Kisspeptin plays an important role in hypothalamic regulation of gonadotropin releasing hormone secretion. Specific receptor of this neuropeptide (GPR54) is expressed in brain areas other than the hypothalamus. Lateral Habenula (LHb) is a brain area where direct and indirect effects on dopaminergic and serotonergic systems are processed. In the present study, effects of kisspeptin through GPR54 receptors in the LHb on motor activity and anxiety behavior in adult male mice were investigated. Cell-attached recordings were made from adult male Balb-C mice LHb in coronal brain-slice preparation. Following decapitation and quick removal of the brain, 300-µm-thick brain slices were prepared with a vibratome and placed in the recording bath filled with artificial cerebrospinal fluid containing (in mM) 119 NaCl, 2.5 KCl, 2 CaCl2, 1.25 MgCl2, 11 D-glucose 25 NaHCO3 and NaH2PO4. Patch pipettes had 3-5 M Ohm resistances. Signals were acquired using a Multiclamp 700B amplifier connected to a Digidata 1440A. After baseline recording, kisspeptin (100 nM) was applied in the bath. Adult male Balb-C mice were divided into three groups (n=3/group) as control, kisspeptin and kisspeptin antagonist (P234). The mice were placed in the stereotaxic frame under isofluoran anesthesia (2%) and intra-habenular infusions were performed. The animals were observed to return normal feeding-drinking behavior within 30 min after anesthesia. Following the infusions,elevated plus maze and open field tests were performed to evaluate anxiety and locomotor activity respectively.At the end of the experiments, all animals were decapitated. Brains were dissected out and stored at -80°C for confirming LHb infusion region and future analysis.These preliminary findings suggest that 100-nM kisspeptin strongly activates neuronal firing in the LHb.It was determined that stereotypic, ambulatory and horizontal locomotor movements were significantly reduced by P234 compared to the kisspeptin group (p<0.01).In addition, distance parameter was significantly increased by kisspeptin compared to the P234 group, and resting time in this antagonist group was significantly higher than the kisspeptin group (p<0.01).No significant change was observed in the elevated plus maze findings among the groups.Electrophysiological recordings from brain slices showed that kisspeptin increased firing rate of neurons in the LHb.Locomotor activity was increased by kisspeptin and decreased by kisspeptin antagonist following intra-habenular infusion.It’s thought that motor activity inhibition processed by the dopaminergic system in the LHb is disinhibited by kisspeptin, and this inhibition is further increased by P234.Since habenular neurons project to the substantia nigra, kisspeptin-GPR54 system in this brain area may be of importance for neurodegenerative diseases affecting locomotion.