mRNAs encoding muscarinic and substance P receptors in cultured sympathetic neurons are differentially regulated by LIF or CNTF.
mRNAs encoding muscarinic and substance P receptors in cultured sympathetic neurons are differentially regulated by LIF or CNTF.
复制标题
培养的交感神经元中编码毒蕈碱受体和 P 物质受体的 mRNA 受到 LIF 或 CNTF 的差异调节。
DOI：
10.1006/dbio.1994.1221
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发表时间：
1994
影响因子：
2.7
通讯作者：
Kessler,JA
中科院分区：
文献类型：
--
作者：
Ludlam,WH;Zang,Z;McCarson,KE;Krause,JE;Spray,DC;Kessler,JA
白血病抑制因子（LIF）和睫状神经营养因子（CNTF）先前已被证明可以调节神经元对神经递质的选择。在本研究中，这些因素被证明是特异性和差异调节水平的毒蕈碱（亚型m1，m2，m3，m4和m5）和P物质受体（SPR）的大鼠上级颈神经节（SCG）的交感神经元的mRNA使用溶液杂交/RNA酶保护分析。在体内，新生大鼠SCG主要表达m2（10.31 ± 0.43 pg mRNA/μg总RNA）和一些m1（1.54 ± 0.84 pg/μg）M受体mRNA，随着发育，m2 mRNA水平逐渐升高至成年水平（m2 mRNA水平比新生大鼠高60%）。相比之下，m3、m4和m5亚型mRNA在测量的所有时间点的丰度要低得多。在离体培养的SCG神经元中也发现了类似的发育规律。培养16天后，m2 mRNA增加334%，为15.76 ± 0.68 pg/μg，而m1 mRNA变化不大，为2.03 ± 1.00 pg/μg。然而，LIF或CNTF治疗（5 ng/ml，14天）在姐妹文化完全阻断了这种发展的增长。此外，LIF治疗阻断了正常的毒蕈碱受体介导的细胞内钙的增加（Fura-2成像），表明受体表型的功能变化。相比之下，未处理的培养物中SPR mRNA水平较低（0.037 ± 0.025 pg SPR mRNA/μg总RNA），经LIF或CNTF处理后，SPR mRNA水平分别升高至0.866 ± 0.034 pg/μg和0.662 ± 0.148 pg/μg。这些观察结果表明，毒蕈碱和SPR受体的表达差异调节相同的因素在SCG神经元和受体表型的神经元的选择，至少部分，在cellularmilieu细胞因子/生长因子的特异性调节。
Leukemia inhibitory factor (LIF) and ciliary neurotrophic factor (CNTF) have previously been shown to regulate neuronal choice of neurotransmitter. In this present study, these factors were shown to specifically and differentially regulate levels of both muscarinic (subtypes m1, m2, m3, m4, and m5) and substance P receptor (SPR) mRNAs in sympathetic neurons of the rat superior cervical ganglion (SCG) using solution hybridization/RNase protection analysis. In vivo, neonatal rat SCG expressed predominantly m2 (10.31 ± 0.43 pg mRNA/μg total RNA) and some ml (1.54 ± 0.84 pg/μg) muscarinic receptor mRNA, which increased developmentally to adult levels (m2 mRNA levels being 60% higher than those in neonates). By contrast, m3, m4, and m5 subtype mRNAs were much less abundant at all time points measured. A similar developmental regulation was found in dissociated SCG neurons in vitro. After 16 days in culture, m2 mRNA increased 334% to 15.76 ± 0.68 pg/μg, while m1 mRNA changed little (2.03 ± 1.00 pg/μg). However, LIF or CNTF treatment (5 ng/ml, 14 days) in sister cultures completely blocked this developmental increase. Further, LIF treatment blocked the normal muscarinic receptor-mediated increase in intracellular calcium (fura-2 imaging), indicating a functional change in receptor phenotype. By contrast, levels of SPR mRNA, which were low in untreated cultures (0.037 ± 0.025 pg SPR mRNA/μg total RNA), were elevated by LIF or CNTF treatment, to 0.866 ± 0.034 pg/μg and 0.662 ± 0.148 pg/μg, respectively. These observations indicate that muscarinic and SPR receptor expression are differentially regulated by the same factors in SCG neurons and that neuronal choice of receptor phenotype may be, at least in part, specifically regulated by cytokines/growth factors in the cellularmilieu.