The nuclear envelope is composed of two membranes deliminating a perinuclear space which displays functional properties similar to those of a Ca2+-storing compartment. ATP-driven Ca2+ uptake and InsP3-induced Ca2+ release processes have been described in isolated nuclei. Recently, it was reported that cADP-ribose and InsP3 can trigger a nucleoplasmic Ca2+ increase. It was hypothesized that the inner nuclear membrane possesses Ca2+ channels that are regulated by ryanodine or InsP3. Radio-ligand binding assays and Western blot experiments were performed in order to investigate their presence in sheep cardiac and rat liver nuclear envelopes. Ryanodine receptors (RyR) were not detected in liver nuclear envelopes by either binding assay or Western blot analysis. However, cardiac nuclear envelopes were found to retain a very low level of specific ryanodine binding, which was not detected on immuno-blots obtained with three types of isoform-specific RyR antibodies. In contrast, nuclear InsP3-binding sites were consistently detected in both cardiac and liver nuclear envelopes. Altogether, these results provide evidence for the major contributor InsP3-gated Ca2+ channels in control of Ca2+ release from the perinuclear space in liver and cardiac cells.