Bipolar Bunny (A Unique Bunny Book 3)

Pretending that it doesn't exist just makes matters worse for children. This books addresses Bipolar disease in simple terms, to help children begin to understand.

Introduction

It may be a little wordy for very young children. A wonderful story, and a wonderful way to introduce a young child to the daily struggles of living with a mental illness. A great way to end the stigma of living with a mental illness. A must have for all children, especially a child who is coping with a parent or relative who is suffering from a mental illness. A very cute story to explain bipolar to little ones. Very heartwarming and easy to understand.

A great book for kiddos. Great little story to introduce and help children understand what Bipolar is all about. We will send you an SMS containing a verification code. Please double check your mobile number and click on "Send Verification Code". Enter the code below and hit Verify. Free Shipping All orders of Don't have an account?

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After 40 — 50 hour incubations, the rabbits were reanesthetized using the same agents and the eyes were ennucleated, hemisected and the retinas removed from the eye cup while immersed in oxygenated Ames Medium Sigma, St. For retinal wholemounts, tissue was stained immediately with antibodies. Polyclonal antibodies were used to identify the two types of cones in rabbit retina.

In each case, the specificity of the antibody was tested by the manufacturer with a Western blot analysis using retinal extract containing the antibody. Free-floating tissue was processed using a combination of bench and microwave staining techniques. In each case, the tissue was permeablized in 0. Permeablized retinas were incubated with goat antiserum against blue-sensitive opsin, n 1: After rinsing, the retina was incubated in the secondary antibody 1: In one experiment, red-green cones were also labeled using the same procedure described above.

The rabbit polyclonal antibody against recombinant human red-green opsin 1: The tissue was processed to reveal biocytin next. After rinsing, the retinas were incubated overnight in fluorescein anti-avidin D 1: The final step was to label all cones with peanut agglutinin, which labels certain disaccharides on the surface of outer and inner segments of all cones and in punctuate regions at the cone pedicles Blanks and Johnson, Retinal sections were processed using the same procedures except for the biocytin step; incubation in avidin DN was shortened to overnight rather than several days and to two hours in fluorescein anti-avidin.

To verify the topographical distribution of cone types in retinal wholemounts, we sampled the distribution of PNA labeled cones that were also labeled with antibodies against red-green cone opsin and blue cone opsin. Cell counts were made from a series of dorsal d and ventral v locations along the retinal midline with the origin located in the middle of the optic disk. Labeled retinas were imaged with an Olympus Fluoview Confocal microscope equipped with helium, argon and neon lasers and an Olympus 60 X water immersion objective NA 1.

A series of optical sections in a single channel were collected every 0. Each channel was scanned separately to reduce spectral cross-talk between channels. Comparisons of the cones, opsins and the biocytin labeled cells were made by merging images of like focal planes using MetaVue v 6. Biocytin injected into the vitreous body of the rabbit eye labeled an assortment of palely-stained ganglion, amacrine and bipolar cell somas and one population of well-stained wide-field bipolar cells Jeon and Masland, They were variable in shape, sometimes radially symmetric while at other times having one axis significantly longer than the other.

The axons tiled the retina without overlap, but extended thinner processes that made apparent contacts with axons from nearby wide-field cells, sometimes making it difficult to judge when one cell ended and another began.

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5. The biocytin wide-field bipolar cell in the rabbit retina selectively contacts blue cones.
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Vertical A and whole mount views B and C of biocytin labeled wide-field bipolar cells. The cell dendrites were made up of thin processes that possessed clusters of terminal swellings. The asymmetric axons ranged between and um in diameter, contained numerous varicosities and in some instances, contacted axons of adjacent wide-field cells. They were narrowly stratified in layer 5 of the inner plexiform layer. The dendrites of the wide-field cells were relatively thinner in diameter than the axons and arose from 2 - 5 thick, proximal segments Figure 1B. Characteristic features of the dendrites included clusters of terminals occurring at branch points or at transitions from thick to thin segments along the length of the dendrite.

Occasionally, the transition points included a fine process emerging from the cluster that eventually merged with a similar process from an adjacent wide-field cell Figure 2A, arrow. These findings are consistent with those of Famiglietti and Jeon and Masland The dendrites of three wide-field bipolar cells green in a patch of mid-peripheral retina together with the overlying cone pedicles magenta ; areas of closely opposed labeling appear white.

The dendrites covered the retinal surface without overlap and had discrete clusters of varicosities that were coincident with a subset of cone pedicles. The arrow indicates a thin process between two cells that did not appear to contact any cone pedicles while traveling in the outer plexiform layer. The box indicates a region of interest shown at higher magnification below. Note that the terminal varicosities green extend quite a distance from the main dendrites and are opposite the lobes of the PNA labeled cone pedicles magenta.

Processes in the vicinity of a cone pedicle that do not extend varicosities into it, are not likely to be making synaptic contacts arrow. The clusters of synaptic terminals found on the wide-field bipolar cell dendrites were not randomly positioned on the dendrites but instead coincident with a subset of cone pedicles.

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Wherever terminal clusters were present on the wide-field dendrites, the varicosities were observed in through focus series entering the PNA-labeled cone pedicles. Four such clusters can be seen at higher magnification in Figure 2B. The wide-field dendrite, labeled in green, extended tiny grape-like clusters sclerally into the overlying cone pedicle. There was a clear separation between the main dendrites which appear out of focus in Figure 2B and the varicosities as they extended into the base of the cone.

Points of possible contact between the terminal clusters and the PNA labeled cones appear white in the image and suggest a synaptic relationship Haverkamp et al. In contrast, dendrites traveling below a pedicle without extending terminal varicosities into it are not likely to be making synaptic contacts 2B, arrow. To assess the topographical distribution of cone types in wholemounts, we labeled all cones using PNA and the cone subtypes with antibodies against red-green cone opsin and blue cone opsin.

In rabbit retina, PNA binds to the membrane of the cone inner and outer segments as well as the cells bodies and axonal extensions of the cones Blanks and Johnson, However, the staining is brightest at the cone pedicles and outer segments. The distribution pattern of red-green cones closely mirrored the distribution of the PNA labeled cones; the peak density of red-green cones was in the visual streak and then gradually declined with increasing eccentricity Figure 3B, green line.

Wholemounted retina triple-labeled with PNA, red-green cone opsin green and blue cone opsin turquoise.

In dorsal d and ventral v retina, 2 — 10 mm ventral to the middle of the optic disk v2 - v10 , PNA labeled cones stained for red-green opsin upper left or blue opsin upper right but not both. The density of blue cones blue , red-green cones green , and all cones black , were sampled along the dorsal-ventral midline in a retina wholemount. For simplicity, the number of dual opsin cones was included with the numbers of red-green cones. In peripheral retina, we also observed the presence of cones expressing more than one cone opsin. Mixed cones were not seen in dorsal d or ventral v retina superior to v8 8 mm inferior to the optic disk where PNA labeled cones stained for either blue cone opsin or red-green cone opsin, but not both Figure 3A, upper panels.

Since it was not possible to differentiate the function of blue cones from cones expressing dual cone opsins, we restricted our study of cone-to-bipolar cell contacts to retinal eccentricities superior to v8. Cone type-specific contacts of the wide-field bipolar cells were studied in sections and wholemounts that had been double-labeled with PNA and blue cone opsin.

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In sections, it was possible to see both the dendrites of a wide-field cell enter a cone pedicle and its corresponding blue-opsin stained outer segment perfectly in register Figure 4. In some instances however, the sections were not oriented perfectly vertical, causing the outer segment to be slightly offset from its pedicle.

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Confocal stacks taken from wholemounted retina confirmed that pedicles contacted by the wide-field bipolar dendrites corresponded to blue cone opsin stained outer segments. Figure 5A shows the dendrites of a single wide-field bipolar cell with its four terminal clusters indicated by arrows. Each one of these clusters was coincident with a cone pedicle magenta whose outer segment expressed blue cone opsin Figure 5B.

Five of these images are shown in Figure 6 with each cone indicated by an arrow as it traveled through the ONL. The base of each outer segment corresponds to each of the four blue cones in Figure 5B. Vertical section showing a wide-field bipolar cell dendrite entering the base of a blue cone pedicle boxed.

Note that the pedicle is in register with its blue opsin stained outer segment turquoise.

The biocytin wide-field bipolar cell in the rabbit retina selectively contacts blue cones

Higher magnification view of the cone pedicle to show that the dendritic terminal cluster green is within the PNA labeled cone pedicle magenta. A wide-field bipolar cell green and the overlying PNA labeled cones magenta. Cones contacted by the wide-field bipolar cell are identified by arrows and are all positive for blue cone opsin turquoise. Through-focus images of the cones in Figure 5 to show their path through the outer nuclear layer.

The arrows identify the blue cones as they extend from the cone pedicles bottom to the cone outer segments top. The number of cones that each bipolar cell contacted depended on retina eccentricity, but usually ranged between 2 and 7. In mid-peripheral retina, each wide-field bipolar cell contacted a unique set of cones and all blue cones were contacted by only one wide-field bipolar cell Figure 7. As the size of the dendritic fields increased, the number of cones contacted increased as well. In dorsal retina, where the density of blue cones is lower and the diameter of the wide-field cell dendrites is large, each cone was routinely contacted by more than one wide-field cell, while in the far ventral retina, which contains a combination of blue cones and dual opsin cones, only a subset of cones expressing blue cone opsin was contacted by each wide-field bipolar cell not shown.

A composite image in mid-peripheral retina that shows the terminal varicosities on wide-field bipolar dendrites circles and the corresponding blue cone opsin staining turquoise. In mid-peripheral retina, wide-field cells contact more than one blue cone and each blue cone is contacted by only one wide-field bipolar cell.

The biocytin stained wide-field bipolar cell is the rabbit's equivalent of the mammalian ON blue cone bipolar cell that has been described in monkey, mouse and ground squirrel retinas Kouyama and Marshak, ; Haverkamp et al. Like its mammalian counterparts, the rabbit's wide-field cell is more than twice as large as any other bipolar cell type MacNeil et al.

Its axon resides in layer 5 of the the inner plexiform layer and is thus presumed to be an ON bipolar cell. Moreover, the dendrites of the wide-field cell extend throughout the inner plexiform layer and make selective contacts with cones that express blue cone opsin in their outer segments and avoid cones that do not.

We assume that these points of contact represent synaptic junctions because we were able to follow the dendrites in through focus series into blue cone pedicles and see them terminate adjacent to patches of PNA labeling. Invaginating bipolar processes are a hallmark of ON cone bipolar cells and mark the locations where synapses between bipolar cells and cones occur Hopkins and Boycott, This suggests that the proximity of the PNA label with the biocytin stained processes in the blue cone pedicles reflects a synaptic relationship.

Synaptic communication between blue cone bipolar cells and blue cones has been clearly shown by Li and DeVries in ground squirrel retina. They demonstrated that only one type of bipolar cell made exclusive contacts with blue cones and these contacts were functional; depolarization in a presynaptic cone generated a prominent outward current in a coupled bipolar cell. In addition to the contacts that the wide-field cell dendrites made with cones, the dendrites also included thin processes Famiglietti, that merged with the thin dendrites of adjacent cells Jeon and Masland, These processes usually arose from a terminal cluster aligned with a blue cone and were smooth and not associated with any cone pedicles that might suggest a synaptic junction.

It is likely that gap junctions exist between the thin dendrites of adjacent cells, but their existence could not be established in the confocal images. One significant difference observed in the cone contacts of the rabbit wide-field bipolar cell was the degree of divergence of the photoreceptor input. In primates and mice, the density ratio of blue cone bipolar cells to blue cones is approximately 1: In rabbits, the divergence of blue cone input onto each wide-field bipolar cell depends on retinal eccentricity. In dorsal retina, where the density of blue cones is low relative to the numbers of wide-field bipolar cells, blue cones were contacted by dendrites from more than one wide-field cell.

In ventral retina, multiple cones were more likely to converge onto each wide-field bipolar cell with none of them contacting more than one Figure 7. The advantage of this arrangement could be to preserve spatial representation of the blue signal in ventral retina while allowing for better detection of the blue-cone signal in dorsal retina where the number of blue cones is lowest. A small fraction of cones in peripheral ventral rabbit retina were labeled with antibodies against both blue and red-green cone opsins.

In mouse retina, the majority of cones express dual cone opsins Applebury et al. In the rabbit retina, the biocytin wide-field cells contacted all blue cones in their dendritic field except in the ventral rim where mixed pigment cones were observed. At this eccentricity, the cell dendrites contacted a subset of blue cones suggesting that cones expressing more than one cone opsin are functionally different from genuine blue cones.

The advantage of having such mixed cones is not yet understood, but may be an adaptation of diurnal mammals for better detection of predators against a blue sky background Ahnelt and Kolb, ; Peichl, The blue cone selectivity of the biocytin wide-field bipolar cell and its structural similarity to blue cone bipolar cells in monkey, mouse and ground squirrel retinas suggests that it is involved with color processing in rabbit retina.

The idea that wide-field cells would be selective for blue cones was first proposed by Famiglietti