Photobiomodulation therapy (PBMT) has emerged as a promising treatment for neurological disorders, demonstrating improvements in neuroinflammatory processes. However, the optimal frequency of PBMT for targeting specific brain regions involved in cognitive tasks remains unclear, especially in normal conditions. This study investigates the effects of 5 Hz and 40 Hz pulsed wave PBM with an 810 nm wavelength on frontal cortex of healthy rats over five consecutive days (46.50J/cm2 for 12minutes in three cycles of 4 minutes). The study focuses on spatial reference memory assessed in the Morris Water Maze which is hippocampal-dependent. The research explores hippocampal cytochrome c-oxidase (CCO) activity, proto-oncogene expression, and the broader effects on neuronal changes, glial cells, inflammatory cascades, and synaptogenesis in this brain region. The 40 Hz group exhibited better performance than 5 Hz in the reference memory task compared to control group, accompanied by increased c-Fos expression in hippocampal layers. While no differences were found in CCO activity, both frequencies (5 Hz and 40 Hz) showed increased IL-6 expression and decreased IFNA2. Additionally, a differential pattern in IL-1β expression was observed, with 40 Hz increasing both premature and mature forms. These changes were associated with increased GFAP, pre- and post-synaptic protein expression, and alterations in inflammatory cascades. The findings indicate that frequency variations in PBMT differentially modulate synaptogenesis and neuroinflammatory pathways, particularly enhancing hippocampal function and cognitive performance, especially in response to 40 Hz treatment under normal conditions.