CRF and Vasopressin gene expression in Stress Response

应激反应中的 CRF 和加压素基因表达

• 批准号: 08044232
• 负责人: ITOI Keiichi
• 金额: $ 2.75万
• 依托单位: Tohoku University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for international Scientific Research
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 无数据
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08044232/
• 关键词: CRF; Vasopressin; gene; mRNA; intron; in situ hybridization; hypothalamic; norepinephrine; ストレス; hnRNA; 脳内微量注入法

Whereas CRF is localized in the parvocellular division, vasopressin (AVP) is present in both parvo- and magnocellular divisions of the hypothalamic paraventricular nucleus (PVN) . Norepinephrine (NE), as a neurotransmitter, has been proposed to convey stimulatory signals to both divisions of the PVN.To elucidate whether NE regulates CRF and AVP gene expression diffferentially in the PVN we examined the effects of NE on CRF and AVP gene expression in vivo .Stainless steel guide cannulas were implanted stereotaxically into male Wistar rats aiming at 2 mm above the left PVN under pentobarbital anesthesia. One week following surgery, a 31-G needle was inserted into the guide cannula to reach the dorsolateral border of the PVN .NE (50 nmol) dissolved in artificial CSF (100 nl), or CSF alone, was injected into the left PVN of conscious rats. Rats were decapitated at 0,15,30,60,90, and 120 min following NE injection (or, 30 min following CSF injection), and trunk blood was collected for corti … More costerone (CORT) RIA,then the brain was removed and stored at -80C.All experiments were carried out from 0800-1200 h. Hypothalami were sectioned at 10 mum on a crmuyostat and mounted onto slides. Antisense 35S-labeled cRNA probes were synthesized for rat CRF exon 2, CRF intron, AVP exon C, AVP intron 1. In situ hybridization was carried out on sections taken every 70 mum throughout the PVN,and the tissue was exposed to X-ray film.Signal intensity on each side of the PVN was quantitated from 10 sections per animal. We found that plasma CORT levels increased markedly following NE injection, reached a peak at 60 min, and remained high at 120 min. CRF hnRNA increased dramatically at 15 min to levels approximately 70 times as high as those in controls, and returned to baseline by 90 min. CRF mRNA incresed by approximately 30% at 90 min. Both CRF hnRNA and mRNA levels were not significantly different from the controls 30 min following CSF injection. AVP hnRNA as well as mRNA levels, examined at 30 and 120 min, were not significantly different from those in controls. These results suggest that NE regulates CRF and AVP gene transcription differentially in the PVN. Less

CRF局限于细小细胞分裂，而抗利尿激素（AVP）则存在于下丘脑室旁核（PVN）的细小细胞分裂和大细胞分裂中。去甲肾上腺素（NE）作为一种神经递质，被认为可以向PVN的两个部分传递刺激信号。为了阐明NE是否在PVN中调控CRF和AVP基因表达差异，我们在体内检测了NE对CRF和AVP基因表达的影响。在戊巴比妥麻醉下，将不锈钢导管定向植入雄性Wistar大鼠左PVN上方2 mm处。术后一周，将一根31-G针插入引导套管，到达PVN的背外侧边界。将NE （50 nmol）溶于人工脑脊液（100 nl）或单独脑脊液注入清醒大鼠左侧PVN。分别于NE注射后0、15、30、60、90、120 min（或CSF注射后30 min）处死大鼠，取干血检测CORT含量，取co酮（CORT） RIA，取脑组织-80℃保存。所有实验于0800- 1200h进行。下丘脑在10℃时在恒温器上切片并装在载玻片上。合成了大鼠CRF外显子2、CRF内含子、AVP外显子C、AVP内含子1的反义35s标记cRNA探针。在整个PVN上每隔70分钟进行原位杂交，并将组织暴露在x射线胶片上。在每只动物的10个切片上定量测量PVN每侧的信号强度。我们发现血浆CORT水平在注射NE后显著升高，在60分钟达到峰值，并在120分钟保持高位。CRF hnRNA在15分钟内急剧增加，达到对照组的70倍左右，并在90分钟后恢复到基线水平。90 min时，CRF mRNA增加约30%。注射脑脊液30分钟后，CRF hnRNA和mRNA水平与对照组无显著差异。AVP hnRNA和mRNA水平在30和120分钟时检测，与对照组无显著差异。这些结果表明，NE在PVN中对CRF和AVP基因转录的调控存在差异。少

项目成果

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Lepoz-Figueiou MO 等人：“ziucocorticond 的一氧化氮合酶的调节”肠道肽研讨会摘要（1997 年）。

Itoi K.et al: "Norepinephrine (NE) stimulates CRF gane transcription in the hypothalomic paraventyiculas nesdus (PVN) of conscicus rats." Abstracrts of the 79th Annual Mecting of the Encocrins Society. (印刷中). (1997)

Itoi K. 等人：“去甲肾上腺素 (NE) 刺激意识大鼠下丘脑旁室静脉 (PVN) 中的 CRF gane 转录。”第 79 届 Encocrins 协会年度会议摘要（1997 年）。

Itoi k and Watson S.J.: "Neural and intracellulur regulatory mechanisms of CRF gene expression" Endocrine J. (印刷中). (1997)

Itoi k 和 Watson S.J.：“CRF 基因表达的神经和细胞内调节机制”Endocrine J.（出版中）。

Itoi K,Watson SJ: "Neural and in tracellulas regulatory mechanisms of CRF gene expression." Enciocrive J. (in press). (1997)

Itoi K，Watson SJ：“CRF 基因表达的神经和微细胞调节机制。”

Itoi K etal: "Norepinephrine (NE) stimulatcr CRF gene transesiption in the hypothalamic paraventricuiar nucleus (PVN) of conscious rats." Abstr 79th Annual Mecting of the Endocrive Society. (in press). (1997)

Itoi K 等人：“去甲肾上腺素 (NE) 刺激 CRF 基因在清醒大鼠的下丘脑室旁核 (PVN) 中的转染。”