In situ hybridization of arginine vasopressin (AVP) heteronuclear ribonucleic acid reveals increased AVP gene transcription in the rat hypothalamic paraventricular nucleus in response to emotional stress

The regulation of anterior pituitary adrenocorticotropin hormone (ACTH) secretion during stress involves several hypothalamic neurohormones, including arginine vasopressin (AVP). In situ hybridization techniques have been used to study the regulation of neuropeptide messenger ribonucleic acids in the hypothalamus. Owing to the relatively slow time course of the changes in cytoplasmic messenger ribonucleic acid concentrations, rapid alterations in the level of neuropeptide gene transcription could not be detected. Because of its rapid processing, the nuclear level of the heteronuclear ribonucleic acid should reflect the rate of its synthesis, namely the transcription of the gene. We have used in situ hybridization with a probe complementary to a portion of an intronic sequence of the rat AVP gene to study rapid changes in the level of AVP gene transcription during emotional stress. The specificity of our technique was demonstrated by the localization of the hybridization signals in the paraventricular nucleus, the supraoptic nucleus and the suprachiasmatic nucleus, and was confirmed by the nuclear localization of the labeling. Isolation and exposure of male rats to a novel environment induced an activation of the pituitary-adrenal axis and an increase in AVP heteronuclear ribonucleic acid concentrations in the paraventricular nucleus 2 h after the onset of the stress, suggesting that an increased AVP gene transcription may play a role in the activation of the pituitary-adrenal axis in response to emotional stress.

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Effects of acute hypotensive stimuli on arginine vasopressin gene transcription in the rat hypothalamus

AJP Endocrinology and Metabolism ◽

10.1152/ajpendo.2000.279.4.e886 ◽

2000 ◽

Vol 279 (4) ◽

pp. E886-E892 ◽

Cited By ~ 17

Author(s):

Satoshi Kakiya ◽

Hiroshi Arima ◽

Hisashi Yokoi ◽

Takashi Murase ◽

Yuko Yambe ◽

...

Keyword(s):

In Situ Hybridization ◽

Mean Arterial Pressure ◽

Arterial Pressure ◽

Gene Transcription ◽

Arginine Vasopressin ◽

The Other ◽

Paraventricular Nuclei ◽

Vasopressin Gene ◽

Avp Gene

We investigated the baroregulation of arginine vasopressin (AVP) gene transcription in the supraoptic (SON) and paraventricular nuclei (PVN) in conscious rats by use of intronic in situ hybridization. Hemorrhage of 16 ml/kg body wt decreased mean arterial pressure (MAP) by 57% and increased both plasma AVP (control, 1.2 ± 0.3 pg/ml; 16 ml/kg body wt, 38.9 ± 3.2 pg/ml) at 10 min and AVP heteronuclear (hn)RNA levels (SON, 150%; PVN, 140% of control values) at 20 min. On the other hand, hemorrhage of 7 ml/kg body wt had no significant effect on MAP, plasma AVP, or the AVP hnRNA levels. To better understand the baroregulation, we also examined the effects of sodium nitroprusside (SNP), which induces hypotension without a change in blood volume. The subcutaneous injection of 2 mg/kg body wt SNP, which decreased the MAP by 60%, increased both plasma AVP (control, 1.6 ± 0.4 pg/ml; 2 mg/kg body wt, 8.1 ± 0.4 pg/ml) at 10 min and AVP hnRNA levels (SON, 150%; PVN, 140% of control values) at 30 min. The injection of 0.1 mg/kg body wt SNP, which reduced the MAP by 10%, failed to increase either the plasma AVP or AVP hnRNA levels. These results indicate that AVP gene transcription increases rapidly after both hypotensive hemorrhage and normovolemic hypotension. In addition, it is suggested that the set point for AVP synthesis in the baroregulation is similar to that for AVP release.

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Analysis of the vasopressin system and water regulation in genetically polydipsic mice

AJP Endocrinology and Metabolism ◽

10.1152/ajpendo.2000.278.2.e189 ◽

2000 ◽

Vol 278 (2) ◽

pp. E189-E194 ◽

Cited By ~ 2

Author(s):

Yuko Yambe ◽

Yasuko Watanabe-Tomita ◽

Satoshi Kakiya ◽

Hisashi Yokoi ◽

Hiroshi Nagasaki ◽

...

Keyword(s):

Gene Expression ◽

In Situ Hybridization ◽

Arginine Vasopressin ◽

Water Retention ◽

Direct Sequencing ◽

Plasma Sodium ◽

Water Regulation ◽

Excessive Water ◽

Avp Gene

Polydipsic mice, STR/N, which show extreme polydipsia and polyuria, were discovered in 1958. In the STR/N, urine outputs are much higher than in control mice. The possibility of an abnormal regulation of the arginine vasopressin (AVP) system, or an abnormality in the renal susceptibility to AVP, should be considered. In this study we investigated the AVP system and water regulation in STR/N. We sequenced the AVP and the AVP V2-receptor genes of the STR/N by direct sequencing. No mutation was found in either of them. AVP gene expression examined by in situ hybridization and plasma sodium in 8-wk-old STR/N was significantly lower than in control mice, whereas it was significantly higher at 20 wk. Renal sensitivity to injected AVP was attenuated in 20-wk-old STR/N. The suppression of AVP synthesis due to excessive water retention in 8-wk-old STR/N suggests that polydipsia may be the primary cause in this strain. The 20-wk-old STR/N became dehydrated with the acceleration of AVP synthesis, which might have resulted from secondary desensitization to AVP.

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The Effects of Estrogen and Progesterone on Corticotropin-Releasing Hormone and Arginine Vasopressin Messenger Ribonucleic Acid Levels in the Paraventricular Nucleus and Supraoptic Nucleus of the Rhesus Monkey

Endocrinology ◽

10.1210/endo.140.5.6684 ◽

1999 ◽

Vol 140 (5) ◽

pp. 2191-2198 ◽

Cited By ~ 58

Author(s):

Brenda N. Roy ◽

Robert L. Reid ◽

Dean A. Van Vugt

Keyword(s):

In Situ Hybridization ◽

Rhesus Monkey ◽

Hpa Axis ◽

Paraventricular Nucleus ◽

Arginine Vasopressin ◽

Supraoptic Nucleus ◽

Mrna Levels ◽

Ovarian Steroids ◽

Messenger Ribonucleic Acid

Abstract Ovarian steroids increase hypothalamic-pituitary-adrenal (HPA) axis activity and sensitize the hypothalamic-pituitary-ovarian (HPO) axis to stress-induced inhibition. The present study investigated the effect of ovarian steroids on CRH and arginine vasopressin (AVP) messenger RNA (mRNA) levels in the rhesus monkey hypothalamus, as both neuropeptides have been shown to stimulate the HPA axis and inhibit the HPO axis in this species. This was accomplished by measuring CRH and AVP mRNA in the paraventricular nucleus (PVN) and supraoptic nucleus (SON) by in situ hybridization histochemistry. Menstrual cycles were simulated in ovariectomized (OVX) rhesus monkeys by sequential addition and removal of SILASTIC brand (Dow Corning Corp.) tubing containing either 17β-estradiol (E2) or progesterone (P4). On the morning of day 11 of the simulated follicular phase (E2 alone) or day 21 of the luteal phase (E2 + P4), animals were anesthetized, and the brains were perfused with paraformaldehyde via the carotid artery. Coronal sections (30 μm) were cut, and mRNA for CRH and AVP in the paraventricular nucleus (PVN) and supraoptic nucleus (SON) were semiquantified by in situ hybridization. CRH mRNA in the PVN of E2-replaced OVX animals (n = 7) was 2-fold greater than that in untreated OVX controls (n = 4), whereas CRH mRNA after E2 + P4 (n = 4) was no different from that in controls (optical density ± sem, 0.38 ± 0.06, 0.13 ± 0.08, and 0.14 ± 0.09 for OVX + E2, OVX + E2 + P4, and OVX, respectively; P = 0.02). CRH in the SON was undetectable. In contrast to CRH, AVP mRNA in the PVN and the SON was similar in the three treatment groups. We conclude that E2 and E2 + P4 replacement to OVX monkeys exert different effects on CRH and AVP gene expression, as estrogen stimulation of CRH mRNA in the PVN was abrogated by progesterone, whereas no effect of ovarian steroids on AVP mRNA in either the PVN or SON was observed. We postulate that ovarian steroid regulation of CRH synthesis and release may in part explain the central nervous system mechanisms by which ovarian steroids affect the HPA and HPO axes during basal and stress conditions.

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In Situ Hybridization as a Quantitative Autoradiographic Method: Vasopressin and Oxytocin Gene Transcription in the Brattleboro Rat

In Situ Hybridization in Brain ◽

10.1007/978-1-4615-9486-4_5 ◽

1986 ◽

pp. 73-95 ◽

Cited By ~ 10

Author(s):

Joseph T. McCabe ◽

Joan I. Morrell ◽

Donald W. Pfaff

Keyword(s):

In Situ Hybridization ◽

Gene Transcription ◽

Brattleboro Rat ◽

Autoradiographic Method

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CRH mRNA expression in the hypothalamic paraventricular nucleus is inhibited despite the activation of the hypothalamo-pituitary-adrenal axis during starvation

Brain Research ◽

10.1016/j.brainres.2008.06.065 ◽

2008 ◽

Vol 1228 ◽

pp. 107-112 ◽

Cited By ~ 7

Author(s):

Mitsuru Nishiyama ◽

Shinya Makino ◽

Yasumasa Iwasaki ◽

Yasushi Tanaka ◽

Hossein Pournajafi Nazarloo ◽

...

Keyword(s):

Mrna Expression ◽

Paraventricular Nucleus ◽

Hypothalamic Paraventricular Nucleus ◽

Adrenal Axis ◽

Pituitary Adrenal Axis

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Spatial- and temporal-restricted pattern for amelogenin gene expression during mouse molar tooth organogenesis

Development ◽

10.1242/dev.104.1.77 ◽

1988 ◽

Vol 104 (1) ◽

pp. 77-85 ◽

Cited By ~ 2

Author(s):

M.L. Snead ◽

W. Luo ◽

E.C. Lau ◽

H.C. Slavkin

Keyword(s):

Gene Expression ◽

In Situ Hybridization ◽

Gene Transcription ◽

Developmental Stages ◽

Three Dimensional ◽

Molar Tooth ◽

Specific Expression ◽

Amelogenin Gene ◽

Stage Of Development

Position- and time-restricted amelogenin gene transcription was analysed in developing tooth organs using in situ hybridization with asymmetric complementary RNA probes produced from a cDNA specific to the mouse 26 × 10(3) Mr amelogenin. In situ analysis was performed on developmentally staged fetal and neonatal mouse mandibular first (M1) and maxillary first (M1) molar tooth organs using serial sections and three-dimensional reconstruction. Amelogenin mRNA was first detected in a cluster of ameloblasts along one cusp of the M1 molar at the newborn stage of development. In subsequent developmental stages, amelogenin transcripts were detected within foci of ameloblasts lining each of the five cusps comprising the molar crown form. The number of amelogenin transcripts appeared to be position-dependent, being more abundant on one cusp surface while reduced along the opposite surface. Amelogenin gene transcription was found to be bilaterally symmetric between the developing right and left M1 molars, and complementary between the M1 and M1 developing molars; indicating position-restricted gene expression resulting in organ stereoisomerism. The application of in situ hybridization to forming tooth organ geometry provides a novel strategy to define epithelial-mesenchymal signal(s) which are believed to be responsible for organ morphogenesis, as well as for temporal- and spatial-restricted tissue-specific expression of enamel extracellular matrix.

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