On the anatomical organization of the vagal nuclei

The neurons of origin of the right vagus and its components in both the monkey (Macaca fascicularis) and albino rats were localized by the retrograde transport of horseradish peroxidase (HRP) applied to the stomach wall, the vagal trunk and its recurrent laryngeal branch. An attempt was also made to localize the neurons forming the superior laryngeal nerve and those supplying the thoracic organs by a combination of operative procedures. The results showed that the stomach was innervated by neurons distributed throughout the entire rostrocaudal extent of the dorsal motor nucleus (DMN) on both sides of the brain stem. Neurons scattered throughout the entire extent of the DMN and nucleus ambiguus (NA) supplied the thoracic viscera. There did not appear to be any topographic arrangement in the DMN neurons supplying the abdominal and thoracic viscera as reported by other workers, and there was no clear evidence of crossing of vagal fibers in the monkey brain stem, though such crossing was seen in the rat brain stem. Both the superior and inferior ganglia of the vagus nerve were labeled following application of HRP to the vagal trunk. Neurons in the caudal part of the NA gave rise to fibers in the ipsilateral recurrent laryngeal nerve, at least on the right side. The neurons giving rise to the superior laryngeal nerve could not be delineated in this study. In all the experimental procedures described, the hypoglossal nucleus was labeled only after applying HRP to the hypoglossal nerve.     

电刺激大鼠束旁核对底丘脑核和丘脑腹内侧核神经元的影响

本工作旨在探讨电刺激束旁核（parafascicular nucleus,PF）对帕金森病模型（Parkinson’s disease,PD）大鼠神经行为的改善作用及其机制。成年雄性Sprague—Dawley大鼠黑质致密部注射6一羟基多巴胺建立PD大鼠模型。采用行为学方法观察电刺激PF对阿朴吗啡诱发的大鼠旋转行为的作用,并应用在体细胞外记录法观察电刺激PF对大鼠底丘脑核（subthalamic nucleus,STN）及丘脑腹内侧核（ventromedial nucleus,VM）神经元放电的影响。结果发现,高频电刺激（130Hz,0．4mA,5s）PF一周,明显改善PD大鼠旋转行为。细胞外放电记录显示,高频电刺激PF使PD大鼠STN神经元自发放电减少,且该作用具有频率依赖性。另外,高频电刺激PF可使VM神经元兴奋,该作用也是频率依赖性的。我们在实验中同时观察到微电泳谷氨酸（glutamicacid,Glu）受体拮抗剂MK-801使STN神经元放电频率减少或完全抑制,微电泳t氨基丁酸（T-amino butyricacid,GABA）受体拮抗剂印防己毒素（picrotoxin,Pic）则使神经元放电频率增加。以上结果表明,GABA能和GIu能传入纤维可会聚于同-STN神经元,并对后者有紧张性作用。高频刺激PF,使该核团到STN神经元的Glu能兴奋性输出减少,导致STN的失活。这一作用通过基底神经节的间接通路,最终释放了丘脑运动核团VM的活性。高频刺激PF经PF,STN和VM的神经通路而改善PD大鼠神经行为。     

Development of the human hypothalamus

The hypothalamus has been claimed to be involved in a great number of physiological functions in development, such as sexual differentiation (gender, sexual orientation) and birth, as well as in various developmental disorders including mental retardation, sudden infant death syndrome (SIDS), Kallman's syndrome and Prader-Willi syndrome. In this review a number of hypothalamic nuclei have therefore been discussed with respect to their development in health and disease. The suprachiasmatic nucleus (SCN) is the clock of the brain and shows circadian, and seasonal fluctuations in vasopressin-expressing cell numbers. The SCN also seems to be involved in reproduction, adding interest to the sex differences in shape of the vasopressin-containing SCN subnucleus and in its VIP cell number. In addition, differences in relation to sexual orientation can be seen in this perspective. The vasopressin and VIP, neurons of the SCN develop mainly postnatally, but as premature children may have circadian temperature rhythms, a different SCN cell type is probably more mature at birth.Thesexually dimorphic nucleus (SDN, intermediate nucleus, INAH-1), is twice as large in young male adults as in young females. At the moment of birth only 20% of the SDN cell number is present. From birth until two to four years of age cell numbers increase equally rapidly in both sexes. After this age cell numbers start to decrease in girls, creating the sex difference. The size of the SDN does not show any relationship to sexual orientation in men. The large neurosecretory cells of thesupraoptic (SON) andparaventricular nucleus (PVN) project to the neurohypophysis, where they release vasopressin and oxytocin into the blood circulation. In the fetus these hormones play an active role in the birth process. Fetal oxytocin may initiate or accelerate the course of labor. Fetal vasopressin plays a role in the adaptation to stress—caused by the birth process—by redistribution of the fetal blood flow.Corticotropin-releasing hormone (CRH) neurons of the PVN play a central role in stress response. Thus fetal CRH neurons may play a role in the timing of the moment of birth. Recently, alterations have been described in peptidergic, aminergic and cholinergic transmitters in the hypothalamus in SIDS. Future research will have to establish whether these changes are part of the course of SIDS. A large proportion of the SON and PVN neurons also produce tyrosine hydroxylase (TH). In neonates the majority of TH-immunoreactive neurons colocalizes vasopressin, while in the adult the majority of TH-positive neurons colocalizes oxytocin. TH-expression might be a sign of hyperactivation, for example from perinatal hypoxia.Oxytocin neurons also project to the brain stem. These neurons have an inhibitory effect on eating. Interestingly, in the Prader-Willi syndrome, characterized for example by insatiable hunger, we have found that the number of oxytocin-expressing neurons is about half the normal value. It can be concluded that the various hypothalamic nuclei are involved in a great number of functions and show clear and differential changes in development with respect to sexual differentiation, birth and a number of diseases. I believe that only a small proportion of such changes has at present been revealed.Special issue dedicated to Dr. Robert Balázs     

Changes in oxytocin content in the magnocellular neurons of the rat hypothalamus following water deprivation or estrogen treatment

Summary The effect of water deprivation or estrogen treatment on the oxytocin content of rat hypothalamic cells was examined using a quantitative immunohistological technique. Oxytocin-containing cells were visualized using the immunoperoxidase technique of Sternberger and a primary antiserum directed against oxytocin. The optical density of the darkest 3.2 m diameter spot in the cytoplasm of a cell was used as a measure of the oxytocin content of that cell. Water deprivation produced a significant decrease in anti-oxytocin staining in the anterior commissural nucleus of males and females. There was a similar decrease in the paraventricular nucleus of males, but not in the paraventricular nucleus of females or the supraoptic nucleus of either males or females. Estrogen treatment of ovariectomized female rats produced a fall in anti-oxytocin staining in the anterior commissural, but not paraventricular or supraoptic nuclei.     

A CreER mouse to study melanin concentrating hormone signaling in the developing brain

The neuropeptide, melanin concentrating hormone (MCH), and its G protein‐coupled receptor, melanin concentrating hormone receptor 1 (Mchr1), are expressed centrally in adult rodents. MCH signaling has been implicated in diverse behaviors such as feeding, sleep, anxiety, as well as addiction and reward. While a model utilizing the Mchr1 promoter to drive constitutive expression of Cre recombinase (Mchr1‐Cre) exists, there is a need for an inducible Mchr1‐Cre to determine the roles for this signaling pathway in neural development and adult neuronal function. Here, we generated a BAC transgenic mouse where the Mchr1 promotor drives expression of tamoxifen inducible CreER recombinase. Many aspects of the Mchr1‐Cre expression pattern are recapitulated by the Mchr1‐CreER model, though there are also notable differences. Most strikingly, compared to the constitutive model, the new Mchr1‐CreER model shows strong expression in adult animals in hypothalamic brain regions involved in feeding behavior but diminished expression in regions involved in reward, such as the nucleus accumbens. The inducible Mchr1‐CreER allele will help reveal the potential for Mchr1 signaling to impact neural development and subsequent behavioral phenotypes, as well as contribute to the understanding of the MCH signaling pathway in terminally differentiated adult neurons and the diverse behaviors that it influences.     

Biorhythm of arginine-vasopressin in the paraventricular, supraoptic and suprachiasmatic nuclei of rats

The diurnal variations in content of arginine-vasopressin in the supraoptic nucleus, the paraventricular nucleus and the suprachiasmatic nucleus of rats were determined using radioimmunoassay. In the supraoptic nucleus and the paraventricular nucleus the arginine-vasopressin level was relatively constant during the light phase (the inactive phase). When it became dark, the level of arginine-vasopressin lowered during the early and middle dark phase and then increased to the highest level during the late dark phase. In the suprachiasmatic nucleus the level was stable during the light phase, while in the early and the late dark phase it was significantly higher than that in the middle dark phase.     

6-OHDA毁损黑质致密部对大鼠PPN和VL神经元放电活动的影响

目的：观察6-羟多巴胺单侧毁损黑质致密部多巴胺神经元后,脚桥核（PPN）和丘脑腹外侧核（VL）神经元自发放电活动的变化,探讨帕金森病（PD）的发病机制。方法：应用玻璃微电极细胞外记录法,观察对照组和PD组PPN和VL神经元的放电频率和放电形式的变化。结果：对照组和PD组大鼠PPN放电频率分别为（8．31±0．62）Hz和（10．70±0．85）Hz,PD组放电频率明显高于对照组（P〈0．05）。和对照组相比,PD组PPN的不规则和爆发式放电神经元构成比例明显增多（P〈0．01）,同时规则放电频率增加（P〈0．01）。对照组和PD组大鼠VL的放电频率分别为（6．25±0．54）Hz和（5．67±0．46）Hz,两组间没有显著性差异。VL神经元放电形式表现为不规则和爆发式放电,两组间构成比也没有明显差异,但PD组爆发式神经元放电频率明显降低（P〈0．01）。结论：PD状态下,PPN神经元活动增强,PPN可能参与了PD的病理生理过程,VL神经元放电可能受PPN神经元投射的调节。     

电刺激大鼠脚内核对脚桥核神经元放电的影响

目的:观察电刺激大鼠脚内核(EP)对大鼠脚桥核(PPN)神经元自发放电的影响,进一步探讨脑内电刺激治疗帕金森病(PD)的机制。方法:应用细胞外记录的方法观察不同频率电刺激(强度0.6 mA,波宽0.06 ms,时程5 s,频率5 Hz、10Hz、20Hz、50Hz、100Hz、150Hz、200Hz)大鼠EP对PPN神经元放电的影响。结果:实验记录了大鼠33个神经元的自发放电,其放电频率在3.6~52.2Hz之间,平均为(15.95±8.56)Hz;当刺激频率为100Hz时,抑制效应最显著(P<0.05)。结论:高频电刺激大鼠EP对PPN神经元自发放电的影响主要为抑制作用,提示高频刺激EP可通过抑制PPN神经元活动参与PD的治疗。     

Immunocytochemical study of the hypothalamo-neurohypophysial system

Summary An antiserum cross-reactive against ovine neurophysins-I-II and -III has been used in conjunction with the immunoperoxidase histochemical procedure to localize the cells of the sheep paraventricular (PVN) and supraoptic nuclei (SON). In order to describe the topographical distribution of the SON and PVN a study was made on the serial sections cut (a) transversely from rostral to caudal positions and (b) sagittally from lateral to medial positions of the hypothalamus.The cells of the SON, when examined in the transverse aspect, extended approximately 1900 caudally and when examined in the sagittal plane were contained within a lateral-medial distance of 4830 . In each case the SON cells lay adjacent to the optic chiasm.As sections were cut transversely, the cells of the PVN first appeared in a rostral position defined as 0 and close to the ventral lining of the third ventricle. This general ventral and ventro-lateral distribution of cells maintained up to a caudal distance of approximately 840 . From positions 1260–2310 there was a dramatic dorsal shift of the PVN cells which by this time had also extended laterally. The total rostral-caudal distance occupied by the PVN cells was 3150 . That the lateral-medial distance occupied by the PVN was small (1050 ) was determined on examining the magnocellular nuclei in sagittal section.This work was financed by a grant awarded by the Medical Research Council of New Zealand     

Distribution of glial-associated proteins in the developing chick auditory brainstem

In the avian brainstem, nucleus magnocellularis (NM) projects bilaterally to nucleus laminaris (NL) in a pathway that facilitates sound localization. The distribution of glia during the development of this pathway has not previously been characterized. Radial glia, astrocytes, and oligodendrocytes facilitate many processes including axon pathfinding, synaptic development, and maturation. Here we determined the spatiotemporal expression patterns of glial cell types in embryonic development of the chick auditory brainstem using glial-specific antibodies and histological markers. We found that vimentin-positive processes are intercalated throughout the NL cell layer. Astrocytes are found in two domains: one in the ventral neuropil region and the other dorsolateral to NM. GFAP-positive processes are primarily distributed along the ventral margin of NL. Astrocytic processes penetrate the NL cell layer following the onset of synaptogenesis, but before pruning and maturation. The dynamic, nonoverlapping expression patterns of GFAP and vimentin suggest that distinct glial populations are found in dorsal versus ventral regions of NL. Myelination occurs after axons have reached their targets. FluoroMyelin and myelin basic protein (MBP) gradually increase along the mediolateral axis of NL starting at E10. Multiple GFAP-positive processes are directly apposed to NM-NL axons and MBP, which suggests a role in early myelinogenesis. Our results show considerable changes in glial development after initial NM-NL connections are made, suggesting that glia may facilitate maturation of the auditory circuit.     

Cytochemical duality of neurosecretory material in the hypothalamo-posthypophysial system of the rat as related to hormonal content

Summary Cytochemical methods using silver proteinate, silver methenamine and potassium ferrocyanide + OsO₄ for ultrastructural detection of glycoproteins allow, in the posthypophysis and the magnocellular nuclei of the rat, differentiation of two types of fibres and neurons: one type containing negative granules with a homogeneous content of low electron density, the second type containing granules which demonstrate a ring-shaped deposit either of silver or of potassium ferrocyanide-osmium complex, likely to be related to a glycoproteic component. The difference between these two types is increased by prestaining en bloc with uranyl acetate before the silver proteinate reaction. A similar investigation was carried out on the vasopressin deficient Brattleboro rat; the neurosecretory material, present in some endings and neurones only, is of the nonreactive type, so that it appears justified to correlate the reactivity of granules with vasopressin, and consequently to distinguish neurones and fibres containing vasopressin from those in which oxytocin is quantitatively the main hormonal peptide. This conclusion is strongly supported by the fact that percentages of reactive and negative endings, as determined on this basis in the posthypophysis of normal rats from two different strains, are in good agreement with biochemical data reported in the literature.     

细胞核起源研究的意义和研究途径的探讨

细胞核的起源是真核细胞进化形成的关键。回顾了过去几十年国内外对细胞核起源问题的探索历程,通过多年的摸索找到了一个条切实可行的探索细胞核起源问题的途径。其要点：在一系列的进化环节中首先抓住原始性的细胞核这一重要环节,探明原始性细胞核的特性,解决了从原始核到典型细胞核的进化问题,原始性细胞核自身的起源问题也就有了基础,为探源始性细胞核的特性,需要在现存的原生生物中间寻找最原始的类群,然后对它们的细胞核进行尽可能深入地和多方面地研究,对所得结果作进化地分析,以期提出一个原始性细胞核的模型,依据这个模型也就可对典型的细胞核的进化形成和原始核自身的起源作出推论,而这个原始性细胞核的模型,依据这个模型也就可以对典型细胞核的进化形成和原始核自身的起源作出推论,这些推论是可以设法加以检验的,不仅可以检验这些推论的正确性,而且对原始核模型的建立是重要的,可以据之加以发展,修正,甚至否定,沿此途径已经否定了原始性细胞核的涡鞭毛虫核模型,进而提出了双滴虫核模型。     

电损毁大鼠杏仁中央核对脑桥臂旁核味觉神经元的影响

应用细胞外记录的电生理学方法,在乌拉坦麻醉的大鼠观察了电损毁双侧杏仁中央核前后脑桥臂旁核味觉神经元对四种基本味觉刺激(即氯化钠、盐酸、奎宁和蔗糖)反应的变化。根据对味觉刺激的优势反应,29个记录的味觉神经元中,有14个NaCl优势、9个HCl优势、3个QH2SO4优势和3个蔗糖优势反应神经元。损毁杏仁中央核明显增强臂旁核味觉神经元对盐酸和硫酸奎宁的反应(P＜0．01)。氯化钠优势、盐酸优势和奎宁优势反应神经元对盐酸和硫酸奎宁的反应在电损毁杏仁中央核后也明显增强。在破坏杏仁中央核后,臂旁核味觉神经元对氯化钠和硫酸奎宁苦味的分辨能力降低。以上结果提示,杏仁中央核在大鼠脑桥水平的味觉编码中发挥重要作用,它可能是通过参与对味觉的影响来调节机体的摄食行为。     

Ultrastructural investigation of ACTH immunoreactivity in arcuate and supraoptic nuclei of the rat

Summary Fine structural localization of an ACTH-like substance was obtained in neurons of the rat arcuate nucleus using immuno-electron microscopy, whereas it could not be confirmed that ACTH-containing cell bodies are present in the supraoptic nucleus. The immunoreactive cells of the arcuate nucleus appeared to be more numerous than the unreactive neurons. Immunostaining was carried out before embedding in resin. Empty vesicles of irregular shape were found in dendrites of immunoreactive arcuate neurons, but their significance and nature remain enigmatic. The reaction product was distributed uniformly throughout the cytoplasm of the ACTH-positive cells, except that the mitochondria, rough endoplasmic reticulum and Golgi vesicles and cisternae were devoid of PAP molecules. This distribution differed from the localization reported in ACTH-secreting cells of the rat anterior pituitary, where the reaction product was found in the rough endoplasmic reticulum and Golgi complex as well as in secretory granules.     

Immunoelectronmicroscopic localization of vasopressin in the rat suprachiasmatic nucleus

Summary The classical areas for arginine-vasopressin (AVP) synthesis are the magnocellular supraoptic (SON) and paraventricular nuclei. More recently AVP was also demonstrated in neurons of the parvocellular suprachiasmatic nucleus (SCN) of the rat. This result was substantiated in the present study by means of immunoelectron microscopy, by subjecting sections to antivasopressin plasma. Conventional electron microscopy revealed dense-core vesicles in the SCN cell bodies and fibres (mean diameter 94.7±0.9 nm and 84.0±1.1 nm respectively). These vesicles were infrequent within the cell bodies and could not be accumulated by ethanol administration. Immunoelectron microscopy showed a positive reaction in the cell bodies and fibres within vesicles of 93.7±1.1 nm and 98.5±1.2 nm respectively. By comparison, the cell bodies and fibres of the SON showed immunoreactive granules of 143.0±1.8 and 147.3±1.8 nm respectively. The presence in the SCN of AVP in vesicles of different size than those in the SON suggests that synthesis of this substance is indeed occurring within the SCN cells.Supported by The Foundation for Medical Research FUNGOThe authors wish to thank Dr. L.A. Sternberger (Edgewood Arsenal, Md., U.S.A.) for the peroxidase-anti-peroxidase complex, Dr. J.G. Streefkerk (Free University, Amsterdam) and the members of our project group Neuroendocrinology for their suggestions, Mr. P.S. Wolters and Miss A. van der Veiden for their skilled assistance     

Colocalization of tyrosine hydroxylase and Fos in the male Syrian hamster brain following different states of arousal

In an investigation of the role that central tyrosine hydroxylase-(TH) containing neurons play in copulation in the male Syrian hamster, The induction of Fos protein was used as an index of neuronal activation. With a double immunoperoxidase technique, the activation of TH neurons was compared in hamsters from three experimental groups: (1) mated in a new cage; (2) handled controls placed in a new cage, and (3) unhandled controls. Although mating selectively induces Fos production in the medial amygdaloid nucleus (Me), more than half of the TH neurons in Me (a region outside of the classical catecholamine systems) expressed Fos equally in all of the experimental groups. In the paraventricular hypothalamic nucleus (PVN), TH neurons were activated equivalently in mated and handled control animals compared to unhandled controls. TH neurons in the neucleus of the solitary tract (NST) were also activated in handled control animals, and mating further enhanced the level of Fos immunostaining in these neurons above both groups of nonmated animals. Although not quantified, co-localization of Fos and TH was also observed in all experimental groups in the olfactory bulbs and the interfascicular nucleus, and in the horizontal limb of the diagonal band of Broca and the cerebral cortex, regions which contain TH neurons but are not part of the classically described TH cell groups. Few, if any, TH neurons in other catecholaminergic brain regions, such as the substantia nigra and locus coeruleus, produced Fos in any of the experimental groups. These results suggest that TH neurons in the PVN and NST may be activated during different states of arousal, and that nonclassical TH neurons in the amygdala produce high levels of Fos even in unstimulated animals. 1994 John Wiley & Sons, Inc.     

Central distribution of nociceptive intradental afferent nerve fibers in the rat

The central distribution of intradental afferent nerve fibers was investigated by combining electron microscopic observations with a selective method for inducing degeneration of the A delta- and C-type afferent fibers. Degenerating terminals were found on the proprioceptive mesencephalic trigeminal neurons and on dendrites in the neuropil of the trigeminal motor nucleus after application of capsaicin to the rat's lower incisor tooth pulp. The results give anatomical evidence of new sites of central projection of intradental A delta- and C-type fibers whereby the nociceptive information from the tooth pulp can affect jaw muscle activity.     

Localization of the preprosomatostatin-mRNA by in situ hybridization in the ewe hypothalamus

The peptidergic neurohormone somatostatin (SRIF) derives from a precursor called preprosomatostatin (PPS) by proteolysis. We have isolated by RT-PCR and sequenced a partial cDNA coding for the ovine PPS. It contains a 348 base pairs coding sequence that shares strong similarities with previously cloned mammalian cDNAs. The ovine cDNA was used to synthesize radiolabeled cRNA to probe the PPS mRNA in the ewe hypothalamus by in situ hybridization. The PPS mRNA-containing cells are widely distributed in the hypothalamus. According to the number of silver grains over a cell, they show various staining intensities. The distribution of the PPS mRNA is in good agreement with that of the peptide previously determined using immunohistochemistry. The strongest labeled areas include the periventricular region of the paraventricular nucleus and the lateral division of the ventromedial nucleus. The difference in labeling intensity observed in the diverse populations of labeled neurons could reflect various levels of neuronal activity.     

Arginine vasopressin in hypothalamic paraventricular nucleus is transferred to the caudate nucleus to participate in pain modulation

Arginine vasopressin (AVP), which is synthesized and secreted in the hypothalamic paraventricular nucleus (PVN), is the most important bioactive substance in the pain modulation. Our pervious study had shown that AVP plays an important role in pain modulation in caudate nucleus (CdN). The experiment was designed to investigate the source of AVP in CdN by the nucleus push-pull perfusion and radioimmunoassay. The results showed that: (1) pain stimulation increased the AVP concentration in the CdN perfusion liquid, (2) PVN decreased the effect of pain stimulation which was stronger in both sides than in one side of PVN cauterization; and (3) L-glutamate sodium would excited the PVN neurons by the PVN microinjection that could increase the AVP concentration in the CdN perfusion liquid. The data suggested that AVP in the CdN might come from the PVN in the pain process, i.e., AVP in the PVN might be transferred to the CdN to participate in the pain modulation.