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R Booker

Publications and source records attributed to R Booker.

At least 19 recordsLinked to original sources

Imaginal pioneers prefigure the formation of adult thoracic muscles in Drosophila melanogaster.

In insects, specialized mesodermal cells serve as templates to organize myoblasts into distinct muscle fibers during embryogenesis. In the grasshopper embryo, large mesodermal cells called muscle pioneers extend between the epidermal attachment points of future muscle fibers and serve as foci for myoblast fusion. In the Drosophila embryo, muscle founder cells serve a similar function, organizing large numbers of myoblasts into larval muscles. During the metamorphosis of Drosophila, nearly all larval muscles degenerate and are replaced by a set of de novo adult muscles. The extent to which specialized mesodermal cells homologous to the founders and pioneers of the insect embryo are involved in the development of adult-specific muscles has yet to be established. In the larval thorax, the majority of imaginal myoblasts are associated with the imaginal discs. We report here the identification of a morphologically distinct class of disc-associated myoblasts, which we call imaginal pioneers, that prefigures the formation of at least three adult-specific muscles, the tergal depressor of the trochanter and dorsoventral muscles I and II. Like the muscle pioneers of the grasshopper, the imaginal pioneers attach to the epidermis at sites where the future muscle insertions will arise and erect a scaffold for developing adult muscles. These findings suggest that a prior segregation of imaginal myoblasts into at least two populations, one of which may act as pioneers or founders, must occur during development.

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Octopamine mimics the effects of parasitism on the foregut of the tobacco hornworm Manduca sexta.

The parasitic braconid wasp Cotesia congregata lays its eggs inside the body of the larval stage of its host, the moth Manduca sexta. The Cotesia congregata larvae develop within the hemocoel of their host until their third instar, when they emerge and spin cocoons and pupate on the outer surface of the caterpillar. From this time until their death approximately 2 weeks later, the Manduca sexta larvae show striking behavioral changes that include dramatic declines in spontaneous activity and in the time spent feeding. Coincident with these behavioral changes, it is known that octopamine titers in the hemolymph of the host become elevated by approximately 6.5-fold. Octopamine is an important modulator of neural function and behavior in insects, so we examined hosts for neural correlates to the behavioral changes that occur at parasite emergence. We found that, in addition to the changes reported earlier, after parasite emergence (post-emergence), Manduca sexta larvae also showed marked deficits in their ability to ingest food because of a disruption in the function of the frontal ganglion that results in a significant slowing or the absence of peristaltic activity in the foregut. This effect could be produced in unparasitized fifth-instar larvae by application of blood from post-emergence parasitized larvae or of 10(-6)mol l(-1)d,l-octopamine (approximately the level in the hemolymph of post-emergence larvae). In contrast, blood from parasitized larvae before their parasites emerge or from unparasitized fifth-instar larvae typically had no effect on foregut activity. The effects of either post-emergence parasitized blood or 10(-6)mol l(-1) octopamine could be blocked by the octopamine antagonists phentolamine (at 10(-5)mol l(-1)) or mianserin (at 10(-7)mol l(-1)).

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Homeotic gene expression in the wild-type and a homeotic mutant of the moth Manduca sexta.

Antibodies were used to examine the expression patterns of Antennapedia (Antp), Ultrabithorax (Ubx), Ubx and abdominal-A combined (Ubx/abd-A), and Distalless (Dll) in the embryos of the moth Manduca sexta. We found that the spatial and temporal pattern of Antp expression in Manduca was correlated with the anterior migration of two patches of epithelium that include the anterior-most tracheal pits, and with the development of functional spiracles. Ubx expression showed an intricate pattern which suggests complex regulation during development. Throughout Manduca embryogenesis the expression of Ubx/Abd-A and Dll was similar to that reported for other insects. However, there was no apparent reduction in Ubx/Abd-A expression in the Manduca abdominal proleg primordia that expressed Dll. The expression of these four proteins was also examined in embryos of the Manduca homozygous homeotic mutant Octopod (Octo). The Octo mutation results in the transformation of A1 and A2 in the anterior direction, with homeotic legs appearing on A1 and occasionally A2. Our results suggest that in Octo animals there is a reduction in the level of Ubx protein expression throughout its domain. Based on homeotic gene expression in wild-type and mutant Manduca and in other insects, we discuss potential roles of homeotic genes in insect morphological evolution.

Amino Acid Sequence↗

Innervation regulates the metamorphic fates of larval abdominal muscles in the moth, Manduca sexta.

With the onset of metamorphosis, the abdominal muscles of the moth, Manduca sexta, follow one of three developmental fates: maintenance, respecification, or death. The maintained muscles retain their larval size and morphology throughout adult development. The respecified and dying muscles dedifferentiate, which involves regression, nuclear degeneration, and myofibril breakdown. Nuclei in both dying and respecified muscles also proliferate. The amount of nuclear degeneration is greater in the dying muscle fibers, and the amount of nuclear proliferation is greater in the respecified muscles. Four to ten days after pupation, the sizes of the respecified muscles stabilize while the dying muscles are lost. During regression, a subset of the respecified muscle fibers die. The surviving respecified muscle fibers grow and differentiate during the last half of adult development. In respecified muscles, denervation triggers an increased amount of nuclear degeneration and a decreased amount of nuclear proliferation. As a result, denervated respecified fibers experience increased muscle regression including an increased loss of muscle fibers and sometimes muscle death. Surviving respecified fibers still grow and differentiate yet are only 5 to 12% of the control size. Denervation triggers dedifferentiation in maintained muscles, resulting in fiber loss and occasionally muscle death. The percentage of fibers which dedifferentiate varies between different muscles. Denervation also triggers nuclear proliferation, with the amount of nuclear proliferation correlated with the extent of dedifferentiation of the individual muscle fibers. The dedifferentiated maintained fibers subsequently undergo differentiation in the absence of muscle growth.

Abdominal Muscles↗

The role of the frontal ganglion in the feeding and eclosion behavior of the moth manduca sexta

We have examined the musculature and motor patterns of the foregut and the role of the frontal ganglion in the adult moth Manduca sexta. During adult development, the structure of the foregut changes from a simple straight tube to a pump consisting of a flexible-roofed chamber or cibarium, with dilator muscles that raise the roof to draw in fluids and a compressor to push it down and force the fluid down the thin-walled esophagus. The frontal ganglion drives the activity of this cibarial pump during feeding, which is triggered by the application of sucrose solution or water to the proboscis. The feeding motor pattern consists of coupled bursts of the pump dilators and shorter-duration, high-frequency bursts of spikes from the pump compressor. The pump is also activated at the adult molt. At this time, it is used both before the moth emerges from the pupal case for swallowing molting fluid and again after emergence for swallowing air. These behaviors are important for eclosion and are necessary for the expansion of the wings after eclosion. Their motor patterns are similar to the feeding program. Up to 24 h before adult ecdysis, this motor pattern can be triggered by the peptide eclosion hormone. The other eclosion-related peptide, Manduca sexta eclosion-triggering hormone, does not appear to trigger activity of the cibarial pump.

Journal Article↗

Asthma inhalers.

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Asthma↗

Genesis of the adult retina and outer optic lobes of the moth, Manduca sexta. I. patterns of proliferation and cell death.

We have examined the development of the adult retina and the outer optic lobes in the moth Manduca sexta. The adult retina is generated from a group of epithelial cells lying within the larval head capsule between the larval ocelli and antenna. Proliferation of these cells begins during the feeding larval stage but accelerates at the end of the final larval instar. Proliferation occurs in two zones of mitotic activity; these zones flank a furrow in the presumptive retinal epithelium. The furrow and flanking mitotic zones migrate from posterior to anterior across the presumptive retinal epithelium. Posterior to the furrow, presumptive retinal cells from clusters and extend axons into the larval optic nerve. We have also examined the temporal patterns of neuronal proliferation and cell death during genesis of the adult outer optic ganglia, the medulla and the lamina. The medulla and the lamina are generated by distinct populations of neuroblasts in the outer optic analage; the neuroblasts divide asymmetrically to generate ganglion mother cells. Ganglion mother cells later divide symmetrically to generate immature neurons. Generation of the medulla cortex starts with the onset of the final larval instar, and cell death within the medulla cortex begins after the end of the final larval instar. Generation of the lamina cortex is initiated with the arrival of retinal afferents at the optic lobes, and cell death in the lamina cortex begins 1 day later. Generation of the outer optic ganglia terminates with the abrupt cessation of mitotic activity followed by degeneration of the outer optic anlage.

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Genesis of the adult retina and outer optic lobes of the moth, Manduca sexta. II. Effects of deafferentation and developmental hormone manipulation.

We have examined factors affecting neuronal proliferation and cell death during development of the adult outer optic ganglia in the moth Manduca sexta. The role of retinal afferents was addressed by optic nerve section on the first day of pupation, when the retina is only partially formed. Deafferentation by optic nerve section resulted in increased levels of cell death in the medulla cortex but did not affect medulla precursor proliferation. Conversely, division of lamina ganglion mother cells ceased following deafferentation, whereas cell death in the lamina cortex did not increase significantly. Implantation of isolated outer optic anlage into host animals demonstrated that medulla precursor proliferation was independent of centripetal inputs and retinal afferents. The role of developmental hormones in regulating outer optic ganglia development was also investigated. Many events in medulla and lamina development are correlated with changes in the titers of the ecdysteroids, and these events could be prevented by application of a juvenile hormone mimic (JHM). Application of JHM early in the fifth (final) larval instar terminated medulla precursor proliferation and prevented the initiation of lamina precursor proliferation, but application late in the fifth instar (after the commitment peak of ecdysteroids) had little effect on proliferation. Application of JHM on the third or fourth day of pupal life prolonged precursor proliferation and delayed degeneration of the outer optic anlage. Precocious application of 20-hydroxyecdysone on the first day of pupal life accelerated outer optic anlage degeneration but did not cause premature termination of proliferation.

Afferent Pathways↗

Postembryonic neurogenesis in the central nervous system of the tobacco hornworm, Manduca sexta. III. Spatial and temporal patterns of proliferation.

Postembryonic neurogenesis leads to a dramatic increase in the number of functional neurons within the segmental ganglia of the moth, Manduca sexta. These adult-specific neurons are generated during larval life by segment-specific arrays of individually identifiable stem cells, or neuroblasts (Nbs). By the end of the feeding larval stage, each Nb has generated a discrete nest of progeny, which ranges in size from less than 10 to more than 70 progeny. The sizes of these identifiable nests of progeny vary in a segment-specific manner, with the thoracic nests containing a greater number of progeny compared with their homologues in the simpler abdominal ganglia. In order to describe those factors that influence the size of the post-embryonic neuronal lineages, we examined the spatial and temporal pattern of postembryonic neurogenesis in the segmental ganglia of Manduca. The rates at which the identifiable nests accumulated progeny were estimated by counting the number of progeny within the nests, using sectioned material isolated from animals at stages ranging from embryonic hatching until the end of the feeding larval stage. All of the postembryonic Nbs began to generate progeny at around the time of the molt to the third larval instar. Each nest added progeny at a rate that was a characteristic of its identity and segment of origin. Although all of the nests within the thorax continued to accumulate progeny throughout the feeding larval stage, several of the abdominal nests showed little or no growth following the molt to the fifth larval instar. The thymidine analog 5-bromo 2-deoxyuridine (5-BrdU) was used to estimate the mitotic rates of the identifiable Nbs. The number of labeled progeny within a nest 24 h after application of 5-BrdU ranged from a low of 1 to 2 to a high of 11 to 13 labeled cells. In some instances there was a good correlation between the estimated mitotic rate of an identified Nb and the rate of growth of its associated nest of progeny. However, several of the identifiable nests accumulated progeny at a slower rate than predicted based on the estimated mitotic rate of the Nb. Cell death appears to be responsible for slowing the growth of the nests during the feeding larval stage. We estimate that 10% to 70% of the neurons generated during the feeding larval stage degenerate within 24 h of their birth. The level of cell death observed within a nest was dependent on both its identity and its segment of origin.

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Projection pattern of sensory neurons in the central nervous system of a homeotic mutation of the moth Manduca sexta.

Octopod (Octo) is a mutation of the moth Manduca sexta, which transforms the first abdominal segment (A1) in the anterior direction. Mutant animals are characterized by the appearance of homeotic thoracic-like legs on A1. We exploited this mutation to determine what rules might be used in specifying the fates of sensory neurons located on the body surface of larval Manduca. Mechanical stimulation of homeotic leg sensilla did not cause reflexive movements of the homeotic legs, but elicited responses similar to those observed following stimulation of ventral A1 body wall hairs. Intracellular recordings demonstrated that several of the motoneurons in the A1 ganglion received inputs from the homeotic sensory hairs. The responses of these motoneurons to stimulation of homeotic sensilla resembled their responses to stimulation of ventral body wall sensilla. Cobalt fills revealed that the mutation transformed the segmental projection pattern of only the sensory neurons located on the ventral surface of A1, resulting in a greater number with intersegmental projection patterns typical of sensory neurons found on the thoracic body wall. Many of the sensory neurons on the homeotic legs had intersegmental projection patterns typical of abdominal sensory neurons: an anteriorly directed projection terminating in the third thoracic ganglion (T3). Once this projection reached T3, however, it mimicked the projections of the thoracic leg sensory neurons. These results demonstrate that the same rules are not used in the establishment of the intersegmental and leg-specific projection patterns. Segmental identity influences the intersegmental projection pattern of the sensory neurons of Manduca, whereas the leg-specific projections are consistent with a role for positional information in determining their pattern.

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Octopod, a homeotic mutation of the moth Manduca sexta, affects development of both mesodermal and ectodermal structures.

Several aspects of leg development in the moth Manduca sexta were examined using the homeotic mutation Octopod (Octo). This mutation causes a transformation of the ventral epidermis of the first abdominal segment (A1) to that of the third thoracic segment (T3), resulting in the presence of thoracic-like legs on A1. The degree of transformation of A1 is variable, ranging from bumps on the cuticle to fully segmented thoracic-like legs. In the normal thoracic legs, clusters of undifferentiated cells known as differentiation centers are located around the coxal-trochanteral, femoral-tibial, and tibial-tarsal joints. The adult thoracic legs develop from the differentiation centers at metamorphosis. The homeotic legs of the Octopod larvae also have differentiation centers at comparable positions in the homeotic leg. As a result, the number of leg segments in a mutant adult is correlated with the number of segments and differentiation centers that animal had in its larval homeotic leg. Our data suggest that the differentiation center located at the coxal-trochanteral joint forms the adult coxa and trochanter, the center at the larval femoral-tibial joint the adult femur and tibia, and the differentiation center at the larval tibial-tarsal joint the adult tarsus. Homeotic larval legs which include at least a femur have supernumerary muscles, while adult homeotic legs rarely show discrete muscle. The homeotic larval muscles appear to have thoracic identities, based on their attachment points and the timing of their degeneration at the larval-pupal transition. They are innervated by a motoneuron that is normally present in A1 where it innervates the ventral lateral external muscle (VLE). In mutant animals, the same motoneuron innervates all of the homeotic muscles and the VLE. We consider possible mechanisms underlying the development of homeotic muscles and their innervation. At the larval-pupal transition, the VLE in mutant animals degenerates at its normal time, which is 3 days after the degeneration of the homeotic muscles. Thus, despite their common innervation, the two muscle types degenerate according to their own schedules, indicating that the developmental fates of the muscles are not dictated by their innervating neuron but are intrinsic to the muscles themselves.

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Isolation and embryonic expression of an abdominal-A-like gene from the lepidopteran, Manduca sexta.

Using sequence homology to the Drosophila Antennapedia gene, we isolated a homeobox-containing gene from the lepidopteran, Manduca sexta. Sequence analysis and in situ hybridizations to tissue sections suggest that the Manduca gene encodes a lepidopteran homologue of the Drosophila Bithorax complex gene abdominal-A. The predicted amino acid sequence of a 76 amino acid region that includes the homeobox and the regions immediately flanking it are identical between the Manduca and Drosophila genes. Northern blots reveal that the manduca abd-A gene is expressed first in the early embryo and continues to be expressed throughout later embryonic and larval stages. In situ hybridizations show that the posterior half of the first abdominal segment marks the anterior border of the Manduca abd-A expression. This expression pattern demonstrates the conservation of parasegments as domains of gene activity in the lepidopteran embryo. The Manduca abd-A expression extends from the posterior half of the first abdominal segment through the tenth abdominal segment, a domain that is greater than that of the Drosophila abd-A expression, and reflects the difference in visible segment number between the two insects.

Amino Acid Sequence↗

Octopod, a homeotic mutation of the moth Manduca sexta, influences the fate of identifiable pattern elements within the CNS.

Octopod (Octo) is a mutation of the moth Manduca sexta, which results in the homeotic transformation of the ventral surface of the first (A1) and less often the second (A2) abdominal segments in the anterior direction. The extent of the transformation ranges from a slight deformation of the ventral cuticle, up to the formation of miniature thoracic legs on A1. The extent of the transformation is always less within A2 as compared to A1. A genetic analysis revealed that Octo is an autosomal mutation which shows incomplete dominance. The effect of this mutation on the central nervous system (CNS) was assessed by examining the distribution and fate of the postembryonic neuroblasts in the segmental ganglia of Octo larvae. In each of the thoracic ganglia of wild-type larvae, there is a set of 45-47 neuroblasts; a reduced but homologous array of 24 and 10 neuroblasts are found in A1 and A2, respectively. Ganglion A1 of Octo larvae had 1 to 6 supernumerary neuroblasts, and 20% of the A2 ganglia showed a single ectopic neuroblast. The supernumerary neuroblasts corresponded to identifiable neuroblasts normally found in more anterior ganglia. The Octo mutation also influenced the mitotic activity of stem cells normally present in A1. In this case, the neuroblasts generated a lineage of cells that were typical of a thoracic location rather than A1. These data demonstrate that homeotic mutations can influence the fate of identifiable pattern elements within the CNS of an insect.

Animals↗

Postembryonic neurogenesis in the CNS of the tobacco hornworm, Manduca sexta. I. Neuroblast arrays and the fate of their progeny during metamorphosis.

The tobacco hornworm Manduca sexta exhibits dramatic changes in its body morphology and behavior as it is transformed from a larva into an adult during metamorphosis. Accompanying these changes is an extensive reorganization of this moth's central nervous system (CNS), which involves both the death and remodeling of subsets of larval neurons. We report here that the segmental ganglia of the larvae also contain a stereotyped array of identifiable neuronal stem cells (neuroblasts) that contribute over 2,000 cells to each thoracic ganglion and about 40-80 cells to each abdominal ganglion. The distribution of these neuroblasts varies in a segment specific manner. Dormant neuroblasts are found adjacent to the neuropil in late embryos and early first instar larvae. After the molt to the second instar, these cells enlarge and begin to divide. Through a series of asymmetrical divisions, each neuroblast generates a discrete nest of 10-90 progeny by the end of larval life. These progeny (the imaginal nest cells) are developmentally arrested at an early stage of differentiation and remain so until metamorphosis. At the onset of metamorphosis, a wave of cell death sweeps through the nests, the extent of the death being much greater within the abdominal nests than in the thoracic nests. The surviving imaginal nest cells then differentiate to become functional neurons that are incorporated into the adult CNS.

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