A Genetic Approach to Visualization of Multisynaptic Neural Pathways Using Plant Lectin Transgene

Yoshihara, Yoshihiro; Mizuno, Takumi; Nakahira, Masakiyo; Kawasaki, Mikio; Watanabe, Yasuyoshi; Kagamiyama, Hiroyuki; Jishage, Kou Ichi; Ueda, Otoya; Suzuki, Hiroshi; Tabuchi, Katsuhiko; Sawamoto, Kazunobu; Okano, Hideyuki; Noda, Tetsuo; Mori, Kensaku

doi:10.1016/s0896-6273(00)80676-5

Cited by 164 publications

(136 citation statements)

References 35 publications

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“…Our splicing analysis in photoreceptors examined the Rh1‐expressing outer photoreceptors, and so we next asked whether proper splicing was also necessary in these outer photoreceptors for visual behavior. We expressed RNAi using Rh1‐Gal4 (Yoshihara et al, 1999). In contrast, the longGMR‐Gal4 is expressed in all photoreceptors (Wernet et al, 2003).…”

Section: Resultsmentioning

confidence: 99%

Proper splicing contributes to visual function in the aging Drosophila eye

et al. 2018

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Changes in splicing patterns are a characteristic of the aging transcriptome; however, it is unclear whether these age‐related changes in splicing facilitate the progressive functional decline that defines aging. In Drosophila, visual behavior declines with age and correlates with altered gene expression in photoreceptors, including downregulation of genes encoding splicing factors. Here, we characterized the significance of these age‐regulated splicing‐associated genes in both splicing and visual function. To do this, we identified differential splicing events in either the entire eye or photoreceptors of young and old flies. Intriguingly, aging photoreceptors show differential splicing of a large number of visual function genes. In addition, as shown previously for aging photoreceptors, aging eyes showed increased accumulation of circular RNAs, which result from noncanonical splicing events. To test whether proper splicing was necessary for visual behavior, we knocked down age‐regulated splicing factors in photoreceptors in young flies and examined phototaxis. Notably, many of the age‐regulated splicing factors tested were necessary for proper visual behavior. In addition, knockdown of individual splicing factors resulted in changes in both alternative splicing at age‐spliced genes and increased accumulation of circular RNAs. Together, these data suggest that cumulative decreases in splicing factor expression could contribute to the differential splicing, circular RNA accumulation, and defective visual behavior observed in aging photoreceptors.

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Section: Resultsmentioning

confidence: 99%

Proper splicing contributes to visual function in the aging Drosophila eye

et al. 2018

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“…To visualize selective and functional transsynaptic neural pathways, wheat germ agglutinin (WGA)-expressing recombinant adenoviruses were used [24,25]. Twelve weeks after injury, 1 ll of saline containing WGA-expressing virus at a titer of 1 Â 10 11 pfu/ml was injected into the bilateral motor cortices (0.5 ll per injection site).…”

Section: Visualization Of Multisynaptic Neural Pathwaysmentioning

confidence: 99%

“…To determine whether disrupted CSTs were reconstructed by forming a local neuronal relay, we injected WGA-expressing adenoviruses into the motor cortex of the hind limb area at 12 weeks after SCI. WGA, a plant lectin, has been widely used as a tracer of neuronal pathways [24,25] because it is transported in axons and dendrites, and across synapses, to second-and even third-order neurons.…”

Section: Local Neurons Reconstruct Disrupted Cst Neuronal Circuits Inmentioning

confidence: 99%

Treatment of a Mouse Model of Spinal Cord Injury by Transplantation of Human Induced Pluripotent Stem Cell-Derived Long-Term Self-Renewing Neuroepithelial-Like Stem Cells

et al. 2012

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Because of their ability to self-renew, to differentiate into multiple lineages, and to migrate toward a damaged site, neural stem cells (NSCs), which can be derived from various sources such as fetal tissues and embryonic stem cells, are currently considered to be promising components of cell replacement strategies aimed at treating injuries of the central nervous system, including the spinal cord. Despite their efficiency in promoting functional recovery, these NSCs are not homogeneous and possess variable characteristics depending on their derivation protocols. The advent of induced pluripotent stem (iPS) cells has provided new prospects for regenerative medicine. We used a recently developed robust and stable protocol for the generation of long-term, self-renewing, neuroepithelial-like stem cells from human iPS cells (hiPS-lt-NES cells), which can provide a homogeneous and well-defined population of NSCs for standardized analysis. Here, we show that transplanted hiPS-lt-NES cells differentiate into neural lineages in the mouse model of spinal cord injury (SCI) and promote functional recovery of hind limb motor function. Furthermore, using two different neuronal tracers and ablation of the transplanted cells, we revealed that transplanted hiPS-lt-NES cell-derived neurons, together with the surviving endogenous neurons, contributed to restored motor function. Both types of neurons reconstructed the corticospinal tract by forming synaptic connections and integrating neuronal circuits. Our findings indicate that hiPS-lt-NES transplantation represents a promising avenue for effective cell-based treatment of SCI. Disclosure of potential conflicts of interest is found at the end of this article.

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“…The transsynaptic trafficking of TTC fragment fusion proteins and WGA is known to be sensitive to presynaptic neuronal activity (Yoshihara et al, 1999;Miana-Mena et al, 2002). These tracers therefore can be used to reveal the presence of functional synapses between V1 INs and motor neurons.…”

Section: Generation and Analysis Of En1mentioning

confidence: 99%