Background: A deficit in interhemispheric transfer was hypothesized in alexithymia more than 30 years ago, following the observation that split-brain patients manifest certain alexithymic characteristics. However, direct evidence of interhemispheric transfer deficit has never been provided. This study investigated the hypothesis of a transcallosal interhemispheric transfer deficit in alexithymia by means of paired-pulse transcranial magnetic stimulation. Methods: A random sample of 300 students was screened for alexithymia using the Italian version of the 20-item Toronto Alexithymia Scale. Eight right-handed males and eight females with high alexithymic scores and an age- and gender-matched group with low alexithymic scores were selected. A first (conditioning) magnetic stimulus was delivered to one motor cortex followed by a second (test) stimulus to the opposite hemisphere at different interstimulus intervals for both motor cortices. Motor evoked responses were recorded from the abductor digit minimi muscles. Results: High alexithymic subjects showed reduced transcallosal inhibition as compared to low alexithymic subjects at interstimulus intervals of 10, 12 and 14 ms in the left-to-right and right-to-left interhemispheric transfer directions. Conclusions: Results point to functional differences in transcallosal interactions in high alexithymic as compared to low alexithymic subjects, supporting the hypothesis of an interhemispheric transfer deficit in alexithymia.