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Use of a Caspase Multiplexing Assay to Determine Apoptosis in a Hypothalamic Cell Model

Use of a Caspase Multiplexing Assay to Determine Apoptosis in a Hypothalamic Cell Model

Published on: April 16, 2014

Estrogen induces caspase-dependent cell death during hypothalamic development.

Elizabeth M Waters¹, Richard B Simerly

¹Harold and Margaret Milliken Hatch Laboratory of Neuroendocrinology, The Rockefeller University, New York, New York 10065, USA. e.waters@rockefeller.edu

The Journal of Neuroscience : the Official Journal of the Society for Neuroscience

|August 7, 2009

Summary

This summary is machine-generated.

Estrogen regulates dopamine neuron numbers in the brain's AVPV region by activating a cell death pathway. This involves caspases, crucial for controlling these specific neuron populations.

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Area of Science:

Neuroscience
Endocrinology
Developmental Biology

Background:

The anteroventral periventricular nucleus (AVPV) contains sexually dimorphic dopamine neurons.
Sex steroids, particularly estrogen derived from testosterone, influence the development of these neurons.
The role of programmed cell death (apoptosis) in determining AVPV dopamine neuron numbers is not fully understood.

Purpose of the Study:

To investigate whether estrogen-mediated cell death regulates the number of dopamine neurons in the AVPV.
To elucidate the molecular mechanisms underlying estrogen's effect on AVPV dopamine neuron survival.

Main Methods:

In vitro and in vivo experiments using estradiol treatment on the AVPV.
Utilized estrogen receptor antagonists (ICI 182,780) and cell death inhibitors (Cyclosporin A, ZVAD).
Assessed cell death using TUNEL and Hoechst staining to identify apoptotic profiles.

Main Results:

Estradiol treatment reduced the number of tyrosine hydroxylase-immunoreactive (TH-ir) cells in the AVPV.
Estrogen receptor antagonist and cell death inhibitors rescued TH-ir cells from estradiol-induced loss.
Estradiol increased apoptotic profiles in the AVPV, dependent on the alpha estrogen receptor form.
A caspase-dependent mechanism was identified for estradiol-mediated TH-ir cell loss.

Conclusions:

Estrogen activates an intrinsic, caspase-dependent apoptotic pathway to regulate dopamine neuron numbers in the AVPV.
This contrasts with the generally neuroprotective roles of sex steroids, highlighting a specific mechanism in the AVPV.
Estrogen actively sculpts dopamine neuron populations in the AVPV through programmed cell death.