Signaling via tyrosine kinases appears necessary for regulation of synaptic efficacy. Interactions of the src-family kinases with phosphorylated proteins were studied in area CAI of rat hippocampal slices 10 min after induction of long-term potentiation (LTP) by 100 Hz/l s stimulation (HFS). HFS enhanced association of the src-family kinases fyn and c-src with an approximately 120 kDa tyrosine phosphorylated component containing the focal adhesion kinase (FAK) and its homologue PYK2. Association of fyn with FAK and of c-src with PYK2 was increased following the HFS. Further, increase in tyrosine phosphorylation of PYK2 was detected following the HFS. These results suggest that fyn and c-src are involved in distinct signaling pathways and provide evidence for activation of FAK and PYK2 following synaptic stimulation inducing LTP in vitro.