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通过原位杂交和Northern印迹法评估大鼠脱水-再水化过程中视上核中血管加压素和催产素信使核糖核酸的变化。

Alterations in vasopressin and oxytocin messenger RNA in the rat supraoptic nucleus during dehydration-rehydration evaluated by in situ hybridization and northern blotting.

作者信息

Arnauld E, Wattiaux J P, Arsaut J, Rostène W, Vincent J D

机构信息

INSERM, U.176, Bordeaux, France.

出版信息

Neurosci Lett. 1993 Jan 12;149(2):177-81. doi: 10.1016/0304-3940(93)90765-d.

DOI:10.1016/0304-3940(93)90765-d
PMID:8474692
Abstract

Male rats were deprived of water for 5 days, and then given water ad libitum for 3, 7, 10 or 14 days. Plasma osmolarity returned to normal in less than 3 days, while pituitary vasopressin (AVP) and oxytocin (OXT) only returned to control levels after 14 days. Sections of the supraoptic nucleus (SON) were hybridized with 35S-labelled cDNA (OXT) or oligonucleotide (AVP) probes. Relative AVP and OXT mRNA contents were quantitated by counting the number of silver grains on a large standard area of the SON, then extrapolating this value to the volume of the whole SON (deduced from surface areas of all the sections). Dehydration significantly enlarged the volume of the SON (x 1.54) and increased the AVP and OXT mRNAcontent (x 2). During rehydration, both SON volume and density of silver grains were higher than normal for at least 7-10 days, although levels started to fall by day 3. The distribution of individual cells according to their silver grain densities remained unimodal during the dehydration-rehydration sequence with an extension, then a return to normal of the distribution range. Maximum sizes of AVP and OXT mRNAs on Northern blots of RNAs extracted from 5 pooled SONs were observed on dehydration day 5. The size of these species fell progressively, reaching control values by rehydration day 14. We conclude that during rehydration, at a time when most of the putative inducers of gene transcription are no longer activated, the peptidergic deficit was accompanied by an increased level of AVP and OXT mRNAs.(ABSTRACT TRUNCATED AT 250 WORDS)

摘要

雄性大鼠被剥夺水分5天,然后随意给水3、7、10或14天。血浆渗透压在不到3天内恢复正常,而垂体抗利尿激素(AVP)和催产素(OXT)仅在14天后恢复到对照水平。视上核(SON)切片与35S标记的cDNA(OXT)或寡核苷酸(AVP)探针杂交。通过计算SON一个大标准区域上的银粒数量来定量相对AVP和OXT mRNA含量,然后将该值外推到整个SON的体积(从所有切片的表面积推导得出)。脱水显著增大了SON的体积(×1.54)并增加了AVP和OXT mRNA含量(×2)。在补液期间,SON体积和银粒密度至少在7 - 10天内高于正常水平,尽管在第3天开始下降。在脱水 - 补液过程中,根据银粒密度的单个细胞分布保持单峰,分布范围先扩展然后恢复正常。在脱水第5天观察到从5个合并的SON中提取的RNA的Northern印迹上AVP和OXT mRNA的最大尺寸。这些种类的尺寸逐渐下降,到补液第14天达到对照值。我们得出结论,在补液期间,当大多数假定的基因转录诱导剂不再被激活时,肽能缺乏伴随着AVP和OXT mRNA水平的增加。(摘要截短至250字)

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