Changes in biological activity of bone cells in ovariectomized rats revealed by in situ hybridization

2009 ◽  
Vol 11 (6) ◽  
pp. 780-788 ◽  
Author(s):  
Tohru Ikeda ◽  
Akira Yamaguchi ◽  
Satoshi Yokose ◽  
Yumiko Nagai ◽  
Hideyuki Yamato ◽  
...  
Keyword(s):  
Biological Activity ◽  
In Situ Hybridization ◽  
Bone Cells ◽  
Ovariectomized Rats

Analysis of mRNA, miRNA, and DNA in Bone Cells by RT-qPCR and In Situ Hybridization

2019 ◽  
pp. 169-196
Author(s):  
Brice Moukengue ◽  
Jérôme Amiaud ◽  
Camille Jacques ◽  
Céline Charrier ◽  
Benjamin Ory ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Bone Cells

scholarly journals Expression of Vesicular Glutamate Transporter-2 in Gonadotrope and Thyrotrope Cells of the Rat Pituitary. Regulation by Estrogen and Thyroid Hormone Status

Endocrinology ◽  
2006 ◽  
Vol 147 (8) ◽  
pp. 3818-3825 ◽  
Author(s):  
Erik Hrabovszky ◽  
Imre Kalló ◽  
Gergely F. Turi ◽  
Katalin May ◽  
Gábor Wittmann ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Thyroid Hormone ◽  
Glutamate Transporter ◽  
Male Rats ◽  
Ovariectomized Rats ◽  
Mrna Levels ◽  
Vesicular Glutamate Transporter ◽  
Local Synthesis ◽  
Enhanced Secretion

Immunocytochemical studies of the rat adenohypophysis identified a cell population that exhibits immunoreactivity for type-2 vesicular glutamate transporter (VGLUT2), a marker for glutamatergic neuronal phenotype. The in situ hybridization detection of VGLUT2 mRNA expression in adenohypophysial cells verified that VGLUT2 immunoreactivity is due to local synthesis of authentic VGLUT2. Dual-immunofluorescent studies of the hypophyses from male rats showed the presence of VGLUT2 in high percentages of LH (93.3 ± 1.3%)-, FSH (44.7 ± 3.9%)-, and TSH (70.0 ± 5.6%)-immunoreactive cells and its much lower incidence in cells of the prolactin, GH, and ACTH phenotypes. Quantitative in situ hybridization studies have established that the administration of a single dose of 17-β-estradiol (20 μg/kg; sc) to ovariectomized rats significantly elevated VGLUT2 mRNA in the adenohypophysis 16 h postinjection. Thyroid hormone dependence of VGLUT2 expression was addressed by the comparison of hybridization signals in animal models of hypo- and hyperthyroidism to those in euthyroid controls. Although hyperthyroidism had no effect on VGLUT2 mRNA, hypothyroidism increased adenohypophysial VGLUT2 mRNA levels. This coincided with a decreased ratio of VGLUT2-immunoreactive TSH cells, regarded as a sign of enhanced secretion. The presence of the glutamate marker VGLUT2 in gonadotrope and thyrotrope cells, and its up-regulation by estrogen or hypothyroidism, address the possibility that endocrine cells of the adenohypophysis may cosecrete glutamate with peptide hormones in an estrogen- and thyroid status-regulated manner. The exact roles of endogenous glutamate observed primarily in gonadotropes and thyrotropes, including its putative involvement in autocrine/paracrine regulatory mechanisms, will require clarification.

Estrogen Receptor mRNA in Uteri of Normal Estrous Cycling and Ovariectomized Rats by In Situ Hybridization

1999 ◽  
Vol 221 (3) ◽  
pp. 207-214 ◽  
Author(s):  
Shin‐Ichi Katsuda ◽  
Midori Yoshida ◽  
Takao Watanabe ◽  
Hiroyuki Kuroda ◽  
Jin Ando‐Lu ◽  
...  
Keyword(s):  
Estrogen Receptor ◽  
In Situ Hybridization ◽  
Ovariectomized Rats ◽  
Receptor Mrna ◽  
Estrogen Receptor Mrna ◽  
Estrous Cycling

Estrogen Receptor mRNA in Uteri of Normal Estrous Cycling and Ovariectomized Rats by In Situ Hybridization

1999 ◽  
Vol 221 (3) ◽  
pp. 207-214 ◽  
Author(s):  
S.-I. Katsuda ◽  
M. Yoshida ◽  
T. Watanabe ◽  
H. Kuroda ◽  
J. Ando-Lu ◽  
...  
Keyword(s):  
Estrogen Receptor ◽  
In Situ Hybridization ◽  
Ovariectomized Rats ◽  
Receptor Mrna ◽  
Estrogen Receptor Mrna ◽  
Estrous Cycling

scholarly journals Osteoporosis Affects Functional Activity and Gene Expression of Osteoblastic Cells Derived from Rat Alveolar Bone

2020 ◽  
Vol 31 (6) ◽  
pp. 617-622
Author(s):  
Paula Katherine Vargas-Sanchez ◽  
Roger Rodrigo Fernandes ◽  
Flávia Aparecida Chaves Furlaneto ◽  
Luiz Gustavo de Sousa ◽  
Selma Siéssere ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Alveolar Bone ◽  
Bone Cells ◽  
Female Rats ◽  
Ovariectomized Rats ◽  
Osteoblastic Cells ◽  
Expression Of Genes ◽  
In Situ Detection

Abstract Recent studies suggest that osteoporosis, in addition to the damage caused in long bones, may cause deterioration in the jaws, especially in alveolar bone sites, with effects in the progress of periodontal disease as well as in bone healing. The aim of this study was to evaluate the effect of osteoporosis in the metabolism of rat alveolar bone osteoblasts. There were used 10 female rats divided in two experimental groups (Sham and OVX), which were ovariectomized and after 8 weeks euthanized to collect mandibular bone samples in order to isolate osteoblastic cells. The cells were cultured in 24-well plates to perform the in vitro experiments. After 7, 10 and 14 days, there were evaluated cell proliferation by MTT assay, in situ detection of alkaline phosphatase (ALP) as well as mineralized nodules and expression of genes associated to bone remodeling. Results showed that at 7, 10 and 14 days cell proliferation was lower for OVX group. In situ detection of ALP was higher at 7 days and lower at 10 and 14 days in OVX group. At 17 and 21 days, OVX group had a significative decrease of mineralization nodules. There was a downregulation in the expression of Alp, Bglap and Runx2 genes and an upregulation of Opg in OVX group, whereas Opn and Rankl modulation was similar between the evaluated groups. Our results suggest that osteoporosis has a deleterious effect on alveolar bone cells from ovariectomized rats, which might affect the treatment of diseases associated to the jaw bones.

DNA sequence mapping in interphase and metaphase chromosomes by fluorescence in situ hybridization

1992 ◽  
Vol 50 (1) ◽  
pp. 496-497
Author(s):  
Barbara Trask ◽  
Susan Allen ◽  
Anne Bergmann ◽  
Mari Christensen ◽  
Anne Fertitta ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Dna Sequence ◽  
Dna Sequences ◽  
Dual Band ◽  
Nick Translation ◽  
Metaphase Chromosomes ◽  
Band Pass ◽  
Texas Red ◽  
Fluorescent Spot

Using fluorescence in situ hybridization (FISH), the positions of DNA sequences can be discretely marked with a fluorescent spot. The efficiency of marking DNA sequences of the size cloned in cosmids is 90-95%, and the fluorescent spots produced after FISH are ≈0.3 μm in diameter. Sites of two sequences can be distinguished using two-color FISH. Different reporter molecules, such as biotin or digoxigenin, are incorporated into DNA sequence probes by nick translation. These reporter molecules are labeled after hybridization with different fluorochromes, e.g., FITC and Texas Red. The development of dual band pass filters (Chromatechnology) allows these fluorochromes to be photographed simultaneously without registration shift.

Ultrastructural analysis of the spatial distribution of MRNA

1992 ◽  
Vol 50 (1) ◽  
pp. 552-553
Author(s):  
Gary Bassell ◽  
Robert H. Singer
Keyword(s):  
Electron Microscopy ◽  
Spatial Distribution ◽  
In Situ Hybridization ◽  
High Resolution ◽  
Nucleic Acids ◽  
Colloidal Gold ◽  
Dna Probe ◽  
Nucleotide Analogs ◽  
Gold Particles

We have been investigating the spatial distribution of nucleic acids intracellularly using in situ hybridization. The use of non-isotopic nucleotide analogs incorporated into the DNA probe allows the detection of the probe at its site of hybridization within the cell. This approach therefore is compatible with the high resolution available by electron microscopy. Biotinated or digoxigenated probe can be detected by antibodies conjugated to colloidal gold. Because mRNA serves as a template for the probe fragments, the colloidal gold particles are detected as arrays which allow it to be unequivocally distinguished from background.

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