{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,11,7]],"date-time":"2026-11-07T17:27:35Z","timestamp":1794072455704,"version":"4.8.0"},"reference-count":36,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2024,10,10]],"date-time":"2024-10-10T00:00:00Z","timestamp":1728518400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100002322","name":"Coordena\u00e7\u00e3o de Aperfei\u00e7oamento de Pessoal de N\u00edvel Superior","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100002322","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Endocrinol."],"abstract":"<jats:sec>\n                    <jats:title>Introduction<\/jats:title>\n                    <jats:p>\n                      Congenital hypopituitarism (CH) is characterized by the deficiency of pituitary hormones. Among CH patients, 85% lack a molecular diagnosis. Whole Exome Sequencing (WES) identified a homozygous variant (c.865G&amp;gt;A, p.Val289Ile) in the\n                      <jats:italic>CDH2<\/jats:italic>\n                      gene, responsible for N-Cadherin production, crucial for cell-cell adhesion. Predicted to be likely pathogenic, the variant was found in a patient deficient in\n                      <jats:italic>GH<\/jats:italic>\n                      ,\n                      <jats:italic>TSH<\/jats:italic>\n                      ,\n                      <jats:italic>ACTH<\/jats:italic>\n                      , and\n                      <jats:italic>LH\/FSH<\/jats:italic>\n                      . Its impact on cell adhesion was confirmed in L1 fibroblast cell lines.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Objective<\/jats:title>\n                    <jats:p>\n                      Create a\n                      <jats:italic>cdh2<\/jats:italic>\n                      knockdown in zebrafish for investigating its role in pituitary development through growth hormone and transcription factors expression.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>\n                      Utilized pET28B-RfxCas13d-His plasmid for Cas13 mRNA production via\n                      <jats:italic>in vitro<\/jats:italic>\n                      transcription, guiding Cas13 to\n                      <jats:italic>cdh2<\/jats:italic>\n                      with three RNAs. Injected the complex into single-cell embryos for analysis up to 96 hpf. Assessed gene expression of\n                      <jats:italic>cdh2<\/jats:italic>\n                      ,\n                      <jats:italic>prop1<\/jats:italic>\n                      ,\n                      <jats:italic>pit1<\/jats:italic>\n                      , and\n                      <jats:italic>gh1<\/jats:italic>\n                      using RT-qPCR. Evaluated cdh2 protein expression through the western blot technique.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      Knockdown animals displayed developmental delay. The\n                      <jats:italic>cdh2<\/jats:italic>\n                      expression decreased by 75% within 24 hours, rebounded by 48 hours, and reached wild-type levels by 96 hpf.\n                      <jats:italic>gh1<\/jats:italic>\n                      expression decreased at 48h but increased by 96 hpf, aligning with WT. No significant differences in\n                      <jats:italic>prop1<\/jats:italic>\n                      and\n                      <jats:italic>pit1<\/jats:italic>\n                      expression were observed.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusion<\/jats:title>\n                    <jats:p>\n                      Our findings underscore\n                      <jats:italic>cdh2<\/jats:italic>\n                      \u2019s role in pituitary development and hormonal regulation, offering insights for developmental biology research.\n                    <\/jats:p>\n                  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