldberg, L. H. Bowman, D. Steinmetz, and D. Schlessinger, Mol. Cell. Biol. 3:1488-1500, 1983).

scholarly journals Tissue-specific regulation of dipeptidyl peptidase IV expression during development

1991 ◽  
Vol 277 (2) ◽  
pp. 331-334 ◽  
Author(s):  
M Hildebrandt ◽  
W Reutter ◽  
J D Gitlin
Keyword(s):  
Lung Development ◽  
Kidney Development ◽  
Blot Hybridization ◽  
Dipeptidyl Peptidase ◽  
Transcriptional Level ◽  
Northern Blot Hybridization ◽  
Dpp Iv ◽  
Organ Specific ◽  
Specific Regulation ◽  
Lung And Kidney

The patterns of dipeptidyl peptidase (DPP) IV activity and protein amount in different rat organs during development were compared. In order to elucidate the molecular basis for these patterns, total RNA was isolated from lung and kidney at different stages of development and analysed by Northern-blot hybridization using an oligonucleotide derived from the DPP IV cDNA sequence. This oligonucleotide hybridized to two distinct mRNAs of approx. 3.2 and 4.8 kb respectively. During kidney development, the pattern for DPP IV mRNA paralleled that of DPP IV activity and protein amount, suggesting that, in kidney, the expression of DPP IV is primarily controlled at the transcriptional level. In contrast, the magnitude of DPP IV activity during lung development compared with that of DPP IV mRNA in lung suggests that post-transcriptional mechanisms are involved in regulating the expression of DPP IV in lung. Organ-specific regulation of DPP IV expression may provide a useful model for further comparative studies of transcriptional and post-transcriptional mechanisms of DPP IV expression within the same organism.

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