Regulation of Neuroendocrine Peptide Gene Expression

Blum, Mariann

doi:10.1016/b978-0-12-185150-7.50007-5

Cited by 10 publications

(17 citation statements)

References 9 publications

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“…Northwestern University. Evanston 111.. USA) [20], The construct was linearized and RNA probe was labeled with a -i2P-UTP (Amersham) using SP6 RNA polymerase ( 15 units/pl; Bethesda Research Laboratories) to a high specific activity (1.3 x 10'1 cprn/pg) [24], The ribosomal 18S cDNA was labeled with a -,:P-dCTP (Amersham) by random primer labeling method [25]. Details of the hybridization procedure were previously described [19], The membrane was exposed to X-ray film (Fuji) at -7 0°C for 3-5 days.…”

Section: Northern Blot Analysismentioning

confidence: 99%

Acute Increase of GABAergic Neurotransmission Exerts a Stimulatory Effect on GnRH Gene Expression in the Preoptic/Anterior Hypothalamic Area of Ovariectomized, Estrogen- and Progesterone-Treated Adult Female Rats

et al. 1995

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Although γ-aminobutyric acid (GABA) is known to play an important role in the regulation of GnRH release from the hypothalamus, GABAergic action on hypothalamic GnRH gene expression is poorly understood. The present study aims to evaluate the effects of several GABAergic compounds on GnRH mRNA and serum LH levels at the times of LH surge induced by estrogen plus progesterone treatment in long-term ovariectomized adult rats. Animals received either aminooxyacetic acid (AOAA, an inhibitor of GABA catabolism, i.p.), muscimol (GABA-A type agonist, i.c.v.) or baclofen (GABA-B type agonist, i.c.v.) 2 h prior to sacrifice. GnRH mRNA in the preoptic/anterior hypothalamic area and serum LH levels were determined by Northern blot analysis and LH radioimmunoassay, respectively. All of three GABA mimetics blocked the LH surge induced by estrogen plus progesterone in a dose-dependent manner. However, inhibition of GABA catabolism with AOAA in a dose range of 10-100 mg/kg b.w. increased GnRH mRNA level by 30%. Activation of GABA-A receptor with muscimol at a low dose (5 nmol) but not at high doses (10 and 30 nmol) elevated GnRH mRNA levels by 60% over the control value. Activation of GABA-B receptor with baclofen augmented GnRH mRNA levels in a dose-dependent manner. These observations indicate that acute increase of GABAergic neurotransmission may differentially regulate the release and GnRH gene expression depending on its receptor subtypes.

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Section: Northern Blot Analysismentioning

confidence: 99%

Acute Increase of GABAergic Neurotransmission Exerts a Stimulatory Effect on GnRH Gene Expression in the Preoptic/Anterior Hypothalamic Area of Ovariectomized, Estrogen- and Progesterone-Treated Adult Female Rats

et al. 1995

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“…In vitro synthesized pXBexI sense RNA was used to generate a standard curve for quantitation of samples. RNA standards and samples (5 ul), 20 pi of hybridization buffer and 5 pi o f antisense probe (500-600 pg) were allowed to hybri dize as previously described [25]. Following hybridization, samples were digested with 600-700 U SI nuclease (Pharmacia, N.J. USA), the protected hybrids purified and ethanol precipitated as described [25], and then analyzed on 6% acrylamide gels.…”

Section: Rna Protection Assaymentioning

confidence: 99%

“…RNA standards and samples (5 ul), 20 pi of hybridization buffer and 5 pi o f antisense probe (500-600 pg) were allowed to hybri dize as previously described [25]. Following hybridization, samples were digested with 600-700 U SI nuclease (Pharmacia, N.J. USA), the protected hybrids purified and ethanol precipitated as described [25], and then analyzed on 6% acrylamide gels. After exposure to X-ray film, radioactive bands corresponding to standards and samples were cut from the gel, subjected to liquid scintillation counting, and linear re gression analysis (Stat View 512 + , Brain Power, Calif., USA) was used to quantitate unkown samples by comparison to the standard curve.…”

Section: Rna Protection Assaymentioning

confidence: 99%

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Corticotropin-Releasing Factor Differentially Regulates Anterior and Intermediate Pituitary Lobe Proopiomelanocortin Gene Transcription, Nuclear Precursor RNA and Mature mRNA in vivo

et al. 1990

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Theproopiomelanocortin (POMC) gene and its peptide products are under complex regulation in the pituitary by multiple hormonal, neurohormonal and neurotransmitter factors. Corticotropin-releasing factor (CRF) stimulates the release of POMC-derived peptides in both anterior and intermediate lobes of the pituitary, while having differential long-term effects on levels of POMC mRNA in the two pituitary lobes in vivo. In the present study, we have analyzed the release of POMC-derived peptides, as well as changes in POMC gene transcription, primary transcript, nuclear mRNA and cytoplasmic mRNA levels following acute and chronic in vivo CRF administration in an attempt to elucidate the mechanism whereby this tissue-specific differential regulation occurs. Subcutaneous injection of CRF led to a substantial increase in plasma adrenocorticotropin, but only a minor sustained rise in plasma α-melanocyte-stimulating hormone. CRF was shown to induce rapid, time-dependent increases in POMC gene transcription in both anterior and intermediate pituitary lobes, which were reflected by increases in the level of POMC primary transcript in the nucleus. POMC primary transcript remained 2-fold elevated in the anterior lobe for at least 4 h after a single injection of CRF; in contrast, CRF stimulation of POMC primary transcript in the intermediate lobe was short-lived, and had returned to control values by 60 min after injection. After 7 days of repetitive CRF administration, POMC primary transcript, mature nuclear mRNA and cytoplasmic mRNA were 2.5 to 3.0-fold elevated in the anterior pituitary. In the intermediate lobe, long-term CRF administration did not alter the levels of POMC primary transcript, but did cause a slight (25–30%) reduction in nuclear and cytoplasmic POMC mRNA. These studies suggest that the differential regulation of POMC gene expression by CRF in anterior and intermediate pituitary lobes occurs in part due to differences in transcriptional responsiveness of the two cell types to this regulatory factor.

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“…The sodium acetate-ethanol precipitation step was repeated twice, and the sample was suspended in water (50 ,ul). Samples (5 ,I) containing 10 1) was modified by oligonucleotide-directed mutagenesis to eliminate all upstream ATG codons and to place the authentic start codon in the optimal sequence context for efficient translation in eucaryotic cells. The modified polymerase gene was then placed downstream of an SV40 early promoter in plasmid pTG100, which also carries the polyadenylation signal from the SV40 t-antigen region (Fig.…”

Section: Methodsmentioning

confidence: 99%

Synthesis of functional mRNA in mammalian cells by bacteriophage T3 RNA polymerase.

Zhou¹,

Giordano²,

Durbin³

et al. 1990

Mol. Cell. Biol.

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We found that the 5' nontranslated leader sequence from encephalomyocarditis virus (EMCV) allowed transcripts that were synthesized by the T3 RNA polymerase in mammalian cells to be translated in a cap-independent fashion. Stable mouse cell lines that carry the T3 RNA polymerase gene expressed the chloramphenicol acetyltransferase (CAT) It has been shown that the bacteriophage T3 and T7 RNA polymerases can function in mammalian cells and can transcribe DNA that is located in either the nucleus or the cytoplasm

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Regulation of Neuroendocrine Peptide Gene Expression

Cited by 10 publications

References 9 publications

Acute Increase of GABAergic Neurotransmission Exerts a Stimulatory Effect on GnRH Gene Expression in the Preoptic/Anterior Hypothalamic Area of Ovariectomized, Estrogen- and Progesterone-Treated Adult Female Rats

Acute Increase of GABAergic Neurotransmission Exerts a Stimulatory Effect on GnRH Gene Expression in the Preoptic/Anterior Hypothalamic Area of Ovariectomized, Estrogen- and Progesterone-Treated Adult Female Rats

Corticotropin-Releasing Factor Differentially Regulates Anterior and Intermediate Pituitary Lobe Proopiomelanocortin Gene Transcription, Nuclear Precursor RNA and Mature mRNA in vivo

Synthesis of functional mRNA in mammalian cells by bacteriophage T3 RNA polymerase.

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