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Quantitative PCR-based Assay to Measure Sonic Hedgehog Signaling in Cellular Model of Ciliogenesis
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Sonic hedgehog expression in the postnatal brain.

Aileen Rivell1, Ronald S Petralia2, Ya-Xian Wang2

  • 1Laboratory of Neurosciences, NIA/NIH, Baltimore, MD 21224, USA.

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|March 7, 2019
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Summary

Sonic hedgehog (Shh) levels in rat brains and hippocampal neurons increase significantly during development. This study tracks Shh expression from birth to adulthood, revealing its dynamic role in the maturing brain.

Keywords:
EndogenousHippocampusNeuronSonic hedgehog

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Area of Science:

  • Neuroscience
  • Developmental Biology
  • Molecular Biology

Background:

  • Sonic hedgehog (Shh) is crucial for embryonic neural tube patterning.
  • The role and expression levels of endogenous Shh in post-embryonic and mature brains remain largely uncharacterized.
  • Understanding Shh dynamics is key to elucidating its functions beyond embryogenesis.

Purpose of the Study:

  • To investigate the developmental expression patterns of Sonic hedgehog (Shh) transcript and protein in postnatal and adult rat brains.
  • To examine the changes in Shh production by hippocampal neurons in culture as they mature and form synapses.
  • To characterize the presence and localization of different Shh fragments within neurons.

Main Methods:

  • Quantification of Shh transcript and protein levels in rat brains at different developmental stages.
  • Analysis of Shh protein production in cultured hippocampal neurons over time.
  • Immunolabeling techniques using antibodies against specific Shh residues to determine fragment presence and co-localization.

Main Results:

  • Shh transcript and protein levels are minimal at birth but increase progressively throughout postnatal development in rat brains.
  • Cultured hippocampal neurons show a significant increase in Shh protein production as they mature and establish synaptic connections.
  • Both N-terminal and C-terminal fragments of Shh are detected in hippocampal neurons and are co-localized within cellular compartments.

Conclusions:

  • Endogenous Sonic hedgehog (Shh) expression is developmentally regulated in the postnatal brain and in maturing neurons.
  • Shh protein fragments are present and co-exist within neurons, suggesting complex biogenesis and function.
  • These findings provide foundational insights into Shh expression dynamics in the developing and mature nervous system.