A new sex dimorphism in the Harderian gland of the frog Rana esculenta

2007 ◽  
Vol 85 (8) ◽  
pp. 909-915 ◽  
Author(s):  
Ismene Serino ◽  
Gaia Izzo ◽  
Diana Ferrara ◽  
Michela d’Istria ◽  
Sergio Minucci
Keyword(s):  
Sexual Dimorphism ◽  
Secretory Activity ◽  
Harderian Gland ◽  
Polymerase Chain Reaction Analysis ◽  
Rana Esculenta ◽  
Rt Pcr ◽  
Polymerase Chain ◽  
Two Peaks ◽  
Constant Expression

The Harderian gland (Hg), the only gland found in the orbit of the frog Rana esculenta L., 1758, probably plays a role in orbital lubrication. The secretory activity of the Hg is seasonal, showing the highest activity in summer. There is little information on Hg gene expression; previously, we identified a mRNA named harderin, whose deduced protein has no homology with other proteins. Differential expression of the harderin transcript between the sexes expressed during the annual cycle implies sexual dimorphism. RT–PCR (reverse transcription – polymerase chain reaction) analysis, revealed that harderin is expressed during the entire year in the Hg of both sexes. It shows a higher level of expression in the female glands than that of male glands. Two peaks of expression, in February and in June, were observed in the female glands, while only the February peak was observed in those of males. These observations were supported by in situ hybridization. Experiments involving gonadectomy and (or) hormonal replacement therapy showed a significant decrease in harderin in the Hg of females; this effect is prevented by estradiol (testosterone had no effect), while ICI (antiestrogen) counteracts the hormonal prevention, suggesting that this sexual dimorphism is under estradiol control. The constant expression of harderin mRNA during the year suggests a probable constitutive role for this molecule.

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.

The bovine sex-determining region Y (Sry) gene and its mRNA transcript are present in Y sperm but not X sperm of bulls

2018 ◽  
Vol 68 (3) ◽  
pp. 321-332 ◽  
Author(s):  
Chun-Mei Han ◽  
Rong Chen ◽  
Tao Li ◽  
Xiao-Li Chen ◽  
Yong-Fu Zheng ◽  
...  
Keyword(s):  
Cellular Localization ◽  
Fish Analysis ◽  
Mrna Transcript ◽  
Rt Pcr ◽  
Sry Gene ◽  
Chain Reaction ◽  
Dna And Rna ◽  
Positive Rate ◽  
Polymerase Chain

AbstractThe aims of this study were to establish whether the sex-determining region Y gene and its mRNA transcript are present in the Y sperm and X sperm of bulls and, if present, determine their cellular localization. Semen was collected from three bulls and sorted by flow cytometry into X- and Y-chromosome populations. Reverse transcription-polymerase chain reaction (RT-PCR) was used to determineSrymRNA expression in X sperm and Y sperm. The presence and localization ofSryDNA and RNA were investigated by fluorescence in situ hybridization (FISH). RT-PCR detected a singleSrytranscript of 142 bp in Y sperm but not in X sperm. In Y sperm, the FISH-positive rates forSryDNA andSryRNA did not differ significantly from the re-analyzed Y sperm purity. In further experiments, there were no significant differences between the FISH-positive rate forSryRNA and the re-analyzed Y sperm purity for X-sorted, Y-sorted, or unsorted sperm. In conclusion, FISH analysis revealed thatSrytranscripts are present at the edges of the sperm heads of Y sperm but are absent from X sperm.

Expression of MMP-8 and MMP-13 Genes in the Periodontal Ligament during Tooth Movement in Rats

2003 ◽  
Vol 82 (8) ◽  
pp. 646-651 ◽  
Author(s):  
I. Takahashi ◽  
M. Nishimura ◽  
K. Onodera ◽  
J.-W. Bae ◽  
H. Mitani ◽  
...  
Keyword(s):  
Gene Expression ◽  
Periodontal Ligament ◽  
Tooth Movement ◽  
Orthodontic Tooth Movement ◽  
Polymerase Chain Reaction Analysis ◽  
Tension And Compression ◽  
Polymerase Chain

Periodontal ligament tissue is remodeled on both the tension and compression sides of moving teeth during orthodontic tooth movement. The present study was designed to clarify the hypothesis that the expression of MMP-8 and MMP-13 mRNA is promoted during the remodeling of periodontal ligament tissue in orthodontic tooth movement. We used the in situ hybridization method and semi-quantitative reverse-transcription/polymerase chain-reaction analysis to elucidate the gene expression of MMP-8 and MMP-13 mRNA. Expression of MMP-8 and MMP-13 mRNA transiently increased on both the compression and tension sides during active tooth movement in vivo. The gene expression of MMP-8 and MMP-13 was induced by tension, while compression indirectly promoted the gene expression of MMP-8 and MMP-13 through soluble factors in vitro. Thus, we concluded that the expression of MMP-8 and MMP-13 is differentially regulated by tension and compression, and plays an important role in the remodeling of the periodontal ligament.

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.

scholarly journals Quantification of hepatitis C virus-infected peripheral blood mononuclear cells by in situ reverse transcriptase-polymerase chain reaction

Blood ◽  
1996 ◽  
Vol 88 (7) ◽  
pp. 2768-2774 ◽  
Author(s):  
L Muratori ◽  
D Gibellini ◽  
M Lenzi ◽  
M Cataleta ◽  
P Muratori ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Chronic Hepatitis ◽  
Hepatitis C ◽  
Mononuclear Cells ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Peripheral Blood Mononuclear ◽  
Blood Mononuclear Cells ◽  
Polymerase Chain

Hepatitis C virus (HCV) is known to infect peripheral blood mononuclear cells (PBMC) of patients with chronic hepatitis C, but the proportion of HCV-infected circulating cells is not detectable by conventional reverse transcriptase-polymerase chain reaction (RT-PCR) and the pathogenic significance of HCV lymphotropism is still unclear. Therefore, we have devised an in situ RT-PCR technique using fluorescein-labeled HCV-specific primers revealed by flow cytometry. PBMC were isolated from 28 patients with chronic HCV-related liver disease; of these, 6 had previously received an orthotopic liver transplantation (OLT) and were on immuno-suppressive treatment. Fourteen patients (50%) were found positive for HCV genome within PBMC by in situ RT-PCR, the proportion of HCV-infected cells ranging from 0.2% to 8.1%. All 6 OLT patients tested positive. The fluorescent signal, corresponding to the HCV-specific 340-bp amplicon, was confined to part of the cytoplasmic compartment of scattered PBMC. Of these 14 patients, 12 had also negativestrand HCV RNA within PBMC detected by “tagged” RT-PCR. We conclude that HCV may infect a significant proportion of PBMC in chronic hepatitis C patients, especially immunosuppressed OLT cases, and that viral replication within PBMC is a common occurrence. Over time, the persistence of HCV-infected immune system cells might interfere with normal immunologic mechanisms and play a role in the pathogenic processes leading to extrahepatic disorders such as mixed cryoglobulinemia and B-cell malignant lymphoma.

Novel and sensitive detection systems for the vitamin D receptor — in situ-reverse transcriptase-polymerase chain reaction and immunogold cytochemistry

1996 ◽  
Vol 16 (2) ◽  
pp. 183-195 ◽  
Author(s):  
A P Mee ◽  
L K Davenport ◽  
J A Hoyland ◽  
M Davies ◽  
E B Mawer
Keyword(s):  
Vitamin D ◽  
Polymerase Chain Reaction ◽  
Cell Line ◽  
Vitamin D Receptor ◽  
Human Kidney ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Low Levels

ABSTRACT The receptor for the active metabolite of vitamin D, 1,25(OH)2D3, known as the vitamin D receptor (VDR), belongs to the steroid hormone nuclear receptor superfamily. We have developed novel methods for detection of VDR mRNA and protein within a human promyelomonocytic cell line, HL-60. Using the newly developed technique of in situ-reverse transcriptase-polymerase chain reaction (IS-RT-PCR), low levels of VDR mRNA could be amplified and demonstrated unequivocally within these cells, and also within a human kidney proximal tubule cell line, CL-8. Use of a novel immunogold cytochemical technique has allowed clear and sensitive detection of VDR protein expression within the HL-60 cells. Further development of IS-RT-PCR has allowed us to apply this technique to tissue sections. We have shown clear amplification of VDR transcripts within sections of formalin-fixed paraffin-embedded human kidney and liver. These techniques will be useful to localise specifically the VDR within cell types that contain low levels of mRNA and protein, and will permit further investigation of the role played by 1,25(OH)2D3 in cellular regulatory mechanisms.

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