The Research Of Estrogen Effect On L-type Calcium Channels

Objective:To explore estrogen effect on L-type Calcium channel current and theexpression of its protein Cav1.2.Methods:1. Cortical neurons from newborn（0-1d） Sprague-Dawley（SD）Rats were cultured by modified enzyme and mechanical digestionmethod. Fluorescent inverted microscope was used to observe theirmorphology and neuronal characteristics were identified byimmunofluorescence on day7.2. The primary cortical neurons were cultured for Seven days beforetreatments without or with estrogen (1umol/L) for72h. L-type calciumchannel current was measured by whole-cell patch clamp recordingtechnique.3. The neurons were divided into four groups: control group,ICI182780-Estrogen receptor antagonist group（100nmol/L）,Estrogengroup （1umol/L）and ICI182780（100nmol/L）+Estrogen（1umol/L）group.72h after treatment, the Cav1.2levels were detected by western blotting.Results:1. The cortical neurons were well developed on the7day.And most ofthe culture cells were identified as neurons.2. Whole-cell Patch clamp recording showed the L-type calciumchannel current was significantly decreased after estrogen treatment.3. The expression of Cav1.2was significantly decreased after estrogentreatment as shown by Western blotting.Conclusion:1. The culture method of primary cortical neurons in vitro issuccessful.2. Estrogen treatment could decrease the L-type calcium current ofcortical neurons.3.Estrogen treatment could downregulate the expression of Cav1.2inprimary cortical neurons.