The brain can eat: Establishing the existence of a central pattern generator for feeding in third instar larvae of Drosophila virilis and Drosophila melanogaster

To establish the existence of a central pattern generator for feeding in the larval central nervous system of two Drosophila species, the gross anatomy of feeding related muscles and their innervation is described, the motor units of the muscles identified and rhythmic motor output recorded from the isolated CNS. The cibarial dilator muscles that mediate food ingestion are innervated by the frontal nerve. Their motor pathway projects from the brain through the antennal nerves, the frontal connectives and the frontal nerve junction. The mouth hook elevator and depressor system is innervated by side branches of the maxillary nerve. The motor units of the two muscle groups differ in amplitude: the elevator is always activated by a small unit, the depressor by a large one. The dorsal protractors span the cephalopharyngeal skeleton and the body wall hence mediating an extension of the CPS. These muscles are innervated by the prothoracic accessory nerve. Rhythmic motor output produced by the isolated central nervous system can simultaneously be recorded from all three nerves. The temporal pattern of the identified motor units resembles the sequence of muscle contractions deduced from natural feeding behavior and is therefore considered as fictive feeding. Phase diagrams show an almost identical fictive feeding pattern is in both species.     

Metamorphosis of Wing Motor System in the Silk Moth, Bombyx mori: Origin of Wing Motor Neurons

The origin of the peripheral nerve and motor neurons that innervate the adult mesothoracic dorsal longitudinal muscles (DLMs) was examined in the silk moth, Bombyx mori . The anatomical features of the peripheral nerve and motor neurons were investigated by dissection, electron microscopy, and cobalt back-fill staining at different pupal stages. These studies showed that the peripheral nerve (IIN1c) that innervates the adult DLMs originates from a branch (db branch) of the larval mesothoracic dorsal nerve that innervates the larval DLMs. During metamorphosis the larval nerve shortens or lengthens locally without change in its basic branching pattern, and the db branch moves towards the mesothoracic ganglion to become the IIN1c. All the adult DLM motor neurons are from larval ones. Nine of the 14 larval DLM motor neurons survive during metamorphosis to become adult DLM motor neurons, and 5 disappear in early pupal stages.     

Innervation pattern of the gill arches and gills of the carp (Cyprinus carpio)

The innervation pattern of the respiratory gill arches of the carp (Cyprinus carpio) is described. The gill region is innervated by the branchial branches of the glossopharyngeal and vagal nerves. Each branchial nerve divides at the level of or just distal to the epibranchial ganglion into: 1) a pretrematic branch, 2) a dorsal pharyngeal branch, and 3) a posttrematic branch. The dorsal pharyngeal branch innervates the palatal organ in the roof of the buccal cavity. The pretrematic and posttrematic branches innervate the posterior and anterior halves, respectively, of the gill arches bordering a gill slit. Each branch splits into an internal and an external part. The internal bundle innervates the buccal side of the gill arch, including the gill rakers. The external bundle terminates in the gill filaments. The epibranchial motor branch, a small nerve bundle containing only motor fibers, circumvents the ganglion and anastomoses distally with the posttrematic branch. The detailed course and branching patterns of these branches are described.     

Cranial nerves in the Australian lungfish,Neoceratodus forsteri,and in fossil relatives (Osteichthyes: Dipnoi)

Three systems, two sensory and one protective, are present in the skin of the living Australian lungfish, Neoceratodus forsteri, and in fossil lungfish, and the arrangement and innervation of the sense organs is peculiar to lungfish. Peripheral branches of nerves that innervate the sense organs are slender and unprotected, and form before any skeletal structures appear. When the olfactory capsule develops, it traps some of the anterior branches of cranial nerve V, which emerged from the chondrocranium from the lateral sphenotic foramen. Cranial nerve I innervates the olfactory organ enclosed within the olfactory capsule and cranial nerve II innervates the eye. Cranial nerve V innervates the sense organs of the snout and upper lip, and, in conjunction with nerve IX and X, the sense organs of the posterior and lateral head. Cranial nerve VII is primarily a motor nerve, and a single branch innervates sense organs in the mandible. There are no connections between nerves V and VII, although both emerge from the brain close to each other. The third associated system consists of lymphatic vessels covered by an extracellular matrix of collagen, mineralised as tubules in fossils. Innervation of the sensory organs is separate from the lymphatic system and from the tubule system of fossil lungfish.     

Anatomical partitioning of three multiarticular human muscles.

To examine neuromuscular partitioning within human muscles, the innervation patterns and muscle fiber architecture of the flexor carpi radialis (FCR), extensor carpi radialis longus (ECRL) and lateral gastrocnemius (LG) muscles were examined. Consistent patterns of innervation between specimens were found within each of the three muscles. The nerve to the FCR clearly innervates three major architectural divisions of the muscle. The ECRL is innervated by two different muscle nerves. Branches of these nerves innervate at least two distinct anatomical subvolumes. However, the subvolumes of the ECRL defined by muscle architecture are not totally congruent with those defined by the innervation pattern. In the LG, the single muscle nerve branches into two main divisions, and these subsequently divide into branches which supply the three heads. However, each head does not receive a completely private nerve. These results indicate that human muscles are partitioned in a manner roughly similar to the divisions of the same muscles in cats and rats, but with less congruency of architecture and innervation.     

Complex innervation of three neck muscles by motor and dorsal unpaired median neurons in crickets

Summary In the crickets, Gryllus campestris and Gryllus bimaculatus, the innervation of the dorso-ventral neck muscles M62, M57, and M59 was examined using cobalt staining via peripheral nerves and electrophysiological methods. M62 and M57 are each innervated by two motoneurons in the suboesophageal ganglion. The four motoneurons project into the median nerve to bifurcate into the transverse nerves of both sides. M62 and M57 are the only neck muscles innervated via this route. These bifurcating axon-projections are identical to those of the spiracular motoneurons in the prothoracic ganglion innervating the opener and closer muscle of the first thoracic spiracle in the cricket. The morphology of their branching pattern is described. The neck muscle M57 and the opener muscle of the first thoracic spiracle are additionally innervated by one mesothoracic motoneuron each, with similar morphology. These results suggest, that in crickets, the neck muscles M57 and M62 are homologous to spiracular muscles in the thoracic segments. The two neck muscles M62 and M59 (the posterior neighbour of M57) receive projections from a prothoracic dorsal unpaired median (DUM) neuron that also innervates dorsal-longitudinal neck muscles but not M57. In addition, one or two mesothoracic DUM neurons send axon collaterals intersegmentally to M59. This is the first demonstration of the innervation of neck muscles by DUM neurons.     

Serial homologies of the motor neurons of the dorsal intersegmental muscles of the cockroach,Periplaneta americana (L.)

The types and locations of serially homologous motor neurons of the dorsal muscles in the cockroach Periplaneta americana remain rather constant regardless of the various adaptations of their muscles or the fusion of ganglia. However, the size and number of neurons do vary according to the development of the muscles they innervate. Neurons in four distinctive locations, two ipsisegmental and two antesegmental, innervate the dorsal longitudinal (DL) muscles in most segments. One of the ipsisegmental neurons (DLC) is common to all of the DL muscles of a segment and probably has a modulatory function. The dorsal oblique (DO) muscles of most segments have neurons in two antesegmental positions. One of these, an antesegmental, contralateral neuron, innervates both DO and DL muscles in each segment and is also probably modulatory. One neuron (DOC) of the prothoracic ganglion is the principal exception to the constancy of these serially homologous neurons. This neuron appears to be homologous to the DLC neurons of other segments but innervates the DO rather than the DL muscles.     

The oculomotor system of Daphnia magna

Summary The highly mobile cyclopic compound eye of Daphnia magna is rotated by six muscles arranged as three bilateral pairs. The three muscles on each side of the head share a common origin on the carapace and insert dorsally, laterally and ventrally on the eye. The dorsal and ventral muscles are each composed of two muscle fibers and the lateral muscle is composed of from two to five fibers, with three the most common number. Individual muscle fibers are spindle-shaped mononucleated cells with organized bundles of myofilaments. Lateral eye-muscle fibers are thinner than those of the other muscles but are otherwise similar in ultrastructure. Two motor neurons innervate each dorsal and each ventral muscle and one motor neuron innervates each lateral muscle. The cell bodies of the motor neurons are situated dorsally in the supraesophageal ganglion (SEG) and are ipsilateral to the muscles they innervate. The dendritic fields of the dorsal-muscle motor neurons are ipsilateral to their cell bodies; those of the ventral-muscle motor neurons are bilateral though predominantly contralateral. The central projections of the lateral-muscle motor neurons are unknown. In the dorsal and ventral muscles one motor axon synapses principally with one muscle fiber; in each lateral muscle the single motor axon branches to, and forms synapses with, all the fibers. The neuromuscular junctions, characterized by pre- and postsynaptic densities and clear vesicles, are similar in all the eye muscles.     

The structure and function of serially homologous leg motor neurons in the locust. I. Anatomy

Twenty-one prothoracic and 17 mesothoracic motor neurons innervating leg muscles have been identified physiologically and subsequently injected with dye from a microelectrode. A tract containing the primary neurites of motor neurons innervating the retractor unquis, levator and depressor tarsus, flexor tibiae, and reductor femora is described. All motor neurons studied have regions in which their dendritic branches overlap with those of other leg motor neurons. Identified, serially homologous motor neurons in the three thoracic ganglia were found to have: (1) cell bodies at similar locations and morphologically similar primary neurites (e.g., flexor tibiae motor neurons), (2) cell bodies at different locations in each ganglion and morphologically different primary neurites in each ganglion (e.g., fast retractor unguis motor neurons), or (3) cell bodies at similar locations and morphologically similar primary neurites but with a functional switch in one ganglion relative to the function of the neurons in the other two ganglia. As an example of the latter, the morphology of the metathoracic slow extensor tibiae (SETi) motor neurons was similar to that of pro- and mesothoracic fast extensor tibiae (FETi) motor neurons. Similarly the metathoracic FETi bears a striking resemblance to the pro- and the mesothoracic SETi. It is proposed that in the metathoracic ganglion the two extensor tibiae motor neurons have switched functions while retaining similar morphologies relative to the structure and function of their pro- and mesothoracic serial homologues.     

Central projections of the thympanic fibres in noctuid moths

The receptor mechanism mediating the avoidance behaviour of flying noctuid moths in response to brief ultrasonic pulses may require only a single pair of acoustic sense cells, one A1 cell in each tympanic organ (Roeder, 1966c). Introduction of the fluorescent dye, procion yellow, into the nerve fibres leaving the tympanic organ has allowed the reconstruction of the central morphology of A1, the more sensitive of the two acoustic cells. The A1 axon follows a superficial course for the first ～100 μ auterior to its dorsal root of entry (3N1) into the thoracic ganglia, then plunges ventrally into the posterior mesothoracic neuropil where it branches. The posterior part reaches through two-thirds of the metathoracic ganglion. The anterior branch bifurcates in the anterior mesothoracic ganglion to give rise to a posteriorly directed branch extending through the ventral mesothoracic neuropil and an anterior branch which passes through the connective into the posterior half of the prothoracic ganglion. Here it ramifies along the midline. The cell remains strictly ipsi-lateral with numerous processes extending right up to the midline in the ventral neuropil of all three ganglia. This morphology correlates well with the map of sites from which A1 acoustic responses can be recorded in the central nervous system.     

Immunohistological localization of serotonin in the CNS and feeding system of the stable fly Stomoxys calcitrans L. (Diptera: Muscidae)

Serotonin, or 5-hydroxytryptamine (5-HT), plays critical roles as a neurotransmitter and neuromodulator that control or modulate many behaviors in insects, such as feeding. Neurons immunoreactive (IR) to 5-HT were detected in the central nervous system (CNS) of the larval and adult stages of the stable fly, Stomoxys calcitrans, using an immunohistological technique. The location and pattern of the 5-HT IR neurons are described and compared for these two different developmental stages. Anatomical features of the fly feeding system were analyzed in third instar larvae and adult flies using a combination of histological and immunohistological techniques. In third instar larvae, the cibarial dilator muscles were observed within the cibarial pump skeleton and innervated by 5-HT IR neurons in nerves arising from the brain. There were four pairs of nerves arising from the frontal surface of the larval brain that innervate the cibarial pump muscles, pharynx, and muscles controlling the mouth hooks. A strong serotoninergic innervation of the anterior stomatogastric system was observed, which suggests 5-HT may play a role in the coordination of different phases of food ingestion by larvae. Similarly, many 5-HT IR neurons were found in both the brain and the thoracico-abdominal ganglia in the adult, some of which innervate the cibarial pump dilator muscles and the stomatogastric muscles. This is tnhe first report describing neuromuscular structures of the stable fly feeding system. The results reported here suggest 5-HT may play a critical role in feeding behaviors of stable fly larvae and adults.     

Central projections of fibers in the auditory and tensor nerves of cicadas (Homoptera: Cicadidae)

Summary The auditory and tensor nerves of cicadas are mixed nerves containing both afferent and efferent elements. In 17-year cicadas, and in Okanagana rimosa, the auditory nerve contains afferents from body hairs, from the detensor tympani-chordotonal organ, and some 1300–1500 afferents from the hearing organ. Within the fused metathoracic-abdominal ganglionic complex the receptors from both the auditory and tensor nerves form a neuropilar structure that reveals the metameric organization of this complex. A few fibers run anteriorly, projecting into the meso and prothoracic ganglia. Within the ganglionic complex a division of auditory nerve afferents into a dense intermediate and a more diffuse ventral neuropile is observed. In addition, a dorsal motor neuropile is outlined by arborizations of the timbal motor neuron. This neuron is one of several efferent cell types associated with the auditory nerve, and there is an indication that several efferent fibers innervate the timbal muscle. There is anatomical evidence for a possible neuronal coupling between the bilaterally symmetrical large timbal motor neurons. In general, central projections from the auditory and tensor nerves support evidence of a structural layering within the CNS of insects.     

The dorsomedial nuclear group of cranial nerves in the frog.

The dorsomedial motor nuclei were demonstrated by the cobalt-labeling technique applied to the so-called somatic motor cranial nerves. The motoneurons constituting these nuclei are oval-shaped and smaller than the motoneurons in the ventrolateral motor nuclei. They give rise to ventral and dorsal dendrite groups which have extensive arborization areas. A dorsolateral cell group in the rostral three quarters of the oculomotorius nucleus innervates ipsilateral eye muscles (m.obl.inf., m.rect.inf., m.rect.med.) and a ventromedial cell group innervates the contralateral m. rectus superior. Ipsilateral axons originate from ventral dendrites, contralateral axons emerge from the medial aspect of cell bodies, or from dorsal dendrites, and form a "knee" as they turn around the nucleus on their way to join the ipsilateral axons. A few labeled small cells found dorsal and lateral to the main nucleus in the central gray matter are regarded as representing the nucleus of Edinger-Westphal. The trochlearis nucleus is continuous with the ventromedial cell group of the oculomotorius nucleus. The axons originate in dorsal dendrites, run dorsally along the border of the gray matter and pierce the velum medullare on the contralateral side. A compact dendritic bundle of oculomotorius neurons traverse the nucleus, and side branches appear to be in close apposition to the trochlearis neurons. A dorsomedial and a ventrolateral cell group becomes labeled via the abducens nerve. The former supplies the m. rectus lateralis, while the latter corresponds to the accessorius abducens nucleus which innervates the mm. rectractores. Neurons in this latter nucleus are large and multipolar, resembling the neurons in the ventrolateral motor nuclei. Their axons originate from dorsal dendrites and form a "knee" around the dorsomedial aspect of the abducens nucleus. Cobalt applied to the hypoglossus nerve reaches a dorsomedial cell group (the nucleus proper), spinal motoneurons and sympathetic preganglionic neurons. Of the dorsomedial motor cells, the hypoglossus neurons are the largest, and a branch of their ventral dendrites terminates on the contralateral side. Some functional and developmental biological aspects of the morphological findings, such as the crossing axons and the peculiar morphology of the accessory abducens nucleus, are discussed.     

Anatomy of the stomatogastric nervous system associated with the foregut in Drosophila melanogaster and Calliphora vicina third instar larvae

The stomatogastric nervous system (SNS) associated with the foregut was studied in 3rd instar larvae of Drosophila melanogaster and Calliphora vicina (blowfly). In both species, the foregut comprises pharynx, esophagus, and proventriculus. Only in Calliphora does the esophagus form a crop. The position of nerves and neurons was investigated with neuronal tracers in both species and GFP expression in Drosophila. The SNS is nearly identical in both species. Neurons are located in the proventricular and the hypocerebral ganglion (HCG), which are connected to each other by the proventricular nerve. Motor neurons for pharyngeal muscles are located in the brain not, as in other insect groups, in the frontal ganglion. The position of the frontal ganglion is taken by a nerve junction devoid of neurons. The junction is composed of four nerves: the frontal connectives that fuse with the antennal nerves (ANs), the frontal nerve innervating the cibarial dilator muscles and the recurrent nerve that innervates the esophagus and projects to the HCG. Differences in the SNS are restricted to a crop nerve only present in Calliphora and an esophageal ganglion that only exists in Drosophila. The ganglia of the dorsal organs give rise to the ANs, which project to the brain. The extensive conformity of the SNS of both species suggests functional parallels. Future electrophysiological studies of the motor circuits in the SNS of Drosophila will profit from parallel studies of the homologous but more accessible structures in Calliphora.     

The central projection of cephalic mechanosensory axons in the haematophagous bug Triatoma infestans

The projections of mechanosensory hairs located on the dorsal and lateral head of the adult haematophagous bug Triatoma infestans were analyzed by means of cobalt filling. Axons run into the anterior and posterior tegumentary nerve and project through the brain to the ventral nerve cord. The fibres are small in diameter and run as a fascicle. Some branches run into suboesophageal and prothoracic centres; others run as far as to the mesothoracic ganglion. These sensory projections resemble that of wind-sensitive head hairs of the locust. The functional role of this sensory system in this species is discussed.     

Suboesophageal neurons involved in head movements and feeding in locusts.

The projections of nerves 6 and 7 of the locust suboesophageal ganglion (SOG) were stained by axonal filling with cobalt chloride. Nerve 6 contains two motoneurons which innervate neck muscles 50 and 51. Sensory neurons innervating hairs on the dorso-occipital region of the head also enter the ganglion through nerve 6 and terminate in a small bilateral plexus. The projections of the head hairs in nerve 6 do not overlap the arborizations of the motoneurons or the neurons of nerve 7, but lie in the same area as descending sensory neurons from wind-sensitive hairs of the front of the head. One branch of nerve 7 (7B) contains two fibres which innervate the salivary gland. These 'salivary' neurons (labelled SN1 and SN2) have their cell bodies in the ganglion. The second branch, 7A, contains sensory neurons from the submentum of the labium, which form four sensory plexuses, two dorsal and two ventral. The sensory plexuses from the submentum have specific regions of overlap with the salivary neurons and with the neck muscle motoneurons. We interpret these as indicating a flow of information from labial receptors signalling head and mouthpart movement to neurons involved in salivation and head movement. We further postulate that the anatomical separation of the various sensory plexuses is indicative of functional localization within the ganglion.