Response properties of auditory activated cells in the occipital cortex of the blind mole rat: an electrophysiological study.

Previous studies have demonstrated that despite its blindness, the subterranean blind mole rat (Spalax ehrenbergi) possesses a noticeable lateral geniculate nucleus and a typical cyto-architectural occipital cortex that are reciprocally connected. These two areas, as revealed by the metabolic tracer 2-deoxyglucose, are activated by auditory stimuli. Using single unit recordings, we show that about 57% of 325 cells located within the occipital cortex of anesthetized mole rats responded to at least one of the following auditory stimuli--white noise, pure tones, clicks, and amplitude modulated tones--with the latter two being the most effective. About 85% of cells driven by either contralateral or ipsilateral stimulation also responded to binaural stimulation; about 13% responded only to binaural stimulation; and 2% were driven exclusively by contralateral stimulation. Comparing responsiveness and response strength to these three modes of stimulation revealed a contralateral predominance. Mean latency (+/-SD) of ipsilateral and contralateral responses were 48.5+/-32.6 ms and 33.5+/-9.4 ms, respectively. Characteristic frequencies could be divided into two distinct subgroups ranging between 80 and 125 Hz and between 2,500 and 4,400 Hz, corresponding to the most intensive spectral components of the vibratory intraspecific communication signals and airborne vocalizations.

Recovery from sarafotoxin-b induced cardiopathological effects in mice following low energy laser irradiation.

OBJECTIVE: Low energy laser irradiation has been shown to accelerate various biological processes, including regeneration of injured tissues. In the present work we studied the effect of low energy laser irradiation on ischemic mice hearts, following administration of sarafotoxin-b, a powerful vasoconstrictor peptide of the endothelin/sarafotoxin family. METHODS: Immediately after injection of the toxin and two days later, hearts were exposed to low energy laser irradiation. Eight days after the injection the mice were sacrificed and their hearts were processed for light and electron microscopy. RESULTS: Sarafotoxin-b induced an approximate 2-fold increase in the relative cavity volume of the left ventricle. Low energy laser irradiation of the sarafotoxin-injected mice caused a significant decrease (62%) in the left ventricular dilatation. Electron microscopy of the sarafotoxin-injected mice hearts revealed that the extent of formation of large vacuoles in the cytoplasm of the cardiomyocytes as well as disorganization of the myofibrils were much higher than in the laser irradiated mice. CONCLUSIONS: Our study indicates that low energy laser irradiation enhanced recovery of the damaged cardiomyocytes from the ischemia induced by sarafotoxin-b.

[Visual, auditory, and bimodal activity in the banks of the lateral suprasylvian sulcus in the cat]. / Zritel'naia, slukhovaia i bimodal'naia aktivnost' na kraiakh lateral'noi suprasil'vievoi borozdy u koshki.

In addition to visually driven cells we found within the lateral suprasylvian visual cortex of cats a considerable number of auditory and/or bimodal cells. Most of the visually driven cells were direction and orientation selective with responses that were neither highly stimulus time locked nor very stable. Most of the auditory responses were also not very stable, had relatively high thresholds and were readily habituated. Previous studies have suggested that populations of cells within the lateral suprasylvian area are specialized for the analysis of optic flow fields. Given that a remarkable proportion of cells within this area can be also driven by auditory stimuli we hypothesize that the "optic flow" model may be extended to the bimodal domain rather than restricted to visual clues only. This, however, remains to be corroborated experimentally.

Pathological and experimentally induced blindness induces auditory activity in the cat primary visual cortex.

Early blindness in humans and experimental visual deprivation in animal models are known to induce compensatory somatosensory and/or auditory activation of the visual cortex. An abnormal hydrocephalic cat with extreme malformation of the visual system, born in our breeding colony, rendered a good model system for investigating possible cross-modal compensation in such a pathological case. For comparison, we used normal and neonatally enucleated cats. When introduced to a novel environment, the abnormal cat behaved as if it was completely blind, yet it responded normally to auditory stimuli. As anticipated, single cells in the visual cortex of normal cats responded to visual, but not to auditory stimuli. In the visual cortex of enucleated cats, flashes of light did not elicit field-evoked potentials or single-unit responses. However, several cells did respond to various auditory stimuli. In the remnant visual cortex of the abnormal cat, auditory stimuli evoked field potentials and single-cell responses. Unexpectedly, however, unlike the enucleated cats, in the abnormal cat, flashes of light also elicited field-evoked potentials. Judging by its behavior, it is very likely that this deformed cat had completely lost its ability to perceive images, but had probably retained some sensitivity to light.

Auditory activation of cortical visual areas in cats after early visual deprivation.

Auditory activation of the primary visual cortex (area 17) and two extrastriate visual cortical areas - the anterolateral lateral suprasylvian area (ALLS) and anteromedial lateral suprasylvian area (AMLS), was investigated in visually impaired cats. Impairment was accomplished shortly after birth by bilateral eyelid suturing (binocularly deprived cats, BD) or bilateral enucleation (binocularly enucleated cats, BE). In BE cats, the optic nerve and chiasm were entirely degenerated. No cortical atrophy or cytoarchitectural malformation was noticed in either BD or BE cats. In both normal and impaired cats we found auditory-responsive cells in the ALLS and AMLS, areas that are considered strictly visual. The most remarkable finding was an increase in the relative number of these auditory cells in the BD and BE cats, which was more prominent in the latter. Some auditory-responsive cells were also found in area 17 of BE cats. On the basis of formal calculation, it is tempting to suggest that the increase in relative number of auditory cells in these areas reflects the transformation of all the visual cells in the ALLS of BD and BE cats into auditory cells. In BE cats, all bimodal cells and an appreciable percentage of non-responsive cells also had transformed to auditory cells. In the AMLS of BD cats, it is primarily the bimodal cells that become auditory cells, whereas in BE cats all the visual and bimodal cells as well as non-responsive cells undergo this transformation. This assumption, however, is one possible interpretation of our results but not the only one. Other modes of neuronal plasticity that might yield similar results in the visually deprived cats can not be ruled out.

Seismic communication signals in the blind mole-rat (Spalax ehrenbergi): electrophysiological and behavioral evidence for their processing by the auditory system.

Based on morphological and behavioral findings we suggest that the seismic vibratory signals that blind mole-rats (Spalax ehrenbergi) use for intraspecific communication are picked up from the substrate by bone conduction and processed by the auditory system. An alternative hypothesis, raised by others, suggest that these signals are processed by the somatosensory system. We show here that brain stem and middle latency responses evoked by vibrations are similar to those evoked by high-intensity airborne clicks but are larger in their amplitudes, especially when the lower jaw is in close contact with the vibrating substrate. Bilateral deafening of the mole-rat or high-intensity masking noise almost completely eliminated these responses. Deafening also gradually reduced head-drumming behavior until its complete elimination about 4-6 weeks after surgery. Successive vibrations, at a rate of 0.5 vibrations/s, elicited prominent responses. At rates higher than 2 vibrations/s the amplitude of the brain stem response did not change, yet the middle latency response disappeared almost completely. It is concluded that the seismic signals that mole rate use for long distance communication are indeed processed primarily by the auditory system.

Resistance of the Egyptian mongoose to sarafotoxins.

The Egyptian mongoose (Herpestes ichneumon) is known for its resistance to viperid and elapid venoms. The current work demonstrates that it is also resistant to the venom of Atractaspis and its most toxic component, sarafotoxin-b. Intravenous administration of this toxin, at a dose of about 13 times LD100 for mice, resulted in disturbance in electrocardiograms in the mongoose, which returned to normal after several hours. Sarafotoxin-b failed to induce contraction of mongoose aortal preparations. Endothelin-1, which was demonstrated in tissue extracts of the mongoose by immunological methods, induced contraction of the isolated mongoose aorta. This contraction, however, was greatly reduced when endothelin-1 was applied on top of sarafotoxin-b. Binding studies revealed endothelin/sarafotoxin-specific binding sites in brain and cardiovascular preparations of the mongoose. It is suggested that some structural features of endothelin/sarafotoxin receptors in the mongoose enable them to differentiate between the two peptides.

Cross-modal neuroplasticity in the blind mole rat Spalax ehrenbergi: a WGA-HRP tracing study.

Spalax ehrenbergi is a subterranean rodent possessing extremely degenerated eyes but a distinguishable dorsal lateral geniculate nucleus (dLGN) which projects ipsilaterally to a fair sized striate cortex. 2-Deoxyglucose and electrophysiological studies have shown that these areas are vigorously activated by auditory input. We show here, using the neuroanatomical tracer WGA-HRP, that the major source of the auditory input to the visual system is the inferior colliculus that, in addition to all its typical auditory targets, also projects to the dLGN. The visual thalamo-cortical pathway thereupon processes auditory information. This is the first naturally blind mammal in which the origin of auditory compensatory projections have been traced and described.

Biological activities of [Thr2]sarafotoxin-b, a synthetic analogue of sarafotoxin-b.

The 21 amino acid sarafotoxins (SRTX) c and d/e as well as endothelin-3 (ET-3) are known to be less toxic and weaker pharmacologically than the other isopeptides SRTX-a, SRTX-b and ET-1. Since SRTX-c, SRTX-d/e and ET-3 possess a Thr instead of a Ser at position 2, we investigated the possibility that this mutation could be responsible for the observed biological differences. Here we show that the synthetic [Thr2]SRTX-b has indeed a lower vasoconstriction efficacy (approximately 35%) in the rabbit aorta, but it is nearly as potent as SRTX-b in toxicity tests and in influencing contraction of the rat uterus. Using monoclonal antibodies directed against the structurally related endothelin-1, we also show that the antigenicity of the analogue is comparable to that of SRTX-b, suggesting that the overall structure of the two peptides is similar, despite the substitution at position 2. We suggest that the Thr2 substitution contributes to the lower activity of the 'weak' peptides in some systems; however, additional substitutions found in the 'weak' peptides of the ET/SRTX family most probably contribute to their low pharmacological activity.

Sarafotoxins and endothelins: evolution, structure and function.

The venom of the burrowing asp Atractaspis engaddensis contains several 21 amino acid residue peptides known as sarafotoxins. The sarafotoxins are homologous to the mammalian endothelin family, and they have similar biological activities. This review covers recent advances in the study of the chemical and biological properties of the sarafotoxins and endothelins.

Cloning and sequence analysis of cDNAs encoding precursors of sarafotoxins. Evidence for an unusual "rosary-type" organization.

Sarafotoxins (SRTXs) are 21-amino acid peptides structurally and functionally similar to endothelins (ETs). To understand how SRTXs are overproduced in venom glands of the snakes Atractaspis engaddensis and hence used as toxins, we cloned cDNAs encoding SRTXs and elucidated their nucleotide sequences. We predict that SRTX precursors are large prepropolypeptide chains with an unusual "rosary-type" structure made of 12 successive similar stretches of 40 residues (39 in the first stretch). Each stretch begins with a "spacer" of 19 invariant residues (18 in the first stretch) immediately followed by the sequence of one SRTX isoform. Six different isoforms are identified within a single precursor molecule. Maturation of the precursor may require endopeptidases that cleave the Leu-Cys bond and the Trp-Arg/Lys bond invariably found at the SRTX N and C termini, respectively.

Equithesin: a hibernation-inducing drug?

1. Hibernating insectivore species (hedgehogs) and non-hibernating rodents (guinea pig and rat) were anaesthetized with 'equithesin' (a mixture of chloral hydrate, magnesium sulphate, and pentobarbitone sodium). 2. The physiological responses shown by the hedgehogs were similar to those observed in hedgehogs during a natural or cold-induced hibernation. 3. These responses included a strong reduction in body temperature, heart rate, respiration and oxygen consumption, and brain activity. 4. Such responses to equithesin were not observed in the non-hibernating rodent species. 5. These results suggest that equithesin is a potential tool for hibernation research.

Activity of sarafotoxin/endothelin peptides in the heart and brain of lower vertebrates.

The effects of sarafotoxin-b (SRTX-b) and endothelin-1 (ET-1) were tested in the fish tilapia (Ore niloticus x O. aureus hybrids) and torpedo (Torpedo ocellata), the toad (Bufo viridis), the agama lizard (Agama stellio) and water snake (Natrix tessellata). In isolated heart preparations of the fish and agama, peptide doses of 0.05-0.5 micrograms/ml induced positive inotropic effects, reduction of the contraction rate and arrhythmia, leading to cardiac arrest. In the toad, a negative inotropic effect and a reduction of the contraction rate were observed, whereas the water snake was hardly affected by either SRTX-b or ET-1. In the agama, an i.v. injection of 15 micrograms of SRTX-b caused changes in the ECG, culminating in A-V block that led to cardiac arrest, while in the toad an injection of 45 micrograms induced only transient disturbances in the ECG. Binding studies with 125I-SRTX-b revealed specific binding sites for SRTX-b and ET-1 in the heart and brain preparations of fish (tilapia and torpedo) and agama, whereas no specific binding could be demonstrated in the toad or in the snake. These results suggest that most vertebrates tested are sensitive to SRTX/ET, while the snake may possess receptors that are of a different structure.

Auditory cortex of the long-eared hedgehog (Hemiechinus auritus): II. Tuning properties.

The forebrain of hedgehogs is considered by many investigators as one of the simplest and most primitive among extant placental mammals. In a recent study we have shown that the auditory cortex of the long-eared hedgehog (Hemiechinus auritus) comprises two distinct auditory fields, which are tonotopically organized. In this study, we describe tuning properties of single cells in these two fields. Application of the Q10dB and Square Root measures for determining sharpness of tuning revealed that, although most of the cells in the more anterior field, which is considered primary, are sharply tuned, on the average they are more broadly tuned than cells in the primary auditory cortex of other mammals. In the posterior field, the distribution of narrowly and broadly tuned cells is equal. Narrowly tuned cells in both fields are equally narrow, as are the broadly tuned cells. Latencies of single cells in both fields are frequency and intensity dependent and are somewhat longer than these found in other mammals. The distribution of BFs vs. threshold intensity matches fairly well the behavioral audiogram previously described. Our findings suggest that, in spite of the view that the isocortex of hedgehogs represents a 'primitive' condition, some basic tuning properties of their auditory cortex cells are comparable to those of other mammals.

Endothelins and sarafotoxins: effects on motility, binding properties and phosphoinositide hydrolysis during the estrous cycle of the rat uterus.

The effects of four peptides of the endothelin/sarafotoxin (ET/SRTX) family on the motility of the rat uterus were examined during the different stages of the estrous cycle. ET-1, ET-3, SRTX-b and SRTX-c showed similar effects on the contraction of the uterus: a slight increase in the maximum tension of the spontaneous rhythmic contractions, a suppression of the relaxation phase of these contractions and an increase in their rate. All three effects were concentration dependent. Of the four peptides, ET-1 and SRTX-b showed the highest potency and efficacy, suggesting that among the various peptides of this family so far studied, ET-1 and SRTX-b are the two full agonists. The rank order of susceptibility of the different stages was, in most cases: proestrus greater than estrus greater than metestrus. Freshly excised diestrus uteri showed no spontaneous contractions and did not respond to any of the peptides. The binding potency of ET-1 and SRTX-b to uterine membranes was similar at the various estrous stages, but their maximal binding decreased gradually from proestrus to diestrus. All four peptides induced phosphoinositide (PI) hydrolysis in uterine slices at all four different stages, with ET-1 and SRTX-b again being more potent than ET-3 or SRTX-c. The maximal PI hydrolysis correlated with the increased rate of the rhythmic contractions. It is suggested that the reaction of the rat uterus to the ET/SRTX peptides depends on its hormonal status and that ET may act in concert with steroid hormones in the modulation of the estrous cycle.

Endothelin and sarafotoxin: influence on steroid-regulated motility of rat uterus.

The sarafotoxins (SRTX) and endothelins (ET) were shown to influence the motility of the isolated rat uterus by inducing an increase in the rate and in the maximum tension of the spontaneous rhythmic contractions and a suppression of the relaxation phase of these contractions. Ovariectomized rats, 24 weeks post-operation, show no spontaneous motility of their uteri and the SRTX/ET peptides induce only a slight tonic increase in the uterine tension. Treatment with 17 beta estradiol restores spontaneous motility and sensitivity to the SRTX/ET peptides in all three contraction modes. It is concluded that the influence of the SRTXs and ETs on uterine motility depends on the hormonal status of the animal.

Invasion of visual cortex by the auditory system in the naturally blind mole rat.

Previously we have shown that the dorsal lateral geniculate body (LGB), which is strictly visual in sighted mammals, receives a strong auditory input in the naturally blind mole rat (Spalax ehrenbergi). Here we show with the 2-deoxyglucose technique and with single-unit recordings that in this species the initially non-degenerated visual cortex, as defined by its connection with LGB, is also activated by the auditory modality. These findings suggest that cross-modal compensation may occur as a natural consequence of the degeneration of a sense organ.

Discrimination of communication calls in the squirrel monkey: "call detectors" or "cell ensembles"?

Single cells in the primary auditory cortex of the awake squirrel monkey were tested for their responses to intraspecific communication calls presented to the monkey normally ("calls") and backwards ("llacs"). These two groups of signals were similarly effective in eliciting responses, and response patterns were of the same nature and equally diverse. In about 2% of the cells the time structure of a response to at least one "llac" was virtually a "mirror image" of the response to the corresponding "call". In about 34% of the cells, for at least one vocalization, at one intensity or other, the time distribution of response peaks closely approximated in time with the envelope of a particular spectral component of the call, corresponding with the cell's best frequency. These results suggest that complex sounds may be represented in the auditory cortex by the synchronized activity of functional cell ensembles in which differently tuned individual members are distributed throughout the cochleotopic space according to their best frequencies.

Contractile effects and binding properties of endothelins/sarafotoxins in the guinea pig ileum.

Seven of the eight known isopeptides of the endothelin/sarafotoxin (ET/SRTX) family were tested on the isolated guinea pig ileum and found to cause a concentration-dependent increase in basal tone. The rate or the amplitude of the spontaneous rhythmic contractions of the ileal smooth muscle were essentially not affected by any of the peptides. The maximum contraction elicited by vasoactive intestinal contractor (VIC) was slightly stronger than that induced by endothelin-1 (ET-1) or sarafotoxin-b (SRTX-b), and significantly stronger than the maximal contractions elicited by sarafotoxin-a (SRTX-a), sarafotoxin-c (SRTX-c), or endothelin-3 (ET-3). Sarafotoxin-d (SRTX-d) caused, essentially, no contraction but a rather marked relaxation. The potencies of the various peptides to induce the increase in tension, in terms of EC50 values (cumulative effective concentrations that induce half-maximum response), ranged between 6 and 95 nM depending on the peptide. VIC, ET-1, SRTX-b and SRTX-a had similar potencies and were significantly more potent than SRTX-c and ET-3. A high concentration of SRTX-b elicited no additional response when applied to the organ bath after one of the other peptides had shown a maximal effect. Binding experiments with ileal membranes revealed similar binding properties for the various peptides. Competition with iodinated SRTX-b showed no meaningful differences between the various peptides. It is concluded that all the ET/SRTX peptides compete for the same receptor subtype in the ileum. In terms of efficacy, VIC can be considered as a full agonist of this receptor, SRTX-d is probably an antagonist, while all the other peptides behave as partial agonists.