Leptin Receptor in Human Term Placenta: in Situ Hybridization and Immunohistochemical Localization

Placenta ◽  
1999 ◽  
Vol 20 (8) ◽  
pp. 677-682 ◽  
Author(s):  
J Bodner ◽  
C.F Ebenbichler ◽  
H.J Wolf ◽  
E Müller-Holzner ◽  
U Stanzl ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Leptin Receptor ◽  
Immunohistochemical Localization ◽  
Human Term Placenta ◽  
Term Placenta

scholarly journals Galanin-Like Peptide (GALP) Is a Target for Regulation by Leptin in the Hypothalamus of the Rat

Endocrinology ◽  
2000 ◽  
Vol 141 (7) ◽  
pp. 2703-2706 ◽  
Author(s):  
Anders Juréus ◽  
Matthew J. Cunningham ◽  
Molly E. McClain ◽  
Donald K. Clifton ◽  
Robert A. Steiner
Keyword(s):  
In Situ Hybridization ◽  
Leptin Receptor ◽  
Fold Increase ◽  
Female Rats ◽  
Dorsomedial Nucleus ◽  
Physiological Regulation ◽  
Tissue Sections ◽  
Galanin Receptors ◽  
Neuroendocrine Functions

Galanin-like peptide (GALP), which was recently isolated from the porcine hypothalamus, shares sequence homology with galanin and binds with high affinity to galanin receptors. To study the distribution and regulation of GALP-expressing cells in the brain, we cloned a 120 base-pair cDNA fragment of rat GALP and produced an antisense riboprobe. In situ hybridization for GALP mRNA was then performed on tissue sections throughout the forebrain of adult ovariectomized female rats. We found GALP mRNA-containing cells in the arcuate nucleus (Arc), caudal dorsomedial nucleus, median eminence and the pituitary. Because GALP mRNA in the Arc appeared to overlap with the known distribution of leptin receptor mRNA, we tested the hypothesis that GALP expression is regulated by leptin. Using in situ hybridization, we compared the number of GALP mRNA-containing cells among groups of rats that were fed ad lib or fasted for 48 h and treated with either leptin or vehicle. Fasting reduced the number of identifiable cells containing GALP mRNA in the Arc, whereas the treatment of fasted animals with leptin produced a 4-fold increase in the number of cells expressing GALP message. The presence of GALP mRNA in the hypothalamus and pituitary and its regulation by leptin suggests that GALP may have important neuroendocrine functions, including the physiological regulation of feeding, metabolism, and reproduction.

Polar Expression and Phosphorylation of Human Leptin Receptor Isoforms in Paired, Syncytial, Microvillous and Basal Membranes from Human Term Placenta

Placenta ◽  
2002 ◽  
Vol 23 (6) ◽  
pp. 516-521 ◽  
Author(s):  
C.F. Ebenbichler ◽  
S. Kaser ◽  
M. Laimer ◽  
H.J. Wolf ◽  
J.R. Patsch ◽  
...  
Keyword(s):  
Leptin Receptor ◽  
Human Term Placenta ◽  
Term Placenta ◽  
Receptor Isoforms

scholarly journals In situ localization of amylase mRNA and protein. An investigation of amylase gene activity in normal human parotid gland.

1987 ◽  
Vol 35 (1) ◽  
pp. 9-14 ◽  
Author(s):  
D J Morley ◽  
M E Hodes
Keyword(s):  
In Situ Hybridization ◽  
Cellular Localization ◽  
Cdna Probe ◽  
Immunohistochemical Localization ◽  
Frozen Sections ◽  
Parotid Glands ◽  
Ductal Cells ◽  
Ethanol Acetic Acid ◽  
Mrna And Protein Expression

The distribution of human salivary amylase mRNA was studied by in situ hybridization to a [32P]-labeled amylase cDNA probe. Amylase mRNA was localized to the apical portion of acinar cells in frozen sections of human parotid salivary gland. No hybridization was noted in ductal cells, skeletal muscle, or in connective tissue. These results were consistent with immunohistochemical localization of amylase. The technique of in situ hybridization was modified to permit localization of amylase mRNA in variously fixed, paraffin-embedded parotid glands. Although the hybridization signal decreased with all fixatives, the pattern of localization paralleled that obtained with frozen sections. No advantage was noted in fixation with ethanol-acetic acid or Bouin solution over routine fixation with formalin. These results have important implications for researchers interested in studies of gene expression. We have demonstrated that routinely fixed paraffin blocks of human tissue can be used for cellular localization of specific mRNA. In coordination with immunocytochemistry, in situ hybridization offers a powerful tool for studies of mRNA and protein expression in individual cells.

scholarly journals Detection of calcitonin-encoding mRNA by radioactive and non-radioactive in situ hybridization: improved colorimetric detection and cellular localization of mRNA in thyroid sections.

1990 ◽  
Vol 38 (3) ◽  
pp. 351-358 ◽  
Author(s):  
M Denijn ◽  
R A De Weger ◽  
M J Berends ◽  
P I Compier-Spies ◽  
H Jansz ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Cellular Localization ◽  
Colorimetric Detection ◽  
Thyroid Tissue ◽  
Immunohistochemical Localization ◽  
Dna Probes ◽  
Peroxidase Reaction ◽  
Gold Silver ◽  
Labeled Rna

The localization of mRNA encoding calcitonin was studied by in situ hybridization using 35S-labeled RNA probes and biotin-labeled DNA probes. Radiolabeled probes were detected by autoradiography and biotin-labeled probes by streptavidin-biotin-peroxidase. To intensify the colorimetric signal, the indirect avidin-biotin complex (ABC) method was performed. However, the results were often variable. To improve the sensitivity, the peroxidase reaction signal was enhanced with a gold-silver deposit intensification reaction. To shorten the incubation times and to enhance the colorimetric reaction, several reaction steps were performed in a microwave oven. The localization of calcitonin mRNA in thyroid tissue, as detected with in situ hybridization, was confirmed by immunohistochemical localization of the calcitonin polypeptide. The results of in situ hybridization using biotinylated probes were compared to in situ hybridization using radioactive probes. Our data show that the results of in situ hybridization applied on frozen and paraffin-embedded sections using biotinylated DNA probes, detected with an indirect streptavidin-biotin-peroxidase reaction and intensified by silver-gold enhancement, were comparable to those obtained with radioactive probes. The localization of calcitonin encoding mRNA was in agreement with the localization of the calcitonin polypeptide.

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