Pereiro Xandra, Beriain Sandra, Rodriguez Lara, Roiz-Valle David, Ruzafa Noelia, Vecino Elena
Experimental Ophthalmo-Biology Group, Department of Cell Biology and Histology, University of Basque Country UPV/EHU, Leioa, Spain.
Begiker-Ophthalmology Research Group, BioCruces Health Research Institute, Cruces Hospital, Barakaldo, Spain.
Front Neurosci. 2022 Mar 14;16:854278. doi: 10.3389/fnins.2022.854278. eCollection 2022.
Müller cells are the principal glial cells in the retina and they assume many of the functions carried out by astrocytes, oligodendrocytes and ependymal cells in other regions of the central nervous system. Müller cells express growth factors, neurotransmitter transporters and antioxidant agents that could fulfill important roles in preventing excitotoxic damage to retinal neurons. Vertebrate Müller cells are well-defined cells, characterized by a common set of features throughout the phylum. Nevertheless, several major differences have been observed among the Müller cells in distinct vertebrates, such as neurogenesis, the capacity to reprogram fish Müller glia to neurons. Here, the Müller glia of the largest adult mammal in the world, the whale, have been analyzed, and given the difficulties in obtaining cetacean cells for study, these whale glia were analyzed both in primary cultures and as immortalized whale Müller cells. After isolating the retina from the eye of a beached sei whale (), primary Müller cell cultures were established and once the cultures reached confluence, half of the cultures were immortalized with the simian virus 40 (SV40) large T-antigen commonly used to immortalize human cell lines. The primary cell cultures were grown until cells reached senescence. Expression of the principal molecular markers of Müller cells (GFAP, Vimentin and Glutamine synthetase) was studied in both primary and immortalized cells at each culture passage. Proliferation kinetics of the cells were analyzed by time-lapse microscopy: the time between divisions, the time that cells take to divide, and the proportion of dividing cells in the same field. The karyotypes of the primary and immortalized whale Müller cells were also characterized. Our results shown that W21M proliferate more rapidly and they have a stable karyotype. W21M cells display a heterogeneous cell morphology, less motility and a distinctive expression of some typical molecular markers of Müller cells, with an increase in dedifferentiation markers like α-SMA and β-III tubulin, while they preserve their GS expression depending on the culture passage. Here we also discuss the possible influence of the animal's age and size on these cells, and on their senescence.
穆勒细胞是视网膜中的主要神经胶质细胞，它们承担着中枢神经系统其他区域中星形胶质细胞、少突胶质细胞和室管膜细胞所执行的许多功能。穆勒细胞表达生长因子、神经递质转运体和抗氧化剂，这些物质在预防视网膜神经元的兴奋性毒性损伤中可能发挥重要作用。脊椎动物的穆勒细胞是定义明确的细胞，其特征是在整个门中具有一组共同的特征。然而，在不同的脊椎动物的穆勒细胞之间观察到了几个主要差异，例如神经发生，即将鱼类穆勒胶质细胞重编程为神经元的能力。在这里，对世界上最大的成年哺乳动物鲸鱼的穆勒胶质细胞进行了分析，鉴于获取鲸类细胞进行研究存在困难，这些鲸鱼胶质细胞在原代培养物中和作为永生化鲸鱼穆勒细胞进行了分析。从搁浅的塞鲸（）的眼睛中分离出视网膜后，建立了原代穆勒细胞培养物，一旦培养物达到汇合状态，一半的培养物用通常用于永生化人类细胞系的猿猴病毒40（SV40）大T抗原进行永生化。原代细胞培养物生长至细胞达到衰老。在每个培养传代时，在原代细胞和永生化细胞中研究了穆勒细胞的主要分子标志物（胶质纤维酸性蛋白、波形蛋白和谷氨酰胺合成酶）的表达。通过延时显微镜分析细胞的增殖动力学：分裂之间的时间、细胞分裂所需的时间以及同一视野中分裂细胞的比例。还对原代和永生化鲸鱼穆勒细胞的核型进行了表征。我们的结果表明，W21M增殖更快，并且它们具有稳定的核型。W21M细胞表现出异质的细胞形态、较低的运动性以及穆勒细胞一些典型分子标志物的独特表达，同时像α-平滑肌肌动蛋白和β-III微管蛋白这样的去分化标志物增加，而它们根据培养传代保留其GS表达。在这里，我们还讨论了动物的年龄和大小对这些细胞及其衰老的可能影响。
