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PubMed1976

Dendritic responses of frog motoneurons produced by antidromic activation.

Responses to ventral root stimulation were studied in spinal cords in situ in unanesthetized frogs. Extracellular as well as intracellular recordings from motoneurons indicated that considerable depolarization of the dendrites occurred in the response to ventral root volley. Active components of this dendritic depolarization could also be observed. Extracellularly, negative field potentials were recorded both in the vicinity of motoneuron cell bodies and in areas occupied mostly by motoneuron dendrites. The refractory period of the negativity recorded at the vicinity of dendrites was longer than in the vicinity of somata. Changes in antidromic excitability were studied by the double volley technique. Augmentation of the field potential to a test volley was observed during the period of dendritic depolarization, followed by a longer-lasting period of depression of the test response. It is concluded that an action potential in frog motoneurons induces depolarization of the dendrites. The depolarized dendrites can generate local action potentials and can produce negative field potentials remotely from the somatic pool. The response of dendrites to stimulation of the ventral root has particular importance in the recurrent facilitation of the frog motoneurons.

Action Potentials↗
PubMed1984

Visual (optokinetic) and somesthetic inputs to the cerebellum of bilaterally labyrinthectomized frogs.

Unitary responses to visual (optokinetic) and somesthetic (cutaneous and propioceptive ) stimulation were recorded from the Purkinje cell layer of the cerebellum of acute (up to 30 h) and chronic (30-90 days) bilaterially labyrinthectomized frogs. Simple spikes from Purkinje cells as well as activity from cells without complex spikes were considered in this paper. The properties of the response and the distribution (restricted to the dorsal rim and auricular lobes) of the units sensitive to optokinetic stimulation of labyrinthectomized frogs, both acute and chronic, were similar to those previously reported for normal animals. The properties of the responses to somesthetic neck and limb stimulation remained similar to those of normal animals. However, there was an increase in the number of units responsive to somesthetic stimulation within the dorsal half of the corpus cerebelli (including the dorsal rim), a region experimentally deprived of vestibular afferents, neck responsive units were 33% of the total in acutely and 61% in chronically labyrinthectomized animals (compared to 5% in normal). Limb responsive units were 49% in acute and 65% in chronic animals (compared to 12% in normal). A consequence of the increase in somesthetic input was convergence of optokinetic and somesthetic inputs at the level of single units within the dorsal rim, totally absent before the lesion. The results suggest that the somesthetic spinal input might substitute for at least some features of the vestibular input to the cerebellum in bilaterally labyrinthectomized frogs.

Action Potentials↗
PubMed1985

Rapid axonal transport of three molecular forms of acetylcholinesterase in the frog sciatic nerve.

Acetylcholinesterase occurs in the frog sciatic nerve under five stable molecular forms with distinct sedimentation coefficients in sucrose gradients: 3 globular forms (3.6S, 6S and 10.5S) and two asymmetric ones (14S and 18S). Whereas in birds and mammals, the asymmetric tailed forms of acetylcholinesterase are present in trace amounts in peripheral nerves and account for only a small part of the enzyme activity submitted to a rapid axonal transport, the two asymmetric 14S and 18S forms represent nearly 50% of total activity in the frog sciatic nerve and account for 60-70% of the acetylcholinesterase activity accumulated at both sides of a nerve transection, the rest being due to an accumulation of globular molecules. We showed that the three forms, 10.5S, 14S and 18S, are all carried with the fast phase of axonal transport at a velocity of 100-120 mm/day in the anterograde direction and 20-30 mm/day in the retrograde direction. The velocity of transport for the light molecular forms 3.6S and 6S could not be calculated. In addition, we observed that large amounts not only of the 10.5S but also of the asymmetric 14S and 18S forms appear to be stationary along the frog sciatic nerve, contrary to the situation described for peripheral nerves in birds or mammals. Our results thus reveal that some axonal transport parameters for the asymmetric forms of acetylcholinesterase greatly differ in the peripheral nerves of amphibians on the one hand and of birds and mammals on the other, suggesting that these heavy molecular forms might have distinct functions in the nerves of lower and higher vertebrates.

Acetylcholinesterase↗
PubMed1990

Nucleus isthmi: its contribution to tectal acetylcholinesterase and choline acetyltransferase in the frog Rana pipiens.

The distribution of acetylcholinesterase and the activity of choline acetyltransferase was studied in the tecta of normal frogs and frogs without retinal and/or nucleus (n.) isthmi inputs. In normal animals acetylcholinesterase activity is found primarily in three bands in the outer layers of the tectum-lamina A, laminae C-F, and lamina G. After retinal and contralateral n. isthmi deafferentation three distinct bands of tectal acetylcholinesterase activity are still present. After bilateral n. isthmi deafferentation there is loss of activity in lamina G and reduced activity in lamina A. With retinal and ipsilateral n. isthmi deafferentation, activity is seen only in lamina A. With retinal and bilateral n. isthmi deafferentation there is virtually no acetylcholinesterase activity in the outer tectal layers. Following unilateral retinal deafferentation there is no statistically significant difference in choline acetyltransferase specific activity between intact and deafferented tectal lobes after two, four and nine weeks. With unilateral nucleus isthmi lesions and survival times of between 10 and 40 days, choline acetyltransferase specific activity in the tectal lobe ipsilateral to the ablation is approximately 38% of the specific activity of the contralateral lobe. With bilateral n. isthmi lesions there is a strong correlation between amount of n. isthmi ablated and reduction of choline acetyltransferase activity. In vitro tectal acetylcholine synthesis was also determined in animals with unilateral n. isthmi ablation. On average, tectal lobes ipsilateral to the ablated n. isthmi synthesize acetylcholine at a rate which is approximately 58% of that of contralateral tecta. Collectively, these results imply that n. isthmi is the sole cholinergic input to the frog optic tectum, with ipsilaterally projecting isthmotectal fibers accounting for the greater share.

Acetylcholine↗
PubMed1993

Distribution of melatonin receptors in the brain of the frog Rana pipiens as revealed by in vitro autoradiography.

Melatonin binding sites were identified in the brain of the frog Rana pipiens using in vitro autoradiography. Coronal sections were incubated for 1 h in 100 pM 2-125I-iodomelatonin. Specific binding was displaced with 1 microM nonradioactive melatonin. Autoradiographic labeling of 3H-Hyperfilm was observed in areas that receive primary, secondary, and tertiary visual input: the superficial layers of the optic tectum, anterior and posterior thalamic nuclei, striatum, medial pallium, and interpeduncular nucleus. Other areas that demonstrated binding included the medial and lateral septal nuclei, medial preoptic area, suprachiasmatic region, and anterodorsal tegmental nucleus. Binding was also apparent in the distribution of the lateral olfactory tract (lateral pallium), and in tracts associated with visual pathways: optic nerve, chiasm and tract, and lateral and medial forebrain bundles. A high degree of melatonin binding was observed in the left habenular nucleus, but not in the habenulum of the right side of the brain. Radioreceptor binding assays of frog whole-brain homogenate demonstrated specific saturable melatonin binding (Kd = 70 pM, Bmax = 0.80 fmol/mg protein). Melatonin and 6-chloromelatonin were potent displacers of 2-125I-iodomelatonin, while 5-hydroxytryptamine, 5-methoxytryptamine, and N-acetylserotonin were much less potent. Melatonin inhibited the forskolin-stimulated increase in cAMP synthesis in optic tectum explants. These results suggest that high-affinity melatonin receptor binding sites are widely distributed in the telencephalon, diencephalon, and mesencephalon and are very prominent in areas of the frog brain that are associated with visual processing.

Animals↗
PubMedDate not supplied

Cutaneous receptive field enlargement following skin-grafting in the frog, D. pictus.

Misdirected wiping responses occur in Discoglossus pictus frogs when skin-grafted early in larval life. Cutaneous receptive fields (CRF) for various peripheral nerve branches were mapped electrophysiologically in control and skin-grafted frogs. The ventrolateral nerve branch (VL) frequently provided innervation for significant areas of dorsal skin in both control and skin-grafted animals. Although the VL crossed over onto dorsal skin areas in some skin-grafted frogs giving rise to misdirected reflexes, these dorsal extensions frequently failed to include all skin areas from which the maladaptive reflexes could be elicited. It was concluded that misdirected reflexes are mediated via dorsal nerve branches occupying normal mid-dorsal areas of the back.

Abdomen↗
PubMed1981

Peripheral reinnervation patterns and dorsal root ganglion cell topography in skin-grafted Rana pipiens frogs.

Behavioral responses to mechanical stimulation of the skin were observed in unoperated, 180 degrees skin-rotated and sham-grafted Rana pipiens frogs before and after crushing various cutaneous nerve bundles. In the two control groups, wiping responses directed towards the animal's dorsum were mediated solely via dorsomedial (DM) and dorsolateral (DL) nerve trunks. In skin-rotated frogs, DM and DL nerve trunks were responsible for almost all of the misdirected responses elicited from dorsal cutaneous areas. Six frogs (2 sham- and 4 skin-rotated) possessed some areas of plical skin from which responses could only have been mediated via more ventrally located nerve trunks. However, such dorsal expansions of ventral receptive fields never included the midline skin areas from which misdirected responses had been elicited in the intact animal. The topographic arrangement of cells within the DRG appears to be weakly polarized in cobalt-stained sections, but with large individual variations in size and observed number of neuronal profiles. For the skin-rotated group the ratio of cobalt-filled profiles in the dorsal as compared with the ventral half of the ganglion was 1.6, which did not differ significantly from the control value of 1.9. Taken as a whole, the results of the present study indicate that selective (re)innervation of displaced skin by the original nerve fibers cannot explain the origin of misdirected responses in Rana pipiens.

Animals↗
PubMedDate not supplied

Changes in formaldehyde-induced fluorescence of the hypothalamus and pars intermedia in the frog, Rana temporaria, following background adaptation.

Adaptation of the frog, Rana temporaria, to a white background for 12 hr has resulted in an intense formaldehyde-induced fluorescence (FIF) in the neurons of the preoptic recess organ (PRO), paraventricular organ (PVO), nucleus infundibularis dorsalis (NID) and their basal processes permitting visualization of the PRO- and PVO-hypophysial tracts that extend into the median eminence (ME) and pars intermedia (PI); the FIF is reduced in all the structures by 3 days. In frogs adapted to a black background, for 12 hr and 3 days, there was a general reduction in the FIF of the PRO neurons and PRO-hypophysial tract. By 12 hr black background adaptation, the PVO/NID neurons and only their adjacent basal processes show FIF which was sharply reduced by 3 days, making the PVO-hypophysial tract undetectable. In the PI fibers the fluorescence was more intense in black-adapted frogs than in white-adapted ones at both the intervals studied. The simultaneous changes in the FIF of the hypothalamic nuclei, tracts and PI suggest that the PRO and PVO/NID neurons participate in PI control through release of neurotransmitter(s) at the axonal ends.

Animals↗
PubMed1986

Effects of glycosylation inhibitors on the frog retina.

Effects of the intraocular injection of three inhibitors of glycosylation (tunicamycin, castanospermine, and swainsonine) on the rhodopsin content and the integrity of disc membranes in frog retina were studied. The administration of 10 or 100 micrograms of tunicamycin resulted in a 78% loss of rhodopsin in the frog retina which also exhibited a significant reduction in the length of photoreceptor outer segments (as examined under light microscope). This suggests that the synthesis and/or insertion of rhodopsin into the disc membrane is inhibited by tunicamycin. In contrast, injections of up to 250 micrograms of castanospermine and swainsonine resulted in neither a decrease in rhodopsin content nor a change in the length of photoreceptor outer segments. Examination of retinal tissue homogenates for alpha Mannosidase and alpha-Glucosidase activities revealed homogenates for alpha Mannosidase and alpha-Glucosidase activities revealed these hydrolase activities to be significantly decreased (70%). We suggest the possibility that specific oligosaccharide processing reactions may not be required for the insertion of rhodopsin and subsequent assembly of disc membranes in frog photoreceptors.

Alkaloids↗
PubMed1986

GABA-ergic control of alpha-melanocyte-stimulating hormone (alpha-MSH) release by frog neurointermediate lobe in vitro.

Measurement of glutamate decarboxylase (GAD) activity in the intermediate lobe of the frog pituitary and brain showed that neurointermediate lobe extracts represented 12% of the GAD activity detected in the whole brain. No significant activity was measured in distal lobe extracts. Immunocytochemical studies revealed GAD-containing fibers among the parenchymal cells of the pars intermedia. The localization of GAD-like material in the intermediate lobe of the frog pituitary suggested a possible role of gamma-aminobutyric acid (GABA) in the regulation of melanotropic cell secretion. Administration of GABA (10(-6) to 10(-4) M), to perifused neurointermediate lobes caused a brief stimulation of alpha-melanocyte stimulating hormone (alpha-MSH) release followed by an inhibition. Picrotoxin (10(-4) M), a Cl- channel blocker, abolished only the stimulatory effect of GABA (10(-4) M), whereas bicuculline (10(-4) M), a specific antagonist of GABAA receptors, totally inhibited the effects of GABA (both stimulatory and inhibitory phases). Bicuculline induced by itself a slight stimulation of alpha-MSH release, suggesting that GABA-ergic nerve fibers present in the intermediate lobe are functionally active in vitro. The GABAA agonist muscimol (10(-7) to 10(-4) M) mimicked the biphasic effect of GABA on alpha-MSH release. Administration of baclofen, a specific GABAB agonist (10(-7) to 10(-4) M) induced a dose-dependent inhibition of alpha-MSH secretion. In contrast to GABA or muscimol, baclofen did not cause any stimulatory effect whatever the dose. Taken together these result suggested that GABAA and GABAB receptors were present on frog melanotrophs.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1992

Cloning and sequencing of the cDNA encoding tyrosinase of the Japanese pond frog, Rana nigromaculata.

We cloned and sequenced the cDNA encoding tyrosinase (TYN) of the Japanese pond frog, Rana nigromaculata. The 3511-bp cDNA contained a 54-bp 5'-noncoding region, a 1596-bp open reading frame encoding TYN of 532 amino acids (aa), and a 1861-bp 3'-noncoding region. The aa sequence of frog TYN predicted from the cDNA sequence was homologous to that of mouse and human TYNs. The aa sequence including the copper-binding domain, which is likely the active center of TYN, was highly conserved among these three species and Neurospora crassa, Streptomyces antibioticus, and S. glaucescens. The frog TYN also contains possible glycosylation sites and conserved Cys at sites similar to those in the mouse and human TYNs. There are two hydrophobic regions at the N-terminus and near the C-terminus, which are likely the signal (leader) peptide and a transmembrane domain, respectively.

Amino Acid Sequence↗
PubMed1984

Detection of amplitude-modulated tones by frogs: implications for temporal processing mechanisms.

The whole nerve action potential (AP) from the auditory nerve and midbrain averaged evoked potential (AEP) were recorded in Hyla chrysoscelis and H. versicolor in response to synthesized amplitude-modulated stimuli with variable modulation frequencies (Fm). The AP from these frogs is similar to the potential described for mammals and showed a bandpass characteristic in its ability to follow sinusoidally amplitude-modulated (AM) sound stimuli. A lesioning study suggests that the midbrain AEP is a localized neural response of neurons near the ventral border of the torus semicircularis. The AEP is a complex waveform consisting of fast and slow components. The fast component encodes the temporal structure of acoustic stimuli and is used to measure temporal sensitivity in these two species. The AEP behaves like a low-pass filter with a cutoff frequency of 250 Hz when tracking AM signals. Threshold for detection requires a modulation depth of 8-12% of the total stimulus amplitude (delta I = 1.5-2.0 dB). Relative to the eighth nerve AP, the AEP displays an enhanced coding of AM signals when Fm less than 100 Hz, and a slightly inferior ability to code Fm above 250 Hz. The AEP reflects only that portion of the neural response that encodes amplitude fluctuations. In comparison to the range of amplitude fluctuations coded by single units in the rat inferior colliculus or by human evoked potential, the frog AEP codes higher rates of Fm. The proposal that these frogs process AM stimuli solely on the basis of amplitude fluctuations, and do not use spectral cues at higher modulation frequencies is considered. The AM sensitivity of the AEP, which encompasses most biologically relevant rates of amplitude fluctuation for the animal, and the limited frequency resolution of the periphery, lend support to this proposal. However, convergent spectral processing at higher auditory centers cannot be excluded by this study. Psychophysical tests will be required to determine whether both of these mechanisms may be operating during temporal information processing in anurans.

Action Potentials↗
PubMed1989

Electrophysiological properties and morphology of hair cells isolated from the semicircular canal of the frog.

Hair cells isolated from the crista ampullaris of the frog (Rana pipiens) remained viable for up to 5 h and were studied using whole cell voltage- and current clamp recordings. Morphological characteristics of isolated crista hair cells were compared with hair cells studied in situ using light- and electron microscopy. While other labyrinthine hair cells such as mammalian inner and outer hair cells of the cochlea, saccular hair cells of the frog, and cochlear hair cells of the turtle typically have a cylindrical shape, the crista hair cells in the frog are predominantly bulbous, having a thin elongated trunk projecting from a spherical base just large enough to enclose the nucleus. This shape correlates well with the compressed packing configuration of hair cells of the crista ampullaris observed in situ in the histological material. The support cells often failed to separate adjacent hair cells, particularly the apical ends of the hair cells. Maximal cell density on the sensory epithelial ridge appears to be achieved by this arrangement. The mean resting membrane potential (Vz) of isolated crista hair cells was -44.8 mV. Cells with smooth surfaces and apparent opacity had the most negative Vz potentials. As the cells appeared to deteriorate, there was development of transparency and cell surface granulation. Such cells had more positive initial Vz values. Cells with Vz values more positive than -15 mV exhibited a distinct, contoured nucleus. Cells lacking these indicators of deterioration were characterized by input resistances of 1.9 +/- 0.31 G omega and membrane time constants of 13 +/- 2.5 ms. A large complex outwardly rectifying current was identified which was abolished by substituting Cs+ for K+ in the internal solution. The outward K+ current had two major components: a fast tetraethylammonium (TEA)-insensitive, voltage dependent I(A)-type current which showed voltage dependent inactivation; and a TEA sensitive current which had characteristics of a calcium dependent IK(Ca)-type current. Transient changes (20 ms duration) in membrane potential mimicking that which could be produced by the transduction current during cilial displacement potently modulated the I(A) current. Depolarizing current pulses of greater than 63800 pA were required to elicit membrane voltage oscillations. The resulting membrane potential offset of at least 40 mV is well beyond the magnitude of hair cell receptor potentials making it unlikely that these oscillations would play a role in enhancing frequency selectivity.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1989

Spontaneous otoacoustic emissions in the European edible frog (Rana esculenta): spectral details and temperature dependence.

Spontaneous otoacoustic emissions were recorded in 41 ears of 29 European edible frogs (Rana esculenta). Emission frequencies ranged from 450 to 1350 Hz. The distribution of frequencies shows two distinct populations: one above and one below 1 kHz. With one exception, a maximum number of two emissions were recorded per ear, each in a different population. An amplitude distribution of a frog emission was sampled, from which it was concluded that the emission is generated by an active oscillator. The spectral width of an emission ranged from 1 to 200 Hz (average 38 Hz). There was negative correlation between sound pressure level of an emission and spectral width. In 4 frogs the dependence of emission power and frequency on temperature was investigated. An emission could be 'switched on and off' within a few degrees centigrade. At temperatures below the switching interval no emission was recorded; for higher temperatures emission power showed no dependence on temperature. Frequency increased with temperature (Q10 = 1.1 to 1.3). This yields a mismatch with temperature dependence of best frequencies of auditory fibers. The consequences of this mismatch are discussed.

Animals↗
PubMed1990

Basic properties of auditory-nerve responses from a "simple' ear: the basilar papilla of the frog.

Spike discharges initiated by mammalian inner hair cells are produced by a complicated system involving both mechanical and neural components that normally operate in a bi-directional configuration involving multiple feedback loops. In contrast, the frog basilar papilla has the equivalent of inner hair cells, but lacks outer hair cells; it has no efferent system, and no basilar membrane. This suggests that the frog basilar papilla lacks some of the mechanical and neural feedback paths characteristic of the mammalian system. Detailed measurements of tuning curves, spontaneous activity and responses to tones an clicks reveal large parametric differences between frog and mammals in spontaneous rate, absolute refractory time, long-term adaptation and phase locking. Responses to tone bursts are qualitatively similar, but parametrically quite different. More focused examinations of these effects will be able to exploit the differences in adaptation to long- versus short-duration stimuli could be caused by depletion of afferent neurotransmitter or by activation of feedback loops involving the efferent system. In the basilar papilla, any differences in adaptation must result from changes in the afferent pathway alone.

Acoustic Stimulation↗
PubMed1992

Evidence for L-glutamate release in frog vestibular organs.

The present study was devised in order to ascertain whether L-glutamate (Glu) is the neurotransmitter at the primary afferent synapse in frog vestibular organs. To this end different groups of frog isolated semicircular canals were stimulated by means of solutions slightly enriched in K+ (5 mM K(+)-rich solutions are sufficient to produce a strong, long-lasting, transmitter release from the basal pole of sensory cells) both in normal conditions and after low-Ca(2+)-high-Mg2+ impairment of the synaptic transmission. The concentration of Glu in the surrounding medium, determined by means of a bioluminescence-enzymatic method, was evaluated in two different experimental conditions: a) when the canals (5 canals placed inside little net bags) were immersed in a 5 mM K(+)-stimulating solution; b) during the superfusion of the canals (25 canals placed into a little perfusion chamber) with a 5 mM K(+)-stimulating solution. The net bag experiments demonstrated that K(+)-rich solutions can provoke an outflow of Glu from canal organs only if the crista ampullaris is present and functioning. Glu fluctuations were in fact suppressed by employing canals deprived of the ampulla or after low-Ca2(+)-high-Mg2+ synaptic blockade. The superfusion experiments demonstrated that the time course of 5 mM K(+)-induced release of Glu from the sensory organ strictly parallels the time course of 5 mM K(+)-induced EPSPs and spike discharge in afferent axons. These results strongly support the hypothesis that Glu is, or is released with, the afferent transmitter in frog inner ear sensory organs.

Animals↗
PubMed1995

Classification of the temporal discharge patterns of single auditory neurons in the frog superior olivary nucleus.

Temporal discharge patterns of neurons in the superior olivary nucleus (SON) of the northern leopard frog (Rana pipiens pipiens) were studied by evaluating peri-stimulus time histograms and interspike interval histograms generated from responses to tone bursts at the neuron's characteristic frequency and at 10 dB above neuron's characteristic threshold. Four basic discharge patterns were observed, i.e., primary-like, phasic-burst, phasic, and pauser. Additionally, within each class, different neurons exhibited further subtle differences in temporal discharge patterns and thus subclasses were distinguishable. For most SON neurons, changes in signal level did not affect the discharge pattern. However, there were a few notable exceptions. The firing pattern of 12% of SON neurons changed from one class to another with a change in signal level. Two-thirds of phasic-burst neurons showed increased regularity (i.e., chopping pattern) in their discharges as the signal level was increased. The majority of SON neurons showed monotonic rate-level functions, but one-fourth gave non-monotonic rate-level functions. The presence of a fairly large number of non-monotonic units in the frog SON suggests an additional role of this nucleus in information processing beyond the simple coding of stimulus level. The discharge patterns displayed by SON neurons resembled those seen in the frog cochlear nucleus and in the mammalian superior olivary complex.

Acoustic Stimulation↗
PubMed1984

Metallothionein present or induced in the three species of frogs Bombina orientalis, Bufo bufo japonicus and Hyla arborea japonica.

A low molecular weight copper (Cu)- and/or zinc (Zn)-binding protein was present in the livers (and kidneys) of the three species of frogs Bombina orientalis, Bufo bufo japonicus and Hyla arborea japonica. Cadmium (Cd) accumulated in the livers (and kidneys) after repeated i.m. injections of Cd2+ was mostly bound to the same metal-binding protein as Cd, Zn- or Cd, Zn, Cu-thionein. Concentrations of Cd and several essential elements in the livers and kidneys of the three species of frogs were determined by inductively coupled plasma-atomic emission spectrophotometry. Metallothionein in the three species of frogs consists of a single isoform both in control and Cd-injected animals.

Animals↗