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PubMed1992

Allatostatins in the nerves of the antennal pulsatile organ muscle of the cockroach Diploptera punctata.

The presence of allatostatins in the nerves of the antennal pulsatile organ muscle of the cockroach Diploptera punctata was confirmed by immunocytochemistry, bioassay, and HPLC. Immunocytochemical reactivity with monoclonal antibody against allatostatin I showed strong allatostatin immunoreactivity in the antennal heart nerve which innervates this muscle with varicosities along the muscle fibers and in the insertion of the muscle on the pulsatile ampullae. Bioassay of Sep-Pak purified muscle extract demonstrated inhibition of juvenile hormone synthesis by corpora allata in vitro. A dose-response curve showed maximum inhibition of juvenile hormone synthesis was achieved with 10-20 pulsatile organ muscle eq/corpora allata, and 50% inhibition achieved with an estimated 2.6 pulsatile organ muscle eq. Two successive HPLC separations of the Sep-Pak purified extract yielded bioactive fractions corresponding to the elution times of the five known allatostatins.

Amino Acid Sequence↗
PubMed1995

In vitro bioassay for hypertrehalosemic hormone-dependent trehalose biosynthesis by fat body from adult Blaberus discoidalis cockroaches.

An in vitro bioassay suitable for routine use to investigate hypertrehalosemic hormone (HTH)-dependent trehalose biosynthesis was developed for the cockroach fat body. Blaberus discoidalis fat bodies were isolated and divided so that eight matched pieces from a single tissue could be compared for multiple control and experimental treatments. Optimum incubation conditions and the properties of HTH-dependent trehalose synthesis were determined. Dose-response studies determined an EC50 of 0.044 nM. HTH for male fat body and 0.16 nM HTH for female tissue. HTH increased trehalose production by male fat body 3-fold compared to only a 67% maximum increase by the female tissue, and only the male tissue was used in subsequent studies. Fat body required only 5-min exposure to HTH for maximum trehalose production for 1 h. Trehalose synthesis was inhibited by > or = 15 mM trehalose in the incubation medium. The fat body showed a developmental increase in trehalose synthesis in vitro that was reflected by hemolymph trehalose in vivo. Basal and HTH-related trehalose synthesis were low between days 0 and 10, increased 3-fold by day 20, and were high thereafter. These studies have established baseline data for future investigations to identify the signal transduction mechanisms involved in HTH regulation of fat body metabolism.

Animals↗
PubMed2001

Candidates for extraocular photoreceptors in the cockroach suggest homology to the lamina and lobula organs in beetles.

Using light- and electron microscopic methods, we describe two novel putative extraocular photoreceptor organs in the optic lobes of the cockroaches Leucophaea maderae and Blaberus craniifer. The lamina organ is an elongated structure distal to the first optic chiasm, adjacent to the anterior edge of the lamina. The lobula organ is situated on the anterior distal surface of the lobula. In cross sections through the pigment-free organs, cell bodies are arranged in a closed or open circle and are interconnected by desmosomes. They send protrusions with rhabdom-like microvilli into a common, central space apparently filled with extracellular matrix. A different cell type gives rise to electron-dense lamellae, which also extend into the central space and partly join to form a common lamellar bundle. Axonal processes extend from the microvillar cells and run along the outer surface of the organs to the neighboring optic neuropils. The organs receive multiple efferent innervation from neurosecretory axons. Both organs show strong immunostaining with an antiserum against Arabidopsis cryptochrome 2 that is associated with the lamellated structure in the central lumen. The specific features of the organs suggest that they are homologous to similar organs in the optic lobe of beetles and may serve a role as extraocular photoreceptors for light entrainment of the circadian system.

Animals↗
PubMed2001

Anatomy and physiology of neurons with processes in the accessory medulla of the cockroach Leucophaea maderae.

The accessory medulla (AMe), a small neuropil in the insect optic lobe, has been proposed to serve a circadian pacemaker function analogous to the role of the suprachiasmatic nucleus in mammals. Building upon considerable knowledge of the circadian system of the cockroach Leucophaea maderae, we investigated the properties of AMe neurons in this insect with intracellular recordings combined with dye injections. Responses of neurons with processes in the AMe to visual stimuli, including stationary white light, moving objects, and polarized light were compared with the responses of adjacent medulla tangential neurons. Neurons with processes in the AMe and additional ramifications in the medulla strongly responded to stationary light stimuli and might, therefore, be part of photic entrainment pathways to the clock. Accessory medulla neurons lacking significant processes in the medulla but with projections to the midbrain or to the contralateral optic lobe, in contrast, responded weakly or not at all to light and, thus, seem to be part of the clock's output pathway. Two types of commissural neurons with tangential arborizations in both medullae were sensitive to polarized light, suggesting a role of these neurons in celestial navigation. Sidebranches in the AMae of one of the two cell types are discussed with respect to a possible involvement of the AMe in polarization vision. Finally, neurons responding to movement stimuli did not arborize in the AMe. The results show that the AMe receives photic input and support a role of this neuropil in circadian timekeeping functions.

Action Potentials↗
PubMed2005

Octopamine-like immunoreactivity in the honey bee and cockroach: comparable organization in the brain and subesophageal ganglion.

A serum raised against octopamine reveals in cockroaches and honey bees structurally comparable systems of perikarya and their extensive yet discrete systems of arborizations in neuropils. Numerous and prominent clusters of lateral cell bodies in the brain as well as many midline perikarya provide octopamine-like immunoreactive processes to circumscribed regions of the subesophageal ganglion, antennal lobe glomeruli, optic neuropils, and neuropils of the protocerebrum. There is dense octopaminergic innervation in the protocerebral bridge and ellipsoid body of the central complex. The antennal lobes are supplied by at least three octopamine-immunoreactive neurons. In contrast, the mushroom bodies show the fewest immunoreactive elements. In Apis a single axon supplies sparse immunoreactive processes to the calyces' basal ring, collar, and lip. A diffuse arrangement of immunoreactive processes invades all zones of the mushroom body calyces in Periplaneta. These processes derive from an ascending axon ascribed to a dorsal unpaired median neuron at the maxillary segment of the subesophageal ganglion. In both taxa octopamine-immunoreactive processes invade only the gamma lobes of the mushroom bodies, omitting their other divisions. The present observations are discussed with respect to possible roles of octopamine in sensory integration and association.

Animals↗
PubMed2006

Development-dependent and -independent ubiquitin expression in divisions of the cockroach mushroom body.

It has been proposed that the alpha and beta divisions of the mushroom bodies support intermediate and long-term memory whereas the gamma lobes support short-term memory. Here we investigate developmentally dependent versus developmentally independent alterations of mushroom body structure with special emphasis on its lobes. We show that in the cockroach mushroom bodies there are two types of plastic remodeling. One is developmental, in which episodic addition of new circuitry to the alpha and beta lobes is accomplished by newly born Kenyon cells. The second is revealed as a persistent alteration of structure within the gamma lobe. In the alpha/beta lobes, newly generated Kenyon cell axons extend glutamate-immunoreactive collaterals across layers of the axons of mature Kenyon cells. At specific times in each developmental episode (instar) these collaterals express ubiquitin, undergo localized degeneration, and are scavenged by glial cells. In contrast, the mature Kenyon cells that comprise the gamma lobe express detectable ubiquitin throughout each developmental episode. This pattern of ubiquitin expression suggests that the gamma lobe circuitry undergoes continuous modification independent of development.

Animals↗
PubMed1976

Formation of cockroach interganglionic connectives: an in vitro analysis.

Individual ganglia of the cockroach embryo (Periplaneta americana) were explanted on clean glass coverslips immersed in a chemically defined liquid medium and incubated for periods up to eight weeks. Substantial, straight interganglionic connections were formed between: (1) rows of ganglia arranged in the normal in vivo configuration; (2) rows of ganglia placed in abnormal orders; (3) rows of ganglia which never form connections in vivo because they occur singly in the embryo; and (4) rows of ganglia in natural sequences but which have had their rostro-caudal axes rotated 90 degrees in relation to the line of the row. Therefore fascicles and interganglionic connectives were formed without regard to normal in vivo relationships. Daily observations with a Nomarski microscope indicated that several processes are involved in connective formation. (1) Initial outgrowth is in a random, radial pattern. (2) Intersecting fibers from adjacent ganglia are deflected toward each others' perikarya. (3) Initially bowed fiber connectives are straightened, perhaps by increases in fiber tension or by fiber shortening which may be brought about by neuronal or extraneuronal (glial) processes. (4) Outgrowing fibers follow already established fiber pathways. The present results indicate that fiber-fiber and fiber-target interactions play a significant role in the formation of interganglionic connectives. In this system, the spatial relationships between ganglia determine the patterns and varieties of permissible neuronal connections. Thus, major, straight nerve trunks may be formed between adjacent ganglia which are growing out fibers on a glass surface submerged in a liquid medium which offers minimal orientation cues and provides a growth substrate vastly different and simpler than that encountered by outgrowing fibers in vivo.

Animals↗
PubMed1979

The specificity of innervation of regenerating motor neurons in the cockroach.

The pattern of neuromuscular connections of cockroach motor neurons regenerating to an intact mesothoracic leg after crushing nerve 5 has been investigated. While a number of anatomically identified motor neurons reestablished functional connections with their correct target muscles, some of the same and other motor neurons were found to functionally innervate inappropriate muscles. Muscles, and in some cases individual fibres, were often simultaneously innervated by both correct and foreign motor neurons. The frequency of innervation errors decreased with increasing post-operative times, although some errors persisted to the latest stages examined (227 days).

Animals↗
PubMed1981

Morphology of the giant interneurons and cercal nerve projections of the American cockroach.

We have investigated the morphology of the giant interneurons (GIs) and the main sensory projections to these interneurons in the American cockroach. These neurons are thought to mediate the animal's escape behavior. We describe here the dendritic branching pattern of each of the 14 GIs (7 bilateral pairs) in the terminal ganglion, the pattern of projection of the cercal sensory nerve, and the overlap of the cercal projections with the dendrites of the GIs. Visualization of the GIs and cercal nerve projection was accomplished by single cell injection and axonal backfilling with cobalt. Comparisons of the same identified GI in different animals show the position of the soma and the locations and orientations of the major processes are characteristic for each GI. The axons of the cercal nerve project to a well-defined ipsilateral region of the terminal ganglion. After entering the terminal ganglion, the cercal afferents split into lateral and medial tracts. The projections of the lateral cercal tract overlap extensively with the dendritic fields of the GIs. In contrast, the medial tract does not overlap the dendritic fields of the GIs in the posterior portion of the ganglion and shows only a small degree of overlap in the anterior portion. Correlations between physiological properties of the GIs and cercal afferents are discussed in relation to our anatomical findings.

Afferent Pathways↗
PubMed1982

Neuropeptide proctolin (H-Arg-Try-Leu-Pro-Thr-OH): immunocytochemical mapping of neurons in the central nervous system of the cockroach.

Proctolinlike immunoreactivity was mapped in the central nervous system of the cockroach, Periplaneta americana, by using whole-ganglion immunoreacted preparations. The procedure for this immunohistochemical staining of whole-mounts is described. Immunoreactivity was confined to neuronal cell bodies and and processes. These were found in all ganglia of the CNS. The cells varied in the consistency and intensity of their staining. The occurrence and variability of staining is described in detail. Cell bodies were found in the dorsal, ventral, and lateral regions of the ganglia. The highest number of cell bodies was found in the terminal ganglion and the lowest number in the cerebral ganglion. Those in the cerebral ganglion occurred mainly in the tritocerebral lobes. The distribution of immunoreactive cell bodies correlated with results previously obtained by radioimmunoassay. Immunoreactive processes were detected in all interganglionic connectives and many ganglionic nerve roots. Dense ramifications of immunoreactive processes and variocosities were detected in many of the ganglia. The widespread presence of immunoreactivity suggests that proctolin has diverse central and peripheral functions. The mapping immunoreactive neuronal somata provides a valuable step in the identification of putative proctolin-containing neurons suitable for further biochemical, anatomical, and physiological analysis.

Animals↗
PubMed1982

Identified motor neurons in the cockroach exhibit stereotypic peripheral branching and nerve terminal structure.

The peripheral branching of the identified motor neurons Df and Ds which innervate the coxal depressor muscles in the legs of cockroaches was examined using the anterograde transport of a cobaltic-lysine complex and the retrograde transport of cobaltous ion or horseradish peroxidase. In all animals the axons of these motor neurons leave the ganglion in the dorsal portion of nerve root 5 and undergo a 180 degree spiral that brings them directly to the first branch. They leave the nerve root at this branch to innervate the muscles. The pattern of branching of Df within muscles 178 and 179 was highly stereotyped with little variability being observed among identical and homologous muscles within the same and different individuals. The great extent of the neuronal branching within the muscles suggests that a relatively large percentage of the surface area of the muscle fibers is involved in intercellular interactions with the motor neurons. The two motor neurons could also be distinguished by the structure of their axon terminals. Those of Df were beaded and extended either parallel or perpendicular to the muscle fiber. those of Ds were thinner, wavy, less regular, and appeared with a greater frequency. Identified neurons may therefore be further characterized by their stereotypical peripheral branching and axon terminal structure.

Animals↗
PubMed1984

Distribution of serotonin-containing neurons and their pathways in the supraoesophageal ganglion of the cockroach Periplaneta americana (L.) as revealed by immunocytochemistry.

The distribution of serotonin (5-HT)-containing neurons in the supraoesophageal (cerebral) ganglion of the cockroach Periplaneta americana was studied using immunocytochemistry and the formaldehyde histofluorescence method ( Klemm , '83). In this material immunocytochemistry was more sensitive than the formaldehyde histofluorescence procedure. A relatively small number of 5-HT-immunoreactive cell bodies (220-280) were found. For the first time, their processes could be followed. They highly arborize and innervate many brain regions. Three patterns of monoamine innervation have been demonstrated: (1) 5-HT and catecholamine fibres ( Klemm , '83) occurring in the same region (e.g., outer lateral protocerebral neuropil, stratum caudale , mushroom body, fan-shaped body, olfactory lobe), but having certain differences with respect to the organization of their projection fields; (2) 5-HT fibres innervating a region lacking catecholamine-containing fibres (pons); and (3) catecholamine neurons innervating a region lacking 5-HT fibres (ellipsoid body). In the mushroom body only the extrinsic neurons contain 5-HT immunoreactivity. They form a commissural fibre system linking the left- and right-hand mushroom bodies and other brain regions. The pons is part of a 5-HT-neuron fibre system innervating many areas including the mushroom bodies. The present study demonstrates novel, complex, and widely distributed connections within the insect brain.

Animals↗
PubMed1986

Monoclonal antibodies to the cockroach nervous system.

In the cockroach nervous system individual motor neurons may be identified with respect to their position in the thoracic ganglia and to the muscles they innervate. When their axons are cut they have the ability to regrow such that when regeneration is completed they have specifically reinnervated their normal target muscles. This suggests the existence of a specific intercellular recognition process between motor neurons and muscles, and that neurons innervating different muscles are biochemically distinct from one another. The goal of this study was to use hybridoma techniques to obtain monoclonal antibodies that bind to some motor neurons and not others. Mice were injected with whole nerve cord and hybridoma supernatants were screened immunohistochemically on sections of ganglion and leg muscle. The monoclonal antibodies were categorized according to four types of specificity: tissue, regional, cell-type, and neuron-subset specificities. Antibodies expressing neuron-subset specificity were obtained very rarely. The probability of their occurrence could be increased by treating the mice with immunosuppressant drugs after initial administration of immunogen or by fixing the immunogen with paraformaldehyde in a manner similar to that of the tissue sections used in the screening process. Two of the neuron-subset specific monoclonal antibodies (MAbs) are of particular interest with respect to the goals of this study. They bind to axon terminals in the muscles of some neurons and not others. They do not bind to neuronal cell bodies in the ganglion, which makes identification of the neurons difficult. However, from the known innervation pattern of the coxal depressor muscles it appears that one of these MAbs selectively binds to axon terminals from either the inhibitory motor neurons or the dorsal unpaired median cells. Other antibodies of interest bind selectively to the synapse-rich neuropile in the ganglia or to peripheral parts of the nervous system like the nerve roots.

Animals↗
PubMed1987

Anatomy and electrophysiology of neurons terminating in the corpora allata of the cockroach Diploptera punctata.

Intracellular recording and dye injection were used to study the structure and electrophysiological properties of individual neurons that project to the corpora allata of the cockroach, Diploptera punctata. Neurons in the pars intercerebralis generate long-duration, tetrodotoxin-sensitive action potentials. Dye injection revealed two cell types. One type extends axons to the contralateral nervi corporis cardiaci I, some of which innervate the corpora allata, and another type extends a major axon down each of the circumoesophageal connectives. Neurons in the pars lateralis also generate long-duration action potentials. These neurons extend axons to the ipsilateral nervi corporis cardiaci II, which continue on to terminate in the corpora cardiaca and the corpora allata. Small groups of all the above neuronal types are dye and electrically coupled. Penetration and dye injection into nerve terminals in the corpora allata and corpora cardiaca confirmed the innervation of the corpora allata by neurons located in the pars intercerebralis and pars lateralis and revealed a third class of neurons that have terminals in the corpora allata: intrinsic neurons of the corpora cardiaca.

Action Potentials↗
PubMed1988

Postembryonic development of the dorsal ocellus of the American cockroach.

Postembryonic development of the dorsal ocellus of the cockroach Periplaneta americana was examined. Small ocelli (20 micron in diameter) with less than 100 cells in the newly hatched nymph become adult ocelli (more than 500 micron in diameter) with more than 10,000 retinular cells, through ten to 11 nymphal stages. Thus, sequential steps of morphogenesis of rhabdomeres from loose interdigitations of apposed cell membranes to regularly arranged microvilli can be seen in the nymphal ocelli. Some retinular axons appear to extend into the brain as a bundle in the first-instar nymph. Retinular axons differentiated later also extend toward the brain, but there is no evidence that they enter it. The ultrastructure of the ocellus and component analysis of the ocellar electroretinogram suggest that functional connections between retinular axons and ocellar interneurons do not occur in the ocellar capsule until mid (the fifth or sixth)-instar nymphs. The ocellar diameter increases linearly with increase in body length during the nymphal stages, but it increases by a factor of 1.5-2.0 during the final molting: the body length of the adult is almost the same as that of the last-instar nymph. These data suggest that the function of the dorsal ocelli may be closely related to specific adult behavior such as flight.

Animals↗
PubMed1992

A morphological correlate of target recognition by regenerating motor axons in the cockroach.

A specific cell recognition process during regeneration of severed axons of identified cockroach motor neurons eventually leads to the reformation of the original innervation pattern of target muscles in the leg. This occurs even though, at early times after nerve crush, the multiple branches of each regenerating axon grow into both appropriate and inappropriate muscles. In this study, we sought to examine whether there are any structural differences between regenerating axon branches in appropriate and inappropriate muscles that could lead to an understanding of why only those in inappropriate muscles are eliminated. A neuron subset-specific monoclonal antibody, NSS-2A, which labels the inhibitory motor neurons, was used to make their axon branches visible at various times after nerve crush. In inappropriate muscles, these axons grow primarily parallel to the muscle fibers and are later eliminated. In the appropriate muscles, these axon branches initially also grow parallel to the muscle fibers, but subsequently grow many interstitial collaterals. The formation of the collateral branches is a morphological correlate of the specific interaction of a neuron with its appropriate muscle. The simultaneous occurrence of axonal elimination and collateral sprouting supports the idea that the two processes are causally related, as suggested by the sibling neurite bias hypothesis.

Animals↗
PubMed1992

A new specific antibody reveals octopamine-like immunoreactivity in cockroach ventral nerve cord.

An antiserum was raised in rabbits immunized with octopamine conjugated to thyroglobulin. The specificity of this antiserum for octopamine is shown by dot blot immunoassay analysis. The antiserum does not crossreact with dopamine, noradrenaline, and serotonin, but slight crossreactivity with the amine tyramine at high concentrations was observed. The tyramine crossreactivity could be eliminated by preabsorption with a tyramine-glutaraldehyde-BSA conjugate. Using this antiserum, we describe the topographical distribution of octopamine-immunoreactive (ir) neuronal elements in wholemounts and paraffin sections of the ventral nerve cord of the American cockroach. The pattern of octopamine immunostaining is completely different from that obtained with an antidopamine serum, and can be blocked by preabsorbing the antioctopamine serum with BSA-conjugated octopamine. Cell bodies and dendritic processes of putatively octopaminergic dorsal (DUM) and ventral (VUM) unpaired median neurons were clearly octopamine-ir in all ganglia examined. The numbers of stained DUM somata in the mesothoracic, metathoracic, and terminal ganglion of females correspond to those of peripherally projecting DUM cells revealed previously by retrograde tracing (Gregory, Philos Trans R Soc Lond [Biol] 306:191, 1984; Tanaka and Washio, Comp Biochem Physiol 91A:37, 1988; Stoya et al., Zool Jb Physiol 93:75, 1989). In addition, various, previously unknown, paired cells with octopamine-like immunoreactivity were found in all ventral ganglia except abdominal ganglia 3-6. Some of these probably project intersegmentally.

Animals↗
PubMed1992

Neurons in the cockroach nervous system reacting with antisera to the neuropeptide leucokinin I.

Antisera were raised against the myotropic neuropeptide leucokinin I, originally isolated from head extracts of the cockroach Leucophaea maderae. Processes of leucokinin I immunoreactive (LKIR) neurons were distributed throughout the nervous system, but immunoreactive cell bodies were not found in all neuromeres. In the brain, about 160 LKIR cell bodies were distributed in the protocerebrum and optic lobes (no LKIR cell bodies were found in the deuto- and tritocerebrum). In the ventral ganglia, LKIR cell bodies were seen distributed as follows: eight (weakly immunoreactive) in the subesophageal ganglion; about six larger and bilateral clusters of 5 smaller in each of the three thoracic ganglia, and in each of the abdominal ganglia, two pairs of strongly immunoreactive cell bodies were resolved. Many of the LKIR neurons could be described in detail. In the optic lobes, immunoreactive neurons innervate the medulla and accessory medulla. In the brain, three pairs of bilateral LKIR neurons supply branches to distinct sets of nonglomerular neuropil, and two pairs of descending neurons connect the posterior protocerebrum to the antennal lobes and all the ventral ganglia. Other brain neurons innervate the central body, tritocerebrum, and nonglomerular neuropil in protocerebrum. LKIR neurons of the median and lateral neurosecretory cell groups send axons to the corpora cardiaca, frontal ganglion, and tritocerebrum. In the muscle layer of the foregut (crop), bi- and multipolar LKIR neurons with axons running to the retrocerebral complex were resolved. The LKIR neurons in the abdominal ganglia form efferent axons supplying the lateral cardiac nerves, spiracles, and the segmental perivisceral organs. The distribution of immunoreactivity indicates roles for leucokinins as neuromodulators or neurotransmitters in central interneurons arborizing in different portions of the brain, visual system, and ventral ganglia. Also, a function in circuits regulating feeding can be presumed. Furthermore, a role in regulation of heart and possibly respiration can be suggested, and probably leucokinins are released from corpora cardiaca as neurohormones. Leucokinins were isolated by their myotropic action on the Leucophaea hindgut, but no innervation of this portion of the gut could be demonstrated. The distribution of leucokinin immunoreactivity was compared to immunolabeling with antisera against vertebrate tachykinins and lysine vasopressin.

Animals↗