INTRODUCTION: Theta rhythm typically occurs during memory processes, REM sleep, and spatial navigation but also in epilepsy, migraines, or even mild Alzheimer’s disease (AD). Recent evidence shows that well-synchronized theta rhythm can successfully be recorded locally from the posterior hypothalamic area (PHa), specifically from the supramammillary nucleus (SuM) and the posterior hypothalamic nuclei (PH). The population of theta-related cells in the PHa were found to be similar types to those found in the hippocampal formation. In addition, a new type of cells has been found in the posterior hypothalamic region and based on its regular firing pattern and possible pacemaker role these cells were termed “timing”. AIM(S): The aim of the present study was to investigate the timing of cell populations in both in vivo and in vitro PHa after theta rhythm induction by kainic acid (KA) application. METHOD(S): Twenty in vivo experiments were performed on 20 urethanized rats and 22 in vitro experiments were performed on 40 PHa slices obtained from 22 rats. Theta rhythm and single unit activity were evoked by intra-PHa microinjection of KA (in vivo) or by bath perfusion of PHa slices with KA-containing artificial cerebrospinal fluid (in vitro). RESULTS: A total number of 123 posterior hypothalamic neurons were recorded during both in vivo and in vitro experiments. Among them, 28 neurons were classified as “timing cells” according to their very regular pattern of discharges in a steady frequency in the theta band (3‑12 Hz). Eight timing cells were recorded in in vivo PHa and 20 timing cells were recorded in PHa slices. CONCLUSIONS: The present data show that glutamatergic stimulation of PHa neuronal network with kainic acid results in the activation of specific subpopulation of neutrons, characterized by regular firing pattern in theta frequency range. The role of PHa “timing cell” activity is discussed regarding hippocampal theta rhythm frequency programing. FINANCIAL SUPPORT: Supported by NCN grant No. 2017/25/B/NZ4/01476.