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Effects of lifelong moderate caloric restriction on levels of neuropeptide Y, proopiomelanocortin, and galanin mRNA

JOURNALS OF GERONTOLOGY SERIES A-BIOLOGICAL SCIENCES AND MEDICAL SCIENCES 1999年第1期54卷 B14-B21页

作者： McShane, TM Wilson, ME Wise, PM Univ Kentucky Coll Med Dept Physiol Lexington KY 40536 USA

We are interested in how neuropeptides that regulate both food intake and reproductive function change with age and how life-prolonging moderate caloric restriction may influence the expression of these neuropeptides. We measured neuropeptide Y (NPY) and proopiomelanocortin (POMC) mRNA in the arcuate nucleus (AN) and galanin (GAL) mRNA in the AN, paraventricular nucleus (PVN) and medial septum-diagonal band of Broca in young, middle-aged and old female rats on a controlled feeding regimen. Female Sprague-Dawley rats (7 weeks old) were placed on caloric restriction (CR;n = 70) which was 60% of ad libitum (AL) intake measured in control rats (n = 70). Rats were rapidly decapitated 2.5 weeks following ovariectomy, when they were 4, 12, or 18 months old. Brains were frozen and coronal sections (12 microns) were cut at -20 degrees C using a cryostat. Relative levels of NPY, POMC and GAL mRNA were measured using in situ hybridization histochemistry, cDNA clones complementary to rat NPY, mouse POMC and rat GAL were used to synthesize S-35-UTP-labeled cRNA probes. Slides were dipped in photographic emulsion. Silver grains were quantified using computer-assisted image analysis. Caloric restriction increased NPY mRNA/cell, decreased POMC mRNA/cell in the AN, and did not influence GAL mRNA/cell. Age and caloric restriction did not influence the number of cells with detectable NPY mRNA, POMC mRNA or GAL mRNA. Reproductive status at the time of decapitation influenced both the level and the number of cells expressing GAL mRNA in the PVN. In summary, CR differentially affects levels of NPY, POMC and GAL mRNA in the AN. Zn this animal model, there was no statistically significant effect of age nor any interaction between age and diet on expression of any of these genes between 4 and 18 months of age.

关键词：衰老/遗传学衰老/代谢方差分析弓状核/细胞学弓状核/代谢细胞计数着色剂/诊断应用 DNA 互补/诊断应用疾病模型动物能量摄取额叶/细胞学额叶/代谢加兰肽/分析加兰肽/遗传学基因表达调控组织细胞化学图像处理计算机辅助原位杂交神经肽Y/分析神经肽Y/遗传学卵巢切除术下丘脑室旁核/细胞学下丘脑室旁核/代谢前阿片黑素细胞皮质激素/分析前阿片黑素细胞皮质激素/遗传学 RNA 互补/诊断应用 RNA 信使/分析 RNA 信使/遗传学大鼠 Sprague-Dawley 银/诊断应用动物女(雌)性小鼠大鼠

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Somatostatin modulation of excitatory synaptic transmission between periventricular and arcuate hypothalamic nuclei in vitro

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JOURNAL OF NEUROPHYSIOLOGY 2000年第3期84卷 1464-1474页

作者： Lanneau, C Peineau, S Petit, F Epelbaum, J Gardette, R INSERM U159 Ctr Paul Broca F-75014 Paris France

Hypophysiotropic somatostatin (SRIF) and growth hormone-releasing hormone (GHRH) neurons are primarily involved in the neurohormonal control of growth hormone (GH) secretion. They are located in periventricular (PEV) and arcuate (ARC) hypothalamic nuclei, respectively, but their connectivity is not well defined. To better understand the neuronal network involved in the control of GH secretion, connections from PEV to ARC neurons were reconstructed in vitro and neuronal phenotypes assessed by single-cell multiplex RT-PCR. Of 814 stimulated PEV neurons, monosynaptic responses were detected in only 45 ARC neurons. Monosynaptic excitatory currents were detected in 29 ARC neurons and inhibitory currents in 16, indicating a 2/1 ratio for excitatory versus inhibitory connections. Galanin (GAL), NPY, pro-opiomelanocortin (POMC), and SRIF mRNAs were detected in neurons from both nuclei but GHRH mRNA almost exclusively in ARC. Among the five SRIF receptors, only sst1 and sst2 were expressed, in 94% of ARC and 59% of PEV neurons, respectively. Of 128 theoritical combinations between neuropeptides and sst receptors, only 22 were represented in PEV and 25 in ARC. For PEV neurons, neuropeptide phenotypes did not influence excitatory connections. However, the occurrence of presynaptic sst receptors on GAL and SRIF PEV neurons significantly increased their probability of connection to ARC neurons. GHRH ARC neurons expressing sst2, but not sst1, receptors were always connected with PEV neurons. Physiological responses to sst1 (CH-275) or sst2 (Octreotide) agonists were always correlated with the detection of respective sst mRNAs. In conclusion, 1) SRIF-modulated excitatory transmission develops in vitro from PEV to ARC neurons, 2) ARC GHRH neurons bearing sst2 receptors appears directly controlled by fast glutamatergic transmission from PEV neurons simultaneously expressing one to four neuropeptides, 3) GHRH neurons bearing sst1 receptors lack this control, and 4) these results sugg

关键词：细胞分离共同培养技术电刺激兴奋性突触后电位/生理学加兰肽/分析加兰肽/遗传学生长激素/生物合成生长激素释放激素/分析生长激素释放激素/遗传学神经网/代谢神经元/化学神经元/细胞学神经元/代谢神经肽Y/分析神经肽Y/遗传学下丘脑室旁核/细胞学下丘脑室旁核/胚胎学下丘脑室旁核/代谢表型前阿片黑素细胞皮质激素/分析前阿片黑素细胞皮质激素/遗传学 RNA 信使/分析受体生长抑素/激动剂受体生长抑素/分析受体生长抑素/遗传学逆转录聚合酶链反应/方法生长抑素/分析生长抑素/遗传学生长抑素/代谢突触传递/生理学动物小鼠

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