Ординатура / Офтальмология / Английские материалы / Eye, Retina, and Visual System of the Mouse_Chalupa, Williams_2008

Plate 6 Morphology and projections of mRGCs in mouse. A, Retina flat mount stained with a polyclonal antimouse melanopsin antibody showing specific staining of dendrites, somata, and axons of a small subset of retinal ganglion cells. B, Schematic drawing showing direct axonal projections of mRGCs to several brain regions. (Results from Hattar et al., 2006, are redrawn here.) Brain regions receiving significant projections are represented in large, bold letters. AH, anterior hypothalamic nucleus; IGL, inter-

geniculate leaflet; LGd, lateral geniculate nucleus, dorsal division; LGv, lateral geniculate nucleus, ventral division; LH, lateral hypothalamus; LHb, lateral habenula; MA, median amygdaloid nucleus; OPN, olivary pretectal nucleus; PAG, periaqueductal gray; PO, preoptic; pSON, perisupraoptic nucleus; SC, superior colliculus; SCN, suprachiasmatic nucleus; SPZ, subparaventricular zone. (See figure 17.2.)

Plate 7 Different maps of mouse visual cortex. A, Flat map of cytoarchitectonic areas (black outlines) in the left hemisphere of mouse cerebral cortex published by Caviness and Frost (1980). Red shaded regions represent schematic outlines of visuotopically organized areas, identified by Wang and Burkhalter (2006). Note that areas 18a and 18b contain multiple visuotopic areas. B, Flat map of cytoarchitectonic areas of the left mouse cerebral cortex constructed by David C. Van Essen by unfolding coronal sections taken from the atlas of Paxinos and Franklin (2001). Red outlines represent schematic borders of visuotopically defined areas identified by Wang and Burkhalter (2007). Note that areas V2L and

V2ML contain multiple visuotopically organized areas. C, Visuotopic organization of the left mouse visual cortex derived by mapping of receptive fields, published by Wagor et al. (1980). In this map, cytoarchitectonic area 18a contains areas V2 and V3 and cytoarchitectonic area 18b contains the rostral and caudal medial visual areas, Vm-r and Vm-c. D, Area map of left mouse visual cortex derived by topographic mapping of V1 connections and receptive field mapping (Wang and Burkhalter, 2007). Blue shading represents the distribution of callosal connections in superficial layers. (See figure 20.1.)

Plate 8 Topographic maps of V1 connections in mouse visual cortex. Representation of azimuth in extrastriate visual cortex is shown in horizontal sections of left occipital cortex. The maps were generated by making three simultaneous injections of fluororuby (FR, red), fluoroemerald (FE, green), and biotinylated dextran amine (BDA, yellow) into V1, followed by triple-anterograde tracing of intracortical connections. A, Darkfield image showing heavy myelination in primary visual cortex (V1) and the barrel field of primary somatosensory cortex (S1). Arrowheads indicate myeloarchitectonic borders. Arrows indicate injection sites in V1.

B, Fluorescently labeled axonal connections after injections of FR, FE, and BDA at different nasotemporal locations (azimuth) of the upper visual field representation in V1. Dashed lines indicate areal borders, which were determined by mapping inputs from the perimeter of V1. Solid lines indicate myeloarchitectonic borders. C, Overlay of BDA-labeled axonal projections shown in B and bisbenzimide-labeled callosally projecting neurons (blue). D, Higher magnification image of axonal labeling shown in area A (inset in B). A, anterior; L, lateral; M, medial; P, posterior. Scale bar = 1 mm (A–C), 0.1 mm (B, inset), and 0.3 mm (D). (See figure 20.2.)

Plate 9 Intracortical connections of the extrastriate lateromedial area (LM) in mouse visual cortex, shown in tangential sections through the flattened posterior cerebral cortex. A, Fluorescence image showing the distribution of retrogradely labeled callosal connections. Dashed lines represent the myeloarchitectonic borders of V1, S1, and RSA. B, Darkfield image of biotinylated dextran amine

(BDA)–labeled axonal connections of area LM. Asterisk indicates the injection site. Note the strong connections to areas POR and 36p. C, Superimposition of BDA-labeled LM connections (gold) with callosal connections (blue somata). A, anterior; L, lateral; M, medial; P, posterior. Scale bar = 1 mm in all images. (See figure 20.3.)

Plate 10 Intracortical connections of the extrastriate lateromedial area (AL) in mouse visual cortex, shown in tangential sections through the flattened posterior cerebral cortex. A, Fluorescence image showing the distribution of retrogradely labeled callosal connections. Dashed lines represent the myeloarchitectonic borders of V1, S1, and RSA. B, Darkfield image of biotinylated dextran amine

(BDA)–labeled axonal connections of area AL. Asterisk indicates injection site. Note the strong connections to areas PL, A, and Cg/RS. C, Superimposition of BDA-labeled LM connections (gold) with callosal connections (blue somata). A, anterior; L, lateral; M, medial; P, posterior. Scale bar = 1 mm in all images. (See figure 20.4.)

Plate 11 Laminar organization of inter-areal feedforward and feedback connections in mouse visual cortex labeled by anterograde tracing with BDA. A, Coronal section showing feedforward axons that originate from the lower area V1 and terminate in the higher extrastriate area LM. The projection column includes layers

Plate 12 Cre expression patterns in three different transgenic strains: Le-Cre (A–C), MLR10 (D–F ), and MLR39 (G–I ). Cre expression in whole-mount (A and B ) or tissue sections (C–I ) is indicated by blue (B and D–I ) or purple (A and C) staining following histochemical detection of a Cre-activated reporter allele. Arrows in

2/3 to 6, and inputs to layer 1 are sparse. B, Coronal section showing feedback axons that originate from area LM and terminate in V1. The projections to layer 1, 2/3, and 5 are dense, whereas inputs to layer 4 are sparse. Scale bar = 0.2 mm. (See figure 20.5.)

A and B indicate Cre expression in the developing pancreas of LeCre mice. Developmental time points are indicated. (See figure 22.2.) (Adapted from Ashery-Padan et al., 2000, and Zhao et al., 2004.)

Plate 13 Visualization of vessels in P17 retinal flat-mount preparations stained with GS-lectin from a mouse model of OIR. A, Low-power (magnification ×25) image of the retina shows areas of central vaso-obliteration, as well as neovascular tufts. B, Higher

magnification (×200) shows neovascular tuft formation at the transition zone between vascular and central avascular retina. C, At even greater magnification (×400), multiple neovascular tufts are evident. (See figure 23.4.)

Plate 14 Developing and mature mouse retinas. A, An embryonic retina at E12.5. B, A postnatal retina at P16. Nuclei are labeled in red with propidium iodide and represent the positions of the cell soma. Neurite processes are labeled in green and blue with antineurofilament antibody and anticholine acetyltransferase antibody, respectively, and represent axons and dendrites. C, Schematic representation of the mature retina. RGCs and displaced

amacrine cells are found within the ganglion cell layer (GCL). Intermediate neurons (horizontal, bipolar, and amacrine) are found within the inner nuclear layer (INL), and rod and cone photoreceptor cells are found within the outer nuclear layer (ONL). IPL and OPL are inner plexiform and outer plexiform layers where axons and dendrites synapse. RGC, retinal ganglion cell. (See figure 26.1.)

Plate 15 A gene regulatory network model for RGC development. Genes are depicted such that the right side of the bent arrow for each gene indicates the gene product and the left side indicates the transcriptional control region. Solid lines connecting genes indicate established upstream-downstream relationships, and dashed lines suggest inferred relationships. Downward-pointing arrows indicate gene activation, and downward perpendicular lines indicate gene repression. In most cases the connections have not been shown to be direct. Genes boxed in yellow represent regulators of general retinal competence and RPC proliferation; genes boxed in orange are proneural bHLH genes associated with establishing competence in

RPCs for specific retinal cell fates. The light brown-purple box represents the general repression of RGC genes hypothesized to be mediated by the neural differentiation transcriptional repressor NSRF/REST (Mu et al., 2005b); the blue and purple-blue boxes represent genes encoding RGC-specific upstream (blue) and downstream (purple-blue) transcription factors. Genes boxed in green are those that encode proteins associated with RGC maturation and function. Genes boxed in purple encode secreted signaling molecules. In some cases only representative examples are shown for each box. A more detailed description can be found in Mu et al., 2005b. (See figure 26.2.)

Plate 16 Segregated expression of Math5 and Pou4f2 in retinogenesis. The image represents an E14.5 retina from a knock-in mouse in which a gene encoding an HA-epitope-tagged Math5 replaces the endogenous Math5 allele. Math5-HA expression is

Plate 17 Three functional stages of retinal wave–generating circuits. A, Schematic of circuits that mediate retinal waves. (Modified from Catsicas and Mobbs, 1995.) B, Summary of development of the synaptic circuitry that mediates waves in mice. Each color corresponds to a different wave-generating circuit. Yellow corresponds to non-nAChR circuitry, which mediates the nonpropagating events in embryonic mice. There is pharmacological evidence that stage I waves in other species are mediated by gap junctions, but this has not been directly demonstrated in mouse retina. In addi-

detected by immunostaining with an anti-HA antibody (red). Pou4f2 expression is detected by immunostaining with an antiPou4f2 antibody (green). Yellow staining shows the overlap in expression of Math5 and Pou4f2. (See figure 26.3.)

tion, it is not known which gap junction–coupled networks mediate stage I waves. Red corresponds to stage II circuits, which require activation of nAChRs. Stage II waves are initiated and propagate through a network of starburst amacrine cells. Blue corresponds to stage III circuits, which require activation of ionotropic glutamate receptors. The source of stage III wave initiation and the location of the horizontal coupling that drives coordinated release of glutamate during this stage are not yet known. (See figure 28.1.) (Modified from Bansal et al., 2000.)

Plate 18 Calcium imaging reveals spatial and temporal properties of stage I and stage II waves. A, Time evolution of a single stage II retinal wave visualized with fluorescence imaging of the calcium indicator fura-2. Decreases in fura-2 fluorescence associated with the increased calcium evoked by waves are shown at successive 0.5 s intervals. The final frame represents the total area of tissue covered by a single wave. B, Retinal waves of embryonic day 17 (E17) and postnatal day 2 (P2) retinas. Each frame summa-

rizes 90 s of activity in control ACSF (top row) and in 100 μM d-tubocurarine, a general nAChR antagonist (bottom row). Gray background represents the total retinal surface labeled with fura2AM. Each color corresponds to individual domains, with a colorcoded time bar below each frame to indicate the time of occurrence of each wave. Scale bar = 100 μm. (See figure 28.2.) (Modified from Bansal et al., 2000.)

Plate 19 Dendritic ramification patterns of YFP expressing RGCs in the IPL can be determined using confocal microscopy from Thy1-YFP transgenic mouse retina. A, View from vitreal side of a flat-mounted retina harvested from a Thy1-YFP-expressing mouse. B, Enlarged view of the area inside the box in A. Axons from individual RGCs cross the retina from each soma to the optic nerve head. C, Four frames taken from a representative stack of confocal images of a bistratified RGC showing the soma and axon, the dendrites ramified in sublamina b (blue), the dendrites ramified in sublamina a (green) of the RGC, and immunolabeling of dopami-

nergic amacrine cells (red). D, A stacked image of the same cell as shown in panel C. E, The 90° rotation view of the cell in D. Three dashed lines indicate the inner border of the IPL, the boundary of sublaminae a and b, and the outer border of the IPL. F, Normalized pixel intensity of the dendrites of each frame (open circles) plotted as a function of IPL depth of the cell in panel C. The data were fitted with two Gaussian distributions (green and blue lines). Doublearrow lines indicate widths. Single arrows indicate the locations of the two peaks of dendritic density. (See figure 29.3.)