Contralateral degeneration in the cat spinal trigeminal nucleus following unilateral retrogasserian trigeminal rhizotomy.

Retrogasserian trigeminal rhizotomy was used to study the central projections and patterns of degeneration in the spinal trigeminal nucleus (STN). At survival times of 3-20 days, reduced silver stains show extensive degeneration throughout the ipsilateral STN and in addition, well delineated degeneration was identified in the periobex region of the contralateral STN that varied with survival time. The results suggest that primary afferents may contribute to this contralateral projection.     

Acetylcholinesterase activity of synaptic structures in the spinal trigeminal nucleus

Summary The electron microscope has been used to study the localization of acetylcholinesterase (AChE) activity in the spinal trigeminal nucleus of normal cats with special emphasis on the distribution near synaptic structures. Reaction product is found around both round and flattened synaptic vesicle-containing axon terminals, particularly in synaptic clefts and often specifically associated with the presynaptic, or less frequently the postsynaptic membrane. The presence of reaction product at these specific sites suggests that these are areas of high AChE activity and that acetylcholine may be important in neurotransmission in these regions.     

Quantitative ultrastructure of synapses on functionally identified primary afferent neurons in the cat trigeminal mesencephalic nucleus

Though a number of studies have reported the presence of synapses on neurons in the trigeminal mesencephalic nucleus (Vmes), there have been no quantitative studies of either the density of innervation, or the ultrastructure, of the synapses on single, physiologically identified neurons in this nucleus. In this study we recorded from single neurons in the Vmes, identified them as being either muscle spindle afferents (MS) or periodontal ligament mechanoreceptor afferents (PL), and then labeled the neurons by intra-axonal injection of horseradish peroxidase (HRP). The material was first processed to reveal the HRP activity, following which ultrathin sections through the labeled somata were cut and examined under the electron microscope. Complete serial reconstructions were made through the soma of one MS neuron and one PL neuron, and the contacts on the neurons reconstructed. Boutons were found on the soma, spines, appendages and the axon hillock and the initial segment of the axon. The numbers of boutons terminating on the two neurons were 198 (PL) and 424 (MS), giving a packing density of 4.4 and 10.7 boutons respectively (i.e., number of boutons/100 micron 2 of the postsynaptic membrane). Boutons could be separated into two types on the basis of their vesicles: those containing clear, round vesicles (i.e., S-type) and those containing a mixture of round, oval and flattened vesicles (P-type). Ninety-five (PL neuron) and 99% (MS neuron) of terminals on the two neurons were P-type. All the S-type boutons and 80% of the P-type boutons formed asymmetric synaptic contacts while 10% of the P-type boutons made symmetric contacts. Quantitative measurements of the P-type boutons on the labeled neurons, in which the data of MS and PL neurons were pooled, revealed that bouton volume was highly correlated with bouton surface area, active zone number, total active zone area, vesicle number, and mitochondrial volume. However, comparing the quantitative measurements of the P-type boutons with those of previously reported vibrissa afferent terminals and their associated axon terminals revealed that all the parameters were smaller for the P-type boutons (on Vmes neurons) than those of the vibrissa afferent terminals but similar to those of axon terminals presynaptic to the vibrissa afferents. Taken together, our results emphasize the wide scope for synaptic interactions in the Vmes and suggest that it may be more fruitful to view the Vmes as an integrating center.     

Early forms of terminal degeneration in the spinal trigeminal nucleus following rhizotomy

Summary Pars interpolaris of the spinal trigeminal nucleus in adult cats has been studied with the electron microscope at several early and consecutive postoperative survival times following retrogasserian rhizotomy. The results show that several different forms of degenerative changes occur in the axon terminals and some of these forms may be interpreted as stages in certain sequences of terminal degeneration.Synaptic vesicle depletion and mitochondrial alterations are probably the earliest changes in some terminals, but the occurrence of completely different forms also at very early survival times suggest that other processes may be involved. Electron-dense forms of terminal degeneration of later survivals are suggested as arising from any of at least four possible types or classes of earlier alterations based upon presence or absence of vesicles and neurofilaments and on the mitochondrial changes. Electron-lucent forms probably progress to dense types with time but may account for only a proportion of the later dense forms.All types of neuroglia, or reactive glia, and a possible third glia are involved in early and rapid phagocytosis and denudation of post-synaptic sites at times earlier than previously reported.The study also confirms a previous study of a time-related degeneration pattern in the same area and offers an explanation for this pattern.     

Dependence of electrical activity of neurons in rostral parts of spinal trigeminal nucleus on functional state of trigeminal Gasser ganglion

Unilateral injection of 100 μl 1% lidocaine into the trigeminal Gasser ganglion of narcotized rats produced a long-term moderation of the discharge rate of neurons in the ipsilateral (relative to the side of injection) rostral area of the spinal trigeminal nucleus. Activity of neurons in the contralateral rostral area of the spinal trigeminal nucleus was not blocked. Functional state of neurons in the trigeminal ganglion determines discharge activity of ipsilateral neurons of the spinal trigeminal nucleus. Activity of neurons in the contralateral rostral area of spinal trigeminal nucleus was not inhibited. Functional state of the cells in the trigeminal ganglion determines the character of electrical activity of neurons in the ipsilateral rostral area of spinal trigeminal nucleus. __________ Translated from Byulleten’ Eksperimental’noi Biologii i Meditsiny, Vol. 144, No. 7, pp. 4–6, July, 2007     

Morphological interrelationship between astrocytes and nerve endings in the rat spinal trigeminal nucleus caudalis

It has been reported that the spinal trigeminal nucleus caudalis (Sp5C), which receives nociceptive information from the oro-facial regions, has four laminae. To clarify the role of glial cells in the transmission of the nociceptive information, the present study was conducted to examine the detailed distribution of astrocytes in each lamina and also to investigate a morphological interrelationship between the astrocytes and nerve endings in the rat Sp5C. After the preparation of the serial cryostat sections, immunohistochemistry for glial fibrillary acidic protein (GFAP) was employed to identify the astrocytes, and immunohistochemistry for substance P (SP), calcitonin gene-related peptide (CGRP), was used for the nerve endings. We also employed double-labeling immunofluorescence and electron microscopic immunohistochemistry for the GFAP/SP or GFAP/CGRP. GFAP-positive reactions were observed in all laminae of the Sp5C, and SP- or CGRP-positive nerve endings were observed in the lamina I and II. Additionally, we clarified the presence of GFAP/SP- or GFAP/CGRP-positive reactions by the double-labeling immunofluorescence and demonstrated the morphological interrelationship between the astrocytes and nerve endings by the double-labeling electron microscopic immunohistochemistry. These findings suggest that astrocytes might play some roles in the transmission of nociceptive information from the oro-facial region.     

Drebrin (developmentally regulated brain protein) is associated with axo-somatic synapses and neuronal gap junctions in rat mesencephalic trigeminal nucleus

Drebrin (developmentally regulated brain protein)-like immunoreactivity was investigated in the adult rat mesencephalic trigeminal nucleus (MTN) using light and electron microscope. Intense immunoreactive puncta were observed on the cytoplasmic membrane and within the cytoplasm. The cytoplasm was also faintly immunopositive for drebrin, and thus MTN somata other than multipolar cells were distinguishable from non-MTN somata. These immunoreactive cell bodies were localized from the level of the superior colliculus to the pons. Electron microscopic observation showed that the post-synaptic cytoplasmic membrane at axo-somatic synapses was immunoreactive for drebrin. Drebrin-like immunoreactivity was also observed on spine-like processes emanating from MTN somata. In addition, the post-synaptic cytoplasmic membrane at axo-somatic synapses was also immunopositive for drebrin. Within the cytoplasm of MTN cell bodies, a part of the rough endoplasmic reticulum and neighboring structures were also immunopositive. Further, both ends of the somato-somatic close appositions that contained neuronal gap junctions harbored immunoreactive structures. We can infer from the results that drebrin is an ideal marker protein for MTN cell bodies. The abundance of drebrin-like immunoreactivity in the MTN neurons suggests that the MTN has highly flexible synaptogenesis.     

Electron microscopy of degenerating axons and terminals in spinal trigeminal nucleus after tooth pulp extirpations

Following multiple tooth pulp extirpations, electron microscopic preparations show degenerating axons and synaptic terminals in the same region of the brain stem trigeminal nucleus previously demonstrating degeneration by light-optical methods. The observations confirm the phenomenon of transganglionic degeneration in this system and identify the class of central nervous system axons and synapses specifically related to innervation of the teeth.     

Interferon-gamma receptors are expressed at synapses in the rat superficial dorsal horn and lateral spinal nucleus

Interferon-gamma can facilitate the spinal nociceptive flexor reflex and may elicit neuropathic pain-related behavior in rats and mice. Immunoreactivity for the interferon-gamma receptor (IFN-gamma R) occurs in the superficial layers of the dorsal horn and the lateral spinal nucleus in the rat and mouse spinal cord, as well as in subsets of neurons in the dorsal root ganglia. The aim of the present study was to examine the cellular localization and origin of the IFN-gamma R in the spinal cord. As viewed by confocal microscopy, the immunopositivity for the IFN-gamma R was co-localized with that of the presynaptic marker synaptophysin and with neuronal nitric oxide synthase in the lateral spinal nucleus, whereas only a minor overlap with these molecules was observed in laminae I and II of the dorsal horn. There was no co-localization of the IFN-gamma R with markers for astrocytes and microglial cells. Ultrastructurally, the IFN-gamma R was found predominantly in axon terminals in the lateral spinal nucleus, but at postsynaptic sites in dendrites in laminae I and II. The IFN-gamma R expressed in neurons in dorsal root ganglia was transported in axons both centrally and peripherally. Hemisection of the spinal cord caused no reduction in immunolabelling of the IFN-gamma R in the dorsal horn or the lateral spinal nucleus. Since rhizotomy does not affect the immunolabelling in the lateral spinal nucleus, our observation indicates that the presynaptic receptors in this nucleus are derived from intrinsic neurons. The localization of the IFN-gamma R in the spinal cord differed from that of the AMPA glutamate receptor subunits 2 and 3 and the substance P receptor (NK1). Our results, showing localization of IFN-gamma R to pre- and postsynaptic sites in the dorsal horn and lateral spinal nucleus indicate that IFN-gamma can modulate nociception at the spinal cord level.     

体外培养条件下兔脊柱运动节段髓核组织的变化

背景：椎间盘器官体外培养模型作为体内试验和细胞培养的联系,能够在原生基质中维持细胞活性,从而保留了重要的细胞与细胞及细胞与基质的相互作用。 目的：观察体外培养兔脊柱运动节段模型髓核组织的变化。 方法：将13只新西兰白兔处死后在无菌条件下取出脊柱运动节段50个,在高渗培养基中进行整体培养  (410 mOsm/kg),于培养前及培养后第3,7,14,21天,各取10个椎间盘分别进行苏木精-伊红染色、Ⅱ型胶原免疫组织化学、蛋白多糖含量和髓核细胞活力测定。 结果与结论：培养14 d苏木精-伊红显示椎间盘组织结构基本保持完整,21 d椎间盘组织形态学破坏;Ⅱ型胶原免疫组织化学染色强度14 d无显著差异(P > 0.05),21 d染色变浅,与之前各时间点相比有显著差异(P <   0.05);蛋白多糖PAS/AB染色7 d内强度无降低,14 d强度有所减弱,21 d染色强度进一步减弱;髓核细胞荧光检测21 d髓核细胞荧光强度减弱,细胞活性降低,与之前各时间点比较差异明显(P < 0.05)。结果表明,培养14 d兔脊柱运动节段椎间盘髓核组织退变不显著,培养21 d模型髓核退变明显,脊柱运动节段目前在14 d内可作为研究生物力学对椎间盘影响的体外实验模型。 中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程      

FRAP-positive and capsaicin-sensitive terminals in the substantia gelatinosa of the mouse spinal trigeminal nucleus caudalis.

FRAP (fluoride-resistant acid phosphatase)-reactivity in the substantia gelatinosa of the mouse spinal trigeminal nucleus caudalis (STNC) was examined by light and electron microscopy. Degenerated figures of terminals caused by capsaicin were compared with the FRAP-positive terminals. Scalloped (fan-like) or indented, sinuous, slender, and cap-like figures with closely packed agranular synaptic vesicles of various sizes were common to both FRAP-positive and capsaicin-sensitive terminals. These terminals had glomerular or nonglomerular endings. Sometimes FRAP-positive and capsaicin-sensitive glomerular terminals made presynapses with surrounding dendrites. Frequently, both nonglomerular terminals were in direct contact with the neuronal soma. The terminal features of FRAP-positive and capsaicin-sensitive ones in the mouse STNC are the same as those seen in the superficial dorsal horn of the spinal cord. These findings suggest that some of the FRAP-positive terminals are capsaicin-sensitive, thereby indicating their nociceptive primary afferent.     

Increases in PKC gamma expression in trigeminal spinal nucleus is associated with orofacial thermal hyperalgesia in streptozotocin-induced diabetic mice

Painful diabetic polyneuropathy (PDN) at the early phrase of diabetes frequently exhibits increased responsiveness to nociception. In diabetic patients and animal models, alterations in the transmission of orofacial sensory information have been demonstrated in trigeminal system. Herein, we examined the changes of protein kinase Cγ subunit (PKCγ) in trigeminal spinal nucleus (Sp5C) and observed the development of orofacial thermal sensitivity in streptozotocin (STZ)-induced type 1 diabetic mice. With hyperglycemia and body weight loss, STZ mice exhibited orofacial thermal hyperalgesia, along with increased PKCγ expression in Sp5C. Insulin treatment at the early stage of diabetes could alleviate the orofacial thermal hyperalgesia and impaired increased PKCγ in Sp5C in diabetic mice. In summary, our results demonstrate that PKCγ might be involved in orofacial thermal hyperalgesia of diabetes, and early insulin treatment might be effective way to treat orofacial PDN.     

Localization of serotonin- and substance P-like immunofluorescence in the caudal spinal trigeminal nucleus of the rat

A double immunofluorescence labeling method was used to study the localization of serotonin (5-HT)- and substance P (SP)-like immunoreactivities within neuronal fibers in the caudal spinal trigeminal nucleus of the rat. The 5-HT- and SP-immunoreactive fibers share extensive topographical overlap within the superficial laminae of the caudal trigeminal nucleus; however, the majority of stained fibers are immunoreactive for either 5-HT or SP, but not both simultaneously. A small population of fibers in which 5-HT and SP are co-localized is present and restricted to the marginal zone as well as the inner and outer layers of the substantia gelatinosa. The results of the present study suggest that fibers containing coexistent 5-HT and SP may be involved in the processing of nociceptive somatic sensation in the medullary dorsal horn of rat.     

GABAA receptor-mediated tonic currents in substantia gelatinosa neurons of rat spinal trigeminal nucleus pars caudalis

In the present study, we describe GABAA receptor-mediated tonic inhibitory currents in the substantia gelatinosa (SG) region of rat spinal trigeminal nucleus pars caudalis (Vc). The GABA(A) receptor-mediated tonic currents were identified by bath-application of the GABAA receptor antagonists, picrotoxin (1mM), SR95531 (100microM) and bicuculline (100microM). All three antagonists completely blocked outward spontaneous (phasic) inhibitory postsynaptic currents, but only picrotoxin and bicuculline induced a significant (>5pA) inward shift of holding currents at a holding potential (Vh) of 0mV in 60-70% of SG neurons, revealing the existence of tonic outward currents. The tonic currents were resistant to further the blockades of glycine receptors or those in addition to glutamate receptors and voltage-dependent sodium channels. An acute bath-application of THDOC (0.1microM), the stress-related neurosteroid, did enhance tonic currents, but only in a small population of SG neurons. In addition, slices incubated with THDOC for 30min increased the probability of neurons with significant tonic currents. The GABAergic tonic inhibition demonstrated in this study may play a significant role in the sensory processing system of the Vc.     

Loss of substance P immunoreactivity in the nucleus of the spinal trigeminal tract after intradural tumour compression of the trigeminal nerve

The peptide, substance P (SP), is suspected of being a mediator of nociception. Immunoreactive SP is conspicuously present in areas of the mammalian central nervous system which receive nociceptive input. We have observed that compression of the trigeminal nerve by an intradural tumour deposit results in the ipsilateral depletion of SP immunoreactivity in human spinal trigeminal nucleus. These results expand previous experimental animal studies which associate this peptide with a subpopulation of primary sensory neurones and, in addition, demonstrate a correlation between the levels of SP immunoreactivity and premortem clinical sensory alterations.