Experimental study on hepatic reinnervation after orthotopic liver transplantation in rats

BACKGROUND/AIMS: The present study examined whether extrinsic hepatic reinnervation occurred after orthotopic liver transplantation (OLT) in rats. METHODS: Inbred male Lewis rats were the recipients and females the donors. Tissue specimens were obtained postoperatively from the stump of a recipient's hepatoduodenal ligament (A), and the hepatic hilus (B) and peripheral parenchyma (C) of liver allografts, up to 6 months post-operation. Specimens were subjected to immunohistochemical examination using growth-associated protein (GAP)-43 as an axonal marker and transmission electron microscopy (TEM) for observing regenerating axons, as well as the polymerase chain reaction assay to detect the rat sex-determining region Y (SRY) protein gene of the regenerating nerves. RESULTS: At site A, GAP-43-positive nerve axons were identified from day 1 to 1 month post-OLT and SRY protein genes were expressed at and after 3 days post-OLT. At site B, GAP-43-positive axons were observed between 3 days and 1 month, and SRY protein genes were detected at 1 month post-OLT and thereafter. TEM confirmed the presence of regenerating axons at and after 3 days post-OLT. CONCLUSIONS: The results demonstrated that regenerating nerve fibers originating from the recipients reinnervated liver allografts. This extrinsic innervation occurred shortly after OLT, and most likely terminated after about 3 months.     

Dendrites of cerebellar granule cells correctly recognize their target axons for synaptogenesis in vitro

Neural circuits are generated by precisely ordered synaptic connections among neurons, and this process is thought to rely on the ability of neurons to recognize specific partners. However, it is also known that neurons promiscuously form synapses with nonspecific partners, in particular when cultured in vitro, causing controversies about neural recognition mechanisms. Here we reexamined whether neurons can or cannot select particular partners in vitro. In the cerebellum, granule cell (GC) dendrites form synaptic connections specifically with mossy fibers, but not with climbing fibers. We cocultured GC neurons with pontine or inferior olivary axons, the major sources for mossy and climbing fibers, respectively, as well as with hippocampal axons as a control. The GC neurons formed synapses with pontine axons predominantly at the distal ends of their dendrites, reproducing the characteristic morphology of their synapses observed in vivo, whereas they failed to do so when combined with other axons. In the latter case, synaptic proteins could accumulate between axons and dendrites, but these synapses were randomly distributed throughout the contact sites, and also their synaptic vesicle recycling was anomalous. These observations suggest that GC dendrites can select their authentic partners for synaptogenesis even in vitro, forming the synapses with a GC-specific nature only with them.     

Netrin-4 regulates thalamocortical axon branching in an activity-dependent fashion

Axon branching is remodeled by sensory-evoked and spontaneous neuronal activity. However, the underlying molecular mechanism is largely unknown. Here, we demonstrate that the netrin family member netrin-4 (NTN4) contributes to activity-dependent thalamocortical (TC) axon branching. In the postnatal developmental stages of rodents, ntn4 expression was abundant in and around the TC recipient layers of sensory cortices. Neuronal activity dramatically altered the ntn4 expression level in the cortex in vitro and in vivo. TC axon branching was promoted by exogenous NTN4 and suppressed by depletion of the endogenous protein. Moreover, unc-5 homolog B (Unc5B), which strongly bound to NTN4, was expressed in the sensory thalamus, and knockdown of Unc5B in thalamic cells markedly reduced TC axon branching. These results suggest that NTN4 acts as a positive regulator for TC axon branching through activity-dependent expression.Axon branching is an essential process to determine the final pattern of neuronal connections. Previous studies have demonstrated that axon branching is controlled not only by axon guidance-related molecules (1–6) but also by neuronal activity, such as firing and synaptic activity (7–10). However, how neuronal activity is converted into the molecular signals that underlie axon branching is still largely unknown.The thalamocortical (TC) projection is a well-characterized system in which to address this issue. TC axons originating from sensory thalamic nuclei form elaborate arbors, primarily in layer 4 of the neocortex (11). Lamina-specific axon branching occurs from the onset of development and is universal in the mammalian cortex (12–17), indicating that a rigid developmental program is predominant for laminar specificity. TC axon branching is also known to be modified by neuronal activity. In the visual system, geniculocortical axon arbors can be remodeled drastically by manipulating visual experience and cortical-cell activity (18–22). A similar feature has been demonstrated in the somatosensory system. In mutant mice in which synaptic transmission or downstream signaling mechanisms are disrupted, TC axon arbors are affected primarily along the tangential axis whereas their laminar pattern is not obviously influenced (23–26). We have also shown in vitro that the loss of firing and synaptic activities substantially suppresses TC axon branching in the target layer (27). All of these findings imply the existence of a target-derived, branch-promoting molecule whose expression is regulated by neuronal activity. In this study, we attempted to identify this hypothetical molecule and to reveal the molecular mechanism of its action.     

The incorporation of growth factor and chondroitinase ABC into an electrospun scaffold to promote axon regrowth following spinal cord injury

Spinal cord injury results in tissue necrosis in and around the lesion site, commonly leading to the formation of a fluid‐filled cyst. This pathological end point represents a physical gap that impedes axonal regeneration. To overcome the obstacle of the cavity, we have explored the extent to which axonal substrates can be bioengineered through electrospinning, a process that uses an electrical field to produce fine fibres of synthetic or biological molecules. Recently, we demonstrated the potential of electrospinning to generate an aligned matrix that can influence the directionality and growth of axons. Here, we show that this matrix can be supplemented with nerve growth factor and chondroitinase ABC to provide trophic support and neutralize glial‐derived inhibitory proteins. Moreover, we show how air‐gap electrospinning can be used to generate a cylindrical matrix that matches the shape of the cord. Upon implantation in a completely transected rat spinal cord, matrices supplemented with NGF and chondroitinase ABC promote significant functional recovery. An examination of these matrices post‐implantation shows that electrospun aligned monofilaments induce a more robust cellular infiltration than unaligned monofilaments. Further, a vascular network is generated in these matrices, with some endothelial cells using the electrospun fibres as a growth substrate. The presence of axons within these implanted matrices demonstrates that they facilitate axon regeneration following spinal cord injury. Collectively, these results demonstrate the potential of electrospinning to generate an aligned substrate that can provide trophic support, directional guidance cues and regeneration‐inhibitory neutralizing compounds to regenerating axons following spinal cord injury. Copyright © 2016 John Wiley & Sons, Ltd.     

白质内的神经递质信号

脑白质是由许多有髓鞘的轴突组成,白质和灰质共同组成中枢神经系统,白质是中枢神经系统内信息快速传递的基础。有髓神经通路主要是由少突胶质细胞、星形胶质细胞及少量的小胶质细胞和少突胶质前体细胞构成。大部分白质内的神经递质信号主要存在于神经细胞胞体外,这提示这些神经递质除了具有完成神经元与神经元之间信息传递的功能外,还有其他生理功能。白质中的神经递质信号种类很多,已经证实的有谷氨酸能、嘌呤能(ATP和腺苷)、GABA能、甘氨酸能、肾上腺素能、胆碱能、多巴胺能、血清素能等信号递质,通过与各种离子型或代谢型受体结合发挥作用。轴突和胶质细胞都可以释放神经递质,也可以表达相应的受体。白质内神经递质信号的生理功能还需进一步研究,但研究已经证实谷氨酸和ATP介导的信号可激活胶质细胞上的钙离子通道,并调节轴突的传导功能。某项研究显示,在动作电位传播的过程中,轴突释放神经递质并与胶质细胞上的受体结合,通过少突胶质细胞来调节星形胶质细胞的稳态和髓鞘形成。星形胶质细胞也释放神经递质,与轴突上的受体相结合,增强动作电位的传播,维持信号电位沿长的轴突传播。白质内神经递质种类的多样性,提示它们有多种功能,对信号的传递有重要作用。白质内的神经递质信号现象很有可能也存在于大脑皮质和灰质,在这些部位的神经递质对于大脑的高级认知功能有更重要的作用。     

17例潜突型、哑铃型舌下腺囊肿行口内进路改良手术＋口内开窗＋颌下区加压术治疗体会

目的探讨潜突型、哑铃型舌下腺囊肿的临床诊断及治疗方法。方法对17例潜突型、哑铃型舌下腺囊肿患者临床诊断及治疗进行回顾性分析总结。结果采用口内进路改良手术＋口内开窗＋颌下区加压术治疗17例潜突型、哑铃型舌下腺囊肿均痊愈,疗程短,无1例复发。结论口内进路改良手术＋口内开窗＋颌下区加压术是治疗潜突型、哑铃型舌下腺囊肿的有效方法,创伤小、疗程短、不影响美观。     

Relationship Between GAP-43 Expression in the Dentate Gyrus and Synaptic Reorganization of Hippocampal Mossy Fibres in Rats Treated with Kainic Acid

Kainic acid-induced seizures in adult rats produce neurodegeneration in the hippocampus followed by sprouting of the mossy fibres in the inner molecular layer of the dentate gyrus and changes in GAP-43 expression in the granule cells. In the present study we observed that 4 days after kainic acid injection a dense plexus of silver impregnated degenerating terminals detected by Gallyas's method and a decrease of GAP-43 immunostaining was observed in the inner molecular layer of the dentate gyrus indicating deafferentiation of this region. This was associated with the formation of an intense GAP-43 immunostained band in the supragranular layer. MK-801, a non-competitive inhibitor of the NMDA receptor, which partially inhibited the behavioural seizures induced by KA, also protected from the inner molecular layer deafferentation and markedly reduced the expression of GAP-43 mRNA in the granule cells and the intense GAP-43 immunostained band in the supragranular layer, suggesting a relationship among these events. Two months after kainic acid injection the intense supragranular GAP-43 positive band was no longer evident but the whole inner molecular layer appeared more labelled in association with the formation of the collateral sprouting of the mossy fibres in the inner molecular layer as detected by Timm's staining. These effects were also markedly reduced by the pretreatment with MK-801. Taken together, these experiments indicate for the first time a direct relationship between the increase of GAP-43 immunostaining in the inner molecular layer of the dentate gyrus and the collateral sprouting of mossy fibres in this district in response to kainic acid induced seizures. This further supports the hypothesis that the early induction of GAP-43 in granule cells may be one of the molecular mechanisms required for the synaptic reorganization of the mossy fibres.     

The lack of effect of baclofen on action potentials of spinal motor axon collateral terminations in vivo

 In the lumbar ventral horn of pentobarbitone-anaesthetised cats, (-)-baclofen reduces both the synaptic release of excitatory transmitter from muscle group Ia afferent terminations and the duration of the presynaptic action potentials of these terminations, presumably by interfering with the influx of calcium ions through voltage-activated channels. Baclofen, however, has little or no effect on cholinergic excitation at motor axon collateral synapses on spinal Renshaw cells and, in the present study, was found not to reduce the duration of the action potential of axon collateral terminations located in the vicinity of Renshaw cells in pentobarbitone-anaesthetised cats. Furthermore, in contrast to group Ia terminations, a 4-aminopyridine-sensitive potassium conductance could not be detected as contributing to axon collateral termination action potentials. These results suggest that there may be differences in presynaptic ion fluxes associated with transmitter release at the intraspinal terminations of group Ia afferent fibres and motor axon collaterals in the cat spinal cord. Received: 16 October 1996 / Accepted: 5 December 1996     

Expression of the surface antigen A2B7 in adult and developing honeybee olfactory pathway

In order to identify molecules involved in the development of the honeybee olfactory pathway, hybridoma technology has been used. Among different cell lines, A2B7 has been selected. It produces a specific antibody for a surface glycoprotein of 91 kDa. This protein is mainly expressed by both the antennal receptor cells and mushroom body neurons. Based on (i) the spatio-temporal pattern of expression during pupal development; (ii) the cell surface location of the antigen; and (iii) the partial molecular characterization of the antigen, a putative role for this protein in axonal fasciculation and guidance is discussed.