大鼠中缝隐核增强dPAG诱导的vPAG的c－Fos表达及对vPAG神经元放电的影响

本实验通过Ｐｏｓ免疫细胞化学、电生理及微量注射法对中缝隐核（ＮＲＯ）的交感抑制作用的相关途径进行探讨。实验在成巴比妥钠或α－氯醛糖和氨基甲酸乙脂麻醉的Ｓｐｒａｇｕｅ－Ｄａｗｌｅｙ（ＳＤ）大鼠上进行。同时予以ＮＲＯ,中脑导水管周围灰质背侧部（ｄＰＡＧ）方波脉冲串刺激,诱导中脑和延髓的ｃ－Ｆｏｓ表达。刺激ＮＲＯ过程中,基础血压升高（Ｐ＜０．０５）,刺激ｄＰＡＧ引起的防御性升压反应则减少（Ｐ＜０．０１）;中脑导水管周围灰质腹侧部（ｖＰＡＧ）、巨细胞旁外侧核（ＰＧＬ）的Ｆｏｓ样免疫阳性反应（ＦＬＩ）细胞计数分别为６６．５±８．３和１０．８±１．５（刺激ＮＲＯ＋ｄＰＡＧ组）,较单独刺激ｄＰＡＧ组明显增加,Ｐ值分别小于０．０１和０．００１;单或双脉冲刺激中缝隐核在ｖＰＡＧ可以记录到相关单位,其中８４％为兴奋单位,抑制单位占１６％。双侧ｖＰＡＧ内微量注射利多卡因（每侧２μｇ／０．１μｌ）,基础血压无明显变化,而刺激ＮＲＯ引起的降压反应幅度减小（Ｐ＜０．０１）,提示,延髓腹外侧区（ＶＬＭ）、ＮＲＯ存在不同功能分化的神经元;ＮＲＯ可能有向ｖＰＡＧ的兴奋性投射,此投射可加强ＮＲＯ的交感抑制效应。     

神经激肽A注入大鼠外侧网状核或中缝大核产生的镇痛效应

本工作采用核团内微量注射的方法,以甩尾反射潜伏期（ＴＦＬ）为痛阈指标,在浅麻状态下的大鼠上,对神经激肽Ａ（ＮＫＡ）在外侧网状核（ＬＲＮ）和中缝大核（ＮＲＭ）中的作用作了探讨。研究结果表明,ＮＫＡ（０．５μｇ／０．５μｌ）注入ＬＲＮ后的１０ｍｉｎ内大鼠ＴＦＬ明显延长（ｎ＝１２,Ｐ＜０．００１）,同样剂量ＮＫＡ注入ＮＲＭ后的５ｍｉｎ内,ＴＦＬ也明显延长（ｎ＝１３,Ｐ＜０．００１）,提示ＮＫＡ可能是ＬＲＮ和ＮＲＭ内参与痛觉的一种神经调质。     

促肾上腺皮质激素对慢痛大鼠脊髓生长抑素、c-fos表达及痛反应的影响

本研究应用免疫组化、原位杂交和痛级均数测定法,探讨鞘内注射促肾上腺皮质激素（ＡＣＴＨ）对甲醛引起大鼠脊髓内生长抑素（Ｓｏｍ）、ｃｆｏｓ表达及痛反应的影响。结果表明,足底注射甲醛可使大鼠脊髓内ｃｆｏｓ样免疫反应（ＦＬＩ）、Ｓｏｍ样免疫反应（ＳｏｍＬＩ）、ＦＬＩ／ＳｏｍＬＩ及前Ｓｏｍ原ｍＲＮＡ（ＰＰＳｍＲＮＡ）神经元数目显著增多以及痛级均数（ＰＩＲ）显著升高。而鞘内注射ＡＣＴＨ可显著抑制甲醛引起的大鼠脊髓内ＦＬＩ、ＳｏｍＬＩ、ＦＬＩ／ＳｏｍＬＩ及ＰＰＳｍＲＮＡ增多和ＰＩＲ升高效应。鞘内预先注射赛庚啶可阻断ＡＣＴＨ的抑制效应,而荷包牡丹碱、纳洛酮则无影响。结果提示,５羟色胺受体可能参与ＡＣＴＨ抑制甲醛引起的痛反应。     

谷氨酸转运体GLAST mRNA在大鼠脑内表达的细胞定位

实验采用荧光双标技术研究谷氨酸转运体ＧＬＡＳＴ ｍ ＲＮＡ 在大鼠脑内表达的细胞定位, 研究表明, 在星形神经胶质细胞和神经元, ＧＬＡＳＴｍ ＲＮＡ 分别与神经胶质纤维蛋白（ＧＦＡＰ） 和神经元特异性烯醇化酶 （ＮＳＥ） 有表达共存, 提示ＧＬＡＳＴ ｍ ＲＮＡ在星形神经胶质细胞和神经元上都有表达。     

神经激肽A注入大鼠外侧风状核或中缝大核产生的镇痛效应

本工作采用核团内微量注射的方法,以甩尾反向潜伏期（ＴＦＬ）为痛阈指标,在线麻状态下的大鼠上,对神经激肽Ａ（ＮＫＡ）在我侧网状核（ＬＲＮ）和中缝大核（ＮＲＭ）中的作用作了探讨。研究结果表明,ＮＫＡ（０．５μｇ／０．５μｌ）注入ＬＲＮ后的１０ｍｉｎ内大鼠ＴＦＬ明显延长（ｎ＝１２,Ｐ〈０．００１）,同样剂量ＮＫＡ注入ＮＲＭ后的５ｍｉｎ内,ＦＴＬ也明显延长ｎ＝１３,Ｐ〈０．００１）,提示ＮＫＡ可能是ＬＲＮ     

中华姬鼠种群生态的初步分析

中华姬鼠种群生态的初步分析ＴＨＥＰＲＥＬＩＭＩＮＡＲＹＡＮＡＬＹＳＩＳＯＮＴＨＥＰＯＰＵＬＡＴＩＯＮＥＣＯＬＯＧＹＯＦＣＨＩＮＥＳＥＦＩＥＬＤＭＯＵＳＥ（ＡＰＯＤＥＭＵＳＤＲＡＣＯ）ＫｅｙＷｏｒｄｓＣｈｉｎｅｓｅＦｉｅｌｄＭｏｕｓｅ（Ａｐｏｄｅｍｕｓ...     

大鼠延髓网状背侧亚核内5-羟色胺能、P物质样和脑啡肽样阳性终末的中枢起源

研究用荧光金（ＦＧ）逆行追踪与免疫荧光组化染色相结合的双标技术对大鼠脑干向延髓网状背侧亚核（ＳＲＤ）的５┐羟色胺（５┐ＨＴ）能、Ｐ物质（ＳＰ）能和亮氨酸┐脑啡肽（Ｌ┐ＥＮＫ）能投射进行了观察。将ＦＧ注入ＳＲＤ后,ＦＧ逆标神经元主要见于中脑导水管周围灰质、脑干中缝核簇（中缝背核、中缝正中核、中缝桥核、中缝大核、中缝隐核和中缝苍白核）、巨细胞网状核α部、延髓网状结构的内侧部和外侧部、延髓外侧网状核、三叉神经脊束核尾侧亚核和孤束核。５┐羟色胺（５┐ＨＴ）样、Ｐ物质（ＳＰ）样和亮氨酸脑啡肽（Ｌ┐ＥＮＫ）样阳性神经元主要见于中脑导水管周围灰质、脑干中缝核簇和巨细胞网状核α部;此外,ＳＰ样和Ｌ┐ＥＮＫ样阳性神经元还见于臂旁核、背外侧被盖核和孤束核。ＦＧ逆标并呈５┐ＨＴ样、ＳＰ样或Ｌ┐ＥＮＫ样阳性的双标神经元也主要见于中脑导水管周围灰质、脑干中缝核簇和巨细胞网状核α部,尤其是位于延髓中缝核团内的双标神经元数量较多。本研究的结果说明ＳＲＤ内的５┐ＨＴ样、ＳＰ样和Ｌ┐ＥＮＫ样阳性终末主要来自中脑导水管周围灰质、脑干中缝核簇和巨细胞网状核α部,向ＳＲＤ发出５┐ＨＴ能、ＳＰ能和Ｌ┐ＥＮＫ能投射的上述核团对ＳＲＤ发挥“弥漫性伤害抑     

脑内血管紧张素Ⅱ系统在穹窿下器升压反应中的作用

文献报道脑内存在血管紧张素Ⅱ系统。与此一致,本工作用氨基甲酸乙脂麻醉、箭毒制动、人工呼吸的大鼠观察到：（１）穹窿下器（ＳＦＯ）、室旁核（ＮＰＶ）或ＮＰＶ的投射区：延髓头端腹外侧区（ＲＶＬＭ）、导水管周围灰质（ＰＡＧ）、蓝斑（ＬＣ）内注入血管紧张素Ⅱ（ＡⅡ）均引起升压反应;（２）ＳＦＯ升压反应可被双侧ＮＰＶ或ＲＶＬＭ内预先注入［Ｓａｒ１,Ｔｈｒ８］ＡⅡ（ＳＴＡⅡ,ＡⅡ拮抗剂）明显衰减,ＮＰＶ升压反应也可被ＲＶＬＭ内注入ＳＴＡⅡ削弱;（３）双侧ＰＡＧ用ＳＴＡⅡ预处理后,ＡⅡ引起的ＮＰＶ或ＳＦＯ升压反应均明显减小;（４）ＮＰＶ升压反应还可被双侧ＬＣ内预先注射ＳＴＡⅡ衰减,但ＳＦＯ升压反应不受影响。结合我们以往工作曾显示兴奋ＰＡＧ或ＬＣ均可作用于ＲＶＬＭ引起升压反应,目前的结果表明：ＳＦＯ内的ＡⅡ能神经元通过ＮＰＶ内ＡⅡ能神经元,不仅可直接作用于ＲＶＬＭ引起升压反应,而且还可间接通过ＰＡＧ作用于ＲＶＬＭ起升压作用,但ＬＣ不参与ＳＦＯ升压反应。     

福尔马林致痛后大鼠中脑导水管周围灰质甲啡肽及脊髓背角P物质含量的变化

本文用免疫组化方法结合计算机图像处理技术观察大鼠后脚掌皮下注射福尔马林后脊髓背角Ｐ物质免疫阳性反应（ＳＰＬＩ）变化的节段性分布及中脑导水管周围灰质（ＰＡＧ）内甲啡肽样免疫阳性反应（ＭＥＬＩ）的变化。结果显示,注射福尔马林后,脊髓腰段（Ｌ１－２,Ｌ４－５）背角ＳＰＬＩ显著增强（Ｐ＜．０５）,３０ｍｉｎ组与６０ｍｉｎ组相比较无显著变化（Ｐ＞０．０５）;胸脊髓（Ｔ８）无显著变化（Ｐ＞０．０５）;颈脊髓背角ＳＰＬＩ有增强趋势（０．０５＜Ｐ＜０．１）;ＰＡＧ中ＭＥＬＩ减弱,腹外侧部３０ｍｉｎ组比６０ｍｉｎ组变化更大（Ｐ＜０．０５）。ＰＡＧ中ＭＥＬＩ与脊髓背角ＳＰＬＩ变化的时相关系提示福尔马林致痛引起的脊髓背角Ｐ物质的增多可能与ＰＡＧ中甲啡肽及阿片受体活动有关。     

EXPRESSION OF cfos IN THE MEDULLA OBLONGATA AFTER CAROTID BARORECEPTOR ACTIVATION BYELEVATED INTRASINUS PRESSURE AND ADENOSINE

在１４只隔离灌流颈动脉窦区的大鼠,观察了窦内压（ＩＳＰ）升高和灌流腺苷（ａｄｅｎｏｓｉｎｅ,Ａｄｏ）激活压力感受器时延髓内ｃｆｏｓ蛋白的表达。结果显示：在孤束核、最后区、延髓腹外侧头端区和中缝苍白核可见Ｆｏｓ蛋白样免疫阳性反应（ＦＬＩ）神经元分布,且其数量随ＩＳＰ升高而增多。在给定ＩＳＰ下,颈动脉窦内灌流Ａｄｏ,可使上述区域中ＦＬＩ表达明显增多。根据以上结果,得出如下结论：ｃｆｏｓ在压力感受器反射延髓通路中的表达,可由ＩＳＰ增高和灌流Ａｄｏ而增强,表明Ａｄｏ对压力感受器反射有易化作用。     

颈动脉注射辣椒素对脑干心血管相关核团一氧化氮合酶和Fos表达的影响

实验采用NADPH－d组化技术和Fos蛋白免疫组化技术相结合的方法,观察了颈动脉注射辣椒不时,大鼠脑干心血管相关核团内NOS和Fos蛋白的分布以及两者的共存关系。结果显示：（1）颈动脉注射辣椒不可诱发脑干中最后区（AP）、孤束核（NTS）、巨细胞旁外侧核（PGL）和蓝斑（LC）等多个部位Fos样免疫反应（FLI)神经元显著增加 中脑中央灰质（PAG）和中缝核群（RN）的FLI神经元无明显改变。（2）PGL和NTS内NO合成神经元以及PGL内双标神经元数量也明显增加,而AG和RN中NO合成神经元无明显变化,在LC和AP仅偶见或未见NO合成神经元。（3）预先应用辣椒素受体阻断剂钌红或NMDA受体阻断剂MK－801,则明显减弱辣椒素的上述效应,以上结果表明,颈动脉注射辣椒素可兴奋脑干心血管活动相关核团神经元,NO在脑干核团对辣椒素的反应中发挥间接的调制作用,辣椒素的效应由香草酸受体（辣椒素受体）介导并有谷氨酸参与。     

辣椒素对大鼠延髓腹外侧头端区神经元电活动的影响

在35只切断两侧缓冲神经的麻醉大鼠,应用细胞外记录的电生理学方法,观察颈总动脉注射辣椒素(capsaicin)对延髓腹外侧头端区(RVLM)巨细胞旁外侧核(PGL)自发电活动的影响。所得结果如下:(1)颈动脉注射辣椒素(10μmol,01ml),MAP由1074±013升至1256±021kPa(P<0001);HR由374±4增至395±5bpm(P<0001);30个PGL神经元自发放电单位的放电频率由126±07增至209±11spikes/s(P<0001)。(2)在10个放电单位,应用辣椒素受体阻断剂钌红(rutheniumred;200mmol,01ml)后,明显抑制辣椒素的上述效应。以上结果提示,辣椒素可能通过激活RVLM神经元上的辣椒素受体,进而兴奋PGL神经元     

Brainstem neuronal populations activated in the model of ovalbumine induced allergic rhinitis in guinea pigs — the c-Fos study

Central neuronal interactions may contribute to up regulation of cough in subjects with rhinitis. Previously we have shown that noxious stimulation of the nose induces considerable Fos-like immunoreactivity (FLI) in the solitary nuclei and the region of ventral respiratory group and these neurons are involved in the cough pattern generator. Recent study addressed the question, which additional groups are activated in model of trigeminal hyperresponsiveness and whether some of them might also cooperate with cough pattern gating areas. 24 guinea pigs were sensitized with intraperitoneal ovalbumin (OVA) and later were once weekly challenged with intranasal OVA to develop the neural hyperresponsiveness, 12 animals were left unsensitized. The nasal symptom score was evaluated after each challenge. Finally, animals were anaesthetized and the latest challenges with nasal OVA, capsaicin and saline were applied to induce c-Fos expression in designed groups. Following the survival time animals were deeply anaesthetized, exsanguinated, and transcardially perfused with heparinised saline (200 ml) and paraformaldehyde fixation (200 ml). The brainstems were removed, postfixed, and brainstem slices were processed immunohistochemically (c-Fos, Calbiochem, SR). FLI at the level of obex and areas relative to the obex was analyzed. In all groups (excluding the saline group) the FLI was detected bilaterally in the trigeminal complex, nuclei of solitary tract, lateral reticular and nucleus ambiguus. There were no differences between the OVA and capsaicin groups. Count of Fos-positive neurons within the trigeminal complex does not correlate with the magnitude of clinical symptoms, which gradually increased each week in OVA induced model of hyperresponsiveness. Whereas trigeminal hyperresponsiveness contributes to the up-regulation of cough in animal models, it does not induce any additional neuronal FLI at the middle medulla than observed in naive animals.     

心外膜应用腺苷时c—fos在脊髓延髓和丘脑中的表达

在１２只切断两侧缓冲神经和迷走神经的麻醉大鼠,观察了心外膜应用腺苷对脊髓,延髓和丘脑ｃ－ｆｏｓ原部基因表达的影响。结果显示：心外膜应用腺苷组大鼠,动脉血压和心率无明显变化;脊髓Ｔ３节段背角,延髓巨细胞旁外侧核以及丘脑的腹后外侧核,后核,中央外侧核和束旁核等部位Ｆｏｓ蛋白样免疫阳性反应神经元显著增加;而在溶剂对照组大鼠,仅见少数ＦＬＩ细胞。     

c-Fos expression in the midbrain periaqueductal gray during static muscle contraction

The periaqueductal gray (PAG) of the midbrain is involved in the autonomic regulation of the cardiovascular system. The purpose of this study was to determine if static contraction of the skeletal muscle, which increases arterial blood pressure and heart rate, activates neuronal cells in the PAG by examining Fos-like immunoreactivity (FLI). Muscle contraction was induced by electrical stimulation of the L7 and S1 ventral roots of the spinal cord in anesthetized cats. An intravenous infusion of phenylephrine (PE) was used to selectively activate arterial baroreceptors. Extensive FLI was observed within the ventromedial region (VM) of the rostral PAG, the dorsolateral (DL), lateral (L), and ventrolateral (VL) regions of the middle and caudal PAG in barointact animals with muscle contractions, and in barointact animals with PE infusion. However, muscle contraction caused a lesser number of FLI in the VM region of the rostral PAG, the DL, L, and VL regions of the middle PAG and the L and VL regions of the caudal PAG after barodenervation compared with barointact animals. Additionally, the number of FLI in the DL and L regions of the middle PAG was greater in barodenervated animals with muscle contraction than in barodenervated control animals. Thus these results indicated that both muscle receptor and baroreceptor afferent inputs activate neuronal cells in regions of the PAG during muscle contraction. Furthermore, afferents from skeletal muscle activate neurons in specific regions of the PAG independent of arterial baroreceptor input. Therefore, neuronal cells in the PAG may play a role in determining the cardiovascular responses during the exercise pressor reflex.     

Nitric oxide synthase (NOS) coexists with activated neurons by skeletal muscle contraction in the brainstem of cats

Contraction of skeletal muscle evokes increases in arterial blood pressure and heart rate. Some regions of the brainstem have been implicated for expression of the cardiovascular responses to muscle contraction. Previous studies have reported that static muscle contraction induced c-Fos protein in the nucleus of tractus solitarii (NTS), lateral reticular nucleus (LRN), lateral tegmental field (FTL), subretrofacial nucleus (SRF), A1 region and periaqueductal gray (PAG) of the brainstem. Furthermore, neuronal NADPH-diaphorase (NADPH-d), which is considered as a marker of neuronal nitric oxide synthase (nNOS), has been localized in those same regions. In this study, static muscle contraction was induced by electrical stimulation of the L7 and S1 ventral roots in anaesthetized cats. Distribution of c-Fos protein within neurons containing nNOS was evaluated by double labeling methods in order to determine if nNOS containing neurons in the brainstem were activated during muscle contraction. The results indicate that c-Fos protein colocalized with NADPH-d positive staining within the neurons of the SRF and PAG, but not within the NTS neurons. Distinct number of neurons with c-Fos protein was in close proximity to NADPH-d positive staining in the NTS, SRF, and PAG. Coexisting of c-Fos protein and NADPH-d positive staining was not observed in the LRN, FTL and A1 region. These findings demonstrate that nNOS containing neurons were activated by muscle contraction in the selective regions of the brainstem, and nNOS positive staining had close anatomic contacts with the neurons activated by contraction. This result provides neuroanatomic evidence suggesting that nitric oxide modulates the cardiovascular responses to muscle contraction within the NTS, SRF and PAG of the brainstem.     

Hemodynamic responses and c-Fos changes associated with hypotensive hemorrhage: standardizing a protocol for severe hemorrhage in conscious rats

The central mechanisms underlying the transition from compensation to decompensation during severe hemorrhage (HEM) are poorly understood. Furthermore, a lack of consistency in HEM protocols exists in the current literature. This study assessed the cardiovascular response and Fos-like immunoreactivity (FLI) in specific brain regions following severe HEM at three rates (2, 1, or 0.5 ml.kg(-1).min(-1)) in conscious rats. Heart rate (HR) and arterial pressure were recorded during the withdrawal of 30% of total blood volume (TBV). Data from animals hemorrhaged at the fast (F-HEM, n = 6), intermediate (I-HEM, n = 7), or slow (S-HEM, n = 7) rates were compared with saline (SAL, n = 5) and hypotensive (hydrazaline-induced, HYDRAZ, n = 5) controls. All HEM rates produced similar degrees of hypotension at the time of 30% TBV withdrawal. All HEM rates also produced bradycardia, but the change in HR was only significant in the F-HEM and I-HEM groups. Associated with I-HEM and F-HEM, but not HYDRAZ treatment were significant increases in FLI in the caudal ventrolateral periaqueductal gray (PAG), the central lateral nucleus of the rostral parabrachial nucleus, and locus coeruleus compared with SAL treatment. I-HEM also induced significant increases in FLI in the dorsomedial PAG, A7 region, and the cuneiform nucleus compared with SAL. S-HEM did not induce any significant change in FLI. Our results suggest that HEM at a rate of 1 ml.kg(-1).min(-1) may be most useful for investigating the potential role of the rostral brainstem regions in mediating hemorrhagic decompensation in conscious rats.     

Different pharmacological anatomy in the paraventricular hypothalamic nucleus, supraoptic nucleus, and suprachiasmatic nucleus of rats: quantitative autoradiography on angiotensin II receptor binding sites

Angiotensin II (AII) and vasopressin (VP) play important roles in cardiovascular function. Using 125I-[Sar1,Ile8]-angiotensin II (125I-SI-AII), a potent AII antagonist, AII receptor binding sites were autoradiographically localized in three VP-producing areas of the hypothalamus and compared in hypertensive and normotensive rats. Within three major VP-producing areas, AII receptor binding was highest in the paraventricular hypothalamic nucleus and lowest in the supraoptic nucleus, suggesting that a differential AII regulation of separate VP systems exists in the brainstem. No statistical difference in 125I-SI-AII receptor binding was found between WKY and SHR rats in each of the three major VP-producing nuclei studied. These results are consistent with a role of AII receptors in a subtle and complicated regulation of VP in cardiovascular function.     

Brainstem areas involved in the aspiration reflex: c-Fos study in anesthetized cats

Expression of the immediate-early gene c-fos, a marker of neuronal activation was employed in adult anesthetized non-decerebrate cats, in order to localize the brainstem neuronal populations functionally related to sniff-like (gasp-like) aspiration reflex (AR). Tissues were immunoprocessed using an antibody raised against amino acids of Fos and the avidin-biotin peroxidase complex method. The level of Fos-like immunoreactivity (FLI) was identified and counted in particular brainstem sections under light microscopy using PC software evaluations in control, unstimulated cats and in cats where the AR was elicited by repeated mechanical stimulation of the nasopharyngeal region. Fourteen brainstem regions with FLI labeling, including thirty-seven nuclei were compared for the number of labeled cells. Compared to the control, a significantly enhanced FLI was determined bilaterally in animals with the AR, at various medullary levels. The areas included the nuclei of the solitary tract (especially the dorsal, interstitial and ventrolateral subnuclei), the ventromedial part of the parvocellular tegmental field (FTL -- lateral nuclei of reticular formation), the lateral reticular nucleus, the ambigual and para-ambigual regions, and the retrofacial nucleus. FLI was also observed in the gigantocellular tegmental field (FTG -- medial nuclei of reticular formation), the spinal trigeminal nucleus, in the medullar raphe nuclei (ncl. raphealis magnus and parvus), and in the medial and lateral vestibular nuclei. Within the pons, a significant FLI was observed bilaterally in the parabrachial nucleus (especially in its lateral subnucleus), the Kolliker-Fuse nucleus, the nucleus coeruleus, within the medial region of brachium conjunctivum, in the ventrolateral part of the pontine FTG and the FTL. Within the mesencephalon a significantly enhanced FLI was found at the central tegmental field (area ventralis tegmenti Tsai), bilaterally. Positive FLI found in columns extending from the caudal medulla oblongata, through the pons up to the mid-mesencephalon suggests that the aspiration reflex is thus co-ordinated by a long loop of medullary-pontine-mesencephalic control circuit rather than by a unique "center".