Bueno D, Baguñà J, Romero R
Department de Genètica, Facultat de Biologia, Universitat de Barcelona, Catalonia, Spain.
Histochem Cell Biol. 1997 Feb;107(2):139-49. doi: 10.1007/s004180050098.
To obtain specific immunological probes for studying molecular mechanisms involved in cell renewal, cell differentiation, and pattern formation in intact and regenerating planarians, we have produced a hybridoma library specific for the asexual race of the fresh-water planarian Dugesia (Girardia) tigrina. Among the 276 monoclonal antibodies showing tissue-, cell-, cell subtype-, subcellular- and position-specific staining, we have found monoclonal antibodies against all tissues and cell types with the exception of neoblasts, the undifferentiated totipotent stem-cells in planarians. We have also detected position-specific antigens that label anterior, central, and posterior regions. Patterns of expression uncovered an unexpected heterogeneity among previously thought single cell types, as well as interesting cross-reactivities that deserve further study. Characterization of some of these monoclonal antibodies suggests they may be extremely useful as molecular markers for studying cell renewal and cell differentiation in the intact and regenerating organism, tracing the origin, lineage, and differentiation of blastema cells, and characterizing the stages and mechanisms of early pattern formation. Moreover, two position-specific monoclonals, the first ones isolated in planarians, will be instrumental in describing in molecular terms how the new pattern unfolds during regeneration and in devising the pattern formation model that best fits classical data on regeneration in planarians.
为了获得用于研究完整和再生涡虫中细胞更新、细胞分化及模式形成所涉及分子机制的特异性免疫探针,我们构建了一个针对淡水涡虫杜氏(吉氏)虎纹涡虫无性繁殖品系的杂交瘤文库。在276种显示出组织、细胞、细胞亚型、亚细胞及位置特异性染色的单克隆抗体中,我们发现了针对除新生细胞(涡虫中未分化的全能干细胞)之外的所有组织和细胞类型的单克隆抗体。我们还检测到了标记前、中、后区域的位置特异性抗原。表达模式揭示了先前认为的单一细胞类型中存在意外的异质性,以及值得进一步研究的有趣交叉反应性。对其中一些单克隆抗体的表征表明,它们可能作为分子标记物非常有用,可用于研究完整和再生生物体中的细胞更新和细胞分化、追踪芽基细胞的起源、谱系和分化,以及表征早期模式形成的阶段和机制。此外,两种位置特异性单克隆抗体是在涡虫中首次分离得到的,它们将有助于从分子层面描述再生过程中新模式如何展开,并有助于设计最符合涡虫再生经典数据的模式形成模型。