Experiment 1: Effect of central (ICV) administration of senktide on LH release in male and female WT and Kiss1 KO mice with or without the presence of sex steroids.

ICV Injection Protocol

2-3 days before starting the experiment

• Anesthetise mice in a chamber with 2.5% isoflurane. 
• When animal loses its toe reflex make a small hole was bored in the skull 1 mm lateral and 0.5 mm posterior to bregma.
• Use a Hamilton syringe attached to a 27-gauge needle fitted with polyethylene tubing, leaving 2.0 mm of the needle tip exposed.
• Bregma can be identified by running the tip of the needle on top of a skull and feeling a small prominence. This is bregma.

On the day of the experiment

• Anesthetise mice in a chamber with 2.5% isoflurane. 
• A anaesthesia cone was used to supplement isoflurane to the animal as needed.
• All subsequent injections were made at the same site
• 5 μl of solution were slowly and continuously injected into the lateral ventricle. 
• The needle remained inserted for approximately 30 s after the injection to minimize backflow up the needle track. 
• Mice were typically anesthetised for a total of 10-15 min and recovered from the anaesthesia within 3 min after the injection.

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