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Cyclic adenosine 3',5'-monophosphate (cAMP) has been implicated as an intracellular messenger mediating osmotic regulation of expression of the gene encoding the neuropeptide vasopressin (VP) in the hypothalamus. We have used a heterologous transient transfection system to demonstrate that cAMP regulates the bovine VP gene promoter following transfection(More)
POMC mRNA levels in the intermediate lobe of the pituitary are known to be regulated by osmotic stimuli. Salt loading with 2% saline causes a decrease in POMC mRNA levels in the IL without affecting levels in the anterior lobe. We have investigated possible mediators of this effect and attempted to elucidate the mechanism by which the changes in mRNA levels(More)
In thyroid hormone-depleted rats, the rate of transcription of the growth hormone (GH) gene in the anterior pituitary gland is lower than the rate in euthyroid controls, and there is a corresponding reduction in the abundance of the GH mRNA. Concomitantly, the poly(A) tail of the GH mRNA increases in length. Examination of nuclear RNA from anterior(More)
There is growing evidence of interactions between the central nervous system and the immune system. We present evidence that the cytokine interleukin-2 (IL-2) influences expression of the genes encoding the neuropeptides vasopressin (VP) and oxytocin (OT) in the hypothalamus of the nude mouse. A single injection of recombinant mouse IL-2 (rmIL-2) caused a(More)
This article describes the use of three reporter enzymes used to study promoter activity in transgenic animals. Chloramphenicol acetyl transferase may be assayed by a nonchromatographic method that is rapid and sensitive. beta-Galactosidase is measured by a photometric assay and luciferase is assayed by measuring the emission of light using a luminometer.(More)
Studies on vasoactive intestinal peptide (VIP) in the anterior pituitary gland have shown that it is synthesized locally, physiologically regulated, and may act as a paracrine/autocrine factor. We have now investigated the regulation of anterior pituitary VIP gene expression in rats during osmotic stimulation. Both salt-loading and dehydration resulted in a(More)
  • K Pardy
  • Methods in molecular biology
  • 1993
Prior to the introduction of transgene constructs into animals, it is useful first to determine the presence of a functional promoter and transcription unit. This can be achieved by transient transfection of the construct into heterologous cell lines and subsequent measurement of reporter enzyme activity in cell extracts. Following ligation of the chosen(More)
  • K Pardy
  • Methods in molecular biology
  • 1993
Reporter genes code for proteins that have a unique enzymatic activity and are used to assess the transcriptional properties of DNA elements. The use of reporter genes in transgenic animals provides a rapid method for the detection of transgene expression, which is easily distinguishable from expression of the corresponding endogenous gene of the animal.(More)
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