CCRL1 marks heterogeneity in cortical and medullary thymic epithelial cells

Cortical thymic epithelial cells (cTECs) and medullary thymic epithelial cells (mTECs), which play essential roles in the establishment of a functionally competent and self‐tolerant repertoire of T cells, are derived from common thymic epithelial progenitor cells (pTECs). Recent findings indicate that mTECs are derived from cells that express molecules that are abundant in cTECs rather than mTECs, and provide fresh insight into the characteristics of pTECs and their diversification pathways into TEC subpopulations. In this issue of the European Journal of Immunology, Ribeiro et al. [Eur. J. Immunol. 2014. 44: 2918–2924] focus on CCRL1, an atypical chemokine receptor that is highly expressed by cTECs rather than mTECs, and show that CCRL1‐expressing embryonic TECs can give rise to mTECs. Interestingly, Ribeiro et al. further report that a fraction of postnatal mTECs express CCRL1 at a low level, suggesting novel complexity in mTECs.     

Effects of the administration of a catalase inhibitor into the fourth cerebral ventricle on cardiovascular responses in spontaneously hypertensive rats exposed to sidestream cigarette smoke

OBJECTIVE:

Previous studies have demonstrated a relationship between brain oxidative stress and cardiovascular regulation. We evaluated the effects of central catalase inhibition on cardiovascular responses in spontaneously hypertensive rats exposed to sidestream cigarette smoke.

METHODS:

Male Wistar Kyoto (WKY) rats and spontaneously hypertensive rats (SH) (16 weeks old) were implanted with a stainless steel guide cannula leading into the fourth cerebral ventricle (4^th V). The femoral artery and vein were cannulated for arterial pressure and heart rate measurement and drug infusion, respectively. The rats were exposed to sidestream cigarette smoke for 180 minutes/day, 5 days/week for 3 weeks (CO: 100-300 ppm). The baroreflex was tested using a pressor dose of phenylephrine (8 μg/kg, bolus) and a depressor dose of sodium nitroprusside (50 μg/kg, bolus). Cardiovascular responses were evaluated before and 5, 15, 30 and 60 minutes after injection of a catalase inhibitor (3-amino-1,2,4-triazole, 0.001 g/100 μL) into the 4^th V.

RESULTS:

Vehicle administration into the 4^th V did not affect the cardiovascular response, whereas administration of the central catalase inhibitor increased the basal HR and attenuated the bradycardic peak (p<0.05) to a greater extent in WKY rats exposed to sidestream cigarette smoke than in WKY rats exposed to fresh air. However, in spontaneously hypertensive rats, the effect of the catalase inhibitor treatment was stronger in the fresh air condition (p<0.05).

CONCLUSION:

Administration of a catalase inhibitor into the 4^th V combined with exposure to sidestream cigarette smoke has a stronger effect in WKY rats than in SH rats.     

The dorsal motor nucleus of the vagus (DMNV) in sudden infant death syndrome (SIDS): Pathways leading to apoptosis

Sudden infant death syndrome (SIDS) remains the commonest cause of death in the post-neonatal period in the developed world. A leading hypothesis is that an abnormality in the brainstem of infants who succumb to SIDS, either causes or predisposes to failure to respond appropriately to an exogenous stressor. Neuronal apoptosis can lead to loss of cardiorespiratory reflexes, compromise of the infant's ability to respond to stressors such as hypoxia, and ultimately a sleep-related death. The dorsal motor nucleus of the vagus (DMNV) is a medullary autonomic nucleus where abnormalities have regularly been identified in SIDS research. This review collates neurochemical findings documented over the last 30 years, including data from our laboratory focusing on neuronal apoptosis and the DMNV, and provides potential therapeutic interventions targeting neurotransmitters, growth factors and/or genes.     

Inhibitory Effects of Ginsenoside-Rb2 on Nicotinic Stimulation-Evoked Catecholamine Secretion

The aim of the present study was to investigate whether ginsenoside-Rb2 (Rb2) can affect the secretion of catecholamines (CA) in the perfused model of the rat adrenal medulla. Rb2 (3~30 µM), perfused into an adrenal vein for 90 min, inhibited ACh (5.32 mM)-evoked CA secretory response in a dose- and time-dependent fashion. Rb2 (10 µM) also time-dependently inhibited the CA secretion evoked by DMPP (100 µM, a selective neuronal nicotinic receptor agonist) and high K⁺ (56 mM, a direct membrane depolarizer). Rb2 itself did not affect basal CA secretion (data not shown). Also, in the presence of Rb2 (50 µg/mL), the secretory responses of CA evoked by veratridine (a selective Na⁺ channel activator (50 µM), Bay-K-8644 (an L-type dihydropyridine Ca²⁺ channel activator, 10 µM), and cyclopiazonic acid (a cytoplasmic Ca²⁺-ATPase inhibitor, 10 µM) were significantly reduced, respectively. Interestingly, in the simultaneous presence of Rb2 (10 µM) and L-NAME (an inhibitor of NO synthase, 30 µM), the inhibitory responses of Rb2 on ACh-evoked CA secretory response was considerably recovered to the extent of the corresponding control secretion compared with the inhibitory effect of Rb2-treatment alone. Practically, the level of NO released from adrenal medulla after the treatment of Rb2 (10 µM) was greatly elevated compared to the corresponding basal released level. Collectively, these results demonstrate that Rb2 inhibits the CA secretory responses evoked by nicotinic stimulation as well as by direct membrane-depolarization from the isolated perfused rat adrenal medulla. It seems that this inhibitory effect of Rb2 is mediated by inhibiting both the influx of Ca²⁺ and Na⁺ into the adrenomedullary chromaffin cells and also by suppressing the release of Ca²⁺ from the cytoplasmic calcium store, at least partly through the increased NO production due to the activation of nitric oxide synthase, which is relevant to neuronal nicotinic receptor blockade.     

应用GQI技术对小脑髓质的初步研究

目的　应用基于GQI的弥散成像方法,对小脑的纤维束进行重建和显示,为小脑疾病的影像诊断进行解剖学和技术上的探索。  方法　首先把10例脑连接组项目的磁共振弥散数据导入DSI-studio软件,然后依次用该软件中的GQI（Generalized Q-sampling imaging）成像技术和流线算法（streamline algorithm）对与小脑有关的纤维束进行重建和显示。  结果　（1）在DSI-studio软件的二维和三维界面中显示了小脑三对脚的位置和交叉情况;（2）显示了小脑3对脚与脊髓、延髓、脑桥、中脑、端脑等的连接情况及其主要纤维成分。  结论　应用GQI技术可以对小脑白质进行重建和显示,为小脑疾病的影像学诊断提供技术和结构学上的帮助。     

大鼠腰骶髓和延髓星形胶质细胞及神经元对慢性结肠炎的反应

目的探讨大鼠腰骶髓和延髓星形胶质细胞及神经元对慢性结肠炎的反应,及反应性星形胶质细胞和反应性神经元之间的关系.方法成年雄性SD大鼠,实验组（n=17）给予三硝基苯磺酸（TNBS）灌肠诱导结肠炎;对照组（n=16）给予生理盐水灌肠.免疫组织化学法显示大鼠腰骶髓和延髓内胶质原纤维酸性蛋白（GFAP）阳性星形胶质细胞和Fos阳性神经元.结果TNBS灌肠后,GFAP阳性星形胶质细胞主要分布在脊髓背角浅层（Ⅰ～Ⅱ层）、中间外侧核（Ⅴ层）、后连合核（Ⅹ层）和腹角外侧核（Ⅸ层）.Fos阳性神经元集中分布在背角深层（Ⅲ～Ⅳ,Ⅴ～Ⅵ层）.在延髓,两者均主要分布在由孤束核、中间网状带和腹外侧区组成的延髓内脏带（MVZ）.TNBS灌肠后3、7、14 d,脊髓中GFAP阳性细胞密度明显高于对照组（P＜0.05）.TNBS灌肠后3 d,延髓中GFAP阳性细胞密度明显高于对照组（P＜0.05）.TNBS灌肠后28 d,脊髓和延髓中GFAP阳性细胞密度下降,与对照组无显著性差异（P＞0.05）.结论结肠炎性刺激引起脊髓和延髓中星形胶质细胞激活.随着结肠炎的恢复,星形胶质细胞的反应性下降.在延髓内脏带,反应性星形胶质细胞与反应性神经元关系密切.     

心肌缺血对大鼠延髓利钠肽受体表达的影响

 目的:探讨大鼠延髓腹侧部利钠肽受体A(NPR-A)、利钠肽受体C（NPR-C）、胆碱乙酰转移酶（ChAT）和酪氨酸羟化酶（TH）的表达在心肌缺血后不同时程的动态变化。方法:大鼠分为空白对照组、假手术组和模型组。建立心肌缺血模型后,于术后3、7、14、18和28 d取材,用Western blotting方法检测延髓腹侧部吻段、尾段内NPR-A、NPR-C、ChAT和TH表达。结果:在模型复制后的上述时点,延髓腹侧部NPR-A的表达显著升高（P<0.05）,NPR-C表达的升高晚于NPR-A,TH的表达显著低于空白对照组（P<0.05）,在延髓腹侧部吻段ChAT的表达低于空白对照组（P<0.05）,而在尾段与对照组无显著差异。结论:延髓心血管中枢内NPR-A和NPR-C的表达在心肌缺血后均显著升高,而ChAT和TH的表达则显著降低。利钠肽、ChAT和TH可能都参与了心肌缺血后心血管系统的植物神经调控。     

Intermediate expression of CCRL1 reveals novel subpopulations of medullary thymic epithelial cells that emerge in the postnatal thymus

Cortical and medullary thymic epithelial cells (cTECs and mTECs, respectively) provide inductive microenvironments for T‐cell development and selection. The differentiation pathway of cTEC/mTEC lineages downstream of common bipotent progenitors at discrete stages of development remains unresolved. Using IL‐7/CCRL1 dual reporter mice that identify specialized TEC subsets, we show that the stepwise acquisition of chemokine (C–C motif) receptor‐like 1 (CCRL1) is a late determinant of cTEC differentiation. Although cTECs expressing high CCRL1 levels (CCRL1^hi) develop normally in immunocompetent and Rag2^?/?thymi, their differentiation is partially blocked in Rag2^?/?Il2rg^?/? counterparts. These results unravel a novel checkpoint in cTEC maturation that is regulated by the cross‐talk between TECs and immature thymocytes. Additionally, we identify new Ulex europaeus agglutinin 1 (UEA)⁺ mTEC subtypes expressing intermediate CCRL1 levels (CCRL1^int) that conspicuously emerge in the postnatal thymus and differentially express Tnfrsf11a, Ccl21, and Aire. While rare in fetal and in Rag2^?/? thymi, CCRL1^int mTECs are restored in Rag2^?/?Marilyn TCR‐Tg mice, indicating that the appearance of postnatal‐restricted mTECs is closely linked with T‐cell selection. Our findings suggest that alternative temporally restricted routes of new mTEC differentiation contribute to the establishment of the medullary niche in the postnatal thymus.     

Methods of single unit recording from medullary neural substrates in awake, behaving guinea pigs

An electrophysiological procedure that permits extracellular single neuronal recording in the medulla of unanesthetized, freely behaving animals is described. System components consist of 1) a flexible, single-strand microwire as recording electrode; 2) a miniature, skull-mounted miniature microdrive used for isolation of single unit activity; and 3) a head-mounted voltage follower that conditions and stabilizes the neuronal signals prior to amplification and transmission for further processing. A unique advantage of this procedure is the latitude and provision for microwire replacement in the event that multiple penetrations are desired or the tip of the microwire recording electrode is insulated as a result of gliomatosis. With minor modification, this technique can be used for single unit recording virtually anywhere along the supra-segmental neuraxis. Technical aspects of this procedure, together with details of design and fabrication of implantable devices, chronic instrumentation procedure, and potentials for other chronic applications, are discussed.     

吗啡处理对大鼠延髓头端腹内侧区的L-型电压门控钙通道蛋白与电流的影响

[目的] 探讨在吗啡引起的痛觉超敏状态下,SD大鼠延髓头端腹内侧区电压门控钙通道(voltage-gated calcium channels, VGCCs)蛋白的表达变化和功能变化.[方法] 采用western blot方法检测α1C和α1H亚基蛋白在大鼠延髓头端腹内侧区(the rostral ventromedial medulla,RVM)的表达,采用脑片全细胞膜片钳技术记录大鼠RVM神经元的VGCCs电流.[结果] 在吗啡诱导的痛觉超敏状态下,SD大鼠RVM的α1C亚单位蛋白表达显著增加,同时神经元的VGCCs总电流和L-型VGCCs电流也显著增加.[结论] 延髓RVM的L-型电压门控钙通道的可塑性变化,可能是导致吗啡痛觉超敏的重要原因.