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  • 1
    Publication Date: 2013-04-12
    Description: Obtaining high-resolution information from a complex system, while maintaining the global perspective needed to understand system function, represents a key challenge in biology. Here we address this challenge with a method (termed CLARITY) for the transformation of intact tissue into a nanoporous hydrogel-hybridized form (crosslinked to a three-dimensional network of hydrophilic polymers) that is fully assembled but optically transparent and macromolecule-permeable. Using mouse brains, we show intact-tissue imaging of long-range projections, local circuit wiring, cellular relationships, subcellular structures, protein complexes, nucleic acids and neurotransmitters. CLARITY also enables intact-tissue in situ hybridization, immunohistochemistry with multiple rounds of staining and de-staining in non-sectioned tissue, and antibody labelling throughout the intact adult mouse brain. Finally, we show that CLARITY enables fine structural analysis of clinical samples, including non-sectioned human tissue from a neuropsychiatric-disease setting, establishing a path for the transmutation of human tissue into a stable, intact and accessible form suitable for probing structural and molecular underpinnings of physiological function and disease.〈br /〉〈br /〉〈a href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4092167/" target="_blank"〉〈img src="https://static.pubmed.gov/portal/portal3rc.fcgi/4089621/img/3977009" border="0"〉〈/a〉   〈a href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4092167/" target="_blank"〉This paper as free author manuscript - peer-reviewed and accepted for publication〈/a〉〈br /〉〈br /〉〈span class="detail_caption"〉Notes: 〈/span〉Chung, Kwanghun -- Wallace, Jenelle -- Kim, Sung-Yon -- Kalyanasundaram, Sandhiya -- Andalman, Aaron S -- Davidson, Thomas J -- Mirzabekov, Julie J -- Zalocusky, Kelly A -- Mattis, Joanna -- Denisin, Aleksandra K -- Pak, Sally -- Bernstein, Hannah -- Ramakrishnan, Charu -- Grosenick, Logan -- Gradinaru, Viviana -- Deisseroth, Karl -- DP1 OD000616/OD/NIH HHS/ -- R01 DA020794/DA/NIDA NIH HHS/ -- R01 MH099647/MH/NIMH NIH HHS/ -- Howard Hughes Medical Institute/ -- England -- Nature. 2013 May 16;497(7449):332-7. doi: 10.1038/nature12107. Epub 2013 Apr 10.〈br /〉〈span class="detail_caption"〉Author address: 〈/span〉Department of Bioengineering, Stanford University, Stanford, California 94305, USA.〈br /〉〈span class="detail_caption"〉Record origin:〈/span〉 〈a href="http://www.ncbi.nlm.nih.gov/pubmed/23575631" target="_blank"〉PubMed〈/a〉
    Keywords: Animals ; Brain/*anatomy & histology ; Cross-Linking Reagents/chemistry ; Formaldehyde/chemistry ; Humans ; Hydrogel/chemistry ; Imaging, Three-Dimensional/*methods ; In Situ Hybridization/methods ; Lipids/isolation & purification ; Mice ; Molecular Imaging/*methods ; Permeability ; Phenotype ; Scattering, Radiation
    Print ISSN: 0028-0836
    Electronic ISSN: 1476-4687
    Topics: Biology , Chemistry and Pharmacology , Medicine , Natural Sciences in General , Physics
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  • 2
    ISSN: 0022-0248
    Source: Elsevier Journal Backfiles on ScienceDirect 1907 - 2002
    Topics: Chemistry and Pharmacology , Geosciences , Physics
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Abstract: Rats born to well-fed mothers (20% protein diet ad libitum), protein-restricted mothers (7.5% protein diet ad libitum) or pair-fed with protein-restricted mothers were killed on days 0, 7, 14, 21, 28 and 35 and activities of the two enzymes of neurotransmitter synthesis, tryptophan-5-hydroxylase (EC 1.14.16.4) and tyrosine hydroxylase (EC 1.14.16.2) were assayed. Enzyme activities in normal animals were low at birth and progressively increased to reach adult levels by day 15. Protein-restricted and pair-fed animals also showed a similar pattern. However, significantly higher activities were observed from day 15 onwards in both experimental groups.
    Type of Medium: Electronic Resource
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  • 4
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Journal of neurochemistry 27 (1976), S. 0 
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: The pattern of development of brain tryptophan in the rat was studied in the progeny of mothers fed a 7.5%; protein diet ad lib., a 20% protein diet ad lib. and those fed a 20%; protein diet pair-fed with mothers who received the 7.5% protein. The pattern of development was similar in all three groups. Starting with a high brain tryptophan content at birth, all animals showed a progressive reduction during the next 3 weeks. However, tryptophan levels at birth were several fold higher in the brains of pups born to mothers receiving either the low protein diet fed ad lib. or those born to mothers who received the 20% protein diet in restricted amounts. From the 14th day after birth, tryptophan concentration of brain in undernourished pups was significantly lower until the 35th day. The implications of this finding are discussed.
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 1573-6903
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Incorporation of tracer doses of radiolabeled tryptophan and tyrosine into brain proteins was investigated in rats malnourished during gestation and lactation. Age and time dependent increases in the radioactivity was observed in the whole homogenate and in the TCA insoluble fraction. Protein malnourished rats showed increased incorporation of tryptophan and tyrosine. However the diet restricted (Pair-fed) animals showed increased incorporation of tyrosine only. The increased incorporation may probably be due to changes in the pool size of the amino acids and effective recycling of the amino acids. The enhanced utilization in protein synthesis may also probably offer a mechanism for conservation of these amino acids.
    Type of Medium: Electronic Resource
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  • 6
    ISSN: 1573-6903
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Free amino acid levels in the brains of young ones born to mothers fed a 20% protein diet ad libitum (well nourished), 7.5% protein diet ad libitum (protein restricted) and a 20% protein diet in restricted amounts (pair-fed) were investigated during brain development in the present study. The dietary protein was obtained from a cereal-legume mixture. Protein restricted animals showed increases in the levels of taurine, glycine and glutamic acid and decreases in the concentrations of methionine, leucine, isoleucine, and GABA. The pair-fed animal showed increases only in glutamic acid and glycine and a decrease only in the levels of GABA. The significance of these observations is discussed.
    Type of Medium: Electronic Resource
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