Stepanyan Ruben, Day Kristen, Urban Jason, Hardin Debra L, Shetty Ranjit S, Derby Charles D, Ache Barry W, McClintock Timothy S
Department of Physiology, Cellular and Molecular Neuroscience of Sensory Systems Training Program, University of Kentucky, Lexington, Kentucky 40536-0298, USA.
Physiol Genomics. 2006 Apr 13;25(2):224-33. doi: 10.1152/physiolgenomics.00276.2005.
The lobster olfactory organ is an important model for investigating many aspects of the olfactory system. To facilitate study of the molecular basis of olfaction in lobsters, we made a subtracted cDNA library from the mature zone of the olfactory organ of Homarus americanus, the American lobster. Sequencing of the 5'-end of 5,184 cDNA clones produced 2,389 distinct high-quality sequences consisting of 1,944 singlets and 445 contigs. Matches to known sequences corresponded with the types of cells present in the olfactory organ, including specific markers of olfactory sensory neurons, auxiliary cells, secretory cells of the aesthetasc tegumental gland, and epithelial cells. The wealth of neuronal mRNAs represented among the sequences reflected the preponderance of neurons in the tissue. The sequences identified candidate genes responsible for known functions and suggested new functions not previously recognized in the olfactory organ. A cDNA microarray was designed and tested by assessing mRNA abundance differences between two of the lobster's major chemosensory structures: the mature zone of the olfactory organ and the dactyl of the walking legs, a taste organ. The 115 differences detected again emphasized the abundance of neurons in the olfactory organ, especially a cluster of mRNAs encoding cytoskeletal-associated proteins and cell adhesion molecules such as 14-3-3zeta, actins, tubulins, trophinin, Fax, Yel077cp, suppressor of profilin 2, and gelsolin.
龙虾嗅觉器官是研究嗅觉系统诸多方面的重要模型。为便于研究龙虾嗅觉的分子基础，我们从美洲龙虾（Homarus americanus）嗅觉器官的成熟区构建了一个消减cDNA文库。对5184个cDNA克隆的5'端进行测序，产生了2389个不同的高质量序列，包括1944个单序列和445个重叠群。与已知序列的匹配对应于嗅觉器官中存在的细胞类型，包括嗅觉感觉神经元、辅助细胞、触角表皮腺分泌细胞和上皮细胞的特异性标记物。序列中丰富的神经元mRNA反映了该组织中神经元的优势。这些序列鉴定出了负责已知功能的候选基因，并提示了嗅觉器官中以前未被认识的新功能。通过评估龙虾两个主要化学感觉结构之间的mRNA丰度差异，设计并测试了一个cDNA微阵列：嗅觉器官的成熟区和步行足的指节，一个味觉器官。检测到的115个差异再次强调了嗅觉器官中神经元的丰富性，特别是一组编码细胞骨架相关蛋白和细胞粘附分子的mRNA，如14-3-3ζ、肌动蛋白、微管蛋白、滋养素、Fax、Yel077cp、profilin 2抑制因子和凝溶胶蛋白。
