Courtship song in Drosophila is produced by the male's wing vibration and consists of pulses of tone produced at intervals of approximately 34 shortens the circadian period, perl, which lengthens it, and pero, which abolishes it-strikingly affects the 60-sec song rhythm in a parallel fashion. Therefore, both circadian rhythms and a very short, noncircadian oscillation appear to be influenced by the same gene.Courtship in Drosophila melanogaster consists of a series of highly stereotyped behavioral patterns, the most conspicuous of which is the male's wing display (1, 2). The male extends and vibrates his wing, producing a patterned acoustic signal or "song" (3). The song is illustrated in Fig. 1 and has several acoustic components, including a train of pulses with an interpulse interval (ipi) of approximately 34 msec together with a sinusoidal hum, which has a frequency of 160 Hz (4). The ipi is species specific; males of D. simulans, a closely related species, sing with an ipi of approximately 48 msec (5). Von Schilcher (6) has reported that D. melanogaster females mate faster with wingless, deafened conspecific males when they are exposed to an artificial song with an ipi of 34 msec compared to one having an ipi of 48 msec. Therefore, the ipi of the courtship song may play an important role as a recognition factor in preserving the sexual isolation between the two species.This paper describes some experiments that demonstrate that the variation in the ipi values produced by males of both D. melanogaster and D. simulans is greater than previously believed. However, we will show that this variation conforms to a highly structured temporal pattern which is dramatically affected in D. melanogaster by three circadian rhythm mutations isolated by behavioral criteria unrelated to courtship (7). 5-day-old male with a 3-day-old virgin attached-X female inside a transparent mating cell 2 X 1 X 0.3 cm high. The floor of the chamber was composed of Nytex (stiff nylon mesh), and the unit was placed 2 mm above the ribbon of a Reslo (Sheffield, England) ribbon microphone, which was then encased within a screened aluminum box (12 X 8 X 8 inches) which served as an anechoic chamber. A porthole was cut in the box to enable the experimenter to observe the flies. The song of the male was amplified and recorded on a Tandberg loX reel-to-reel tape recorder. A visual record of the song was obtained by tracing the recording, together with a time reference, on light-sensitive paper (Dupont, Type 5B) with a multichannel oscillograph (Consolidated Electrodynamics, Newark, NJ, type 5-124). The paper was exposed, and ipi were measured peak to peak with a ruler.The courtship was divided into 10-sec fractions, and the mean and SEM of all ipi falling within each successive time period was computed. The minimum number of ipi used to obtain a mean was 10, and most means were based on between 25 and 70 individual ipi. These means were then used to compute a nonlinear regression of ipi against the successive time periods for each court...