scholarly journals Localization of neuropeptide Y mRNA in neurons of human cerebral cortex by means of in situ hybridization with a complementary RNA probe.

1987 ◽  
Vol 84 (20) ◽  
pp. 7315-7318 ◽  
Author(s):  
G. Terenghi ◽  
J. M. Polak ◽  
Q. Hamid ◽  
E. O'Brien ◽  
P. Denny ◽  
...  
Keyword(s):  
Cerebral Cortex ◽  
In Situ Hybridization ◽  
Neuropeptide Y ◽  
Human Cerebral Cortex ◽  
Rna Probe

Expression of a Novel Neuropeptide Y Receptor Subtype Involved in Food Intake: An in Situ Hybridization Study of Y5 mRNA Distribution in Rat Brain

2000 ◽  
Vol 165 (1) ◽  
pp. 90-100 ◽  
Author(s):  
Margaret M. Durkin ◽  
Mary W. Walker ◽  
Kelli E. Smith ◽  
Eric L. Gustafson ◽  
Christophe Gerald ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Food Intake ◽  
Neuropeptide Y ◽  
Rat Brain ◽  
Receptor Subtype ◽  
Hybridization Study ◽  
Neuropeptide Y Receptor

scholarly journals LGR4 and Its Ligands, R-Spondin 1 and R-Spondin 3, Regulate Food Intake in the Hypothalamus of Male Rats

Endocrinology ◽  
2014 ◽  
Vol 155 (2) ◽  
pp. 429-440 ◽  
Author(s):  
Ji-Yao Li ◽  
Biaoxin Chai ◽  
Weizhen Zhang ◽  
Danielle M. Fritze ◽  
Chao Zhang ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Food Intake ◽  
Neuropeptide Y ◽  
Paraventricular Nucleus ◽  
Median Eminence ◽  
Energy Homeostasis ◽  
Male Rats ◽  
The Third ◽  
The Brain

The hypothalamus plays a key role in the regulation of feeding behavior. Several hypothalamic nuclei, including the arcuate nucleus (ARC), paraventricular nucleus, and ventromedial nucleus of the hypothalamus (VMH), are involved in energy homeostasis. Analysis of microarray data derived from ARC revealed that leucine-rich repeat-containing G protein-coupled receptor 4 (LGR4) is highly expressed. LGR4, LGR5, and LGR6 form a subfamily of closely related receptors. Recently, R-spondin (Rspo) family proteins were identified as ligands of the LGR4 subfamily. In the present study, we investigated the distribution and function of LGR4–LGR6 and Rspos (1–4) in the brain of male rat. In situ hybridization showed that LGR4 is expressed in the ARC, VMH, and median eminence of the hypothalamus. LGR4 colocalizes with neuropeptide Y, proopiomelanocortin, and brain-derived neurotrophic factor neurons. LGR5 is not detectable with in situ hybridization; LGR6 is only expressed in the epithelial lining of the lower portion of the third ventricle and median eminence. Rspo1 is expressed in the VMH and down-regulated with fasting. Rspo3 is expressed in the paraventricular nucleus and also down-regulated with fasting. Rspos 1 and 3 colocalize with the neuronal marker HuD, indicating that they are expressed by neurons. Injection of Rspo1 or Rspo3 into the third brain ventricle inhibited food intake. Rspo1 decreased neuropeptide Y and increased proopiomelanocortin expression in the ARC. Rspo1 and Rspo3 mRNA is up-regulated by insulin. These data indicate that Rspo1 and Rspo3 and their receptor LGR4 form novel circuits in the brain to regulate energy homeostasis.

scholarly journals PCR-derived ssDNA Probes for Fluorescent In Situ Hybridization to HIV-1 RNA

2000 ◽  
Vol 48 (2) ◽  
pp. 285-293 ◽  
Author(s):  
Marlyse C. Knuchel ◽  
Brigit Graf ◽  
Erika Schlaepfer ◽  
Herbert Kuster ◽  
Marek Fischer ◽  
...  
Keyword(s):  
T Cells ◽  
In Situ Hybridization ◽  
Fluorescent In Situ Hybridization ◽  
Uracil Dna Glycosylase ◽  
Rna Probes ◽  
Immunodeficiency Virus ◽  
Polymerase Chain ◽  
Rna Probe ◽  
Hiv 1

We developed a simple and rapid technique to synthesize single-stranded DNA (ssDNA) probes for fluorescent in situ hybridization (ISH) to human immunodeficiency virus 1 (HIV-1) RNA. The target HIV-1 regions were amplified by the polymerase chain reaction (PCR) and were simultaneously labeled with dUTP. This product served as template for an optimized asymmetric PCR (one-primer PCR) that incorporated digoxigenin (dig)-labeled dUTP. The input DNA was subsequently digested by uracil DNA glycosylase, leaving intact, single-stranded, digoxigenin-labeled DNA probe. A cocktail of ssDNA probes representing 55% of the HIV-1 genome was hybridized to HIV-1-infected 8E5 T-cells and uninfected H9 T-cells. For comparison, parallel hybridizations were done with a plasmid-derived RNA probe mix covering 85% of the genome and a PCR-derived RNA probe mix covering 63% of the genome. All three probe types produced bright signals, but the best signal-to-noise ratios and the highest sensitivities were obtained with the ssDNA probe. In addition, the ssDNA probe syntheses generated large amounts of probe (0.5 to 1 μg ssDNA probe per synthesis) and were easier to perform than the RNA probe syntheses. These results suggest that ssDNA probes may be preferable to RNA probes for fluorescent ISH.

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