Biomedical subjects

B Mulloney

Publications and source records attributed to B Mulloney.

At least 19 recordsLinked to original sources

PubMed1995

Coordination in the crayfish swimmeret system: differential excitation causes changes in intersegmental phase.

1. Gradients of excitation in the swimmeret system were created by applying either pilocarpine or carbachol to selected ganglia in isolated abdominal nerve cords. The state of the system was monitored in each segment with extracellular electrodes on nerves that innervated swimmerets. In preparations that were quiescent before drugs were applied, these cholinergic agonists elicited well-coordinated swimmeret motor patterns from the entire system, including ganglia that were not directly treated with pilocarpine or carbachol. 2. The periods of these patterns depended on the number of ganglia that were directly excited. As this number increased, period decreased. When the same numbers of ganglia were excited by direct application of a drug, the mean period of the swimmeret activity elicited by pilocarpine was greater than that elicited by carbachol. 3. Selective excitation of anterior or posterior ganglia caused significant changes in intersegmental phase at the boundary between excited and nonexcited regions of the nerve cord. When only anterior ganglia were excited directly, the phases of their power-stroke activity relative to the most posterior ganglion were advanced. When only posterior ganglia were excited directly, the phases of power-stroke activity in more anterior ganglia were retarded. Neither pilocarpine nor carbachol caused a complete reversal of the normal phase relations of the swimmeret motor patterns. 4. These results are consistent with an asymmetric-coupling model of the intersegmental coordinating circuit of the swimmeret system but contradict an alternative excitability-gradient model.

Abdomen↗

PubMed1994

Acetylcholinesterase activity in neurons of crayfish abdominal ganglia.

Acetylcholine is known to be a neurotransmitter in crustacean central nervous systems, but the numbers and distribution of cholinergic neurons in the segmental ganglia have not been described. To begin a census of cholinergic neurons in these ganglia, we used a histochemical assay for acetylcholinesterase to map neurons that contained this enzyme in the six abdominal ganglia of crayfish. In each abdominal ganglion, about 47 cell bodies were stained. The distributions of these stained cells in individual ganglia were similar, and the numbers were not significantly different. None of these stained cell bodies could be identified from their structures or locations as previously identified motor neurons or sensory neurons with central cell bodies. The process of one unpaired midline neuron that occurred only in the first three abdominal ganglia divided to send a pair of axons anteriorly into both halves of the connective. The central projections of afferent axons from many peripheral sensory neurons stained clearly as they entered each ganglion. Terminals of these axons were heavily stained in the horseshoe neuropil and the lateral neuropils. We labeled both gamma-aminobutyric acid (GABA) and acetylcholinesterase in individual ganglia. Only a few neurons in each ganglion were double-labeled. The unpaired midline neurons in the three anterior ganglia that stained for acetylcholinesterase did not show GABA-like immunoreactivity, but cells with similar shapes did label with the GABA antiserum. Acetylcholinesterase is not a definitive marker of cholinergic neurons, but its presence is often associated with the cholinergic phenotype. These stained cells should be considered as putative cholinergic neurons.

Abdomen↗

PubMed1994

Proctolin and excitation of the crayfish swimmeret system.

The ventral nerve cord of crayfish contains axons of five pairs of excitatory interneurons, each of which can activate the swimmeret system. Perfusion of the ventral nerve cord with the neuropeptide proctolin also activates the swimmeret system. The experiments reported here were conducted to test the hypothesis that one or more of these excitatory interneurons uses proctolin as a transmitter. Each of the five excitatory axons was located and stimulated separately in an individual crayfish, and similar motor activity was elicited by stimulating each of them. Quantitative comparison of spontaneous swimmeret motor patterns with activity caused by stimulating one of these excitatory axons, EC, or by perfusing with proctolin solutions showed that the motor patterns produced under these three conditions were not significantly different (P > 0.05). By using a new, affinity-purified proctolin antiserum, we labeled axons in the connective tissue between the last thoracic and first abdominal ganglion and compared the positions of labeled axons with the previously described positions of the excitatory axons. About 0.3% of the axons in these connective tissues showed proctolin-like immunoreactivity, but heavily labeled pairs of axons did occur bilaterally in the regions of excitatory swimmeret axons. The projections of these labeled axons into the abdominal ganglia were traced in serial plastic sections. Labeled processes were abundant in the lateral neuropils, the loci of the swimmeret pattern-generating circuitry. From this evidence, we propose that three of these excitatory swimmeret interneurons use proctolin as a transmitter, but that a fourth does not. The evidence for the fifth axon is ambiguous.

Amino Acid Sequence↗

PubMed1994

State-dependent responses of two motor systems in the crayfish, Pacifastacus leniusculus.

The expression of both swimmeret and postural motor patterns in crayfish (Pacifastacus leniusculus) were affected by stimulation of a second root of a thoracic ganglion. The response of the swimmeret system depended on the state of the postural system. In most cases, the response of the swimmeret system outlasted the stimulus. Stimulation of a thoracic second root also elicited coordinated responses from the postural system, that outlasted the stimulus. In different preparations, either the flexor excitor motor neurones or the extensor excitor motor neurones were excited by this stimulation. In every case, excitation of one set of motor neurones was accompanied by inhibition of that group's functional antagonists. This stimulation seemed to coordinate the activity of both systems; when stimulation inhibited the flexor motor neurones, then the extensor motor neurones and the swimmeret system were excited. When stimulation excited the flexor motor neurones, then the extensor motor neurones and the swimmeret system were inhibited. Two classes of interneurones that responded to stimulation of a thoracic second root were encountered in the first abdominal ganglion. These interneurones could be the pathway that coordinates the response of the postural and swimmeret systems to stimulation of a thoracic second root.

Abdomen↗

PubMed1993

Cholinergic modulation of the swimmeret motor system in crayfish.

1. The muscarinic agonist pilocarpine induced the swimmeret motor pattern in resting isolated preparations of the crayfish abdominal nerve cord and modulated the burst frequency in a dose-dependent manner. 2. Nicotine did not elicit rhythmic activity in resting isolated preparations but increased the burst frequency in active preparations. Nicotine produced higher burst frequencies than pilocarpine. 3. The acetylcholine (ACh) analogue carbachol combined the effects of pilocarpine and nicotine. It activated isolated resting preparations and increased the burst frequency as effectively as nicotine. The ACh-esterase inhibitor eserine also increased the burst frequency in active preparations. 4. Neither muscarinic nor nicotinic antagonists disrupted the proctolin-induced motor pattern, suggesting that proctolin and cholinergic agonists affect two different pathways for the activation of the swimmeret system. 5. We conclude that cholinergic interneurons participate in initiation of the swimmeret motor pattern and can modulate its burst frequency.

Acetylcholine↗

PubMed1993

A separate local pattern-generating circuit controls the movements of each swimmeret in crayfish.

1. Within an abdominal segment, the motor output from the segmental ganglion to the swimmerets consists of coordinated bursts of impulses in the separate pools of motor neurons innervating the left and right limbs. This coordinated motor pattern features alternating (out-of-phase) bursts of impulses in the power-stroke (PS) and return-stroke (RS) motor axons that innervate each swimmeret. PS bursts on both sides of each segment occur simultaneously (in-phase), and so RS bursts on both sides are also in-phase. 2. With all intersegmental connections interrupted, isolated abdominal ganglia were able to sustain the normal swimmeret motor pattern of alternating PS/RS activity that was bilaterally in-phase. 3. After an isolated ganglion was surgically bisected down the midline, the isolated hemiganglia that resulted could produce stable, coordinated alternation of PS and RS bursts. 4. The neuropeptide proctolin could induce rhythmic oscillations of membrane potential in swimmeret neurons when spiking was blocked by tetrodotoxin (TTX). For neurons within the same hemiganglion, these oscillations retained the same phase relations they displayed in controls, but the oscillations of neurons in different hemiganglia became uncoordinated. 5. Synaptic transmission between swimmeret neurons in the same hemiganglion persisted in the presence of TTX. Swimmeret interneurons that could activate the pattern-generating circuitry under control conditions could induce membrane-potential oscillations in swimmeret neurons of the same hemiganglion when TTX was present. 6. We conclude that a separate hemisegmental pattern-generating circuit controls the rhythmic PS and RS movements of each swimmeret. Each circuit is located in the same hemiganglion as the population of motor neurons that innervates the local swimmeret. Graded transmission is sufficient to coordinate the timing of oscillatory activity within the hemisegmental circuitry. These hemisegmental circuits are coupled by intersegmental and bilateral coordinating pathways that are dependent on sodium action potentials for their operation.

Animals↗

PubMed1991

Neurons with histaminelike immunoreactivity in the segmental and stomatogastric nervous systems of the crayfish Pacifastacus leniusculus and the lobster Homarus americanus.

We used a polyclonal antiserum against histamine to map histaminelike immunoreactivity (HLI) in whole mounts of the segmental ganglia and stomatogastric ganglion of crayfish and lobster. Carbodiimide fixation permitted both HRP-conjugated and FITC-conjugated secondary antibodies to be used effectively to visualize HLI in these whole mounts. Two interneurons that send axons through the inferior ventricular nerve (ivn) and the stomatogastric nerve to the stomatogastric ganglion had strong HLI, both in crayfish and in lobster. These ivn interneurons were known from other evidence to be histaminergic. The neuropil of the stomatogastric ganglion in both crayfish and lobster contained brightly labeled terminals of axons that entered the ganglion from the stomatogastric nerve. No neuronal cell bodies in this ganglion had HLI. Each segmental ganglion contained at least one pair of neurons with HLI. Some neurons in the subesophageal ganglion and in each thoracic ganglion labeled very brightly. Axons of projection interneurons with strong HLI occurred in the dorsal lateral tracts of each segmental ganglion, and sent branches to the lateral neurophils and tract neurophils of each ganglion. All the labeled neurons were interneurons; no HLI was observed in peripheral nerves.

Animals↗

PubMed1991

Red pigment concentrating hormone is a modulator of the crayfish swimmeret system.

The crustacean red pigment concentrating hormone (RPCH) has been localized in neurons of the crayfish abdominal nerve cord and modulates the crayfish swimmeret rhythm. An antibody to RPCH labels a small set of cell bodies and axons in each abdominal ganglion. Physiological experiments in which RPCH was perfused into the ganglia of isolated nerve cords showed that RPCH modulated the swimmeret rhythm. In nerve cords that were spontaneously producing the swimmeret rhythm, RPCH lengthened both the period and the duration of bursts of action potentials, but did not alter the phase relationships between bursts in different segments. RPCH did not initiate the swimmeret rhythm in preparations that showed intermittent or no bursting activity. We believe that RPCH is released as a neurotransmitter in the lateral neuropil, where it exerts its effects on the local swimmeret circuits.

Animals↗

PubMed1990

GABA-ergic neurons in the crayfish nervous system: an immunocytochemical census of the segmental ganglia and stomatogastric system.

We used an antiserum directed against gamma-aminobutyric acid (GABA) fixed with glutaraldehyde (Hoskins et al., Cell Tissue Res. 244:243-252, '86) to label neurons with GABA-like immunoreactivity (GLI) in wholemounts of the stomatogastric ganglion and each segmental ganglion of crayfish, except the brain. Each abdominal ganglion had an average of 63 labeled neurons, or 10% of all their neurons. Each peripheral nerve of each abdominal ganglion except the last contained labeled axons. Within each segment, the first peripheral nerve, N1, had five axons; the second peripheral nerve, N2, had at most four; and the third peripheral nerve, N3, had two. In the last ganglion, N2 had one labeled axon, N3 had two and N6 had two; the other nerves contained no labeled axons. A tabulation of the identified inhibitory neurons in the abdominal ganglia revealed that 40% of these GABA-ergic neurons have been identified. The subesophageal ganglion had many labeled neurons in clusters that formed a repeating pattern; it also had labeled neurons near its dorsal midline. The thoracic ganglia contained more labeled neurons than did the abdominals, but their patterns of labeling were similar. The commissural ganglia contained three clusters of labeled neurons and sent labeled axons to the esophageal ganglion. The esophageal ganglion contained four labeled neurons and many labeled axons. The stomatogastric ganglion contained labeled axon terminals but not labeled neurons.

Animals↗

PubMed1988

The central projections of the stretch receptor neurons of crayfish: structure, variation, and postembryonic growth.

Each stretch receptor neuron (SR) of the crayfish abdomen projects from its peripherally located soma an axon that enters the CNS through the second nerve (N2) of its segmental ganglion. Co2+ backfills of N2 revealed that this axon bifurcates, sending one branch to the brain and the other to the terminal abdominal ganglion. Each axon makes many short lateral branches as it traverses each of the other segmental ganglia, and has 2 major arborizations--one in the tritocerebrum of the brain and the other in the terminal abdominal ganglion, G6. These projections to G6 are described in detail. Individual SR axons from different segments were impaled in G6 with a Lucifer yellow-filled microelectrode, identified physiologically, and then filled to discover their structures. In a sample of more than 50 SR neurons, no systematic structural differences between the projections of phasic SR neurons and tonic SR neurons were observed. Three kinds of structural variability occurred in these projections to G6: unusual primary axon shape, long primary axon, and large terminal varicosities. Comparisons of SR neurons that originated in different segments revealed a segmental gradient in the probability of each structural variant. SR neurons from more anterior segments were more likely to show these structural variations. Comparisons of these variations in crayfish of different sizes showed that unusual primary axon shape and long primary axons were more probable in large crayfish than in small ones. Therefore, these variations are probably the products of postembryonic growth of the nervous system.

Animals↗

PubMed1988

The central projections of the stretch receptor neurons of crayfish: segmental gradients of synaptic probability and strength.

The 20 stretch receptor neurons (SRs) of the crayfish abdomen send axons into the CNS that then project both to the brain and to the last abdominal ganglion, G6 (Bastiani and Mulloney, 1988). In G6, we recorded intracellularly from different kinds of neurons postsynaptic to SR axons. In a sample of 100 postsynaptic neurons, 59 synapsed with both SR1 and SR2 axons, 19 synapsed only with SR1 axons, and 22 synapsed only with SR2 axons. Most monosynaptic connections in G6 were excitatory and behaved like typical chemical synapses. The EPSPs showed moderate facilitation but could be depressed about 50% by protracted stimulation at 20 Hz or more. In individual postsynaptic neurons, comparisons of synapses made by SRs that originated from different abdominal segments and from each side of the abdomen revealed gradients of probability of synaptic connection and of relative sizes of EPSPs; SRs originating in anterior segments were less likely to synapse with most postsynaptic neurons than were SRs originating in posterior segments, and the EPSPs caused by these anterior SRs tended to be smaller. Similarly, SRs contralateral to the postsynaptic neuron were less likely to make a connection, and the EPSPs they caused tended to be smaller than those caused by ipsilateral SRs. Some local interneurons in G6 had reversed anterior-posterior gradients in EPSP amplitude. Calculations of shape indices for PSPs from SRs originating in different segments and measurements of the maximum shunting by preceding PSPs from other SR axons indicated that neither electrotonic decrement in the postsynaptic neurons nor shunting could account fully for the observed gradients in PSP strength.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1987

Functional organization of crayfish abdominal ganglia: II. Sensory afferents and extensor motor neurons.

Abdominal ganglia of crayfish contain identifiable neuropils, commissures, longitudinal tracts, and vertical tracts. To determine the functional significance of this ganglionic framework, we backfilled the following types of neurons with cobalt chloride: sensory hair afferents, slow and fast extensor motor neurons, the segmental stretch receptor neurons, and their inhibitory accessory cells. After the cobalt ions were precipitated and intensified, we studied the central projections of the filled neurons within the ganglionic structures. All of the axons of these neurons exit or enter each of the first five abdominal ganglia through the second pair of nerves. Our description of the central projections of the hair afferents is the first in the literature. These afferents innervate the large ventral horseshoe neuropil (HN) in the core of each ganglion. This neuropil is homologous to the insect ventral association centers, which also process sensory information. Furthermore, we discovered that some of the crayfish afferents innervate glomeruli within the HN. The slow and fast extensor motor neurons, the stretch receptor neurons, and the accessory cells branch mostly in the dorsal part of the ganglion. We reinterpret previous identifications of the extensor neurons that were based largely on soma position. Together with our previous descriptions of the flexor motor neurons, these results allow us to relate both rapid tail-flips and slower postural movements to the structure of the segmental ganglia.

Afferent Pathways↗

PubMed1987

The PD programs: a method for the quantitative description of motor patterns.

We describe a family of 6 computer programs that measure, analyse and create graphic displays of complex motor patterns. The programs create lists of times at which successive bursts of impulses in different nerves started and stopped, and use these lists to calculate the periods and durations of these bursts and to calculate their phases relative to some specified frame of reference. When calculating phases, the programs take into account missing bursts or extra bursts in each reference interval. Individual programs then calculate descriptive statistics for these parameters, select lists of paired data for plotting and regression analysis, and prepare files for graphical display of statistics as boxplots. A final-program plots these files on a digital plotter. These programs are available for non-commercial use.

Action Potentials↗

PubMed1987

Synaptic integration in excitatory and inhibitory crayfish motoneurons.

The passive integrative properties of two crayfish abdominal motoneurons, the fast flexor inhibitor (FI) and a posterior, ipsilateral fast flexor excitor (FE), were studied electrophysiologically and through simulations with multicompartment models of their electrotonic structures. Responses of the models to simulated giant neuron input were quite similar to the motoneurons' responses to giant neuron stimulation, which suggests that differences in the electrotonic structures and the sites of synaptic input to the two cells can account in large part for differences in their responses to a common input. A full action potential created in the initial axon compartment of the FI model produced attenuated potentials in the adjacent integrating segment compartment and contralateral soma compartment. These potentials are similar in amplitude and time course to attenuated antidromic action potentials recorded in the corresponding regions of the FI neuron. A location of the spike initiation zone of the FI at the initial axon segment is consistent with this result. The responses of FI to ipsi- and contralateral inputs are different. Shock of a single abdominal second root produced a larger, faster rising excitatory postsynaptic potential in the ipsilateral FI soma than in the contralateral soma. Second root shock also caused the contralateral FI to produce an action potential either alone or before the ipsilateral FI neuron. Responses of the FI model to ipsilateral and contralateral inputs differ in the same way as the cell's responses. Inputs to the FI model that are ipsilateral to the soma compartment produce larger responses there than do contralateral inputs. Conversely, those contralateral inputs produce larger responses in the initial axon compartment than do ipsilateral inputs. This difference results from the long integrating segment that connects the soma compartment to the initial axon compartment. These results can account for the FI responses to lateralized inputs. Unlike the responses of FIs, the soma responses of contralaterally homologous FEs to ipsilateral and contralateral second root shocks were similar in waveform and amplitude, with the ipsilateral root producing the larger response. This result is consistent with theoretical results from the FE model simulations. We conclude that a smaller size, larger input resistance and shorter membrane time constant allow the FE to respond to giant neuron input before the FI, and so help to achieve the proper timing of flexor contraction and relaxation during a tailflip.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

PubMed1987

Modulation of the crayfish swimmeret rhythm by octopamine and the neuropeptide proctolin.

1. The swimmeret system can be excited by perfusing the neuropeptide proctolin through the isolated ventral nerve cord of the crayfish. Previously silent preparations begin to generate a characteristic motor pattern, the swimmeret rhythm, in the nerves that innervate the swimmerets. The response to proctolin is dose dependent and reversible. The threshold concentration of proctolin perfused through the ventral artery is approximately 10(-8) M. The EC50 is 1.6 X 10(-6) M. 2. Proctolin-induced motor patterns have periods and phases similar to those of spontaneously generated motor patterns. The durations of the bursts of impulses in power-stroke motor neurons generated in the presence of proctolin are, however, significantly longer than those that occur during spontaneous activity. 3. DL-Octopamine inhibits the swimmeret system, both when the system is spontaneously active and when it has been excited by proctolin. The inhibition by octopamine is dose dependent and reversible. The threshold for inhibition is approximately 10(-6) M, and the EC50 is approximately 5 X 10(-5) M. 4. Octopamine's effect is mimicked by its agonists, synephrine and norepinephrine. Synephrine has a lower threshold concentration than does octopamine, but norepinephrine is much less effective than octopamine. 5. Octopamine's inhibition is partially blocked by an antagonist, phentolamine. 6. Phentolamine also blocks inhibition of the swimmeret system by inhibitory command interneurons. This block is dose dependent and can be partially overcome by stimulating the command interneurons at higher frequencies. 7. Perfusion with 11 other suspected crustacean neurotransmitters and transmitter analogues did not similarly excite or inhibit the swimmeret system, so we suggest that proctolin and octopamine are transmitters used by the neurons that normally control expression of the swimmeret rhythm.

Animals↗

PubMed1986

Functional organization of crayfish abdominal ganglia: I. The flexor systems.

For insect ganglia, Altman (Advances in Physiological Science, Vol. 23. Neurobiology of Invertebrates. New York: Pergamon Press, pp. 537-555, '81) proposed that individual neuropils control different motor activities. A corollary of this hypothesis is that motor neurons involved in many behavioral functions should branch in more neuropils than those active in fewer behaviors. In crayfish, the abdominal fast-flexor muscles are active only during the generation of the powerstroke for tailflips, whereas the slow-flexor muscles are involved in the maintenance of body posture. The slow flexors are thus active in many of the crayfish's behavioral activities. To test the generality of Altman's idea, we filled groups of crayfish fast-flexor and slow-flexor motor neurons with cobalt chloride and described their shapes with respect to the ganglionic structures through which they pass. Individual fast flexors were also filled intracellularly with HRP. Ganglia containing well-filled neurons were osmicated, embedded in plastic, and sectioned. Unstained sections were examined by light microscopy and pertinent sections were photographed. We found that the paths of the larger neurites were invariant, that the dendritic domains of fast and slow motor neurons occupied distinctive sets of neuropils, and that dendrites of slow motor neurons branched in more ganglionic structures than did those of fast motor neurons. These results are consistent with Altman's hypothesis.

Animals↗

PubMed1986

The osmium-ethyl gallate procedure is superior to silver impregnations for mapping neuronal pathways.

Ganglia processed through the osmium-ethyl gallate procedure (OEG)19 retain more structural integrity than those processed through various silver impregnation methods. However, the OEG method continues to be neglected by most neuroanatomists. Both types of procedures have been used to trace large neuronal tracts, but during silver impregnation the neuropils lose many of their identifying characteristics. We demonstrate here the advantages of the OEG procedure by comparing it with two silver techniques, Rowell's and Holmes's. The OEG method yields consistent and reliable results and is easier to carry out than silver protocols. Most importantly, the better preservation of the neuropils has led to the discovery and study of regional specializations that were previously undetected from silver preparations.

Animals↗

PubMed1985

Nonspiking local interneuron in the motor pattern generator for the crayfish swimmeret.

We describe a type of nonspiking premotor local interneuron (interneuron IA) in the abdominal nervous system of Pacifasticus leniusculus. All of its branches are restricted to one side of the midline. These interneurons are identifiable and occur as bilateral pairs, one neuron on each side of abdominal ganglia 3, 4, and 5. The membrane potential of interneuron IA oscillated in phase with the swimmeret rhythm, a motor pattern generated in each of these ganglia, because the neuron received postsynaptic potentials in phase with the rhythm. Sustained hyperpolarization of an individual interneuron IA initiated generation of the swimmeret rhythm in all the ganglia of a quiescent nervous system. Sustained depolarization stopped the swimmeret rhythm in all the active ganglia of a nervous system that was generating the rhythm. Currents injected into one interneuron reset the rhythm. Comparisons of the shapes of the IA interneurons in different ganglia showed that they are similar to each other and distinct from other local interneurons in these ganglia. Interneuron IA has a large integrative segment and relatively few branches that are largely restricted to the lateral neuropil, to which all other kinds of swimmeret neurons also project. We conclude that this interneuron occurs only once in each hemiganglion in abdominal segments 3, 4, and 5, and that it is identifiable. Furthermore, this interneuron is an essential component of the circuit in each hemiganglion that generates the swimmeret rhythm. The interneuron was dye coupled to a particular identifiable motor neuron and not to any other neurons. The motor neuron was not dye-coupled to any other local interneurons. The ability of this motor neuron to reset the rhythm is attributed to its being electrically coupled to interneuron IA in its ganglion.

Animals↗