Determining a Test Growth Hormone Effect on Kisspeptin Neurons using a Whole-Cell Patch Clamp

In dit artikel

Samenvatting

Source: Silva, J. d. N., et al., Hypothalamic Kisspeptin Neurons as a Target for Whole-Cell Patch-Clamp Recordings. J. Vis. Exp. (2023)

This video demonstrates the use of the whole-cell patch-clamp technique to measure responses in kisspeptin neurons from transfected mouse hypothalamic slices. The process involves achieving whole-cell configuration and recording membrane potential changes induced by the test hormone using current-clamp mode.

Protocol

All procedures involving animal samples have been reviewed and approved by the appropriate animal ethical review committee.

1. Cell sealing for recording

Ensure that all the pieces of equipment (microscope, amplifier, digitizer, micromanipulator, and others) are turned on before starting the recording.
Fill the recording chamber, from a commercial brand (Table of Materials), attach to the microscope with the aCSF solution for recordings. Use a perfusion pump to constantly perfuse the aCSF at a rate of 2 mL/min.
Transfer a brain slice of interest (one at a time) to the recordi....

Toegang beperkt. Log in of start een proefperiode om deze inhoud te bekijken.

Resultaten

Hypothalamic neuron pathways diagram; GnRH and kisspeptin roles in hormone regulation processes.
Figure 1: Schematic diagram summarizing the whole-cell patch-clamp technique's contribution to the knowledge of the kisspeptin neurons' activity. Kisspeptin neurons (shown in green) are located in the anteroventral periventricular and rostr.......