Journal of pediatric surgery · 2010
Abnormal enteric innervation identified without histopathologic staining in aganglionic colorectum from a mouse model of Hirschsprung's disease.
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Miyahara Katsumi, Kato Yoshifumi, Koga Hiroyuki, Lane Geoffrey J, Inoue Takayoshi, Akazawa Chihiro, Yamataka Atsuyuki
Аннотация
PURPOSE: The piebald lethal mouse with a deletion of endothelin-B receptor gene (EDNRB) is a model for Hirschsprung's disease (HD), whereas the SOX10 gene is vital for the development of intestinal neural crest-derived cells. Recently, we created a SOX10 transgenic mouse with intestinal neural crest-derived cells visible with enhanced green fluorescent protein (VENUS), that is, SOX10-VENUS(+)/EDNRB(sl/sl) to investigate intestinal innervation in HD.
METHODS: SOX10-VENUS(+)/EDNRB(sl/sl) (n = 30) were compared with wild-type littermates as controls (EDNRB(s/s), n = 30). Mice were killed on days 3, 7, or 12 of age. The entire colorectum was excised, fixed with 4% paraformaldehyde, and examined using fluorescence microscopy alone without staining.
RESULTS: In normoganglionic colorectum from controls, a grid network of nerve fibers/glial cells was visualized that connected smoothly with extrinsic nerve fibers running along the colorectal wall. In aganglionic colorectum from SOX10-VENUS(+)/EDNRB(sl/sl) mice, there was no grid network and more extrinsic nerve fibers than controls that invaded the colon wall becoming elongated with branching fibers. Normoganglionic colon from controls and SOX10-VENUS(+)/EDNRB(sl/sl) mice appeared the same. Innervation patterns did not change over time.
CONCLUSION: This is the first time for abnormal enteric innervation in aganglionic colon in a model for HD to be visualized without staining.
METHODS: SOX10-VENUS(+)/EDNRB(sl/sl) (n = 30) were compared with wild-type littermates as controls (EDNRB(s/s), n = 30). Mice were killed on days 3, 7, or 12 of age. The entire colorectum was excised, fixed with 4% paraformaldehyde, and examined using fluorescence microscopy alone without staining.
RESULTS: In normoganglionic colorectum from controls, a grid network of nerve fibers/glial cells was visualized that connected smoothly with extrinsic nerve fibers running along the colorectal wall. In aganglionic colorectum from SOX10-VENUS(+)/EDNRB(sl/sl) mice, there was no grid network and more extrinsic nerve fibers than controls that invaded the colon wall becoming elongated with branching fibers. Normoganglionic colon from controls and SOX10-VENUS(+)/EDNRB(sl/sl) mice appeared the same. Innervation patterns did not change over time.
CONCLUSION: This is the first time for abnormal enteric innervation in aganglionic colon in a model for HD to be visualized without staining.
Предметные рубрики MeSH
AnimalsBacterial ProteinsCell LineageColonDisease Models, AnimalEnteric Nervous SystemFluorescent Antibody TechniqueFluorescent DyesGanglia, ParasympatheticHirschsprung DiseaseLuminescent ProteinsMiceMice, Inbred C57BLMice, Mutant StrainsMice, TransgenicMicroscopy, ConfocalMicroscopy, FluorescenceNerve FibersNeural CrestRNA, MessengerReceptor, Endothelin BRectumReverse Transcriptase Polymerase Chain ReactionSOXE Transcription FactorsUltraviolet Rays