GRF-induced feeding: Evidence for protein selectivity and opiate involvement

Growth hormone-releasing factor (GRF) is a hypothalamic peptide named for its ability to induce release of growth hormone from the anterior pituitary. GRF also acts as a neurotransmitter in the suprachiasmatic nucleus/medial preoptic area (SCN/MPOA) to stimulate food intake. The purpose of this series of experiments was to explore the nature of GRF-induced feeding, with a particular emphasis on macronutrient selectivity, and to examine the role of opiate activity in the paraventricular nucleus of the hypothalamus (PVN). Chow intake stimulated by GRF microinjection (1 pmol/0.5 μl) into the SCN/MPOA was blocked by injection of methyl-naltrexone (3 μg/0.5 μl) into the PVN. In animals habituated to macronutrient diets (Teklad, WI), GRF preferentially stimulated intake of protein at 2 and 4 h postinjection, whereas it had no effect on carbohydrate intake. Further, this effect was blocked by injection of naloxone (40 nmol/0.5 μl) into the PVN. Microinjection of morphine (0, 1, 10, and 17 μg/0.5 μl) into the PVN also specifically stimulated protein intake at 2 and 4 h postinjection. These results suggest that feeding derived from GRF actions in the SCN/MPOA is macronutrient selective, and is dependent on PVN opiate activity for expression.     

Morphological correlates of serotonin-neuropeptide Y interactions in the rat suprachiasmatic nucleus: Combined radioautographic and immunocytochemical data

Summary The morphological substrate of putative serotonin (5-HT)/neuropeptide Y (NPY) interactions in thé suprachiasmatic nucleus (SCN) was investigated by combined radioautography and immunocytochemistry after intraventricular administration of (³H)5-HT in the rat. In the ventral portion of the SCN, the distribution of (³H)5-HT uptake sites overlapped closely the NPY-immunoreactive terminals. Previous investigations have shown that the dense 5-HT and NPY innervations of the SCN originate in different structures, i.e., the midbrain raphe nuclei and the ventral lateral geniculate nucleus, respectively. Accordingly, in the present study, destruction of 5-HT afferents by 5,7-dihydroxytryptamine was not found to induce any modification in NPY staining and, in ultrastructural immuno-radioautographic preparations, two distinct pools of axonal varicosities could be identified. Both 5-HT and NPY terminals established morphologically defined synaptic junctions, sometimes on the same neuronal target. Some cases of direct axo-axonic appositions between the two types of terminals were also encountered. These data constitute additional criteria for characterizing the cytological basis of the multiple transmitter interactions presumably involved in the function of the SCN as a central regulator of circadian biological rhythms.     

Catecholaminergic innervation of GRF-containing neurons in the rat hypothalamus revealed by electron-microscopic cytochemistry

Summary The Catecholaminergic innervation of neurons containing growth hormone-releasing factor (GRF) was examined by use of a method which combined either 5-hydroxydopamine (5-OHDA) uptake or autoradiography after intraventricular injection of ³H-noradrenaline with immunocytochemistry for GRF in the same tissue sections at the electron-microscopic level. In the ventrolateral part of the arcuate nucleus of the rat hypothalamus a large number of immunonegative axon terminals were found to make synaptic contact with GRF-like immunoreactive (GRF-LI) cell bodies and processes. ³H-noradrenaline autoradiography or 5-OHDA-labeling combined with GRF immunocytochemistry revealed that axon terminals labeled with ³H-noradrenaline or 5-OHDA make synaptic contact with the GRF-LI nerve cell bodies and processes. These findings indicate that catecholamine-containing neurons innervate GRF neurons to regulate GRF secretion via synapses in the rat arcuate nucleus.     

视前区微量注射吗啡对大鼠丘脑束旁核痛反应神经元电活动的影响

本实验观察了视前区(POA)内微量注入阿片样物质对丘脑束旁核(Pf)痛反应神经元电活动影响。结果如下:(1)POA 内微量注射高浓度吗啡(10μg/μl)能显著抑制 Pf 内大部分(20/26)痛兴奋神经元(PEN)的痛诱发放电,其中3个神经元注药后对伤害性刺激转变成抑制反应;POA 内微量注射低浓度吗啡(1μg/μl)也显著抑制 Pf 内大部分(19/23)PEN 的痛诱发放电。(2) POA 内微量注射两种浓度的吗啡,均使大多数痛抑制神经元(PIN,共27/33)的完全抑制时程缩短。上述结果提示,POA 内阿片样物质对 Pf 内痛反应神经元的电活动可能具有抑制作用。     

刺激大鼠下丘脑室旁核激活的中脑中央灰质神经元对躯体传入的反应

电刺激大鼠下丘脑室旁核(PVH),在同侧中脑中央灰质(CG)内寻找逆行及顺行反应单位,然后观察它们对躯体感觉刺激的反应。实验结果表明:CG 及邻近网状结构内有10%(32/318)的单位呈逆行反应。逆行传导速度平均为0.37±0.24m/s(均数±标准差);推测这种CG→PVH 投射纤维属于细有髓或无髓神经纤维。这些单位分布于 CG 的腹外侧及背外侧亚核。50%(14/28)的逆行单位对坐骨(胫)神经的强电刺激和夹尾等损伤性刺激起反应,但对触毛或低强度的神经干刺激无明显反应。以上结果表明:外周躯体感觉,特别是损伤性信息传入 PVH 时,CG 是其中枢驿站之一。电刺激 PVH 还能顺行激活7.55(24/318)、抑制0.7%(2/318)的 CG 单位。有69%(18/26)的顺行反应单位对外周躯体神经强电刺激及夹尾起反应。提示 PVH 可能通过影响 CG神经元的活动而参与中枢痛觉的整合。     

Freeze-substitution improves the ultrastructural preservation of legume root hairs

The freeze-substitution method was applied toVicia hirsuta root hairs to test its effectiveness in improving preservation of the cell ultrastruture. Freeze-substitution almost certainly represents more faithfully the structure of the hair cell. A previously unreported ‘pyriformis’ vesicle is described. Also unique to freeze-substituted material are coated secretory vesicles; a smooth plasma membrane profile; mitochondrial ribosomes; long microfilament bundles which are associated with vesicles, mitochondria, coated pits and coated vesicles; microtubule-associated filaments; well-preserved coated vesicles and coated pits with enclosing rings; a pliciform nucleus. The results are discussed in context of previous reports using conventional fixation techniques.     

In situ hybridization of corticotropin-releasing factor-encoding messenger RNA in the hypothalamus of the white sucker,Catostomus commersoni

Summary In situ hybridization procedure with a ³²P-labelled synthetic oligonucleotide probe was used to detect corticotropin-releasing factor-encoding messenger RNA (CRF mRNA) in the hypothalamus of the white sucker, Catostomus commersoni. Adjacent sections were immunostained by a sucker CRF-specific antiserum. CRF mRNA-containing neurons were identified by autoradiography in the magnocellular and parvocellular subdivisions of the preoptic nucleus (PON). Many of these neurons were also immunostained by sucker antiserum, showing the same distribution patterns. These results confirm the presence of CRF mRNA and CRF peptide in the white sucker hypothalamus and support the view that the magnocellular and parvocellular neurons of the PON may be involved in the control of adrenocorticotropic hormone secretion from the pituitary in the white sucker.     

外侧臂旁核神经元对穹隆下器逆向刺激和孤束核顺向刺激的反应

细胞外记录大鼠外侧臂旁核(LPBN)神经元单位放电,观察了刺激穹隆下器(SFO)在记录单位诱发的逆向反应和静脉注射新福林兴奋外周压力感受器和刺激孤束核(NTS)减压区诱发的顺向反应。实验发现:刺激 SFO,9.9%(15/151)的 LPBN 神经元有逆向反应。静脉注射新福林,40.7%(22/54)的 LPBN 神经元有抑制反应,27.8%(17/54)有兴奋反应。刺激 NTS,55.6%(94/169)的 LPBN 神经元呈现顺向兴奋反应;22.5%(38/169)呈现顺向抑制反应。观察静脉注射新福林对 SFO 刺激有逆向反应的 LPBN 神经元自发放电的影响,在两个受试单位均见抑制反应。观察刺激 NTS 对逆向反应单位自发放电的影响,在8个受试单位中,6个呈兴奋反应;2个呈抑制反应。以上结果表明:LPBN 接受来自 NTS 的兴奋性或抑制性压力感受性传入,并把这种信息经 LPBN-SFO 直接通路传输到 SFO。