Search PubMed⌕ Search

Biomedical subjects

Sakiko Shiga

Publications and source records attributed to Sakiko Shiga.

9 recordsLinked to original sources

Synaptic connections between eyelet photoreceptors and pigment dispersing factor-immunoreactive neurons of the blowfly Protophormia terraenovae.

Studies using various mutants of Drosophila melanogaster bearing defects in their visual system, including those of the retinal and extraretinal photoreceptor systems, have indicated that the extraretinal photoreceptor known as the Hofbauer-Buchner (H-B) eyelet plays an active, if subsidiary, role in the entrainment of circadian rhythms. In the present study, in the context of unraveling the function of extraretinal photoreception on circadian rhythms and photoperiodic responses, we searched for extraretinal photoreceptors in the blowfly, Protophormia terraenovae, and found that this fly has a homolog of the H-B eyelet. In addition, we show morphologically direct synaptic connections between the eyelet of P. terraenovae (called here Pt-eyelet, after the species' name) and pigment-dispersing factor (PDF)-immunoreactive neurons, which are putative circadian pacemaker neurons, by immunogold electron microscopy combined with intracellular dye injection. The Pt-eyelet was found to reside in the middle of the posterior surface of the optic lobe between the retina and the lamina, as does the H-B eyelet. This extraretinal photoreceptor was composed of at least four photoreceptor cells equipped with well-organized microvillar rhabdomeres. Rhodopsin 6-like immunoreactivity and also the response to light stimuli clearly showed the Pt-eyelet to be functional. The Pt-eyelet terminals in the accessory medulla exhibited synaptic bouton-like appearances and formed divergent multiple-contact output synapses. Synaptic contacts from the Pt-eyelet terminal to the PDF-immunoreactive neurons were identified by the presence of presynaptic ribbons and accumulated synaptic vesicles. Their possible function is discussed in relation to previous studies on circadian rhythm and photoperiodic response of P. terraenovae.

Animals↗

Synaptic connections between pigment-dispersing factor-immunoreactive neurons and neurons in the pars lateralis of the blow fly Protophormia terraenovae.

In females of the blow fly Protophormia terraenovae, neurons with cell bodies in the pars lateralis (PL) projecting to the retrocerebral complex (designated as PL neurons) are necessary for the induction of reproductive diapause under short-day and low-temperature conditions. In the present study, neural connections between PL neurons and pigment-dispersing factor (PDF)-immunoreactive neurons were examined via immunolight microscopy and immunoelectron microscopy combined with backfills through the cardiac-recurrent nerve. Immunolight microscopy showed that fibers of PL neurons overlapped with PDF-immunoreactive fibers in the dorsolateral region of the superior protocerebral neuropil. Immunoelectron microscopy showed that PDF-immunoreactive fibers formed output synapses with fibers of PL neurons and unlabeled neurons in a region dorsoanteriorly located with respect to the calyx of the mushroom body. The distribution of synaptic connections between PDF-immunoreactive fibers and the fibers of PL neurons was sparse. According to the projection patterns, PDF-immunoreactive fibers with synaptic connections with PL neurons appeared to originate from PDF-immunoreactive neurons with cell bodies at the base of the medulla of the optic lobe (medulla PDF neurons), which are putative circadian clock neurons in P. terraenovae. PDF immunoreactivity was restrictively detected in dense-core vesicles but not in clear synaptic vesicles. The present results suggest that medulla PDF neurons convey time or photoperiodic information to PL neurons for diapause induction through direct synaptic connections.

Animals↗

Roles of the mushroom bodies in olfactory learning and photoperiodism in the blow fly Protophormia terraenovae.

Mushroom bodies (MBs) in Protophormia terraenovae were ablated by hydroxyurea (HU) treatment to larvae just after hatching in order to examine roles of the MBs in olfactory learning and photoperiodism. In all individuals the structures of the alpha, beta, and gamma lobes, the pedunculi, and the calyces of the MB were not found after HU treatment. The other structures of the brain were not obviously damaged. The volumes of both the antennal lobes and the central complex, however, were smaller in the HU-treated flies than those in the control flies. The HU-treated and non-treated flies were tested for their appetitive olfactory learning ability and photoperiodism. In the olfactory learning paradigm, an odor of methylsalicylate or coumarin was paired with a reward of sucrose. The non-treated flies learned to associate both odors with the reward, but the HU-treated flies did not. In the test for photoperiodism, both the HU-treated and non-treated flies responded to photoperiod. Both groups of flies developed ovaries under long-day conditions but entered diapause under short-day conditions. The results imply that the MBs are indispensable for olfactory learning but not for photoperiodism, and that storage of daily cycles of photoperiodic information occurs by a neural system other than the MBs.

Animals↗

Morphology and electrophysiological properties of neurons projecting to the retrocerebral complex in the blow fly, Protophormia terraenovae.

Morphological and electrical properties of neurons with somata in the pars intercerebralis (PI) and pars lateralis (PL) were examined by intracellular recording and staining in the adult blow fly, Protophormia terraenovae. According to the location of somata and fiber distribution, two types of PI neurons (PIa and PIb) and two types of PL neurons (PLa and PLb) were identified. PIb neurons were further divided into two subgroups of PIb1 and PIb2 depending on fiber branching patterns in the retrocerebral complex. PIa neurons projected axons to the contralateral nervi corporis cardiaci, whereas PLa and PLb neurons projected axons to the ipsilateral nervi corporis cardiaci. PIb neurons characteristically showed symmetrical morphology with their somata along the midline. PLb neurons had a large branching area in the subesophageal ganglion. In the retrocerebral complex, PIb2 and PLa neurons sent fibers into the corpus allatum. PIa, PIb1 and PLb neurons projected not to the corpus allatum but to the corpus cardiacum-hypocerebral complex or visceral muscles in their vicinity. PIa, PIb and PLa neurons showed long spike durations (3-10 ms). PLb neurons were immunoreactive with antisera against corazonin, FMRFamide, or beta-pigment-dispersing hormone. This is the first report revealing the morphology of individual neurons with somata residing in PI and PL in the adult fly.

Animals↗

Role of neurosecretory cells in the photoperiodic induction of pupal diapause of the tobacco hornworm Manduca sexta.

In the tobacco hornworm, Manduca sexta, pupal diapause can be induced by exposure of fifth-instar larvae to a short-day photoperiod. We studied the effect of surgical ablation of tissues containing the neurosecretory cells of the brain of fifth-instar larvae on the photoperiodic induction of pupal diapause. At the end of the experiments, we immunostained the neurosecretory cells to determine the success of the ablations. Under long-day conditions (LD 16:8 at 22 degrees C), all intact larvae, most of the sham-operated larvae, and control-operated larvae developed into nondiapausing pupae. Under short-day conditions (LD 10:14 at 22 degrees C), most intact, sham-operated, and control-operated larvae developed into diapausing pupae. Removal of type-II cells did not interfere with the photoperiodic response. Under long-day conditions, elimination of type-Ia(1) cells did not affect the incidence of nondiapausing pupae. When type-Ia(1) cells were removed under short-day conditions, however, the incidence of nondiapausing pupae was higher (51%, n = 41) than that of the intact (16%, n = 75), sham-operated (24%, n = 88), control-operated larvae (5%, n = 40), and larvae with type-II cells removed (11%, n = 27). Thus, removal of type-Ia(1) cells can impede induction of diapause. These results indicate that the type-Ia(1) neurosecretory cells have an important role in the induction of pupal diapause.

Animals↗

Juvenile hormone biosynthesis in diapause and nondiapause females of the adult blow fly Protophormia terraenovae.

In vitro synthetic activities of juvenile hormones (JH) were examined using a radiochemical assay in diapause females and reproductive females of the blow fly, Protophormia terraenovae. Thin layer chromatography showed that products of the corpus allatum (CA) comigrated with a synthetic sample of JH III bisepoxide but neither with JH III nor methylfarnesoate. JH synthetic activities increased in females reared under LD 18:6 at 25 degrees C, as the ovaries developed. The synthetic activities remained low in previtellogenic females reared under LD 12:12 at 20 degrees C. Removal of the pars intercerebralis completely prevented ovaries from development under reproductive conditions, and removal of the pars lateralis caused partial or full development of ovaries under diapause-inducing conditions. In these operated animals, the JH synthetic activities were not significantly different from those of the intact and sham-operated animals. The results indicate that the CA in P. terraenovae produces mainly JH III bisepoxide and a decrease in the JH production rate is a cause of diapause induction. PI neurons and PL neurons in the brain do not directly mediate changes in the JH production rate, but regulate ovarian development cooperatively with some unknown allatostatic and allatotropic factors.

Animals↗

Neural- and endocrine control of flight muscle degeneration in the adult cricket, Gryllus bimaculatus.

Neural- and endocrine mechanisms controlling degeneration of a dorsal longitudinal flight muscle, M112a, have been studied in adult Gryllus bimaculatus (Orthoptera: Gryllidae). Decapitation completely prevented muscle degeneration. Implantation of a pair of corpora allata or injection of juvenile hormone III into decapitated crickets caused muscle degeneration. Denervation of M112a resulted in reduction of muscle mass compared with that in sham-operated crickets. Denervation of M112a in decapitated crickets, however, did not affect muscle mass. Birefringence and ultrastructure of M112a showed an obvious regional difference in the onset of degeneration. Fibrillar structures of M112a always disappeared from the ventral to dorsal part. Distribution of axon terminals of motor neurons and mechanical responses to the motor nerve stimuli showed that M112a is composed of five motor units with similar twitch properties. When M112a was fully denervated, regional differences in degeneration disappeared. Partial denervation resulted in denervated muscle fibers losing birefringence earlier than innervated fibers. These results suggest that juvenile hormone causes breakdown of flight muscles, and neural factors control degeneration of flight muscles to some extent under the presence of the juvenile hormone.

Journal Article↗

Neuroanatomical approaches to the study of insect photoperiodism.

The anatomical locations of three components of insect photoperiodism--the photoperiodic photoreceptor, photoperiodic clock and hormonal effector--are summarized and compared between species. Among photoperiodic photoreceptors, either the retinal or extraretinal types or both are operative, and there is no general relationship between phylogeny and photoreceptor type. The photoperiodic clock comprises time measurement and counter systems. Currently, it is generally accepted that circadian oscillators are involved in the photoperiodic clock. Several recent studies have raised the possibility that timeless, a circadian clock gene, plays a role in the photoperiodic clock in flies. The dorsal protocerebrum has been identified as an important region regulating the endocrine system for adult, pupal and embryonic diapause controlled by photoperiod. In the blow fly Protophormia terraenovae, neural connections between circadian clock neurons and indispensable neurons in the pars lateralis for diapause induction in the dorsal protocerebrum have been demonstrated. This neural network may provide the access needed to investigate the neural components of the photoperiodic clock.

Animals↗