Labra Antonieta, Brann Jessica H, Fadool Debra A
Department of Biological Science, Programs in Neuroscience and Molecular Biophysics, Florida State University, Tallahassee, 32306, USA.
J Neurophysiol. 2005 Oct;94(4):2535-48. doi: 10.1152/jn.00490.2005. Epub 2005 Jun 22.
Liolaemus lizards were explored to ascertain whether they would make an amenable model to study single-cell electrophysiology of neurons in the vomeronasal organ (VNO). Despite a rich array of chemosensory-related behaviors chronicled for this genus, no anatomical or functional data exist for the VNO, the organ mediating these types of behaviors. Two Liolaemus species (L. bellii and L. nigroviridis) were collected in Central Chile in the Farellones Mountains and transported to the United States. Lizards were subjected to hypothermia and then a lethal injection of sodium pentabarbitol prior to all experiments described in the following text. Retrograde dye perfusion combined with histological techniques demonstrated a compartmentalization of the proportionally large VNO from the main olfactory epithelium (MOE) in cryosections of L. bellii. SDS-PAGE analysis of the VNO of both species demonstrated the expression of three G protein subunits, namely, G(alphao), G(alphai2), and G(beta), and the absence of G(alphaolf), G(alpha11), and G(q), the latter of which are traditionally found in the MOE. Vomeronasal (VN) neurons were enzymatically isolated for whole cell voltage-clamp electrophysiology of single neurons. Both species demonstrated a tetrodotoxin (TTX)-sensitive, rapidly inactivating sodium current and a tetraethylammonium (TEA)-sensitive potassium current that had a transient and sustained component. VN neurons were classified into two types dependent on the ratio of sodium over sustained potassium current. VN neurons exhibited outward and inward chemosignal-evoked currents when stimulated with pheromone-containing secretions taken from the feces, skin, and precloacal pores. Fifty-nine percent of the neurons were responsive to at least one compound when presented with a battery of five different secretions. The breadth of responsiveness (H metric) demonstrated a heterogeneous population of tuning with a mean of 0.29.
对鬃狮蜥进行了研究,以确定它们是否会成为研究犁鼻器(VNO)中神经元单细胞电生理学的合适模型。尽管已记录了该属丰富多样的与化学感应相关的行为,但对于介导此类行为的器官——犁鼻器,尚无解剖学或功能数据。在智利中部的法雷洛内斯山脉采集了两种鬃狮蜥(贝利鬃狮蜥和绿黑鬃狮蜥),并运往美国。在进行以下文本所述的所有实验之前,先对蜥蜴进行低温处理,然后注射致死剂量的戊巴比妥钠。逆行染料灌注结合组织学技术表明,在贝利鬃狮蜥的冰冻切片中,相对较大的犁鼻器与主嗅觉上皮(MOE)是分隔开的。对这两个物种的犁鼻器进行的SDS-PAGE分析表明,有三种G蛋白亚基表达,即G(alphao)、G(alphai2)和G(beta),而传统上在主嗅觉上皮中发现的G(alphaolf)、G(alpha11)和G(q)则未表达。通过酶解法分离出犁鼻器(VN)神经元,用于单个神经元的全细胞电压钳电生理学研究。两个物种都表现出对河豚毒素(TTX)敏感、快速失活的钠电流以及对四乙铵(TEA)敏感的钾电流,该钾电流有一个瞬态和持续成分。VN神经元根据钠电流与持续钾电流的比值分为两种类型。当用取自粪便、皮肤和泄殖腔前孔的含信息素分泌物刺激时,VN神经元表现出外向和内向的化学信号诱发电流。当呈现一组五种不同分泌物时,59%的神经元对至少一种化合物有反应。反应广度(H指标)表明存在一个调谐各异的群体,平均值为0.29。