Opposing microtubule- and actin-dependent forces in the development and maintenance of structural polarity in retinal photoreceptors.
We have used embryonic cells grown in vitro to study the roles of microtubules and microfilaments in the development and maintenance of the polarized shape of retinal photoreceptors. After several days in culture, isolated cone photoreceptors displayed a highly elongated, compartmentalized morphology similar to that of photoreceptors in vivo. When treated with the microtubule-depolymerizing agent nocodazole, these elongated photoreceptors became progressively shorter, eventually losing their compartmentalized structure and becoming round. Conversely, treatment with the actin-depolymerizing agent cytochalasin D caused the elongated photoreceptors to lengthen even further. Computer-assisted, quantitative analysis showed that responses of individual cells to both nocodazole and Cytochalasin D were concentration-dependent, graded, and reversible. Immunocytochemical studies suggested the presence of longitudinally oriented actin filaments and microtubules in these photoreceptors, prominent in the region that undergoes the most pronounced length changes in response to cytoskeletal inhibitors. Prior to becoming elongated, photoreceptor precursors could be accurately identified in early retinal cultures. These round cells undergo a stereotyped sequence of morphogenetic transformations during in vitro development, including elongation and compartmentalization of the cell body as well as extension of a single neurite. Treatment with either cytochalasin D or nocodazole completely blocked morphogenesis. In addition, cytochalasin D caused the development of an abnormal, elongated cell process, which formed by a microtubule-dependent mechanism. These nocodazole and cytochalasin D effects also were reversible. Taken together, these data indicate that the complex developmental transformations leading to photoreceptor polarization occur in the absence of intercellular contacts, and are predominantly controlled by intracellular cytoskeletal forces. They suggest the existence of continuously active, oppositely directed, microtubule- and actin-dependent forces, the balance of which is a determining factor in the development as well as the maintenance of the elongated, compartmentalized organization of photoreceptor cells.
Madreperla
S A
SA
Retinal Degenerations Research Center, Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205.
Adler
R
R
D000199:Actins / Q000502:physiology*; D000818:Animals; D001562:Benzimidazoles / Q000494:pharmacology; D002642:Chick Embryo; D015638:Cytochalasin D; D003572:Cytochalasins / Q000494:pharmacology; D005455:Fluorescent Antibody Technique; D006651:Histocytochemistry; D008870:Microtubules / Q000502:physiology*; D009024:Morphogenesis; D015739:Nocodazole; D010786:Photoreceptor Cells / Q000187:drug effects / Q000196:embryology / Q000648:ultrastructure*; D012160:Retina / Q000196:embryology / Q000648:ultrastructure*; D014404:Tubulin / Q000032:analysis / Q000502:physiology
D000199:Actins; D001562:Benzimidazoles; D003572:Cytochalasins; D014404:Tubulin; D015638:Cytochalasin D; D015739:Nocodazole