Auto-spermatophore extrusion in male crickets

The reproductive cycle of the male cricket consists of the mating stage and the sexually refractory stage. The latter is further divided into the first refractory stage (RS1) from spermatophore extrusion in copulation to spermatophore preparation after copulation, and the second refractory stage (RS2) from spermatophore preparation to recommencement of a calling song. RS2 is time-fixed and unaffected by the female or by stress, hence RS2 is assumed to be controlled by the reproductive timer. Previously, we suggested that the timer is located in the terminal abdominal ganglion (TAG), because functional inactivation of the TAG by local cooling lengthened RS2 in proportion to cooling time. To obtain further evidence of timer localization and to examine the operation of the timer in dissected animals, we investigated the characteristics of auto-spermatophore extrusion, a phenomenon in which males eject the mature spermatophore themselves without any prior courtship. The occurrence of auto-spermatophore extrusion was 100% in dissected males with the TAG separated, compared to 1.7% in intact males. The time interval (SPaSE) between spermatophore preparation and autospermatophore extrusion was comparable to RS2 measured by the calling song. Spike recording from a genital motor neurone in the separated TAG indicated that burst discharge associated with auto-spermatophore extrusion occurred with a SPaSE comparable to RS2. Other efferent neurones, some of which were identified as dorsal unpaired median (DUM) neurones, showed a timedependent spike frequency increase during SPaSE. These results strengthen our previous conclusion that the reproductive timer is located within the TAG, and demonstrate that the timer functions normally even when the TAG is separated from the central nervous system.</p

Reproductive behaviour in the male cricket Gryllus bimaculatus DeGEER: I. Structure and function of the genitalia

We have investigated the morphology and physiology of the genitalia of the male cricket to establish a basis for neuroethological study of its reproductive behaviour. First, the structure of the phallic complex, including the dorsal pouch, guiding rod, epiphallus, ventral lobes and median pouch, are described, as are the muscles, cuticle, membranes and biomechanics of copulation. The innervation and sensory receptors have also been examined. Second, the functional role of the muscle in each genital organ has been determined by direct observation of muscle contraction during spontaneous or evoked movements and by analysis of the changes in movements after the ablation of the muscle. Third, for the flexible membranous organs, the ventral lobes and median pouch, the passages for haemolymph and their dynamic properties have been examined using petroleum jelly. Fourth, the sequence of coordinated motor actions performed by the internal and external genital organs, which were induced in both restrained and dissected males using newly developed techniques, has been analyzed during tethered copulation and spermatophore formation. As a result, the mechanisms of copulation and spermatophore formation are now more fully understood

Reproductive behaviour in the male cricket Gryllus bimaculatus DeGEER: II. Neural control of the genitalia

To understand the neural mechanisms of reproductive behaviour in the male cricket, we identified motor neurones innervating the muscles in each genital organ by backfilling with cobalt/nickel and recording their extracellular spike activity from nerve bundles of the terminal abdominal ganglion during tethered copulation and spermatophore formation. During tethered copulation, at least two motor neurones innervating two ipsilateral muscles were activated during projection of the guiding rod of the phallic dorsal pouch. Only one motor neurone, innervating four ipsilateral muscles of the dorsal pouch, was responsible for spermatophore extrusion by deforming the dorsal pouch. For spermatophore transfer, three motor neurones, singly innervating three epiphallus muscles, played a major role in opening passages for haemolymph to enter the ventral lobes and median pouch by bending the epiphallus. Two ventral lobe and 3–5 median pouch motor neurones seemed to play a role in expanding or folding the two membranous structures by relaxing or contracting their muscle fibres. After spermatophore transfer, most of the genital motor neurones exhibited a rhythmic burst of action potentials causing movement of the phallic complex coupled with strong abdominal contractions. For spermatophore formation, the genital motor neurones began to accelerate their rhythmic bursts approximately 30 s prior to subgenital plate opening and then changed their activity to tonic bursting or silence. The results have allowed us to describe the timing of the onset and termination of genital muscle contraction more precisely than before, to examine the neural mechanisms of copulatory motor control and to speculate on the neural organization of the reproductive centre for spermatophore extrusion and protrusion