Rondini Tatiane Araujo, Rodrigues Bruno de Crudis, de Oliveira Anne Pierre, Bittencourt Jackson Cioni, Elias Carol Fuzeti
Laboratory of Chemical Neuroanatomy, Department of Anatomy, Institute of Biomedical Sciences, University of São Paulo, 05508-900 São Paulo, SP, Brazil.
Brain Res Bull. 2007 Sep 14;74(1-3):21-8. doi: 10.1016/j.brainresbull.2007.04.006. Epub 2007 May 21.
Melanin-concentrating hormone (MCH) is a neuropeptide originating from prepro-MCH. In male rats, neurons expressing MCH are found in the lateral hypothalamic area and medial zona incerta, as well as, sparsely, in the olfactory tubercle and pontine reticular formation. The wide distribution of MCH fibers suggests the involvement of this neuropeptide in a variety of functions, including arousal, neuroendocrine control and energy homeostasis. In lactating females, MCH is expressed in the preoptic area, indicating sexual dimorphism in MCH gene activation according to the female reproductive state. We hypothesized that MCH is also expressed differentially in the brainstem of female rats. Adult male rats and female rats (in the afternoon of diestrus and proestrus days; ovariectomized; or on lactation days 5, 12 and 19) were perfused between 2 and 4 p.m., and the brainstems were processed for in situ hybridization using a 35S-labeled prepro-MCH riboprobe. As described in males, prepro-MCH was expressed in the pontine reticular formation of females. We also observed consistent prepro-MCH expression in the caudal laterodorsal tegmental nucleus (LDT) of females but no differential expression comparing the various female reproductive states. Using dual-label immunohistochemistry or dual-label in situ hybridization, we found that brainstem MCH neurons coexpress glutamic acid decarboxylase mRNA, the gamma aminobutyric acid (GABA) processing enzyme, but do not colocalize choline acetyl transferase (acetylcholine processing enzyme). Since changes in LDT GABAergic cell activity are associated with rapid eye movement (REM) sleep, our findings suggest that MCH interacts with LDT GABAergic neurons and plays a role in REM sleep regulation.
黑色素聚集激素(MCH)是一种源自前体MCH的神经肽。在雄性大鼠中,表达MCH的神经元见于下丘脑外侧区和内侧未定带,在嗅结节和脑桥网状结构中也有少量分布。MCH纤维的广泛分布表明这种神经肽参与多种功能,包括觉醒、神经内分泌控制和能量稳态。在哺乳期雌性大鼠中,MCH在前视区表达,这表明根据雌性生殖状态,MCH基因激活存在性别差异。我们推测MCH在雌性大鼠脑干中也有差异表达。成年雄性大鼠和雌性大鼠(处于动情间期和动情前期下午;已切除卵巢;或处于哺乳期第5、12和19天)于下午2点至4点进行灌注,脑干经处理后用35S标记的前体MCH核糖探针进行原位杂交。如在雄性大鼠中所述,前体MCH在雌性大鼠的脑桥网状结构中表达。我们还观察到雌性大鼠尾侧背外侧被盖核(LDT)中前体MCH表达一致,但在比较不同雌性生殖状态时未发现差异表达。使用双标记免疫组织化学或双标记原位杂交,我们发现脑干MCH神经元共表达谷氨酸脱羧酶mRNA(γ-氨基丁酸(GABA)加工酶),但不与胆碱乙酰转移酶(乙酰胆碱加工酶)共定位。由于LDT GABA能细胞活性的变化与快速眼动(REM)睡眠相关,我们的研究结果表明MCH与LDT GABA能神经元相互作用并在REM睡眠调节中发挥作用。