Müller Glia Stabilizes Cell Columns During Retinal Development: Lateral Cell Migration but not Neuropil Growth is Inhibited in Mixed Chick-Quail Retinospheroids

Radial columnar organization of cell clones is a characteristic feature of vertebrate retinae that is structurally not understood. Here we provide in vitro evidence that Müller glia processes stabilize cells within columns. Dissociated embryonic chick retinal plus pigmented cells regenerate in vitro into fully laminated stratospheroids. After reaggregating chick and quail cells, quail-derived spheroid areas are detected as isolated sectors, as shown by a quail-specific antibody. Each sector contains one or multiple cell columns. The radial borders separating chick and quail sectors are fully congruent with the extension of 3A7-labelled Müller glia processes. While cell somata do not show any lateral interspecies mixing, quail-derived neuropil extends within the inner plexiform areas far into chick sectors. After selective damage of Müller cells by the gliotoxin dl -α-aminoadipic acid, the columnar organization is destabilized, as evidenced by a decrease in vimentin expression and by the migration of individual neurons out of their cell column. These data demonstrate that Müller cells actively stabilize cells within their columns, while neuritic growth is not hindered.     

Retinal histogenesis and cell differentiation in an elasmobranch species, the small-spotted catshark Scyliorhinus canicula

Here we present a detailed study of the major events in the retinal histogenesis in a slow-developing elasmobranch species, the small-spotted catshark, during embryonic, postnatal and adult stages using classical histological and immunohistological methods, providing a complete neurochemical characterization of retinal cells. We found that the retina of the small-spotted catshark was fully differentiated prior to birth. The major developmental events in retinal cell differentiation occurred during the second third of the embryonic period. Maturational features described in the present study were first detected in the central retina and, as development progressed, they spread to the rest of the retina following a central-to-peripheral gradient. While the formation of both plexiform layers occurs simultaneously in the retina of the most common fish models, in the small-spotted catshark retina the emergence of the outer plexiform layer was delayed with respect to the inner plexiform layer. According to the expression of the markers used, retinal cell differentiation followed a vitreal-to-scleral gradient, with the exception of Müller cells that were the last cell type generated during retinogenesis. This vitreal-to-scleral progression of neural differentiation seems to be specific to slow-developing fish species.     

Cannabinoid receptor type 1 expression during postnatal development of the rat retina

Cannabinoid receptor type 1 (CB1R) participates in developmental processes in the central nervous system (CNS). The rodent retina represents an interesting and valuable model for studying CNS development, because it contains well-identified cell types with clearly established and distinct developmental timelines. Very little is known about the distribution or function of CB1R in the developing retina. In this study, we investigated the expression pattern of CB1R in the rat retina during all stages of postnatal development. Western blots were performed on retinal tissue at different time points between P1 and adulthood. In order to identify the cells expressing the receptor and the age at which this expression started, immunohistochemical co-staining was carried out for CB1R and markers of the different cell types comprising the retina. CB1R was already present at P1 in various cell types, i.e., ganglion, amacrine, horizontal, and mitotic cells. In the course of development, it appeared in cone photoreceptors and bipolar cells. For some cell types (bipolar, Müller, and some amacrine cells), CB1R was transiently expressed, suggesting a potential role of this receptor in developmental processes, such as migration, morphological changes, sub-identity acquisition, and patterned retinal spontaneous activity. Our results also indicated that CB1R is largely expressed in the adult retina (cone photoreceptors and horizontal, most amacrine, and retinal ganglion cells), and may therefore contribute to retinal functions. Overall these results indicate that, as shown in other structures of the brain, CB1R could play an instrumental role in the development and function of the retina.     

Cell autonomous and nonautonomous requirements for Delltalike1 during early mouse retinal neurogenesis

Background: In the vertebrate retina, six neuronal and one glial cell class are produced from a common progenitor pool. During neurogenesis, adjacent retinal cells use Notch signaling to maintain a pool of progenitors by blocking particular cells from differentiating prematurely. In mice there are multiple Notch pathway ligands and receptors, but the role(s) of each paralogue during retinal histogenesis remains only partially defined. Results: Here we analyzed the cell autonomous and nonautonomous requirements for the Deltalike1(Dll1) ligand during prenatal retinogenesis. We used the α‐Cre driver to simultaneously delete a Dll1 conditional allele and activate the Z/EG reporter, then quantified Dll1 mutant phenotypes within and outside of this α‐Cre GFP‐marked lineage. We found that Dll1 activity is required for Hes1 expression, both autonomously and nonautonomously, but were surprised that retinal ganglion cell differentiation is only blocked cell autonomously. Moreover, Dll1 does not act during cone photoreceptor neurogenesis. Finally, Dll1 mutant adult retinas contained small retinal rosettes and RGC patterning defects but were otherwise normal. Conclusions: Although Dll1 participates in bidirectional (cis + trans) Notch signaling to regulate Hes1 expression, it only acts cell autonomously (in cis) to interpret inhibitory signals from other cells that block RGC neurogenesis. Developmental Dynamics 245:631–640, 2016. © 2016 Wiley Periodicals, Inc.     

The role of cell cycle in retinal development: Cyclin‐dependent kinase inhibitors co‐ordinate cell‐cycle inhibition,cell‐fate determination and differentiation in the developing retina†

The mature retina is formed through multi‐step developmental processes, including eye field specification, optic vesicle evagination, and cell‐fate determination. Co‐ordination of these developmental events with cell‐proliferative activity is essential to achieve formation of proper retinal structure and function. In particular, the molecular and cellular dynamics of the final cell cycle significantly influence the identity that a cell acquires, since cell fate is largely determined at the final cell cycle for the production of postmitotic cells. This review summarizes our current understanding of the cellular mechanisms that underlie the co‐ordination of cell‐cycle and cell‐fate determination, and also describes a molecular role of cyclin‐dependent kinase inhibitors (CDKIs) as co‐ordinators of cell‐cycle arrest, cell‐fate determination and differentiation. Developmental Dynamics 239:727–736, 2010. © 2010 Wiley‐Liss, Inc.     

Relationship between Delta-like and proneural bHLH genes during chick retinal development.

Notch signaling in the retina maintains a pool of progenitor cells throughout retinogenesis. However, two Notch-ligands from the Delta-like gene family, Dll1 and Dll4, are present in the developing retina. To understand their relationship, we characterized Dll1 and Dll4 expression with respect to proliferating progenitor cells and newborn neurons in the chick retina. Dll4 matched the pattern of neural differentiation. By contrast, Dll1 was primarily expressed in progenitor cells. We compared Dll1 and Dll4 kinetic profiles with that of the transiently up-regulated cascade of proneural basic helix-loop-helix (bHLH) genes after synchronized progenitor cell differentiation, which suggested a potential role for Ascl1 in the regulation of Delta-like genes. Gain-of-function assays demonstrate that Ascl1 does influence Delta-like gene expression and Notch signaling activity. These data suggest that multiple sources of Notch signaling from newborn neurons and progenitors themselves coordinate retinal histogenesis.