Progranulin regulates neurogenesis in the developing vertebrate retina

We evaluated the expression and function of the microglia‐specific growth factor, Progranulin‐a (Pgrn‐a) during developmental neurogenesis in the embryonic retina of zebrafish. At 24 hpf pgrn‐a is expressed throughout the forebrain, but by 48 hpf pgrn‐a is exclusively expressed by microglia and/or microglial precursors within the brain and retina. Knockdown of Pgrn‐a does not alter the onset of neurogenic programs or increase cell death, however, in its absence, neurogenesis is significantly delayed—retinal progenitors fail to exit the cell cycle at the appropriate developmental time and postmitotic cells do not acquire markers of terminal differentiation, and microglial precursors do not colonize the retina. Given the link between Progranulin and cell cycle regulation in peripheral tissues and transformed cells, we analyzed cell cycle kinetics among retinal progenitors following Pgrn‐a knockdown. Depleting Pgrn‐a results in a significant lengthening of the cell cycle. These data suggest that Pgrn‐a plays a dual role during nervous system development by governing the rate at which progenitors progress through the cell cycle and attracting microglial progenitors into the embryonic brain and retina. Collectively, these data show that Pgrn‐a governs neurogenesis by regulating cell cycle kinetics and the transition from proliferation to cell cycle exit and differentiation. © 2017 The Authors. Developmental Neurobiology Published by Wiley Periodicals, Inc. Develop Neurobiol 77: 1114–1129, 2017Peer Reviewedhttps://deepblue.lib.umich.edu/bitstream/2027.42/138360/1/dneu22499.pdfhttps://deepblue.lib.umich.edu/bitstream/2027.42/138360/2/dneu22499_am.pd

Dynamic expression of the basic helix-loop-helix transcription factor neuroD in the rod and cone photoreceptor lineages in the retina of the embryonic and larval zebrafish

NeuroD is a basic helix-loop-helix (bHLH) transcription factor critical for determining neuronal cell fate and regulating withdrawal from the cell cycle. We showed previously that, in goldfish, neuroD is expressed in the rod photoreceptor lineage, and we inferred that neuroD is also expressed in a subset of amacrine cells and nascent cone photoreceptors. Here we extended that study by examining the temporal and spatial expression pattern of neuroD in the embryonic and larval zebrafish and by identifying the cell types that express this gene. NeuroD expression in the developing zebrafish retina is dynamic, spanning early retinogenesis and the maturation of cone photoreceptors. In early retinogenesis neuroD expression expands from a small patch in the ventronasal retina, through the remaining retinal neuroepithelium. As retinogenesis progresses, neuroD expression becomes restricted to amacrine cells, immature cones, and cells of rod and cone lineages. This expression achieves an adult pattern by 96 hours postfertilization (hpf), whereupon the temporal pattern of neuroD expression in central retina is spatially recapitulated at the germinative margin. The cellular pattern of expression suggests that neuroD regulates aspects of rod and cone genesis, but through separate cellular lineages. Furthermore, neuroD is coexpressed with the cone-rod-homeobox transcription factor (Crx) in putative cone progenitors and nascent cone photoreceptors, suggesting that, in the zebrafish retina, as in other vertebrate retinas, similar genetic cascades regulate photoreceptor genesis and maturation. J. Comp. Neurol. 501:1–12, 2007. © 2007 Wiley-Liss, Inc.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/55874/1/21150_ftp.pd

Neuronal cell proliferation and ocular enlargement in black moor goldfish

The mechanisms that control cell proliferation in the developing nervous system are not well understood. In larval and adult goldfish addition of new retinal neurons continues as the eye grows, but the factors that modulate the rate of cell proliferation are unknown. The eyes of Black Moors grow excessively during postembryonic life, probably as a direct result of abnormally elevated intraocular pressure. Ocular growth must be partly autonomous in Black Moors because in some individuals the two eyes are very different in size. To determine whether cell proliferation and neuronal cell number in the retina were correlated with size of the eye, we counted dividing neuronal progenitor cells (rod precursors) and mature retinal neurons (ganglion cells) in the retinas of ocularly asymmetric fish. Rod Precursors, which are scattered across the retina in the outer nuclear layer, were labeled with 3 H-thymidine and counted on histological sections processed for autoradiography. Ganglion cells were counted in retinal whole mounts. We found that the total population of dividing rod precursors and the total number of ganglion cells were systematically greater in the large eye compared to the small eye of individual fish. We conclude that control of the rate of neuronal proliferation in the teleost retina is intrinsic to the eye and is probably regulated by the same factors that control ocular growth.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/50040/1/902760207_ftp.pd

Constant dendritic coverage by ganglion cells with growth of the goldfish's retina

This study demonstrates that in the retina of the goldfish a type of ganglion cell, whose dendritic development has been well characterized [Hitchcock P.F. and Easter S.S. Jr (1986) J. Neurosci. 6, 1037-1050], is distributed across the retina in a nonrandom pattern, and the dendritic fields of this cell type overlap to completely cover, or "tile" the retina. Further, it is shown that the dendritic coverage of the retina by this cell type is established when the retina is small and is maintained as the retina grows.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/26849/1/0000412.pd

The MicroRNA, miR‐18a, Regulates NeuroD and Photoreceptor Differentiation in the Retina of Zebrafish

During embryonic retinal development, six types of retinal neurons are generated from multipotent progenitors in a strict spatiotemporal pattern. This pattern requires cell cycle exit (i.e. neurogenesis) and differentiation to be precisely regulated in a lineage‐specific manner. In zebrafish, the bHLH transcription factor NeuroD governs photoreceptor genesis through Notch signaling but also governs photoreceptor differentiation though distinct mechanisms that are currently unknown. Also unknown are the mechanisms that regulate NeuroD and the spatiotemporal pattern of photoreceptor development. Members of the miR‐17‐92 microRNA cluster regulate CNS neurogenesis, and a member of this cluster, miR‐18a, is predicted to target neuroD mRNA. The purpose of this study was to determine if, in the developing zebrafish retina, miR‐18a regulates NeuroD and if it plays a role in photoreceptor development. Quantitative RT‐PCR showed that, of the three miR‐18 family members (miR‐18a, b, and c), miR‐18a expression most closely parallels neuroD expression. Morpholino oligonucleotides and CRISPR/Cas9 gene editing were used for miR‐18a loss‐of‐function (LOF) and both resulted in larvae with more mature photoreceptors at 70 hpf without affecting cell proliferation. Western blot showed that miR‐18a LOF increases NeuroD protein levels and in vitro dual luciferase assay showed that miR‐18a directly interacts with the 3′ UTR of neuroD. Finally, tgif1 mutants have increased miR‐18a expression, less NeuroD protein and fewer mature photoreceptors, and the photoreceptor deficiency is rescued by miR‐18a knockdown. Together, these results show that, independent of neurogenesis, miR‐18a regulates the timing of photoreceptor differentiation and indicate that this occurs through post‐transcriptional regulation of NeuroD.Peer Reviewedhttps://deepblue.lib.umich.edu/bitstream/2027.42/147840/1/dneu22666.pdfhttps://deepblue.lib.umich.edu/bitstream/2027.42/147840/2/dneu22666_am.pd

Southern annular mode dynamics in observations and models, Part II: Eddy feedbacks

Many global climate models (GCMs) have trouble simulating Southern Annular Mode (SAM) variability correctly, particularly in the Southern Hemisphere summer season where it tends to be too persistent. In this two part study, a suite of experiments with the Canadian Middle Atmosphere Model (CMAM) is analyzed to improve our understanding of the dynamics of SAM variability and its deficiencies in GCMs. Here, an examination of the eddy-mean flow feedbacks is presented by quantification of the feedback strength as a function of zonal scale and season using a new methodology that accounts for intraseasonal forcing of the SAM. In the observed atmosphere, in the summer season, a strong negative feedback by planetary scale waves, in particular zonal wavenumber 3, is found in a localized region in the south west Pacific. It cancels a large proportion of the positive feedback by synoptic and smaller scale eddies in the zonal mean, resulting in a very weak overall eddy feedback on the SAM. CMAM is deficient in this negative feedback by planetary scale waves, making a substantial contribution to its bias in summertime SAM persistence. Furthermore, this bias is not alleviated by artificially improving the climatological circulation, suggesting that climatological circulation biases are not the cause of the planetary wave feedback deficiency in the model. Analysis of the summertime eddy feedbacks in the CMIP-5 models confirms that this is indeed a common problem among GCMs, suggesting that understanding this planetary wave feedback and the reason for its deficiency in GCMs is key to improving the fidelity of simulated SAM variability in the summer season

Southern annular mode dynamics in observations and models, Part I: The influence of climatological zonal wind biases in a comprehensive GCM

A common bias among global climate models (GCMs) is that they exhibit tropospheric southern annular mode (SAM) variability that is much too persistent in the Southern Hemisphere (SH) summertime. This is of concern for the ability to accurately predict future SH circulation changes, so it is important that it be understood and alleviated. In this two-part study, specifically targeted experiments with the Canadian Middle Atmosphere Model (CMAM) are used to improve understanding of the enhanced summertime SAM persistence. Given the ubiquity of this bias among comprehensive GCMs, it is likely that the results will be relevant for other climate models. Here, in Part I, the influence of climatological circulation biases on SAM variability is assessed, with a particular focus on two common biases that could enhance summertime SAM persistence: the too-late breakdown of the Antarctic stratospheric vortex and the equatorward bias in the SH tropospheric midlatitude jet. Four simulations are used to investigate the role of each of these biases in CMAM. Nudging and bias correcting procedures are used to systematically remove zonal-mean stratospheric variability and/or remove climatological zonal wind biases. The SAM time-scale bias is not alleviated by improving either the timing of the stratospheric vortex breakdown or the climatological jet structure. Even in the absence of stratospheric variability and with an improved climatological circulation, the model time scales are biased long. This points toward a bias in internal tropospheric dynamics that is not caused by the tropospheric jet structure bias. The underlying cause of this is examined in more detail in Part II of this study

Antibodies against pax6 immunostain amacrine and ganglion cells and neuronal progenitors, but not rod precursors, in the normal and regenerating retina of the goldfish

Pax6 is a developmental regulatory gene that plays a key role in the development of the embryonic brain, eye, and retina. This gene is also expressed in discrete groups of neurons within the adult brain. In this study, antibodies raised against a fusion protein from a zebra fish pax6 cDNA were used to investigate the expression of the pax6 gene in the mature, growing, and regenerating retina of the goldfish. On western blots of retinal proteins, the pax6 antibodies recognize a single band at the approximate size of the zebra fish pax6 protein. In retinal sections, the antibodies label the nuclei of mature amacrine and some ganglion cells. At the retinal margin, where neurogenesis and cellular differentiation continually occur in goldfish, the antibodies label neuronal progenitors and the newly postmitotic neurons. Following injury and during neuronal regeneration, the antibodies label mitotically active progenitors of regenerating neurons. Rod precursors, proliferating cells that normally give rise solely to rod photoreceptors and are the presumed antecedents of the injury-stimulated neuronal progenitors, are not immunostained by antibodies to the pax6 protein. The results of this study document the identity of pax6 -expressing cells in the mature retina and demonstrate that in the goldfish pax6 is expressed in neuronal progenitors during both retinal growth and regeneration. © 1996 John Wiley & Sons, Inc.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/50087/1/10_ftp.pd

The myopic eye of the black moor goldfish

Optical and anatomical methods were used to establish the refractive state of the eye of the Black Moor variety of goldfish. The eye is strongly myopic. The refractive error in 12 eyes from 6 fish ranged from 89 to 268 D.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/26461/1/0000549.pd