Differential expression sites of TGF-β isoforms in chicken limb buds during morphogenesis

2005 ◽  
Vol 83 (4) ◽  
pp. 620-625 ◽  
Author(s):  
Shinya Aramaki ◽  
Fuminori Sato ◽  
Tomoki Soh ◽  
Nobuhiko Yamauchi ◽  
Masa-aki Hattori
Keyword(s):  
Limb Development ◽  
Developmental Stages ◽  
Polymerase Chain Reaction Analysis ◽  
Weak Signal ◽  
Temporal Expression ◽  
Chain Reaction ◽  
White Leghorn Chicken ◽  
Whole Mount ◽  
Polymerase Chain

TGF-β gene is expressed at various developmental stages and its principle role may be an involvement in organogenesis. The present study was performed to investigate the temporal expression of these TGF-β isoforms in the developing limb of White Leghorn Chicken, Gallus gallus (L., 1758). TGF-β isoforms were expressed in the developing limb as revealed by whole-mount in situ hybridization, but each showed a different pattern of expression. TGF-β2 was the dominant isoform compared with the other two isoforms. TGF-β2 first appeared along the proximodistal axis of the limb at stage 24 and condensed at the tip at stage 26. At stages 29–31, expression appeared in digits and then was extended to the interdigital spaces. A weak signal for TGF-β3 was first shown in the developing limb at stage 26, but there was no interdigital expression, unlike for TGF-β2. TGF-β4 was expressed in the developing limb at stage 26 and only in the interdigital spaces at stage 29. Reverse transcription – polymerase chain reaction analysis also showed that the transcript levels of TGF-β isoforms, especially TGF-β2, drastically increased at stage 29. These results suggest that TGF-β isoforms, with their patterns of expression, are specific regulatory factors that participate in limb development and digit morphogenesis.

Contribution of zebrafish-mouse cell hybrids to the mapping of the zebrafish genome

1997 ◽  
Vol 75 (5) ◽  
pp. 641-649 ◽  
Author(s):  
Mario Chevrette ◽  
Lucille Joly ◽  
Patricia Tellis ◽  
Marc Ekker
Keyword(s):  
Chromosomal Location ◽  
Polymerase Chain Reaction Analysis ◽  
Mouse Cell ◽  
Zebrafish Genome ◽  
Vertebrate Development ◽  
Linkage Groups ◽  
Chain Reaction ◽  
Cell Hybrids ◽  
Polymerase Chain

The zebrafish, Danio rerio, is becoming an increasingly popular model for the study of vertebrate development. Indeed, the biology of the fish offers great advantages for such studies. The life cycle of the zebrafish is relatively short (2-3 months) and the embryos develop outside the mother, facilitating the visualization of any mutated phenotype. At present, more than 1000 embryonic mutations have been reported. However, until recently, there was no physical or genetic map for this organism. In an effort to generate such a map, we have produced and characterized a panel of zebrafish-mouse cell hybrids. We have used whole-cell fusion to transfer zebrafish chromosomes from two different zebrafish cell lines into mouse recipient cells, thus generating more than 100 hybrids. Using fluorescence in situ hybridization and polymerase chain reaction analysis, we have determined the zebrafish chromosome composition of these hybrids. Here we report that elements from the 25 linkage groups of the zebrafish genome are present in our hybrids. These hybrids could identify the chromosomal location of genes affected in zebrafish mutants.

Enterovirus is Not Present in Placentas from Cases of Perinatal Depression Using Polymerase Chain Reaction Analysis

2009 ◽  
Vol 12 (3) ◽  
pp. 177-179 ◽  
Author(s):  
Jesse Cover ◽  
Edwina Popek ◽  
Myra Wyckoff ◽  
N. Kristine Leos ◽  
Beverly Barton Rogers
Keyword(s):  
Polymerase Chain Reaction ◽  
Respiratory Insufficiency ◽  
Pcr Amplification ◽  
Polymerase Chain Reaction Analysis ◽  
Chain Reaction ◽  
Apgar Scores ◽  
Specific Patient ◽  
In Situ Pcr ◽  
Polymerase Chain

Enteroviruses have been implicated as a cause of low Apgar scores in conjunction with perinatal seizures and respiratory insufficiency. Using in-situ reverse transcriptase polymerase chain reaction (in-situ PCR), Nuovo et al detected enterovirus in up to 86% of placentas from perinates exhibiting these symptoms. In-situ PCR has been the only method employed to assess for the presence of enterovirus in this specific patient population. The purpose of our study was to use PCR amplification of enterovirus from extracted RNA to confirm these observations. RNA was extracted from 26 placentas of infants with low Apgar scores, perinatal seizures, and respiratory insufficiency. Each extraction was positive for beta-actin RNA, which confirmed that the integrity of RNA was maintained in the sample. Enterovirus RNA was not detected in any of the cases. Our results indicate that enterovirus is not present in placentas from neonates with the combination of low Apgar scores, respiratory insufficiency, and seizures, as previously reported.

scholarly journals Persistent Viral Shedding during Asymptomatic Aleutian Mink Disease Parvoviral Infection in a Ferret

2005 ◽  
Vol 17 (6) ◽  
pp. 594-597 ◽  
Author(s):  
Kate E. Pennick ◽  
Mary Ann M. Stevenson ◽  
Kenneth S. Latimer ◽  
Branson W. Ritchie ◽  
Christopher R. Gregory
Keyword(s):  
Polymerase Chain Reaction ◽  
Polymerase Chain Reaction Analysis ◽  
Chain Reaction ◽  
Viral Dna ◽  
Viral Shedding ◽  
Antibody Titers ◽  
Polymerase Chain ◽  
Cell Fractions ◽  
Observation Period

A 2-year-old domestic ferret that appeared clinically healthy was repeatedly seropositive for Aleutian mink disease parvovirus (ADV) over a 2-year observation period. Antibody titers, determined by counter-immunoelectrophoresis, ranged from 1024 to 4096. Viral DNA also was identified in serum, urine, feces, and blood cell fractions by polymerase chain reaction analysis. Ultimately, DNA in situ hybridization revealed ADV DNA in histologic sections of various tissues and organs. These data indicate that this asymptomatic ferret was persistently infected with ADV.

scholarly journals Expression of tal-1 and GATA-binding proteins during human hematopoiesis

Blood ◽  
1993 ◽  
Vol 81 (3) ◽  
pp. 647-655 ◽  
Author(s):  
MA Mouthon ◽  
O Bernard ◽  
MT Mitjavila ◽  
PH Romeo ◽  
W Vainchenker ◽  
...  
Keyword(s):  
Binding Sites ◽  
Lymphoblastic Leukemia ◽  
Hematopoietic Cells ◽  
Polymerase Chain Reaction Analysis ◽  
Chain Reaction ◽  
Basic Helix Loop Helix ◽  
Helix Loop Helix ◽  
Putative Transcription Factor ◽  
Polymerase Chain

Tal-1 rearrangements are associated with nearly 30% of human T acute lymphoblastic leukemia. Tal-1 gene encodes a putative transcription factor with a basic helix-loop-helix domain and is known to be predominantly expressed in hematopoietic cells. We investigated the pattern of tal-1 expression in purified human hematopoietic cells by in situ hybridization and reverse transcriptase polymerase chain reaction analysis. Both methods demonstrated that the tal-1 gene is expressed in megakaryocytes and erythroblasts as well as in basophilic granulocytes. In addition, our results indicate that the tal-1 1A promoter, which contains two consensus GATA-binding sites, is active mainly in these lineages. Because the GATA-1 gene is known to transactivate several genes specific for the erythroid, megakaryocytic, and mastocytic/basophilic lineages, we studied GATA-1 expression in these purified hematopoietic cells. We found that GATA-1 and tal-1 genes are coexpressed in these three lineages. Remarkably, the expression of both genes is downmodulated during erythroid and megakaryocytic terminal maturation. In immature hematopoietic cells, tal-1 and GATA-1 genes are coexpressed in committed progenitors cells (CD34+/CD38(2+)), whereas they are not detectable in the most primitive cells (CD34(2+)/CD38-). In contrast, GATA-2 is strongly expressed in both most primitive and committed progenitors cells, whereas GATA-3 is mostly detected in most primitive ones. Altogether our results strongly suggest that GATA-1 modulates the transcription of tal-1 during the differentiation of the erythroid, megakaryocytic, and basosophilic lineages.

scholarly journals Expression of tal-1 and GATA-binding proteins during human hematopoiesis

Blood ◽  
1993 ◽  
Vol 81 (3) ◽  
pp. 647-655 ◽  
Author(s):  
MA Mouthon ◽  
O Bernard ◽  
MT Mitjavila ◽  
PH Romeo ◽  
W Vainchenker ◽  
...  
Keyword(s):  
Binding Sites ◽  
Lymphoblastic Leukemia ◽  
Hematopoietic Cells ◽  
Polymerase Chain Reaction Analysis ◽  
Chain Reaction ◽  
Basic Helix Loop Helix ◽  
Helix Loop Helix ◽  
Putative Transcription Factor ◽  
Polymerase Chain

Abstract Tal-1 rearrangements are associated with nearly 30% of human T acute lymphoblastic leukemia. Tal-1 gene encodes a putative transcription factor with a basic helix-loop-helix domain and is known to be predominantly expressed in hematopoietic cells. We investigated the pattern of tal-1 expression in purified human hematopoietic cells by in situ hybridization and reverse transcriptase polymerase chain reaction analysis. Both methods demonstrated that the tal-1 gene is expressed in megakaryocytes and erythroblasts as well as in basophilic granulocytes. In addition, our results indicate that the tal-1 1A promoter, which contains two consensus GATA-binding sites, is active mainly in these lineages. Because the GATA-1 gene is known to transactivate several genes specific for the erythroid, megakaryocytic, and mastocytic/basophilic lineages, we studied GATA-1 expression in these purified hematopoietic cells. We found that GATA-1 and tal-1 genes are coexpressed in these three lineages. Remarkably, the expression of both genes is downmodulated during erythroid and megakaryocytic terminal maturation. In immature hematopoietic cells, tal-1 and GATA-1 genes are coexpressed in committed progenitors cells (CD34+/CD38(2+)), whereas they are not detectable in the most primitive cells (CD34(2+)/CD38-). In contrast, GATA-2 is strongly expressed in both most primitive and committed progenitors cells, whereas GATA-3 is mostly detected in most primitive ones. Altogether our results strongly suggest that GATA-1 modulates the transcription of tal-1 during the differentiation of the erythroid, megakaryocytic, and basosophilic lineages.

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