Beaujean Delphine, Do-Rego Jean-Luc, Galas Ludovic, Mensah-Nyagan Ayikoe G, Fredriksson Robert, Larhammar Dan, Fournier Alain, Luu-The Van, Pelletier Georges, Vaudry Hubert
European Institute for Peptide Research (IFRMP 23), Laboratory of Cellular and Molecular Neuroendocrinology, INSERM U-413, UA Centre National de la Recherche Scientifique, University of Rouen, 76821 Mont-Saint-Aignan, France.
Endocrinology. 2002 May;143(5):1950-63. doi: 10.1210/endo.143.5.8765.
We have recently shown that hydroxysteroid sulfotransferase (HST), the enzyme responsible for the biosynthesis of pregnenolone sulfate (Delta(5)PS) and dehydroepiandrosterone sulfate (DHEAS), is expressed in neurons located in the anterior preoptic area and the dorsal magnocellular nucleus of the frog diencephalon. As these two nuclei are richly innervated by NPY-immunoreactive fibers, we investigated the possible implication of NPY in the control of Delta(5)PS and DHEAS biosynthesis. Double labeling of frog brain sections revealed that 42% of the HST-immunoreactive perikarya in the diencephalon were contacted by NPY-containing fibers. In situ hybridization studies showed that Y(1) and Y(5) receptor mRNAs are expressed in the anterior preoptic area and the dorsal magnocellular nucleus. Pulse-chase experiments with (35)S-labeled 3'-phosphoadenosine 5'-phosphosulfate as a sulfate donor demonstrated that frog NPY (fNPY) inhibited the conversion of [(3)H]Delta(5)P and [(3)H]dehydroepiandrosterone ([(3)H]DHEA) into [(3)H,(35)S]Delta(5)PS and [(3)H,(35)S]DHEAS by diencephalic explants. The inhibitory effect of fNPY on Delta(5)PS and DHEAS formation was mimicked by (pPYY) and [Leu(31),Pro(34)]pNPY, which is an agonist for non-Y(2) receptors in mammals, and was completely suppressed by the Y(1) receptor antagonist BIBP3226. Conversely, the Y(2) receptor agonist pNPY-(13-36) and the Y(5) receptor agonist [D-Trp(32)]pNPY did not significantly modify the biosynthesis of [(3)H,(35)S]Delta(5)PS and [(3)H,(35)S]DHEAS. The present study provides the first evidence for the innervation of neurosteroid-producing neurons by NPY fibers. Our data also demonstrate that NPY, acting via Y(1) receptors, exerts an inhibitory effect on the biosynthesis of sulfated neurosteroids.
我们最近发现，羟类固醇硫酸转移酶（HST），即负责孕烯醇酮硫酸盐（Δ⁵PS）和硫酸脱氢表雄酮（DHEAS）生物合成的酶，在位于青蛙间脑视前区前部和背侧大细胞核的神经元中表达。由于这两个核被神经肽Y（NPY）免疫反应性纤维大量支配，我们研究了NPY在控制Δ⁵PS和DHEAS生物合成中的可能作用。青蛙脑切片的双重标记显示，间脑中42%的HST免疫反应性核周体与含NPY的纤维接触。原位杂交研究表明，Y(1)和Y(5)受体mRNA在视前区前部和背侧大细胞核中表达。用³⁵S标记的3'-磷酸腺苷5'-磷酸硫酸盐作为硫酸盐供体进行脉冲追踪实验表明，青蛙NPY（fNPY）抑制间脑外植体将[³H]Δ⁵P和[³H]脱氢表雄酮（[³H]DHEA）转化为[³H,³⁵S]Δ⁵PS和[³H,³⁵S]DHEAS。fNPY对Δ⁵PS和DHEAS形成的抑制作用被（pPYY）和[Leu³¹,Pro³⁴]pNPY模拟，后者是哺乳动物中非Y(2)受体的激动剂，并且被Y(1)受体拮抗剂BIBP3226完全抑制。相反，Y(2)受体激动剂pNPY-(13 - 36)和Y(5)受体激动剂[D-Trp³²]pNPY并没有显著改变[³H,³⁵S]Δ⁵PS和[³H,³⁵S]DHEAS的生物合成。本研究为NPY纤维对神经甾体生成神经元的支配提供了首个证据。我们的数据还表明，NPY通过Y(1)受体发挥作用，对硫酸化神经甾体的生物合成产生抑制作用。
