RESUMO
Post-transcriptional switches are flexible effectors of dynamic changes in gene expression. Here we report a new post-transcriptional switch that dictates the spatiotemporal and mutually exclusive expression of two alternative gene products from a single transcript. Expression of primate-specific exonic microRNA-198 (miR-198), located in the 3'-untranslated region of follistatin-like 1 (FSTL1) messenger RNA, switches to expression of the linked open reading frame of FSTL1 upon wounding in a human ex vivo organ culture system. We show that binding of a KH-type splicing regulatory protein (KSRP, also known as KHSRP) to the primary transcript determines the fate of the transcript and is essential for the processing of miR-198: transforming growth factor-ß signalling switches off miR-198 expression by downregulating KSRP, and promotes FSTL1 protein expression. We also show that FSTL1 expression promotes keratinocyte migration, whereas miR-198 expression has the opposite effect by targeting and inhibiting DIAPH1, PLAU and LAMC2. A clear inverse correlation between the expression pattern of FSTL1 (pro-migratory) and miR-198 (anti-migratory) highlights the importance of this regulatory switch in controlling context-specific gene expression to orchestrate wound re-epithelialization. The deleterious effect of failure of this switch is apparent in non-healing chronic diabetic ulcers, in which expression of miR-198 persists, FSTL1 is absent, and keratinocyte migration, re-epithelialization and wound healing all fail to occur.
Assuntos
Proteínas Relacionadas à Folistatina/genética , Regulação da Expressão Gênica/genética , MicroRNAs/genética , RNA Mensageiro/genética , Transcrição Gênica/genética , Cicatrização/genética , Proteínas Adaptadoras de Transdução de Sinal/antagonistas & inibidores , Proteínas Adaptadoras de Transdução de Sinal/metabolismo , Movimento Celular , Pé Diabético/genética , Pé Diabético/metabolismo , Pé Diabético/patologia , Éxons/genética , Proteínas Relacionadas à Folistatina/biossíntese , Forminas , Humanos , Técnicas In Vitro , Queratinócitos/citologia , Queratinócitos/metabolismo , Laminina/antagonistas & inibidores , Laminina/metabolismo , Fases de Leitura Aberta/genética , Proteínas de Ligação a RNA/genética , Proteínas de Ligação a RNA/metabolismo , Pele/citologia , Pele/lesões , Pele/metabolismo , Pele/patologia , Fatores de Tempo , Transativadores/genética , Transativadores/metabolismo , Fator de Crescimento Transformador beta1/metabolismo , Ativador de Plasminogênio Tipo Uroquinase/antagonistas & inibidores , Ativador de Plasminogênio Tipo Uroquinase/metabolismoRESUMO
Background The posterior ledge (PL) is a vital structure that supports the implant posteriorly during orbital floor reconstruction. This study describes a technique for mapping the PL in relation to the infraorbital margin (IM) in patients with orbital floor blowout fractures. This study establishes the location of the optic foramen in relation to the PL. Methods Facial computed tomography (FCT) scans of 67 consecutive patients with isolated orbital floor blowout fractures were analyzed using Osirix. Planes of reference for orbital fractures, a standardized technique for performing measurements on FCT, was used. Viewed coronally, the orbit was divided into seven equal sagittal slices (L1 laterally to L7 medially) with reference to the midorbital plane. The distances of PL from IM and location of optic foramen were determined. Results The greatest distance to PL is found at L5 (median: 30.1 mm, range: 13.5-37.1 mm). The median and ranges for each slice are as follows: L1 (median: 0.0 mm, range: 0.0-19.9 mm), L2 (median: 0.0 mm, range: 0.0-21.5 mm), L3 (median: 15.8 mm, range: 0.0-31.7 mm), L4 (median: 26.1 mm, range: 0.0-34.0 mm), L5 (median: 30.1 mm, range: 13.5-37.1 mm), L6 (median: 29.0 mm, range: 0.0-36.3 mm), L7 (median: 20.8 mm, range: 0.0-39.2 mm). The median distance of the optic foramen from IM is 43.7 mm (range: 37.0- 49.1) at L7. Conclusion Distance to PL from IM increases medially until the L5 before decreasing. A reference map of the PL in relation to the IM and optic foramen is generated. The optic foramen is located in close proximity to the PL at the medial orbital floor. This aids in preoperative planning and intraoperative dissection.