Neuroblastic Layer Research Articles

Analysis of Thyroid Response Element Activity during Retinal Development

Thyroid hormone (TH) signaling components are expressed during retinal development in dynamic spatial and temporal patterns. To probe the competence of retinal cells to mount a transcriptional response to TH, reporters that included thyroid response elements (TREs) were introduced into developing retinal tissue. The TREs were placed upstream of a minimal TATA-box and two reporter genes, green fluorescent protein (GFP) and human placental alkaline phosphatase (PLAP). Six of the seven tested TREs were first tested in vitro where they were shown to drive TH-dependent expression. However, when introduced into the developing retina, the TREs reported in different cell types in both a TH-dependent and TH-independent manner, as well as revealed specific spatial patterns in their expression. The role of the known thyroid receptors (TR), TRα and TRβ, was probed using shRNAs, which were co-electroporated into the retina with the TREs. Some TREs were positively activated by TR+TH in the developing outer nuclear layer (ONL), where photoreceptors reside, as well as in the outer neuroblastic layer (ONBL) where cycling progenitor cells are located. Other TREs were actively repressed by TR+TH in cells of the ONBL. These data demonstrate that non-TRs can activate some TREs in a spatially regulated manner, whereas other TREs respond only to the known TRs, which also read out activity in a spatially regulated manner. The transcriptional response to even simple TREs provides a starting point for understanding the regulation of genes by TH, and highlights the complexity of transcriptional regulation within developing tissue.

DCC is specifically required for the survival of retinal ganglion and displaced amacrine cells in the developing mouse retina

Netrin-1 and DCC are well known for their roles in neurite growth, axonal guidance, and neuronal migration. Recently, a number of studies showed that DCC is involved in the induction of apoptosis, and this proapoptotic activity can be blocked in the presence of Netrin-1. However, here, we found that DCC is required for the survival of two types of neurons selectively in the developing mouse retina where DCC is abundantly expressed. Our results showed that the DCC−/− retina displayed a reduced ganglion cell layer with relatively normal neuroblastic layer. Immunostaining assays revealed that in DCC−/− mice, initial neurogenesis within retina was unchanged while the numbers of differentiated retinal ganglion cells and displaced amacrine cells in ganglion cell layer were greatly reduced due to increased apoptosis. By contrast, other neuronal types including horizontal cells, bipolar cells, amacrine cells, photoreceptors, and Müller cells appeared normal in DCC mutant retinas. Moreover, DCCkanga mice that lack the intracellular P3 domain of DCC receptor displayed the same defects as DCC−/− mice. Thus, our findings suggest that DCC is a key regulator for the survival of specific types of neurons during retinal development and that DCC–P3 domain is essential for this developing event.

Expression of classic cadherins and δ-protocadherins in the developing ferret retina

BackgroundCadherins are a superfamily of calcium-dependent adhesion molecules that play multiple roles in morphogenesis, including proliferation, migration, differentiation and cell-cell recognition. The subgroups of classic cadherins and δ-protocadherins are involved in processes of neural development, such as neurite outgrowth, pathfinding, target recognition, synaptogenesis as well as synaptic plasticity. We mapped the expression of 7 classic cadherins (CDH4, CDH6, CDH7, CDH8, CDH11, CDH14, CDH20) and 8 δ-protocadherins (PCDH1, PCDH7, PCDH8, PCDH9, PCDH10, PCDH11, PCDH17, PCDH18) at representative stages of retinal development and in the mature retina of the ferret by in situ hybridization.ResultsAll cadherins investigated by us are expressed differentially by restricted populations of retinal cells during specific periods of the ferret retinogenesis. For example, during embryonic development, some cadherins are exclusively expressed in the outer, proliferative zone of the neuroblast layer, whereas other cadherins mark the prospective ganglion cell layer or cells in the prospective inner nuclear layer. These expression patterns anticipate histogenetic changes that become visible in Nissl or nuclear stainings at later stages. In parallel to the ongoing development of retinal circuits, cadherin expression becomes restricted to specific subpopulations of retinal cell types, especially of ganglion cells, which express most of the investigated cadherins until adulthood. A comparison to previous results in chicken and mouse reveals overall conserved expression patterns of some cadherins but also species differences.ConclusionsThe spatiotemporally restricted expression patterns of 7 classic cadherins and 8 δ-protocadherins indicate that cadherins provide a combinatorial adhesive code that specifies developing retinal cell populations and intraretinal as well as retinofugal neural circuits in the developing ferret retina.

ATP controls cell cycle and induces proliferation in the mouse developing retina

Previous data suggest that nucleotides are important mitogens in the developing chick retina. Here, we extended the study on the mitogenic effect of ATP to newborn mouse retinal explants. Our results showed that P2Y1 receptors were widely distributed in C57bl/6 mice retina and that the majority of PCNA positive cells co-localized with P2Y1 receptor. To evaluate proliferation, retinal explants obtained from newborn mice were incubated with 0.5μCi [3H]-thymidine or 3μM BrDU 1h before the end of culture. Our data showed that ATP induced a dose-dependent increase in [3H]-thymidine incorporation, an effect that was mimicked by ADP but not by UTP and was blocked by the P2 antagonist PPADS in a dose-dependent manner. The increase in [3H]-thymidine incorporation induced by ATP was only observed in explants cultured for 3 days or less and was mimicked by the ectoapyrase inhibitor ARL 67156. It corresponded to an increase in the number of BrdU+ cells in the neuroblastic layer (NL) of the tissue, suggesting that ATP, through activation of P2Y1 receptors, induced proliferation of late developing progenitors in retinal explants of newborn mice. The increase in the number of BrdU+ cells was observed across the whole NL when explants were incubated with ATP for 24h and no increase in the number of p-histone H3 labeled cells could be noticed at this time point. In longer incubations of 48h with ATP or 24h with ATP followed by a period of 24h in fresh medium, an increase in the number of BrdU+ cells promoted by ATP was observed only in the middle and outer, but not in the inner NL. In these conditions, an increase in the number of p-histone H3 labeled cells was detected in the outer NL, suggesting that ATP induced cells to enter S and progress to G2 phase of the cell cycle in the first 24h period of incubation. ATP also induced an increase and a decrease in the expression of cyclin D1and p27kip1, respectively, in retinal progenitors of the NL. While the increase in the expression of cyclin D1 was observed when retinal explants were incubated for 3h or longer periods of time, the decrease in the expression of p27kip1 was noticed only after 6h incubation with ATP. Both effects were blocked by the P2 receptor antagonist PPADS. These data suggest that ATP induces cell proliferation in retinal explants by inducing late developing progenitors to progress from G1 to S phase of cell cycle.

Localization of p27kip1 in the developing avian retina: Sustained expression in the mature tissue

p27kip1 is a cyclin/CDK inhibitor that is expressed in cells that exit cell cycle and turn post-mitotic. Here, we characterized the expression and localization of p27kip1 during the development of the chick embryo retina. Expression of p27kip1 in this tissue begins at embryonic day 5 (E5), increasing as development proceeds. In contrast to the expression in the developing rat retina that markedly decreases after postnatal day 6, expression of p27kip1 in the chick retina decreases only slightly (∼30%) after E12. Thereafter, it remains highly expressed in the tissue. p27kip1 expression increases in an orderly succession. By E5, immunoreactivity was observed over β-tubulin III (TUJ-1) positive cell bodies located in the prospective Ganglion Cell Layer. By E7, p27kip1 was also detected over elongated cell nuclei located in the inner and outer portions of the Neuroblastic Layer and over cell bodies in the middle of the Inner Plexiform Layer. By E12, besides labeled cell bodies, labeled processes from amacrine cells and from cells at the GCL in the IPL were identified. In retinas from post-hatched chicken, immunoreactivity was observed over cell bodies located at all nuclear layers. Several differentiated ChAT positive cholinergic cells were labeled for p27kip1. Our data suggest that, as in the retina of other species, p27kip1 is expressed in cells that are exiting cell cycle and differentiating in the early developing chick embryo retina. However, as opposed to rodents and amphibians, neuronal expression of p27kip1 is sustained in the adult chick retina, indicating that its expression is differently regulated during development in this specie.

Ontogenetic expression of the vanilloid receptors TRPV1 and TRPV2 in the rat retina

The present study aimed to analyze the gene and protein expression and the pattern of distribution of the vanilloid receptors TRPV1 and TRPV2 in the developing rat retina. During the early phases of development, TRPV1 was found mainly in the neuroblastic layer of the retina and in the pigmented epithelium. In the adult, TRPV1 was found in microglial cells, blood vessels, astrocytes and in neuronal structures, namely synaptic boutons of both retinal plexiform layers, as well as in cell bodies of the inner nuclear layer and the ganglion cell layer. The pattern of distribution of TRPV1 was mainly punctate, and there was higher TRPV1 labeling in the peripheral retina than in central regions. TRPV2 expression was quite distinct. Its expression was virtually undetectable by immunoblotting before P1, and that receptor was found by immunohistochemistry only by postnatal day 15 (P15). RNA and protein analysis showed that the adult levels are only reached by P60, which includes small processes in the retinal plexiform layers, and labeled cellular bodies in the inner nuclear layer and the ganglion cell layer. There was no overlapping between the signal observed for both receptors. In conclusion, our results showed that the patterns of distribution of TRPV1 and TRPV2 are different during the development of the rat retina, suggesting that they have specific roles in both visual processing and in providing specific cues to neural development.

Developmental expression of thyroid hormone receptor β2 protein in cone photoreceptors in the mouse

Thyroid hormone receptor beta2 (TRbeta2) controls the patterning of cone opsin photopigments that mediate colour vision. We raised an antiserum against TRbeta2 to study cone photoreceptor development by western blot and immunostaining analyses. TRbeta2-positive cells first appeared between embryonic day 10 (E10) and E12. Numbers increased until near birth, correlating with generation of the cone population. At birth, signals decreased until postnatal day 10, then declined to very low levels in adulthood. TRbeta2-positive cells were initially dispersed but became aligned at the edge of the outer neuroblastic layer by E15. Postnatally, these cells migrated inwardly until postnatal day 10, then outwardly to the edge of the outer nuclear layer, the location of mature cones. TRbeta2 represents a functionally unique marker for cone development.

Pias3-Dependent SUMOylation Directs Rod Photoreceptor Development

Specification of retinal rod photoreceptors is determined by several different transcription factors that activate expression of rod-specific genes and repress expression of cone photoreceptor-specific genes. The mechanism by which this dual regulation occurs is unclear. We have found that Pias3, a transcriptional coregulator and E3 SUMO ligase that is selectively expressed in developing photoreceptors, promotes the differentiation of rod photoreceptors while preventing rods from adopting cone photoreceptor-like characteristics. Pias3 binds the photoreceptor-specific transcription factors Crx and Nr2e3 and is specifically targeted to the promoters of photoreceptor-specific genes. Pias3 SUMOylates Nr2e3, converting it into a potent repressor of cone-specific gene expression. Rod- and cone-specific promoters are bound by hyperSUMOylated proteins in rod photoreceptors, and blocking SUMOylation in photoreceptors results in cells with morphological and molecular features of cones and an absence of rod-specific markers. Our data thus identify Pias3-mediated SUMOylation of photoreceptor-specific transcription factors as a key mechanism of rod specification.

Erythropoietin expresson during the postnatal development of the rat retina

Event Abstract Back to Event Erythropoietin expresson during the postnatal development of the rat retina Arnold Szabó1*, Dora Vegvari1, Attila Magyar1, Gabor Deak2, Akos Lukats1, Agnes I. Berta1 and Agoston Szel1 1 Department of Human Morphology and Developmental Biology, Semmelweis University, Hungary 2 Eye Clinic, Semmelweis University, Hungary Both the erythropoietin (EPO) and its receptor (EPO-R) are expressed in the retina, however, their cellular distribution and involvement in retinal development is not known. In the present study, we examined developing postnatal and adult rat retinas in order to identify the temporal and spatial onset of their expression. In the adult retina both the EPO and its receptor were expressed in the photoreceptor inner segments, in the inner nuclear, ganglion, and both plexiform layers. Their expression patterns changed similarly. At birth, we found strong immunoreactivities in the ganglion cell layer and in the inner cell rows of the neuroblast layer and weak staining in the outer part of the neuroblast layer. During the following days the immunoreactivity of the ganglion cells and presumptive amacrine cells became more prominent. The cells of the ganglion cell layer showed a cytoplasmatic staining to the anti-erythropoietin antibody. From the second half of the first postnatal week strongly stained horizontal cells were observed by both antibodies at the level of the separating outer plexiform layer. During the second week a transient EPO-R expression was found in some cells of the outer nuclear layer, which followed a centroperipheral gradient. At the level of the developing photoreceptor inner and outer segments exclusively the inner segments were positively stained by both antibodies. We assume that the EPO is produced locally by the ganglion cells and its main targets are the amacrine, horizontal and photoreceptor cells and it may act as a regulating factor during the retinal development. Conference: 12th Meeting of the Hungarian Neuroscience Society, Budapest, Hungary, 22 Jan - 24 Jan, 2009. Presentation Type: Poster Presentation Topic: Developmental neurobiology and subcortical functions Citation: Szabó A, Vegvari D, Magyar A, Deak G, Lukats A, Berta AI and Szel A (2009). Erythropoietin expresson during the postnatal development of the rat retina. Front. Syst. Neurosci. Conference Abstract: 12th Meeting of the Hungarian Neuroscience Society. doi: 10.3389/conf.neuro.01.2009.04.076 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 03 Mar 2009; Published Online: 03 Mar 2009. * Correspondence: Arnold Szabó, Department of Human Morphology and Developmental Biology, Semmelweis University, Budapest, Hungary, szabar@ana2.sote.hu Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers Arnold Szabó Dora Vegvari Attila Magyar Gabor Deak Akos Lukats Agnes I Berta Agoston Szel Google Arnold Szabó Dora Vegvari Attila Magyar Gabor Deak Akos Lukats Agnes I Berta Agoston Szel Google Scholar Arnold Szabó Dora Vegvari Attila Magyar Gabor Deak Akos Lukats Agnes I Berta Agoston Szel PubMed Arnold Szabó Dora Vegvari Attila Magyar Gabor Deak Akos Lukats Agnes I Berta Agoston Szel Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page.

Distribution of transferrin and transferrin receptor of the eype 1 in the process of formation of the rat eye retina in early postnatal ontogenesis

By the method of indirect immunohistochemistry, distribution of transferrin and of transferrin receptor of the type (TFR1) was studied in the formed rat eye retina at the period of early postnatal ontogenesis (from birth to opening of eyelids). It has been established that the character of distribution of these proteins and intensity of specific staining change dependent on the retina formation stage. Retina of the newborn rat is characterized by diffuse transferrin distribution in nuclear retina layers (in the neuroblast layer--NBL) and in the ganglionic cell layer (GCL) as well as in the eye pigment epithelium (PE); relative immunoreactivity to transferrin is not high. At the 5th postnatal day, immunoreactivity to transferrin is maximal and is revealed both in nuclear and in plexiform layers of retina and in the eye PE, the greatest signal being characteristic of NBL. At the 10th postnatal day the transferrin signal intensity in retina decreases, specific staining is revealed in GCL, PE, and in the area of formed outer segments of photoreceptors. At the 15th postnatal day, transferrin is revealed in GCL, in outer and inner photoreceptor segments and in the eye PE. TFR1 is present in all retinal layers at all stages of the retina formation; the relative immunoreactivity to TFR1 sharply rises beginning from the 10th postnatal day; correlation between distribution of transferrin and TFR1 is detected in the entire retina of newborn rats as well as in the external retina area at subsequent stages of its development. A possible role of transferrin at various stages of formation of retinal is discussed.

Sox2 Plays a Role in the Induction of Amacrine and Müller Glial Cells in Mouse Retinal Progenitor Cells

The transcription factor Sox2 plays important roles in both human and mouse retinal development. Although loss-of-function mutations in Sox2 have been studied in mice, gain-of-function experiments in the neural retina have been lacking. The detailed expression pattern of Sox2 in the developing mouse retina was examined by immunohistochemistry. Then, Sox2 was expressed in a retinal explant culture prepared from E17 mouse embryos by retrovirus-mediated gene transfer to examine its role in retinal development. In addition, shRNA was used to suppress Sox2 in a retinal explant using a retrovirus-mediated system. Sox2 was expressed throughout the neuroblastic layer in the embryonic retina, but only in the inner nuclear layer in the mature retina. Double immunostaining revealed that Sox2 was expressed in Müller glial cells and in a subset of amacrine cells. Forced expression of Sox2 in a mouse retinal explant culture resulted in the dramatic accumulation of amacrine cells in the inner nuclear layer; in addition, cells expressing amacrine cell markers were also found on the innermost side of the outer nuclear layer. The expression of Pax6, which plays an important role in amacrine cell differentiation, was observed in the Sox2-expressing cells, and Sox2 activated the Pax6 promoter to drive luciferase expression in Y79 cells. A decrease in retinal progenitor cell proliferation accompanied these effects. The suppression of Sox2 expression by shRNA resulted in a decreased number of cells in the inner nuclear layer. Therefore, ectopic Sox2 expression can induce amacrine cells in the mouse retina from stage E17 onward, possibly by facilitating cell cycle exit.

The C. elegans glycosyltransferase BUS-8 has two distinct and essential roles in epidermal morphogenesis

Ventral enclosure in Caenorhabditis elegans involves migration of epidermal cells over a neuroblast substrate and subsequent adhesion at the ventral midline. Organisation of the neuroblast layer by ephrins and their receptors is essential for this migration. We show that bus-8, which encodes a predicted glycosyltransferase, is essential for embryonic enclosure and acts in or with ephrin signalling to mediate neuroblast organisation and to permit epidermal migration. BUS-8 acts non-cell-autonomously in this process, and likely modifies an extracellular regulator of ephrin signalling and cell organisation. Weak and cold-sensitive alleles of bus-8 show that the gene has a separate and distinct post-embryonic role, being essential for epidermal integrity and production of the cuticle surface. This disorganisation of the epidermis and cuticle layers causes increased drug sensitivity, which could aid the growing use of C. elegans in drug screening and chemical genomics. The viable mutants are also resistant to infection by the pathogen Microbacterium nematophilum, due to failure of the bacterium to bind to the host surface. The two separate essential roles of BUS-8 in epidermal morphogenesis add to our growing understanding of the widespread importance of glycobiology in development.

Growth hormone action in the developing neural retina: A proteomic analysis

The possible presence and action of growth hormone (GH) in the neural retina was investigated in newborn mice. The neural retina was found to be a site of GH gene expression, as GH mRNA was abundant in cells of the retinal ganglion cell layer, in which GH was also detected. It was also a site of GH action, since GH receptor (GHR) immunoreactivity mirrored that of GH. Actions of GH within the eye were indicated by a reduction in its axial length and retinal width (its neuroblastic, inner plexiform, and optic fiber layers) in GHR gene disrupted mice (GHR-/-), in comparison with wild type (GHR+/+) littermates. In the absence of GH signaling, four proteins in the retinal proteome of the GHR-/- mice (identified by 2-D gels and MS) differed in abundance with those in the wild type mice. Brain abundant membrane attached signal protein-1 (BASP-1) was down-regulated, whereas protein kinase C inhibitor 1, cyclophilin A, KH domain-containing, RNA-binding, signal transduction-associated protein 3 were up-regulated in GHR-/- mice. These proteins are involved in retinal vascularization, neural proliferation and neurite outgrowth. GH might thus have hitherto unsuspected roles in these processes during retinal development.

Identification of Photoreceptor Precursors in the Pars Plana during Ocular Development and after Retinal Injury

To study the distribution and differentiation of photoreceptor precursors in the ciliary epithelium in mice. Proliferating cells in flat-mount specimens of the ciliary body and retina were studied by bromodeoxyuridine (BRDU; 150 mg/kg) labeling in young C57Bl mice. Immunoreactivity to anti-recoverin, rhodopsin, and Pax6 antibodies and binding to peanut agglutinin were analyzed histologically to assess the distribution and differentiation of photoreceptor progenitors or precursors. Mice injected intraperitoneally with N-methyl-N-nitrosourea (MNU; 60 mg/kg) were also examined. Part of the neuroblast layer composed of BrdU-positive retinal progenitor cells was identified within the ciliary epithelium of the pars plana in continuation of the layer of the peripheral retina during ocular development. In both the ciliary epithelium and the retina, the layer size decreased rapidly and disappeared mostly by postnatal day (P)9. Within the ciliary epithelium of the pars plana, numerous postmitotic rod and cone photoreceptor precursors were identified that rapidly differentiated morphologically and decreased in number with ocular development. Rod precursors were no longer seen in the pars plana at P12, whereas rare presumptive cone precursors persisted even at P120. An increase in the number of presumptive cone precursors (approximately 16-fold) was identified in the pars plana of adult mice with MNU-induced photoreceptor degeneration. Rod and cone precursors were identified in the ciliary epithelium of the murine pars plana during ocular development but nearly disappeared after the completion of histogenesis. However, in response to retinal injury, an increased number of presumptive cone precursors was found even in the adult pars plana.

Expression of PTPIP51 during mouse eye development

The expression pattern of the novel tyrosine phosphatase interacting protein 51 (PTPIP51) was studied during the organogenesis of mouse eye on a transcriptional (RT-PCR and in situ hybridization) and translational level (immunohistochemistry and immunoblotting). Timed developmental stages from day E12 to day E18 were analyzed regarding the distribution of PTPIP51 and compared to the expression patterns observed during postnatal developmental stages and adult eye. PTPIP51 was found to be expressed in all investigated developmental stages in derivatives of mesoderm and ectoderm, such as developing cornea, lens, neuroretina and extraocular muscles. Conjuctiva and corneal epithelia were PTPIP51 reactive during all investigated developmental stages including the mature eye. Embryonic differentiation led to reactive keratocytes of the corneal stroma and remained so in post partal stages, as well as in the adult eye. On day E12, all cells comprising the developing lens body showed PTPIP51 expression. Further development unto the adult eye resulted in a restriction of PTPIP51 expression to the anterior lens epithelium and finally to the equatorial region of the lens epithelium. The developing neuroretina showed a strong PTPIP51 expression in the inner neuroblastic layer and the future receptor cell layer. In the adult eye, the retinal ganglion cells and the inner nuclear layer remained PTPIP51 reactive. The data presented here suggests PTPIP51 to be integrated in signaling cascades regulating differentiation and apoptosis during eye development.

Signal transduction pathways associated with ATP-induced proliferation of cell progenitors in the intact embryonic retina

ATP and ADP induce retinal cell proliferation through activation of PKC and extracellular signal-regulated kinases (ERKs). Here, we characterized the effect of purinergic agonists on the turnover of phosphoinositides and activation of ERKs during development of the chick embryo retina. When intact retinas were incubated with ATP, ADP or UTP, a dose-dependent accumulation of [3H]-phosphoinositides was observed (% of control, EC50: 548±20.5%, 0.18mM; 314±53.8%, 0.51mM; 704±139.9%, 0.018mM, respectively). Only the response promoted by ADP was completely inhibited by the P2 receptor antagonists, PPADS and suramin. All the responses decreased with the progression of retinal development. Western blot assays revealed that ATP, ADP and UTP stimulated the phosphorylation of ERKs in the chick embryo retina very early during development (% of control: 174±16; 199±16.4 and 206±37, respectively). The responses to ADP and UTP were transient and dose-dependent, showing EC50 values of 0.12mM and 0.009mM. The response to ADP was inhibited by the antagonists PPADS and suramin and by U73122 and chelerythrine chloride, which block PLC and PKC, respectively. Conversely, chelerythrine chloride did not block the response induced by UTP. Immunohistochemical analysis revealed that ATP and ADP induced the phosphorylation of ERKs in cells of the neuroblastic layer of retinas from embryos at E8. Our data showed that ATP, ADP and UTP stimulate the turnover of InsPs and promoted the activation of ERKs in the chick embryo retina. ADP, through activation of P2Y1 receptors, activated ERK pathway through PLC and PKC and UTP, via P2Y4-like receptors, induced the phosphorylation of ERKs through a pathway that did not involve PKC.

Hop/STI1 modulates retinal proliferation and cell death independent of PrPC

Hop/STI1 is a co-chaperone adaptor protein for Hsp70/Hsp90 complexes. Hop/STI1 is found extracellularly and modulates cell death and differentiation through interaction with the prion protein (PrPC). Here, we investigated the expression of hop/STI1 and its role upon cell proliferation and cell death in the developing retina. Hop/STI1 is more expressed in developing rat retina than in the mature tissue. Hop/STI1 blocks retinal cell death in the neuroblastic layer (NBL) in a PrPC dependent manner, but failed to protect ganglion cells against axotomy-induced cell death. An antibody raised against hop/STI1 (α-STI1) blocked both ganglion cell and NBL cell death independent of PrPC. cAMP/PKA, ERK, PI3K and PKC signaling pathways were not involved in these effects. Hop/STI1 treatment reduced proliferation, while α-STI1 increased proliferation in the developing retina, both independent of PrPC. We conclude that hop/STI1 can modulate both proliferation and cell death in the developing retina independent of PrPC.

Gene expression patterns in primary neuronal clusters of the Drosophila embryonic brain

The brain of Drosophila is formed by approximately 100 lineages, each lineage being derived from a stem cell-like neuroblast that segregates from the procephalic neurectoderm of the early embryo. A neuroblast map has been established in great detail for the early embryo, and a suite of molecular markers has been defined for all neuroblasts included in this map [Urbach, R., Technau, G.M. (2003a) Molecular markers for identified neuroblasts in the developing brain of Drosophila. Development 130, 3621–3637]. However, the expression of these markers was not followed into later embryonic or larval stages, mainly due to the fact that anatomical landmarks to which expression patterns could be related had not been defined. Such markers, in the form of stereotyped clusters of neurons whose axons project along cohesive bundles (“primary axon bundles” or “PABs”) are now available [Younossi-Hartenstein, A., Nguyen, B., Shy, D., Hartenstein, V. 2006. Embryonic origin of the Drosophila brain neuropile. J. Comp. Neurol. 497, 981–998]. In the present study we have mapped the expression of molecular markers in relationship to primary neuronal clusters and their PABs. The markers we analyzed include many of the genes involved in patterning of the brain along the anteroposterior axis (cephalic gap genes, segment polarity genes) and dorso-ventral axis (columnar patterning genes), as well as genes expressed in the dorsal protocerebrum and visual system (early eye genes). Our analysis represents an important step along the way to identify neuronal lineages of the mature brain with genes expressed in the early embryo in discrete neuroblasts. Furthermore, the analysis helped us to reconstruct the morphogenetic movements that transform the two-dimensional neuroblast layer of the early embryo into the three-dimensional larval brain and provides the basis for deeper understanding of how the embryonic brain develops.

NMDA-receptor blockade enhances cell apoptosis in the developing retina of the postnatal rat

During visual system development, programmed cell death occurs in order to facilitate the establishment of correct connections and synapses. During this period, glutamate plays a very important role as an excitatory neurotransmitter. With a view to evaluating if NMDA glutamate receptors participate in the regulation of apoptosis which occurs during the development of the rat retina, we subcutaneously injected the NMDA receptor antagonist MK-801 into rats at different stages of early postnatal development (P2 to P9). Ensuing cell death in the retina and superior colliculus was analyzed by using the Feulgen method. MK-801 administration had no effect on the survival of photoreceptor cells. In contrast, the presence of this antagonist induced a significant increase in the number of apoptotic cells in the neuroblastic layer (P7 and P8) and ganglion cell layer (P6-P8), as well as in the superior colliculus which receives afferent contacts from retinal ganglion cells during P7-P9. We conclude that during development, specific types of cells in the mammalian retina are critically dependent for their survival on glutamate stimulation through NMDA receptors. These findings thus throw fresh light on the mechanisms of development of the rat visual system by identifying NMDA glutamate receptors as participants in the regulation of apoptotic processes which occur during the initial stages of development.

The initial fetal human retinal vasculature develops by vasculogenesis

There is increasing evidence that the hemangioblast, a common progenitor for hematopoietic cells and endothelial cells, participates in embryonic and extra-embryonic vasculogenesis in some organs. Whether resident angioblasts or endothelial progenitor cells (EPCs) contribute to human retinal vasculogenesis is still a matter of controversy. To address this controversy, fetal human retinas of 6-23 weeks gestation (WG) were examined using immunohistochemistry and a panel of antibodies against endothelial cell markers (CD34, CD31), a marker for retinal angioblasts and endothelium (CD39/ecto-ADPase), and a marker for precursors and hemangioblasts (CXCR4). Confocal microscopic spectral analysis and double labeling with Ki67 was used to identify the proliferating cell types. In the inner neuroblastic layer of the 6-8 WG retina and in the putative ganglion cell layer in avascular regions of older eyes (14 WG-20 WG), scattered CD39+ angioblasts were well in advance of forming vasculature. There was a layer of CXCR4+ cells in the inner retina that was reduced in size with development. As blood vessels formed, CD39+ cells were always well in advance of the vascular front and they expressed CXCR4. This demonstrates that a pool of resident angioblasts express CD39 and CXCR4 as they differentiate and participate in vasculogenesis in the fetal human. They retain expression of CD39 as endothelial cells in the newly formed retinal vasculature but they down-regulate CXCR4 expression.