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通过特定结构域的意大利蜜蜂E-钙黏蛋白和N-钙黏蛋白抗体揭示的蜜蜂性腺中非常规钙黏蛋白定位表明其具有其他功能。

Unconventional Cadherin Localization in Honey Bee Gonads Revealed Through Domain-Specific Apis mellifera E- and N-Cadherin Antibodies Indicates Alternative Functions.

作者信息

Florecki Mônica M, Hartfelder Klaus

机构信息

Departamento de Biologia Celular e Molecular e Bioagentes Patogênicos, Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo, Avenida Bandeirantes 3900, Ribeirão Preto 14049-900, SP, Brazil.

出版信息

Insects. 2012 Nov 22;3(4):1200-19. doi: 10.3390/insects3041200.

Abstract

As key factors in intercellular adhesion processes, cadherins play important roles in a plethora of developmental processes, including gametogenesis. In a previous study on cadherin localization in the gonads of honey bees, performed with heterologous pan-cadherin antibodies, we detected these proteins as (i) associated with cell membranes, (ii) as homogeneously distributed throughout the cytoplasm, and (iii) as nuclear foci in both somatic and germline cells, raising the possibility of alternative functions. To further investigate such unusual intracellular cadherin localization we produced specific antibodies against the N- and C-terminal domains of honey bee N- and E-cadherin. A 160 kDa protein was recognized by the E-cadherin antibodies as well as one of approximately 300 kDa from those raised against N-cadherin. In gonad preparations, both proteins were detected as dispersed throughout the cytoplasm and as nuclear foci in both germline and somatic cells of queen and worker ovarioles, as well as in the testioles of drones. This leads us to infer that cadherins may indeed be involved in certain signaling pathways and/or transcriptional regulation during gametogenesis. In late oogenesis stages, immunolabeling for both proteins was observed at the cell cortex, in conformity with a role in cell adhesion. In testioles, E-cadherin was seen in co-localization with fusomes, indicating a possible role in cyst organization. Taken together, the distribution of N- and E-cadherins in honey bee gonads is suggestive of alternative roles for cadherins in gametogenesis of both sexes.

摘要

作为细胞间黏附过程中的关键因素,钙黏蛋白在包括配子发生在内的众多发育过程中发挥着重要作用。在之前一项使用异源泛钙黏蛋白抗体对蜜蜂性腺中钙黏蛋白定位进行的研究中,我们检测到这些蛋白质:(i)与细胞膜相关;(ii)均匀分布于整个细胞质中;(iii)在体细胞和生殖细胞中均为核内焦点,这增加了其具有其他功能的可能性。为了进一步研究这种不寻常的细胞内钙黏蛋白定位,我们制备了针对蜜蜂N -钙黏蛋白和E -钙黏蛋白N端和C端结构域的特异性抗体。E -钙黏蛋白抗体识别出一种160 kDa的蛋白质,针对N -钙黏蛋白产生的抗体识别出一种约300 kDa的蛋白质。在性腺制剂中,在蜂王和工蜂卵巢的生殖细胞和体细胞以及雄蜂的精巢中,这两种蛋白质均被检测到分散在整个细胞质中且为核内焦点。这使我们推断钙黏蛋白在配子发生过程中可能确实参与了某些信号通路和/或转录调控。在卵子发生后期,在细胞皮层观察到这两种蛋白质的免疫标记,这与它们在细胞黏附中的作用一致。在精巢中,E -钙黏蛋白与纺锤体共定位,表明其在囊肿组织中可能发挥作用。综上所述,N -钙黏蛋白和E -钙黏蛋白在蜜蜂性腺中的分布表明钙黏蛋白在两性配子发生中具有其他作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/32c3/4553572/31d27f59c033/insects-03-01200-g001.jpg

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