Patched1 regulates hedgehog signaling at the primary cilium

Primary cilia are essential for transduction of the Hedgehog (Hh) signal in mammals. We investigated the role of primary cilia in regulation of Patched1 (Ptc1), the receptor for Sonic Hedgehog (Shh). Ptc1 localized to cilia and inhibited Smoothened (Smo) by preventing its accumulation within cilia. When Shh bound to Ptc1, Ptc1 left the cilia, leading to accumulation of Smo and activation of signaling. Thus, primary cilia sense Shh and transduce signals that play critical roles in development, carcinogenesis, and stem cell function.     

Activity-dependent internalization of smoothened mediated by beta-arrestin 2 and GRK2

Binding of Sonic Hedgehog (Shh) to Patched (Ptc) relieves the latter's tonic inhibition of Smoothened (Smo), a receptor that spans the cell membrane seven times. This initiates signaling which, by unknown mechanisms, regulates vertebrate developmental processes. We find that two molecules interact with mammalian Smo in an activation-dependent manner: G protein-coupled receptor kinase 2 (GRK2) leads to phosphorylation of Smo, and beta-arrestin 2 fused to green fluorescent protein interacts with Smo. These two processes promote endocytosis of Smo in clathrin-coated pits. Ptc inhibits association of beta-arrestin 2 with Smo, and this inhibition is relieved in cells treated with Shh. A Smo agonist stimulated and a Smo antagonist (cyclopamine) inhibited both phosphorylation of Smo by GRK2 and interaction of beta-arrestin 2 with Smo. beta-Arrestin 2 and GRK2 are thus potential mediators of signaling by activated Smo.     

奶牛腰荐尾段脊髓灰质板层构筑的研究

本研究取８头初生黑白花奶牛的腰荐尾段脊髓，分节后横切成３ｍｍ厚的脊髓块。其中７例在切片剪用苯胺蓝或尼氏混合液的块染法作细胞染色；１例用Ｗｅｉｌ氏法作髓鞘染色。均作石蜡切片。结果表明：奶牛腰荐尾段脊髓灰质基本上象猫那样分为１０个板层，但在层的范围，形态，结构等方面也有自己的特点。本文还与其它一些动物和人的脊髓灰质板层构筑进行了比较。     

Transmembrane protein GDE2 induces motor neuron differentiation in vivo

During neural development, coordinate regulation of cell-cycle exit and differentiation is essential for cell-fate specification, cell survival, and proper wiring of neuronal circuits. However, the molecules that direct these events remain poorly defined. In the developing spinal cord, the differentiation of motor neuron progenitors into postmitotic motor neurons is regulated by retinoid signaling. Here, we identify a retinoid-inducible gene, GDE2 (glycerophosphodiester phosphodiesterase 2), encoding a six-transmembrane protein that is necessary and sufficient to drive spinal motor neuron differentiation in vivo. A single amino acid mutation in the extracellular catalytic domain abolishes protein function. This reveals a critical role for glycerophosphodiester metabolism in motor neuron differentiation.     

Control of embryonic motoneuron survival in vivo by ciliary neurotrophic factor

During development of the nervous system, neurons in many regions are overproduced by proliferation, after which the excess cells are eliminated by cell death. The survival of only a proportion of neurons during normal development is thought to be regulated by the limited availability of neurotrophic agents. One such putative trophic agent is ciliary neurotrophic factor (CNTF), a polypeptide that promotes the survival of ciliary, sensory, and sympathetic neurons in vitro. In contrast to the results of in vitro studies, however, the daily treatment of chick embryos in vivo with purified human recombinant CNTF failed to rescue any of these cell populations from cell death, whereas CNTF did promote the in vivo survival of spinal motoneurons. Thus, CNTF may not act as a neurotrophic agent in vivo for those embryonic neurons (especially ciliary neurons) on which it acts in vitro. Rather, CNTF may be required for in vivo survival of motoneurons.     

DNA topoisomerase IIbeta and neural development

DNA topoisomerase IIbeta is shown to have an unsuspected and critical role in neural development. Neurogenesis was normal in IIbeta mutant mice, but motor axons failed to contact skeletal muscles, and sensory axons failed to enter the spinal cord. Despite an absence of innervation, clusters of acetylcholine receptors were concentrated in the central region of skeletal muscles, thereby revealing patterning mechanisms that are autonomous to skeletal muscle. The defects in motor axon growth in IIbeta mutant mice resulted in a breathing impairment and death of the pups shortly after birth.     

出生前山羊脑和脊髓的生长发育规律

测量了6～21周山羊胎儿脑和脊髓的相关指标,并绘制生长曲线.结果表明:山羊胎儿脑和脊髓生长发育具有快慢交替的阶段性变化规律.发育早期脑的生长速度要快于同期脊髓的生长速度.出生前,山羊大脑在长、宽、高3个方向上的发育表现出不均衡的特点.脊髓不同节段的生长高峰期出现的时间、数量和增长趋势都不一致,可能与脊髓内不同组份发育先后次序有关.     

Synaptic connections in vitro: modulation of number and efficacy by electrical activity

The functional architecture of synaptic circuits is determined to a crucial degree by the patterns of electrical activity that occur during development. Studies with an in vitro preparation of mammalian sensory neurons projecting to ventral spinal cord neurons slow that electrical activity induces competitive processes that regulate synaptic efficacy so as to favor activated pathways over inactive convergent pathways. At the same time, electrical activity initiates noncompetitive processes that increase the number of axonal connections between these sensory and spinal cord neurons.     

Regional astrocyte allocation regulates CNS synaptogenesis and repair

Astrocytes, the most abundant cell population in the central nervous system (CNS), are essential for normal neurological function. We show that astrocytes are allocated to spatial domains in mouse spinal cord and brain in accordance with their embryonic sites of origin in the ventricular zone. These domains remain stable throughout life without evidence of secondary tangential migration, even after acute CNS injury. Domain-specific depletion of astrocytes in ventral spinal cord resulted in abnormal motor neuron synaptogenesis, which was not rescued by immigration of astrocytes from adjoining regions. Our findings demonstrate that region-restricted astrocyte allocation is a general CNS phenomenon and reveal intrinsic limitations of the astroglial response to injury.     

Astrocytes block axonal regeneration in mammals by activating the physiological stop pathway

Regenerating sensory axons in the dorsal roots of adult mammals are stopped at the junction between the root and spinal cord by reactive astrocytes. Do these cells stop axonal elongation by activating the physiological mechanisms that normally operate to stop axons during development, or do they physically obstruct the elongating axons? In order to distinguish these possibilities, the cytology of the axon tips of regenerating axons that were stopped by astrocytes was compared with the axon tips that were physically obstructed at a cul-de-sac produced by ligating a peripheral nerve. The terminals of the physically obstructed axon tips were distended with neurofilaments and other axonally transported structures that had accumulated when the axons stopped elongating. By contrast, neurofilaments did not accumulate in the tips of regenerating axons that were stopped by spinal cord astrocytes at the dorsal root transitional zone. These axo-glial terminals resembled the terminals that axons make on target neurons during normal development. On the basis of these observations, astrocytes appear to stop axons from regenerating in the mammalian spinal cord by activating the physiological stop pathway that is built into the axon and that normally operates when axons form stable terminals on target cells.     

Cell recognition by neuronal growth cones in a simple vertebrate embryo

The mechanism that guides neuronal growth cones to their targets in vertebrate embryos has been difficult to study primarily because of the complexity and large number of neurons found in many vertebrate nervous systems. The spinal cord of a simple vertebrate, the fish embryo, is used to analyze pathfinding mechanisms. The early embryonic spinal cord consists of a relatively small number of identifiable neurons. From the beginning of axonal outgrowth the growth cones of these identified neurons extend along stereotyped and precise pathways in the spinal cord. Laser ablation experiments (i) support the hypothesis that early growth cones that pioneer specific spinal tracts appear to recognize cues on subsets of longitudinally arrayed neuroepithelial cells and (ii) demonstrate that later growth cones that selectively fasciculate in these spinal tracts appear to recognize cues on specific subsets of axons.     

Repairing the injured spinal cord

Certain cell, molecular, and bioengineering strategies for repairing the injured spinal cord are showing encouraging results (either alone or in combination) in animal models, with limited recovery of mobility being reported in some cases.     

Androgens prevent normally occurring cell death in a sexually dimorphic spinal nucleus

The spinal nucleus of the bulbocavernosus (SNB) contains many more motoneurons in adult male rats than in females. Androgens establish this sex difference during a critical perinatal period, which coincides with normally occurring cell death in the SNB region. Sex differences in SNB motoneuron number arise primarily because motoneuron loss is greater in females than in males during the early postnatal period. Perinatal androgen treatment in females attenuates cell death in the SNB region, reducing motoneuron loss to levels typical of males. The results suggest that steroid hormones determine sex differences in neuron number by regulating normally occurring cell death and that the timing of this cell death may therefore define critical periods for steroid effects on neuron number.     

6种家禽孤束核的脊髓通路：HRP逆行追踪法研究

应用微电泳逆行追踪技术，分别向北京鸭，麻鸭，鹅、鸡、鸽和鹌鹑脊髓的颈、胸、腰段的一侧灰质中导入辣根过氧化物酶系统地研究了其孤束核至脊髓传导通路和细胞构筑。结果如下：在颈髓不同节段单侧灰质导入Ｈ ＲＰ后，６种家禽延髓的双侧孤束核内都出现了大量的标准细胞；     

Heterogeneity of glycine receptors and their messenger RNAs in rat brain and spinal cord

Messenger RNAs isolated from adult or newborn rat spinal cord were fractionated in a sucrose gradient. The fractions were injected into Xenopus oocytes to determine their potencies for expression of glycine receptors (GlyRs), which were then examined electrophysiologically. The sedimentation profiles disclosed two classes of GlyR mRNAs, one heavy and the other light. The adult spinal cord was rich in heavy GlyR mRNA, whereas the light GlyR mRNA was more abundant in neonatal spinal cord and in adult cerebral cortex. Glycine receptors encoded by heavy and light mRNAs of adult spinal cord showed some electrophysiological differences. Thus there are two types of GlyRs encoded by mRNAs of different sizes, and the expression of these mRNAs is developmentally regulated. A tissue- and age-dependent distribution of heterogeneous GlyR mRNAs may imply diverse roles of the GlyRs in neuronal function in the central nervous system.     

Comparison of birth weight and umbilical and placental characteristics of cloned and artificial insemination-derived piglets

Somatic cell nuclear transfer (SCNT)-derived piglets have significantly higher stillbirth rate and postnatal mortality rate than artificial insemination (AI)-generated piglets. The question whether the low survival rate of SCNT piglets was related to birth weight, umbilical cord or placenta development was investigated. In this study, stillbirth rate, neonatal death rate, birth weight, umbilical cord status, placental parameters and placental gene expression patterns were compared between SCNT and AI piglets. Results showed that mortality rates at birth and during the neonatal stage of SCNT piglets were significantly higher than those of AI piglets. The incidence of abnormal umbilical cord in SCNT and SCNT-liveborn (SCNT-LB) piglets was significantly higher than in AI and AI-liveborn (AI-LB) piglets. Birth weight, placental weight, placental surface area and placental efficiency in SCNT and SCNT-LB piglets were significantly lower than those of AI and AI-LB piglets. Placental expression profiles of imprinting, angiopoiesis and nutrient transport-related genes were defective in SCNT-LB piglets compared with those in AI-LB piglets. Thus, the low survival rate of SCNT piglets may be associated with abnormal umbilical cord and placenta development. These characteristics may have resulted from aberrant expression of angiogenesis, nutrient transport, and imprinting-related genes in the placentas.     

北京鸭脊髓动脉的观察

用乳胶灌注北京鸭脊髓动脉10例,肉眼及解剖镜下观察。得到结果如下:供应北京鸭脊髓的动脉有:脊髓腹侧动脉1条、脊髓背外侧动脉2条和脊髓背正中动脉1条。脊髓腹侧动脉最粗大,伸延于脊髓全长,前端接脑基底动脉,伸延途中得到根动脉腹侧支的加强。在颈段,脊髓腹侧动脉为双干。脊髓背外侧动脉较细小,前面接小脑后动脉,途中由背侧根动脉加强,该动脉伸延于脊髓全长。脊髓背正中动脉位于脊髓的背正中沟内,只伸延于 C_(12)～L_1节段范围内。根动脉由枕动脉、椎动脉升支、椎动脉降支、肋间动脉、腰动脉、荐动脉和尾动脉发出。根动脉沿脊神经根入椎管,分为腹侧支和背侧支。根动脉腹侧支与脊髓腹侧动脉相联结的形式在不同节段不同。两条脊髓背外侧动脉间由交通支相连。供应脊髓内部的血液由发自于脊髓腹侧动脉的沟动脉、周缘动脉和发自于脊髓背侧动脉的周缘动脉的穿质支供给。     

颈部脊髓硬膜外电刺激治疗脑性瘫痪的临床观察

目的：观察颈部脊髓硬膜外电刺激治疗脑性瘫痪的临床疗效。方法：采用C2--C5脊髓硬膜外电刺激治疗脑性瘫痪病人18例，具体参数为：频率25—100ppm，强度3—15V，波宽0．3--0．4ms，时间6h／d。结果：18例病人中15例有效。结论：颈部脊髓硬膜外电刺激是一种治疗脑性瘫痪病人有效方法。     

Progression of vertebrate limb development through SHH-mediated counteraction of GLI3

Distal limb development and specification of digit identities in tetrapods are under the control of a mesenchymal organizer called the polarizing region. Sonic Hedgehog (SHH) is the morphogenetic signal produced by the polarizing region in the posterior limb bud. Ectopic anterior SHH signaling induces digit duplications and has been suspected as a major cause underlying congenital malformations that result in digit polydactyly. Here, we report that the polydactyly of Gli3-deficient mice arises independently of SHH signaling. Disruption of one or both Gli3 alleles in mouse embryos lacking Shh progressively restores limb distal development and digit formation. Our genetic analysis indicates that SHH signaling counteracts GLI3-mediated repression of key regulator genes, cell survival, and distal progression of limb bud development.