Jackson Laboratory, Bar Harbor, ME, USA.
Graduate School of Biomedical Science and Engineering, University of Maine, Orono, ME, USA.
Hum Mol Genet. 2018 Oct 1;27(19):3340-3352. doi: 10.1093/hmg/ddy238.
Photoreceptor dysplasia, characterized by formation of folds and (pseudo-)rosettes in the outer retina, is associated with loss of functional nuclear receptor subfamily 2 group E member 3 (NR2E3) and neural retina leucine-zipper (NRL) in both humans and mice. A sensitized chemical mutagenesis study to identify genetic modifiers that suppress photoreceptor dysplasia in Nr2e3rd7mutant mice identified line Tvrm222, which exhibits a normal fundus appearance in the presence of the rd7 mutation. The Tvrm222 modifier of Nr2e3rd7/rd7 was localized to Chromosome 6 and identified as a missense mutation in the FERM domain containing 4B (Frmd4b) gene. The variant is predicted to cause the substitution of a serine residue 938 with proline (S938P). The Frmd4bTvrm222 allele was also found to suppress outer nuclear layer (ONL) rosettes in Nrl-/- mice. Fragmentation of the external limiting membrane (ELM), normally observed in rd7 and Nrl-/-mouse retinas, was absent in the presence of the Frmd4bTvrm222 allele. FRMD4B, a binding partner of cytohesin 3, is proposed to participate in cell junction remodeling. Its biological function in photoreceptor dysplasia has not been previously examined. In vitro experiments showed that the FRMD4B938P variant fails to be efficiently recruited to the cell surface upon insulin stimulation. In addition, we found a reduction in protein kinase B phosphorylation and increased levels of cell junction proteins, Catenin beta 1 and tight junction protein 1, associated with the cell membrane in Tvrm222 retinas. Taken together, this study reveals a critical role of FRMD4B in maintaining ELM integrity and in rescuing morphological abnormalities of the ONL in photoreceptor dysplasia.
光感受器发育不良的特征是在外视网膜形成褶皱和(假)玫瑰花结,与人类和小鼠中功能性核受体亚家族 2 组 E 成员 3(NR2E3)和神经视网膜亮氨酸拉链(NRL)的丧失有关。一项旨在鉴定抑制 Nr2e3rd7 突变小鼠光感受器发育不良的遗传修饰因子的敏感化学诱变研究鉴定出了 Tvrm222 系,该系在存在 rd7 突变的情况下表现出正常的眼底外观。Nr2e3rd7/rd7 的 Tvrm222 修饰因子定位于 6 号染色体上,并被鉴定为 FERM 结构域包含 4B(Frmd4b)基因中的错义突变。该变体预计会导致丝氨酸残基 938 被脯氨酸取代(S938P)。还发现 Frmd4bTvrm222 等位基因也抑制 Nrl-/- 小鼠的外核层(ONL)玫瑰花结。在存在 Frmd4bTvrm222 等位基因的情况下,rd7 和 Nrl-/- 小鼠视网膜中观察到的外节层(ELM)碎裂缺失。FRMD4B 是细胞浆素 3 的结合伴侣,据推测它参与细胞连接重塑。其在光感受器发育不良中的生物学功能尚未被先前研究过。体外实验表明,在胰岛素刺激下,FRMD4B938P 变体不能有效地被招募到细胞膜表面。此外,我们发现 Tvrm222 视网膜中与细胞膜相关的蛋白激酶 B 磷酸化减少,细胞连接蛋白 Catenin beta 1 和紧密连接蛋白 1 水平增加。综上所述,这项研究揭示了 FRMD4B 在维持 ELM 完整性和挽救光感受器发育不良中 ONL 形态异常方面的关键作用。