Selective phonotaxis to advertisement calls in the gray treefrog Hyla versicolor: behavioral experiments and neurophysiological correlates

1. The significance of particular acoustic properties of advertisement calls for selective phonotaxis by the gray treefrog, Hyla versicolor (= HV), was studied behaviorally and neurophysiologically. Most stimuli were played back at 85 dB SPL, a level typically measured at 1–2 m from a calling male.
2. Females preferred stimuli with conspecific pulse shapes at 20° and 24°C, but not at 16°C. Tests with normal and time-reversed pulses indicated the preferences were not influenced by the minor differences in the long-term spectra of pulses of different shape.
3. Pulse shape and rate had synergistic or antagonistic effects on female preferences depending on whether the values of one or both of these properties in alternative stimuli were typical of those in HV or heterospecific (H. chrysoscelis = HC) calls.
4. More auditory neurons in the torus semicircularis were temporally selective to synthetic calls (90%) than to sinusoidally AM tones and noise (< 70%).
5. Band-pass neurons were tuned to AM rates of 15–60 Hz. Neurons were more likely to be tuned to HV AM rates ( < 40 Hz) when stimuli had pulses with HV rather than HC shapes.
6. Sharp temporal tuning was uncommon and found only in neurons with band-pass or low-pass characteristics.
7. Many neurons differed significantly in response to HV and HC stimulus sets. Maximum spike rate was more often elicited by an HV stimulus (74%) than by an HC stimulus (24%).
8. Differences in spike rates elicited by HV and HC stimuli were attributable to combinations of differences in the rise times and shapes of the pulses.

     

Temporal discharge patterns of tectal and medullary neurons chronically recorded during snapping toward prey in toads Bufo bufo spinosus

In freely moving toads, the temporal discharge patterns of tectal and medullary neurons were observed during prey-catching.

1. Tectal T5.2 and T8.1 neurons displayed a premotor warming up firing that in the former was addressed specifically to prey orienting or snapping and in the latter generally to almost any kind of body movement.
2. The temporal discharge patterns of T5.2 neurons during snapping were different from those during orienting toward prey. Snapping started in the peak phase of warming up; firing was immediately terminated during the snap; thereafter some rebound activity was observed. Orienting started after the premotor warming up in the declining phase whilst the neuron kept on firing during orienting and then settled when the orienting movement was completed.
3. In toads which were not motivated to catch prey — comparabl to immobilized ones — the discharge frequency of T5.2 neurons toward a prey stimulus revealed no such warming up.
4. Because it is known that prey-selective T5.2 neurons are controlled by pretectal inhibitory influences, the following experiment was conducted: during recording a T5.2 neuron a pretectal lesion was applied ipsilaterally to the recording site. After a few seconds, the neuron showed a strong premotor wanning up in response to any kind of moving object, followed by prey-catching.
5. In the medulla oblongata, different H-type neurons of the hypoglossal nucleus displayed specific discharge patterns which resembled the tongue protractor and retractor muscle activities; a third type resembled the activity of the genio/sterno-hyoid muscle, which are suggested to stabilize the hyoid bone during snapping.
6. There were medullary M8-type neurons with properties similar to T8.1.
7. Snapping could be triggered by electrical stimulation of the optic tectum in the representation of the frontal visual field, but not by stimulation in the hypoglossal nucleus or the adjacent medial reticular formation.
8. A concept of a neuronal circuit for the coordination of tongue muscle contractions in response to prey is proposed.

     

Descending stridulatory interneurons in the suboesophageal ganglion of two grasshopper species

1. The activity of interneurons in the suboesophageal ganglion of the acridid grasshoppers Omocestus viridulus (L.) and Chorthippus mollis (Charp.), recorded intracellularly during stridulation, was found to conform to the rhythm of the singing movements. The arborizations of these neurons in this ganglion are largely bilaterally symmetrical; the axon descends contralaterally to the soma and passes at least into the metathoracic ganglion.
2. The anatomical and physiological characteristics of these neurons are similar in the two species and of four types. Three of them exhibit a tonic, spontaneous activity in the resting animal, which is modulated in the stridulatory rhythm as soon as singing begins. The fourth type has no resting activity and discharges only during the song, in a stridulation-specific pattern.
3. By transecting the connectives it was shown that the rhythmic activity of the neurons is not determined by input from the brain, nor is it generated in the suboesophageal ganglion itself. It is based on information about the song pattern that ascends from the thoracic ganglia.

     

Effects of prenatal cocaine exposure in the prefrontal cortex of the rat

This work was undertaken in order to assess the organization of the prelimbic area of the medial prefrontal cortex of rats exposed prenatally to cocaine. Pregnant Wistar rats were assigned to the following groups:

1. Cocaine—60 mg/kg body wt/d sc, from gestational days 8–22; 0131
2. Saline;
3. Pair-fed; and
4. Nonmanipulated.

Male offspring were perfused on postnatal days 14 and 30. Six brains per group and per age were embedded in celloidin to calculate the volumes of the prelimbic area; sections from the other six brains were embedded in resin and processed for electron microscopy. Using semithin sections (2 μm) of layers II–III and V–VI, the following parameters were calculated:

1. The fraction of the neuropil occupied by neurons (V_V);
2. The packing (N_A) density; and
3. The numerical (N_V) density.

Qualitative alterations consisted of dispersed profiles of degenerated neurons and dendrites in the medial prefrontal cortex. No significant differences were found in the gross morphometric parameters when the cocaine group was compared with the other groups. A high interanimal variation was shown in the prelimbic volumes of postnatal day (PND) 14 cocaine-treated rats, and a decrease in volumes was detected at PND30. Although there are some alterations in the main afferent cortical target area for dopaminergic input, its gross morphometric parameters do not seem to be sufficiently affected to account for the behavioral alterations referred to as being dependent on this brain region.     

Encoding of plant odours by receptor neurons in the pine weevil (Hylobius abietis) studied by linked gas chromatography-electrophysiology

Receptor neuron responses to plant volatiles, trapped by head-space procedures, were examined in the pine weevil Hylobius abietis, using gas chromatography linked with electrophysiological recordings from single neurons. Seventy-two receptor neurons were tested 173 times for various plant volatile mixtures, either via a polar or a non-polar column.

1. All responses appeared as increased firing rates which followed the concentration profiles of the GC-eluted compounds.
2. The neurons were classified separately for the two column types in 17 and 19 groups respectively, according to the compounds they responded to. It suggests that the plant odour information is encoded by a large, but limited number of receptor neuron types.
3. Most neurons responded to a limited number of compounds (1–5) and showed a marked best response to one of them, whereas additional responses to several other components which seems to be structurally similar, was recorded for some neurons. It suggests that the plant odour receptor neurons are rather narrowly than broadly tuned, and that each neuron is specialized for receiving information about one or a few related compounds.
4. Most neurons responded to monoterpenes, whereas the other neurons responded to compounds of other categories.
5. Both major and minor plant volatile components activated specifically receptor neurons.

     

Geographical distribution of leaf life span and species diversity of trees simulated by a leaf-longevity model

The leaf life span, leaf habit (evergreenness and deciduousness), and species diversity of trees were simulated by a cost-benefit model of leaf longevity (Kikuzawa 1991), using monthly mean temperature values and their decreasing rate with altitude of 6°C with 1000 m of sites of different latitude and altitude in eastern Asia. Numbers of tree species in tropical regions with different lengths of favorable period for photosynthesis were also simulated. The following results were obtained by the model simulation.

1. In tropical areas, evergreen forests predominate from lowlands to the altitudinal limit of forests.
2. However, leaf longevity is shorter in the lowland than that at a higher altitude.
3. Percentages of deciduousness are high in mid latitude, and the percentages of evergreenness again increase in even higher latitude, resulting in a bimodal distribution in percentages of evergreenness with increasing latitudes.
4. Altitudinal distribution of percentages of evergreenness and deciduousness in mid latitude duplicates the latitudinal distribution. In low altitudes, percentages of evergreenness are high. But in mid altitudes, percentages of deciduousness become high, in even higher altitudes, however, evergreenness again predominates.
5. Number of species is highest in the non-seasonal tropical region and decreases towards seasonal tropics and higher altitudes and latitudes.

     

‘Absolute steepness’ of ramps as an essential cue for auditory pattern recognition by a grasshopper (Orthoptera; Acrididae; Chorthippus biguttulus L.)

Male grasshoppers of the species Chorthippus biguttulus react to female songs with a characteristic turn towards the female. The probability of turning towards female song models was used to evaluate those parameters which are essential for a signal to be interpreted as female song.

1. The shape of sound pulses turned out to be most decisive; pulses with ramps rising gradually over 3 and more ms were efficient (Figs. 2, 3), whereas rectangularly modulated pulses evoked only weak responses and only when pulse intervals were between 3 and 5 ms (Fig. 2). The decline of a pulse did not influence its efficiency (Fig. 3). In particular, pulses with sudden onsets and gradual declines were as weakly effective as rectangularly modulated ones and thus remarkably less effective than pulses with ramp-like onsets (Fig. 4).
2. Intensity tuning curves suggest, that the absolute steepness of ramps (expressed as μbar/ms) is detected by the grasshopper nervous system (Figs. 6, 7), possibly by processing the delay in excitation onset of at least two receptor types differing in threshold sensitivity.
3. The sawtooth shape of pulses in female signals is suggested to be adaptive with respect to directional hearing.

     

Model study of muscle spindles subjected to static fusimotor activation

An earlier proposed mechanical model of the de-efferented muscle spindle endings has been developed further in order to study whether the functional effects of activating the static fusimotor fibres may be accounted for by relatively simple mechanical events in the intrafusal muscle fibres. Particular attention has been paid to important controversial problems related to the processes involved in the fusimotor activation of the mammalian muscle spindles. In order to develop an adequate model, preliminary simulation studies of the functional effects of various reasonable modifications of the original “de-efferented” model were first carried out by means of a convenient direct electronic analogue of the mechanical system. The following results apparently reflect pertinent details about the peripheral receptor mechanisms associated with the activation of the static fusimotor fibres:

1. The γ _s-activation is most adequately accounted for by mechanical events occurring in the nuclear chain fibres.
2. Any uniform chain fibre contraction fails to account for the effects of the γ _s-activation.
3. The simulation of a local chain fibre contraction could apparently account for all significant effects of the γ _s-activation.
4. A reasonable increase of the position sensitivity and the vibration sensitivity of both the primary and the secondary endings could be accounted for by increasing the stiffness of about 1/4–1/2 of the length of the simulated nuclear chain fibre by a factor of about 2–10.
5. The powerful modulation of the background discharge of the receptor endings that is associated with the γ _s-activation could be accounted for by an independent contraction force related to the intensity of the static fusimotor activity.
6. The model predicts a decrease of the rate sensitivity of the primary endings (group Ia) and a slight increase of the (moderate) rate sensitivity of the secondary endings.
7. By an adequate selection of the parameters, the simulated γ _s-activation showed a negligible influence on the absolute level of the dynamic phase of the ramp response of the simulated primary endings, in spite of a considerable increase of the steady state (background) discharge before and after the termination of a simulated stretch.

     

Neurotransmitters of cephalopods

1. ACh, dopamine, noradrenaline, 5-HT,l-glutamate, and GABA are widely distributed in cephalopods and probably all function as neurotransmitters; octopamine also occurs and at one site is known to act as a neuromodulator.
2. Several peptides are also present, as well as nitric oxide synthase.
3. In the brain and sense organs cholinergic, aminergic, serotonergic and glutamatergic systems seem to be the most important.
4. ACh is also active in the gut, vascular system and some body muscles: it is generally inhibitory. The ACh receptors are similar to the vertebrate nicotinic type.
5. The catecholamines are important in the gut and vascular system: they are generally excitatory. The NA receptors are like the α-adrenergic subtype of vertebrates, but the nature of the DA and OA receptors is less certain.
6. 5-HT is important in the gut but is endogenous in some chromatophore nerves and acts on receptors that seem like the vertebrate 5-HT₁ type.
7. l-glutamate is an excitatory transmitter at the chromatophore (and probably at other) nerve-muscle junctions and is an extremely strong candidate for being the excitatory transmitter at the squid giant synapse. There are NMDA receptors on Schwann-cells but the receptors on neurons and muscles are like the vertebrate kainate type.
8. Little is known about the mode of action of cephalopod peptides; nor has it ever been shown that they co-exist with conventional transmitters in these animals.
9. The structure of one (FMRFamide) receptor has been elucidated, but apart from this nothing is known of the molecular biology of receptors in cephalopods.

     

A study on the rate of water transport of the clam katelysia opima in relation to environmental conditions

1. The neutral red technique was employed to study the rate of filtration in Katelysia opima.
2. The weight specific water filtration was found to be greater for younger clams compared to the older ones.
3. The rate of water filtration increased with decreasing salinity.
4. Water filtration was found to increase as temperature increased, reaching a maximum at 35°C. but then sharply decreasing at 39°C.
5. Light had no significant effect on the rate of filtration.
6. Suspended matter was found to affect the rate of water filtration.
7. The rate of filtration was low at high pH and high in low pH.
8. The rate of water filtration was found to be faster during high tide than during low tide.
9. The presence of the parasitic crab, Pennotheris sp., in the mantle cavity of clams had a marked effect on the particle filtration.
10. Accidental cut of the siphon tips had no effect on the rate of filtration.

     

Quantitative assessment of song-selectivity in the zebra finch “high vocal center”

1. Auditory responses in the zebra finch (Taenopygia guttata) song-system nucleus HVc were assessed at 54 recording sites by 3 different methods: discriminated action potentials; excitatory summed responses; and excitatory minus inhibitory summed responses. Four standard stimuli were presented at each site: the bird's own song; this song reversed; a conspecific song; and a noise burst. Responses were quantified by calculating a relative response index that partitioned the response, to provide a response profile, across the stimuli.
2. Regardless of analysis method, the strongest response was most often to the bird's own song (78–82%, depending on method). The predominant rank order of response strength across the remaining three stimuli was conspecific song > reversed song > noise.
3. The distribution of relative response magnitude was sensitive to analysis method. Discriminated spikes captured the heterogeneity of HVc neurons, whereas the excitatory summed responses reflected the overall trends more consistently. When inhibition was subtracted from excitation in the summed responses, the variance of the relative responses increased, but this method presented some problems for statistical analysis.
4. A small sample of neurons in other forebrain auditory areas was used for comparative analyses. At these recording sites, the bird's own song did not consistently elicit the best response and there were generally smaller differences in the relative responses to the four stimuli. The smaller degree of stimulus selectivity among these cells resulted in less sensitivity to differences in the assessment methods.

     

The use of a continuous assay system to show the stimulation of phosphoenolpyruvate production in intact mitochondria by valinomycin and by Mn2+

1. A method for the direct recording of the PEP efflux from isolated mitochondria is described.
2. This method has been used to show the stimulation of PEP efflux by externally added Mn⁺⁺ ions.
3. Valinomycin, uncoupler and oleate were also shown to stimulate PEP efflux.
4. Valinomycin caused an increase in the internal concentration of both PEP and citrate.
5. The results indicate that the major pathway of PEP synthesis in isolated mitochondria is via PEP carboxykinase and the results do not call for an unknown pathway of metabolism.
6. Two interactions between PEP and citrate are described; competition for the mitochondrial interior and the stimulation of PEP production by citrate.

     

Phase shifts in heterochromatic flicker photometry and Bidwell's ghost: A model

A simple model of brightness encoding in the retina is proposed that explains:

1. the differential phase shift needed to minimize brightness flicker in a homogeneous red-green counterphase flickering field.
2. the modulation transfer function at low temporal frequencies for flickering lights of one dominant wavelength against the background of light with another dominant wavelength.
3. the occurrence of the Hering, the Purkinje and the Hess afterimages.
4. the ability of monochromatic light to provoke the Purkinje afterimage (Bidwell's ghost).

It is shown that these effects can be treated as pure brightness effects; or that the signal processing of the opponent system is irrelevant to the results. A physiological substrate for the model is tentatively indicated. Some suggestions are made concerning possible experimental checks of the model.     

The use of toxicokinetics for the safety assessment of drugs acting in the brain

Pharmacological and toxicological studies undertaken on drugs that affect the brain are frequently performed in disparate species under various experimental conditions, at doses often greatly in excess of those expected to be administered to humans, and the findings are extrapolated implicitly or explicitly with scant regard to differences in the biodisposition of the drugs. Such considerations are necessary since:

1. Species;
2. Strain;
3. Gender;
4. Route;
5. Dose;
6. Frequency and time of administration;
7. Temperature;
8. Coadministration of drugs; and
9. Surgical manipulation

are but some of the factors that have been shown to influence the kinetics and metabolism of drugs. This article, using MDMA and other phenylethylamines as examples, provides evidence for the need to measure the exoosure of the drugs and their active metabolites in blood and brain (toxicokinetics) in order that conclusions based only on dynamic, biochemical, or histological evidence are more pertinent. Further, the combined use of toxicokinetic-dynamic modeling can lead to a better appreciation of the mechanisms involved and a more useful approach to the calculation of safety margins.     

The timing of activity in motor neurons that produce radula movements distinguishes ingestion from rejection in Aplysia

1. We have studied the neural circuitry mediating ingestion and rejection in Aplysia using a reduced preparation that produces ingestion-like and rejection-like motor patterns in response to physiological stimuli.
2. We have characterized 3 buccal ganglion motor neurons that produce specific movements of the radula and buccal mass. B8a and B8b act to close the radula. B10 acts to close the jaws and retract the radula.
3. The patterns of activity in these neurons can be used to distinguish the ingestion-like and rejection-like motor patterns. B8a, B8b and B10 are active together during the ingestion-like pattern. Activity in B8a and B8b ends prior to the onset of activity in B10 during the rejection-like pattern.
4. Our data suggest that these neurons undergo similar patterns of activity in vivo. During both feeding-like patterns, the activity and peripheral actions of B8a, B8b, and B10 are consistent with radula movements observed during ingestion and rejection. In addition, the extracellular activity produced by these neurons is consistent with neural activity observed in vivo during ingestion and rejection.
5. Our data suggest that the different activity patterns observed in these motor neurons contribute to the different radula movements that distinguish ingestion from rejection.

     

Modulation of swimming in Tritonia: excitatory and inhibitory effects of serotonin

1. In the mollusc Tritonia escape swimming is produced by a network of central pattern generator (CPG) neurons. The purpose of this study was to determine which neurotransmitters might be involved in the swim system.
2. Injection of serotonin (5HT) into whole animals elicited swimming followed by a long-lasting inhibition of swimming. In isolated brain preparations, bath-applied 5HT elicited a swim pattern at short latency and also caused a long-lasting inhibition of the swim pattern. The activation of swimming by 5HT was associated with a tonic depolarization of cerebral cell 2 (C2) and the dorsal swim interneurons (DSI) which form part of the swim CPG network.
3. In isolated brain preparations, bath applied glycine, histamine, proctolin, and FMFRamide had no effect on the swim motor pattern elicited by electrical stimulation of a peripheral nerve. Aspartate, carbacol, dopamine, glutamate, octopamine, pilocarpine, and small cardioactive peptide-B (SCP_B) inhibited the activation of swimming by nerve stimulation.
4. The 5HT antagonists cyproheptidine, tryptamine, and 7-methyltryptamine had no effect on swimming, but methysergide and fenfluramine inhibited swimming to both normal sensory stimuli and exogenously applied 5HT.
5. Staining with a polyclonal antibody indicated that one class of CPG neurons, the dorsal swim interneurons (DSI), was immunoreactive for 5HT.
6. Taken together, the data suggest that pattern generator interneurons, particularly the DSIs, use 5HT as a neurotransmitter.

     

Diurnal variation in Sagar Lake,Sagar (India) I studies in the deep water area

1. A diurnal study of inland fresh water lake has been made with respect to physical and chemical properties and the plankton.
2. Chlorides have followed the total carbonates while dissolved oxygen and pH have shown no relation.
3. Microcystis has followed no definite pattern of diurnal movement.
4. All crustaceans, some of the rotifers andTrachelomonas perform considerable diurnal movement in the course of a twenty four hour period.

     

Relationship between ecg,eeg and sps responses during arousal in the goldfish (Carassius auratus)

• 1.1. Goldfish (Carassius auratus) were fitted with ECG electrodes and intra-cranial stainless steel electrodes to monitor cardiac, EEG and SPS responses, during minimal restraint, to presentation of environmental stimuli (light-on, moving shadow, tap).
• 2.2. All three stimuli evoked a bradycardia and increases in the EEG frequency; correlates of arousal responses in fish.
• 3.3. EEG frequency changes were most evident in the fore- and midbrain regions; in the hindbrain smaller responses nevertheless showed discrimination between stimuli.
• 4.4. EEG amplitude changes were more site- and stimulus-specific than frequency changes.
• 5.5. SPSs occurred on stimulus presentations which were negative in polarity in the midbrain and positive in the forebrain and hindbrain, though the latter were smaller and less consistent.
• 6.6. Principal components analyses and regression analyses were used to examine detailed associations between peripheral and central physiological changes.
• 7.7. It was found that increases in the EEG frequency of fore- and midbrain regions were related to cardiac deceleration on early stimulus presentations.
• 8.8. This was also shown for the negative SPS of the midbrain to the presentation of the tap stimulus.
• 9.9. Positive SPSs of the forebrain were related to the bradycardia on later stimulus presentation i.e. during habituation of the arousal response.
• 10.10. The results indicate that in fish, as in other vertebrates, negative SPSs in the midbrain are associated with arousal and implicate the forebrain in the modulation of arousal by its habituation.

     

Physiological and pharmacological properties of responses to GABA and ACh by abdominal motor neurons in Manduca sexta

1. GABA, ACh, and other agents were applied by pressure ejection to the neuropil of the third abdominal ganglion in the isolated nerve cord of Manduca sexta. Intersegmental muscle motor neurons with dendritic arborizations in the same hemiganglion were inhibited by GABA (Fig. 2) and excited by ACh (Fig. 5).
2. Picrotoxin was a potent antagonist of GABA (Fig. 4A). Bicuculline reduced GABA responses in some motor neurons (Fig. 4C), but had no effect on many other motor neurons. Curare reduced ACh responses (Fig. 6A). Bicuculline was an effective ACh antagonist in most motor neurons tested (Fig. 6B).
3. Motor neurons with dendrites across the ganglion from the ejection pipette exhibited different responses to GABA and ACh. Contralateral motor neurons often showed smaller, delayed hyperpolarizing GABA responses (Fig. 7). On two occasions, contralateral motor neurons had excitatory responses (Fig. 8). Contralateral motor neurons were hyperpolarized by ACh (Fig. 9). The inhibitory responses had only slightly longer latencies than ipsilateral excitatory ACh responses (Fig. 10A). The contralateral inhibitory ACh responses, but not the ipsilateral excitatory ACh responses, were eliminated by TTX (Fig. 10B).
4. A model, which includes inhibitory interneurons that cross the ganglionic midline to inhibit their contralateral homologs and motor neurons (Fig. 11), is proposed to account for contralateral responses to GABA and ACh and antagonistic patterns of activity of motor neurons during mechanosensory reflex responses.