Cooperative Synthesis of Ultra Long-Chain Fatty Acid and Ceramide during Keratinocyte Differentiation
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{"title"=>"Cooperative Synthesis of Ultra Long-Chain Fatty Acid and Ceramide during Keratinocyte Differentiation", "type"=>"journal", "authors"=>[{"first_name"=>"Yukiko", "last_name"=>"Mizutani", "scopus_author_id"=>"8703543700"}, {"first_name"=>"Hui", "last_name"=>"Sun", "scopus_author_id"=>"55729309800"}, {"first_name"=>"Yusuke", "last_name"=>"Ohno", "scopus_author_id"=>"13610456100"}, {"first_name"=>"Takayuki", "last_name"=>"Sassa", "scopus_author_id"=>"7007024709"}, {"first_name"=>"Takeshi", "last_name"=>"Wakashima", "scopus_author_id"=>"55777742400"}, {"first_name"=>"Mari", "last_name"=>"Obara", "scopus_author_id"=>"55777736800"}, {"first_name"=>"Kohei", "last_name"=>"Yuyama", "scopus_author_id"=>"6602207643"}, {"first_name"=>"Akio", "last_name"=>"Kihara", "scopus_author_id"=>"7004864387"}, {"first_name"=>"Yasuyuki", "last_name"=>"Igarashi", "scopus_author_id"=>"7401635178"}], "year"=>2013, "source"=>"PLoS ONE", "identifiers"=>{"pmid"=>"23826266", "sgr"=>"84879515141", "doi"=>"10.1371/journal.pone.0067317", "scopus"=>"2-s2.0-84879515141", "pui"=>"369208870", "isbn"=>"1932-6203 (Electronic)\\r1932-6203 (Linking)", "issn"=>"19326203"}, "id"=>"bcb7797b-0eaa-31cf-bf1a-210951604218", "abstract"=>"The lipid lamellae in the stratum corneum is important for the epidermal permeability barrier. The lipid lamellae component ceramide (CER), comprising an ultra long-chain (ULC) fatty acid (FA) of ≥26 carbons (ULC CER), plays an essential role in barrier formation. ULC acyl-CoAs, produced by the FA elongase ELOVL4, are converted to ULC CERs by the CER synthase CERS3. In the presented study, we observed that ELOVL4 and CERS3 mRNAs increased during keratinocyte differentiation in vivo and in vitro. We also determined that peroxisome proliferator-activated receptor β/δ is involved in the up-regulation of the mRNAs. Knockdown of CERS3 caused a reduction in the elongase activities toward ULC acyl-CoAs, suggesting that CERS3 positively regulates ULCFA. Thus, we reveal that the two key players in ULC CER production in epidermis, CERS3 and ELOVL4, are coordinately regulated at both the transcriptional and enzymatic levels.", "link"=>"http://www.mendeley.com/research/cooperative-synthesis-ultra-longchain-fatty-acid-ceramide-during-keratinocyte-differentiation", "reader_count"=>22, "reader_count_by_academic_status"=>{"Unspecified"=>2, "Researcher"=>6, "Student > Doctoral Student"=>1, "Student > Ph. D. Student"=>6, "Student > Master"=>3, "Student > Bachelor"=>3, "Professor"=>1}, "reader_count_by_user_role"=>{"Unspecified"=>2, "Researcher"=>6, "Student > Doctoral Student"=>1, "Student > Ph. D. Student"=>6, "Student > Master"=>3, "Student > Bachelor"=>3, "Professor"=>1}, "reader_count_by_subject_area"=>{"Unspecified"=>2, "Biochemistry, Genetics and Molecular Biology"=>6, "Medicine and Dentistry"=>4, "Agricultural and Biological Sciences"=>7, "Pharmacology, Toxicology and Pharmaceutical Science"=>1, "Chemistry"=>2}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>4}, "Chemistry"=>{"Chemistry"=>2}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>7}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>6}, "Unspecified"=>{"Unspecified"=>2}, "Pharmacology, Toxicology and Pharmaceutical Science"=>{"Pharmacology, Toxicology and Pharmaceutical Science"=>1}}, "group_count"=>0}

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/1104383"], "description"=>"<p>(A) Total RNA was prepared from keratinocytes differentiated for 0, 3, or 6 days in differentiation medium. SYBR green-based real-time quantitative PCR was performed using primers specific for <i>ELOVL1</i>, <i>ELOVL4</i>, <i>ELOVL5, ELOVL6</i>, or <i>ELOVL7</i>, and for <i>GUSB</i> as an internal control. The expression level of each <i>ELOVL</i> mRNA was calculated using a standard curve and normalized to that of <i>GUSB</i>. Values presented are the amount of each <i>ELVOL</i> mRNA relative to that of <i>ELOVL4</i> at day 0, and represent the mean ± S.D. from three independent reactions. Statistically significant differences from day 0 are indicated (*p<0.05, **p<0.01, ***p<0.001; Student’s t-test). E1, <i>ELOVL1</i>; E4, <i>ELOVL4</i>; E5, <i>ELOVL5</i>; E6, <i>ELOVL6</i>; E7, <i>ELOVL7</i>. (B) Total cell lysates (10 µg protein) prepared from keratinocytes differentiated for 0, 2, 4, or 8 days in differentiation medium were subjected to immunoblotting with an anti-CERS3 antibody, anti-CERS2 antibody, anti-involucrin antibody, or, to demonstrate uniform protein loading, anti-GAPDH antibody. (C) Total membrane proteins (40 µg protein) from keratinocytes differentiated for 0, 2, or 4 days were incubated with the indicated acyl-CoA (50 µM) and 0.075 µCi [<sup>14</sup>C] malonyl-CoA for 30 min at 37°C. After termination of the reactions, acyl-CoAs were converted to FAs and separated by normal-phase TLC, followed by detection and quantification by a BAS-2500 bioimaging analyzer (Fuji Photo Film). Values presented are FA levels and represent the mean ± S.D. from three independent experiments. Statistically significant differences compared to 0 day cells are indicated (*p<0.05; Student’s t-test).</p>", "links"=>[], "tags"=>["Biochemistry", "enzymes", "Enzyme regulation", "lipids", "Fatty acids", "sphingolipids", "proteins", "Molecular cell biology", "Signal transduction", "Nuclear receptor signaling", "gene expression", "up-regulated", "keratinocyte"], "article_id"=>734069, "categories"=>["Biological Sciences"], "users"=>["Yukiko Mizutani", "Hui Sun", "Yusuke Ohno", "Takayuki Sassa", "Takeshi Wakashima", "Mari Obara", "Kohei Yuyama", "Akio Kihara", "Yasuyuki Igarashi"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0067317.g001", "stats"=>{"downloads"=>0, "page_views"=>15, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_ELOVL4_is_up_regulated_during_keratinocyte_differentiation_/734069", "title"=>"ELOVL4 is up-regulated during keratinocyte differentiation.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-06-27 04:03:45"}
  • {"files"=>["https://ndownloader.figshare.com/files/1104384"], "description"=>"<p>Skin samples isolated from E18.5 mouse embryos were fixed with 4% paraformaldehyde, hybridized with a digoxygenin-labeled RNA probe for <i>Elovl4</i>, <i>CerS3</i>, <i>keratin 14</i>, or <i>involucrin</i>, and stained with alkaline phosphatase-conjugated anti-digoxygenin antibody (F<sub>ab</sub> fragment) and nitroblue tetrazolium/5-bromo-4-chloro-3-indolyl phosphate solution. Frozen sections (25 µm) were subjected to microscopic observation under a DM5000B light microscope and photographed. Bar, 20 µm. De, dermis.</p>", "links"=>[], "tags"=>["Biochemistry", "enzymes", "Enzyme regulation", "lipids", "Fatty acids", "sphingolipids", "proteins", "Molecular cell biology", "Signal transduction", "Nuclear receptor signaling", "gene expression", "mrnas", "ss"], "article_id"=>734070, "categories"=>["Biological Sciences"], "users"=>["Yukiko Mizutani", "Hui Sun", "Yusuke Ohno", "Takayuki Sassa", "Takeshi Wakashima", "Mari Obara", "Kohei Yuyama", "Akio Kihara", "Yasuyuki Igarashi"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0067317.g002", "stats"=>{"downloads"=>1, "page_views"=>13, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Expression_of_Elovl4_and_CerS3_mRNAs_in_the_SS_and_SG_/734070", "title"=>"Expression of <i>Elovl4 and CerS3</i> mRNAs in the SS and SG.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-06-27 04:03:45"}
  • {"files"=>["https://ndownloader.figshare.com/files/1104385"], "description"=>"<p>(A–C) Keratinocytes were transfected with control or <i>CERS3</i> siRNA. Twenty four h after transfection, transfection media were changed to differentiation medium, and the cells were incubated for an additional 2 days. (A) Total RNA was prepared from the cells, then subjected to real-time PCR using primers specific for <i>CERS3</i> and <i>GUSB</i> as an internal control. The expression level of <i>CERS3</i> mRNA was calculated by normalizing to that of <i>GUSB</i>. Values presented are the amount of <i>CERS3</i> mRNA relative to that in control cells and represent the mean ± S.D. from three independent experiments. Statistically significant differences to control siRNA are indicated (*p<0.05; the Student’s t-test). (B) Total cell lysates (10 µg protein) prepared from the cells were subjected to immunoblotting with an anti-CERS3 antibody or anti-CERS2 antibody, or with an anti-GAPDH antibody to demonstrate uniform protein loading. (C) Total membrane proteins (40 µg) were prepared and subjected to an <i>in vitro</i> FA elongase assay by incubating with 50 µM C22∶0-CoA or C26∶0-CoA and 0.075 µCi [<sup>14</sup>C]malonyl-CoA, for 30 min at 37°C. After termination of the reactions, lipids were subjected to methanolysis, extraction, separation by reverse-phase TLC, and detection by an FLA7000 bioimaging analyzer. Values indicate the radioactivities of the FA methyl ester products relative to that of control siRNA-transfected cells, and represent the mean ± S.D. of three independent experiments. Statistically significant differences to control siRNA are indicated (*p<0.05, **p<0.01; Student’s t-test).</p>", "links"=>[], "tags"=>["Biochemistry", "enzymes", "Enzyme regulation", "lipids", "Fatty acids", "sphingolipids", "proteins", "Molecular cell biology", "Signal transduction", "Nuclear receptor signaling", "gene expression", "regulates", "activities", "elovl1", "elovl4", "differentiated"], "article_id"=>734071, "categories"=>["Biological Sciences"], "users"=>["Yukiko Mizutani", "Hui Sun", "Yusuke Ohno", "Takayuki Sassa", "Takeshi Wakashima", "Mari Obara", "Kohei Yuyama", "Akio Kihara", "Yasuyuki Igarashi"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0067317.g003", "stats"=>{"downloads"=>0, "page_views"=>33, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_CERS3_regulates_activities_of_ELOVL1_and_ELOVL4_in_differentiated_keratinocyte_/734071", "title"=>"CERS3 regulates activities of ELOVL1 and ELOVL4 in differentiated keratinocyte.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-06-27 04:03:45"}
  • {"files"=>["https://ndownloader.figshare.com/files/1104386"], "description"=>"<p>Keratinocytes were incubated for 24 h with vehicle (DMSO) or with an activator of the indicated transcription factor: LXR (10 µM TO901317), RAR (1 µM all-<i>trans</i>-retinoic acid), RXR (1 µM 9-<i>cis</i>-retinoic acid), PPARα (10 µM WY14643), PPARβ/δ (10 µM L-165,041), PPARγ (7.5 µM troglitazone), or vitamin D receptor (VDR) (0.1 µM 1α, 25-dihydroxyvitamin D3). Total RNA prepared from each culture was subjected to real-time quantitative PCR using primers specific for <i>ELOVL1</i>, <i>ELOVL4</i>, <i>ELOVL6</i>, <i>ELOVL7</i>, <i>CERS2</i>, or <i>CERS3</i> and for <i>GUSB</i> for standardization. The expression level of each mRNA was calculated by normalizing to that of <i>GUSB</i>. Values presented are the amount of the respective mRNA relative to that from cells treated with vehicle, and represent the mean ± S.D. from three independent experiments. Statistically significant differences to DMSO controls are indicated (*p<0.05; **p<0.01; ***p<0.001; Student’s t-test).</p>", "links"=>[], "tags"=>["Biochemistry", "enzymes", "Enzyme regulation", "lipids", "Fatty acids", "sphingolipids", "proteins", "Molecular cell biology", "Signal transduction", "Nuclear receptor signaling", "gene expression", "lxr", "mrna"], "article_id"=>734072, "categories"=>["Biological Sciences"], "users"=>["Yukiko Mizutani", "Hui Sun", "Yusuke Ohno", "Takayuki Sassa", "Takeshi Wakashima", "Mari Obara", "Kohei Yuyama", "Akio Kihara", "Yasuyuki Igarashi"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0067317.g004", "stats"=>{"downloads"=>3, "page_views"=>16, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Activation_of_PPAR_PPAR_or_LXR_increases_CERS3_ELOVL4_and_ELOVL7_mRNA_expression_in_keratinocytes_/734072", "title"=>"Activation of PPARβ/δ, PPARγ, or LXR increases <i>CERS3, ELOVL4,</i> and <i>ELOVL7</i> mRNA expression in keratinocytes.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-06-27 04:03:45"}
  • {"files"=>["https://ndownloader.figshare.com/files/1104387"], "description"=>"<p>(A) Keratinocytes were incubated with either DMSO or activators of the indicated receptor, PPARβ/δ (10 µM L-165,041) or PPARγ (7.5 µM troglitazone), for the indicated times. (B) Keratinocytes were incubated for 24 h with the indicated concentrations of the PPARβ/δ activator L-165,041 or the PPARγ activator troglitazone. (A and B) Total RNA prepared from each culture was subjected to real-time PCR using primers specific for <i>ELOVL4</i> or <i>CERS3</i>, and for <i>GUSB</i> for standardization. The expression level of each mRNA was calculated by normalizing to that of <i>GUSB</i>. Values presented are the amount of the respective mRNA relative to that from cells harvested at 0 h (A) or from cells not treated with activator (B), and represent the mean ± S.D. from three independent experiments. Statistically significant differences to the samples at 0 h (A) or untreated samples (B) are indicated (*p<0.05, **p<0.01; Student’s t-test).</p>", "links"=>[], "tags"=>["Biochemistry", "enzymes", "Enzyme regulation", "lipids", "Fatty acids", "sphingolipids", "proteins", "Molecular cell biology", "Signal transduction", "Nuclear receptor signaling", "gene expression", "ligands", "mrna", "dose-", "time-dependent"], "article_id"=>734073, "categories"=>["Biological Sciences"], "users"=>["Yukiko Mizutani", "Hui Sun", "Yusuke Ohno", "Takayuki Sassa", "Takeshi Wakashima", "Mari Obara", "Kohei Yuyama", "Akio Kihara", "Yasuyuki Igarashi"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0067317.g005", "stats"=>{"downloads"=>0, "page_views"=>30, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Treatment_with_ligands_for_PPAR_or_PPAR_increase_CERS3_and_ELOVL4_mRNA_expression_in_keratinocytes_in_both_a_dose_and_time_dependent_manner_/734073", "title"=>"Treatment with ligands for PPARβ/δ or PPARγ increase <i>CERS3</i> and <i>ELOVL4</i> mRNA expression in keratinocytes, in both a dose- and time-dependent manner.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-06-27 04:03:45"}
  • {"files"=>["https://ndownloader.figshare.com/files/1104388"], "description"=>"<p>Keratinocytes were transfected with control or <i>PPARβ/δ</i> siRNA. Twenty four h after transfection, the transfection media were changed to serum-free keratinocyte growth medium (undifferentiated keratinocytes, UDK) or to differentiation medium (differentiated keratinocytes, DK), and cells were incubated for another 2 days. Total RNA prepared from each of the cell cultures was subjected to real-time quantitative PCR using primers specific for <i>PPARβ/δ</i>, <i>ELOVL4</i>, <i>CERS3</i>, or <i>CERS2</i>, and for <i>GUSB</i> for standardization. The expression level of each mRNA was calculated by normalizing to that of <i>GUSB</i>. Values presented are the amount of the respective mRNA relative to that from undifferentiated keratinocytes transfected with control siRNA, and represent the mean ± S.D. from three independent experiments. Statistically significant differences between undifferentiated keratinocytes and differentiated keratinocytes or between control siRNA and <i>PPARβ/δ</i> siRNAs are indicated (*p<0.05, **p<0.01; Student’s t-test).</p>", "links"=>[], "tags"=>["Biochemistry", "enzymes", "Enzyme regulation", "lipids", "Fatty acids", "sphingolipids", "proteins", "Molecular cell biology", "Signal transduction", "Nuclear receptor signaling", "gene expression", "sirna", "attenuates", "differentiation-induced", "mrna"], "article_id"=>734074, "categories"=>["Biological Sciences"], "users"=>["Yukiko Mizutani", "Hui Sun", "Yusuke Ohno", "Takayuki Sassa", "Takeshi Wakashima", "Mari Obara", "Kohei Yuyama", "Akio Kihara", "Yasuyuki Igarashi"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0067317.g006", "stats"=>{"downloads"=>0, "page_views"=>12, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Knockdown_of_PPAR_946_948_by_siRNA_attenuates_differentiation_induced_ELOVL4_and_CERS3_mRNA_expression_in_keratinocytes_/734074", "title"=>"Knockdown of <i>PPARβ/δ</i> by siRNA attenuates differentiation-induced <i>ELOVL4</i> and <i>CERS3</i> mRNA expression in keratinocytes.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-06-27 04:03:45"}
  • {"files"=>["https://ndownloader.figshare.com/files/1104390", "https://ndownloader.figshare.com/files/1104392", "https://ndownloader.figshare.com/files/1104393", "https://ndownloader.figshare.com/files/1104394"], "description"=>"<div><p>The lipid lamellae in the stratum corneum is important for the epidermal permeability barrier. The lipid lamellae component ceramide (CER), comprising an ultra long-chain (ULC) fatty acid (FA) of ≥26 carbons (ULC CER), plays an essential role in barrier formation. ULC acyl-CoAs, produced by the FA elongase ELOVL4, are converted to ULC CERs by the CER synthase CERS3. In the presented study, we observed that <i>ELOVL4</i> and <i>CERS3</i> mRNAs increased during keratinocyte differentiation <i>in vivo</i> and <i>in vitro</i>. We also determined that peroxisome proliferator-activated receptor β/δ is involved in the up-regulation of the mRNAs. Knockdown of <i>CERS3</i> caused a reduction in the elongase activities toward ULC acyl-CoAs, suggesting that CERS3 positively regulates ULCFA. Thus, we reveal that the two key players in ULC CER production in epidermis, CERS3 and ELOVL4, are coordinately regulated at both the transcriptional and enzymatic levels.</p></div>", "links"=>[], "tags"=>["Biochemistry", "enzymes", "Enzyme regulation", "lipids", "Fatty acids", "sphingolipids", "proteins", "Molecular cell biology", "Signal transduction", "Nuclear receptor signaling", "gene expression", "synthesis", "ultra", "long-chain", "fatty", "ceramide", "keratinocyte"], "article_id"=>734076, "categories"=>["Biological Sciences"], "users"=>["Yukiko Mizutani", "Hui Sun", "Yusuke Ohno", "Takayuki Sassa", "Takeshi Wakashima", "Mari Obara", "Kohei Yuyama", "Akio Kihara", "Yasuyuki Igarashi"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0067317.s001", "https://dx.doi.org/10.1371/journal.pone.0067317.s002", "https://dx.doi.org/10.1371/journal.pone.0067317.s003", "https://dx.doi.org/10.1371/journal.pone.0067317.s004"], "stats"=>{"downloads"=>8, "page_views"=>10, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Cooperative_Synthesis_of_Ultra_Long_Chain_Fatty_Acid_and_Ceramide_during_Keratinocyte_Differentiation_/734076", "title"=>"Cooperative Synthesis of Ultra Long-Chain Fatty Acid and Ceramide during Keratinocyte Differentiation", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2013-06-27 04:03:45"}

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Relative Metric

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