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Related Experiment Video

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Laser Capture Microdissection of Enriched Populations of Neurons or Single Neurons for Gene Expression Analysis After Traumatic Brain Injury
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GnRH and gpcr: laser-captured single cell gene profiling.

Ishwar S Parhar1

  • 1School of Medicine and Health Sciences, Monash University Malaysia, Bandar Sunway, 46150 Petaling Jaya, Malaysia, ishwar.parhar@med.monash.edu.my.

Fish Physiology and Biochemistry
|December 26, 2009
PubMed
Summary

We developed a novel single-cell gene profiling technique to study tilapia. This method revealed G-protein coupled receptors in individual neurons and pituitary cells, uncovering cellular heterogeneity and new hormonal roles in reproduction.

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Single-cell Profiling of Developing and Mature Retinal Neurons
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Area of Science:

  • Endocrinology
  • Neuroscience
  • Molecular Biology

Background:

  • Gonadotropin-releasing hormone (GnRH) neurons regulate reproduction.
  • Understanding the molecular mechanisms of GnRH action requires single-cell resolution.

Purpose of the Study:

  • To develop and apply a novel single-cell gene profiling technique for analyzing GnRH neurons and pituitary cells.
  • To investigate the expression of G-protein coupled receptors (GPCRs) in individual cells of the tilapia (Oreochromis niloticus) reproductive axis.

Main Methods:

  • Developed a single-cell real-time quantitative PCR technique combined with laser-capture microdissection.
  • Analyzed gene expression in individual gonadotropin-releasing hormone (GnRH) neurons and pituitary endocrine cells.

Main Results:

  • First report of GPCRs in individual GnRH neurons and pituitary cells in a vertebrate.
  • Demonstrated differential expression of GnRH receptor types (R1, R2, R3) in tilapia pituitary cells, indicating cellular heterogeneity.
  • Identified a novel nonmammalian GPR54 structure for kisspeptins, conserved across evolution and expressed in GnRH neurons.

Conclusions:

  • Single-cell profiling reveals functional heterogeneity in the GnRH system.
  • GnRH receptors play differential roles in pituitary cell types during reproduction.
  • The novel kisspeptin receptor (GPR54) is crucial for GnRH secretion and pubertal maturation.