Miampamba M, Tan D T, Oliver M R, Sharkey K A, Scott R B
Gastrointestinal Group, Faculty of Medicine, University of Calgary, Alberta, Canada.
Am J Physiol. 1997 Jan;272(1 Pt 1):G181-9. doi: 10.1152/ajpgi.1997.272.1.G181.
Fos immunohistochemistry was used to identify myenteric neurons activated as a consequence of intestinal anaphylaxis in Hooded-Lister rats sensitized to egg albumin (EA 10 micrograms ip). After incubation in test solutions, or after in vivo challenge, jejunal tissues were processed for immunohistochemistry with an anti-Fos antibody (1:500, TF161). The neuronal identity of the Fos-labeled nuclei was confirmed by double labeling with neuron-specific enclose (1:1,000). In in vitro studies, exposure of control tissue to 50 mM K(+)-Krebs-EA (2 x 10(-5) M) solutions significantly increased Fos immunoreactivity in the myenteric plexus, whereas a basal level of Fos was seen in control tissue incubated in Krebs solution, sham-sensitized tissue exposed to bovine serum albumin (BSA, 2 x 10(-5) M), or EA and sensitized tissue exposed to BSA. Pretreatment of sensitized tissue with doxantrazole (10(-4) M) markedly reduced Fos immunoreactivity observed after EA exposure. In in vivo studies, there was negligible Fos immunoreactivity in the myenteric plexus of control, sham-sensitized, or sensitized rats challenged with saline. A low level of Fos was seen in neurons of sham-sensitized rats challenged with BSA or EA and in sensitized rats challenged with BSA. Significantly greater levels of Fos were observed in the myenteric plexus of sensitized animals challenged with EA, even after pretreatment with capsaicin (125 mg/kg). These results suggest a role for myenteric neurons in intestinal anaphylaxis. In sensitized rats, activation of myenteric neurons is dependent on antigen-induced mast cell activation and occurs independently of capsaicin-sensitive afferent nerves.
采用Fos免疫组织化学法鉴定对卵清蛋白(10微克腹腔注射)致敏的帽状利斯特大鼠因肠道过敏反应而被激活的肌间神经元。在试验溶液中孵育后,或在体内激发后,用抗Fos抗体(1:500,TF161)对空肠组织进行免疫组织化学处理。通过与神经元特异性烯醇化酶(1:1000)双重标记来确认Fos标记细胞核的神经元身份。在体外研究中,将对照组织暴露于50 mM K(+)-Krebs-EA(2×10(-5) M)溶液中可显著增加肌间神经丛中的Fos免疫反应性,而在Krebs溶液中孵育的对照组织、暴露于牛血清白蛋白(BSA,2×10(-5) M)的假致敏组织或暴露于BSA的致敏组织中可见基础水平的Fos。用多克唑(10(-4) M)预处理致敏组织可显著降低EA暴露后观察到的Fos免疫反应性。在体内研究中,用盐水激发的对照、假致敏或致敏大鼠的肌间神经丛中Fos免疫反应性可忽略不计。用BSA或EA激发的假致敏大鼠以及用BSA激发的致敏大鼠的神经元中可见低水平的Fos。在用EA激发的致敏动物的肌间神经丛中观察到显著更高水平的Fos,即使在用辣椒素(125 mg/kg)预处理后也是如此。这些结果表明肌间神经元在肠道过敏反应中起作用。在致敏大鼠中,肌间神经元的激活依赖于抗原诱导的肥大细胞激活,且独立于辣椒素敏感的传入神经而发生。