Reversal of hypoglossal dominance in canaries following unilateral syringeal denervation

Summary The song of intact male canaries develops under the almost exclusive control of the left hypoglossus and left syringeal half. Section of the left hypoglossus or of its tracheosyringealis branch induces the right hypoglossus to assume a dominant control over vocal behavior. When this operation is done during the first two weeks after hatching the ensuing song is under sole right hypoglossal control; if done during the third and fourth week after hatching song develops under shared right-left hypoglossal control. In either case the quality of song is close to that of intact birds of comparable age. If the left hypoglossal innervation to the syrinx is cut when song development is already well under way (plastic song) or after stable adult song has been acquired, then the quality of song developed is markedly poorer than that of controls. From these observations we may conclude that: 1) Left or right hypoglossal dominance are not necessary for the production of normal canary song. 2) Soon after hatching either hypoglossus has the potential to assume a dominant role in song development. 3) The ability of the right hypoglossus to develop normal song decreases as birds master song under dominant left hypoglossal control. It is suggested that hypoglossal dominance and dominance reversal may provide convenient material to study neuronal changes related to the learning of new motor tasks.Abbreviations e.l. external labium - i.t.m. internal tympaniform membrane - ts tracheosyringealisSteven Green, Myron C. Baker and Timothy DeVoogd provided invaluable help on statistical analysis. Steven Green also read the text and made useful suggestions.     

Bilateral and unilateral antennal lesions alter orientation abilities of the crayfish, Orconectes rusticus.

Numerous animals use chemical cues within their environments to execute various behaviors. One of these behaviors is orientation to an odor source. Crayfish, in particular, can orient to food sources under a number of different conditions. It has not been determined, however, what kind of search strategy these animals employ to successfully locate a food source. To determine the role of antennae and antennules in this behavior and to investigate different modes of orientation behavior, the orientation patterns of crayfish with complete and partial antennal lesions were examined. Detailed analysis of orientation paths confirmed that crayfish could not locate odor sources with either bilateral or unilateral lesions. This suggests that crayfish are using the spatial information obtained from these appendages to successfully orient. Animals using information from the bilaterally paired appendages in the control group exhibited increased walking speed, increased speed to source and decreased heading angles towards the source compared to these measurements taken from lesioned groups. There was no significant difference in any parameters between animals with unilateral or bilateral lesions. This strongly suggests that these animals are reliant on the spatial comparison of differences between bilaterally paired olfactory appendages for successful orientation.     

Neurochemical evidence for denervation supersensitivity: the effect of unilateral substantia nigra lesions on apomorphine-induced increases in neostriatal acetylcholine levels

Unilateral lesions of the nigro-neostriatal dopaminergic projection were induced by injections of 6-hydroxydopamine into the zona compacta of the substantia nigra. This resulted in a reduction of neostriatal dopamine to less than 6 percent of the control side. Two months later intraperitoneal injections of apomorphine (1 mg/kg) produced contralateral turning and a significant increase in neostriatal acetylcholine levels. The increase was significantly greater in the neostriatum ipsilateral to the lesion than in the intact side. Haloperidol (1 mg/kg) produced a significant decrease in neostriatal acetylcholine but this decrease did not differ between the “denervated” and intact neostriata. The nigral 6-hydroxydopamine lesions did not by themselves affect neostriatal acetylcholine levels. The fact that apomorphine produces a greater increase in neostriatal acetylcholine after lesions of the dopaminergic nigro-neostriatal projection supports earlier behavioral data suggestive of denervation supersensitivity of neostriatal dopaminergic receptors after these lesions.     

Role of renin-angiotensin-aldosterone system in salt-sensitive hypertension induced by sensory denervation

To define the role of the renin-angiotensin-aldosterone system in a novel salt-sensitive model, neonatal Wistar rats were given capsaicin (50 mg/kg sc) on the first and second days of life. After weaning, male rats were divided into the following six groups and treated for 3 wk with: control + normal sodium diet (CON-NS), CON + high-sodium diet (CON-HS), CON + HS + spironolactone (50 mg x kg(-1) x day(-1), CON-HS-SP), capsaicin pretreatment + NS (CAP-NS), CAP-HS, and CAP-HS-SP. Radioimmunoassay shows that plasma renin activity (PRA) and plasma aldosterone level (PAL) were suppressed by HS, but they were higher in CAP-HS than in CON-HS and CON-HS-SP (P < 0.05). Both tail-cuff systolic blood pressure and mean arterial pressure were higher in CAP-HS than in all other groups (P < 0.05). Urine water and sodium excretion were increased with HS intake, but they were lower in CAP-HS than in CON-HS (P < 0.05). Western blot did not detect differences in adrenal AT1 receptor content. Therefore, insufficiently suppressed PRA and PAL in response to HS intake by sensory denervation may contribute to increased salt sensitivity and account for effectiveness of spironolactone in lowering blood pressure in this model.     

Pharmacomorphological study of denervation induced by 6-hydroxydopamine in Porichthys photophores

The effects of 6-hydroxydopamine (6-OHDA) on the bioluminescent response of Porichthys photophores were investigated as part of a pharmacological study of the neural control of luminescence in this fish. Subcutaneous injections of 6-OHDA induce a luminescent response similar to that of norepinephrine (NE), suggesting a sympathomimetic action. The luminescent response to electrical stimulation is almost completely and irreversibly abolished within 24 hours following low-dose treatment of the photophores with 6-OHDA, while the sensitivity of these organs to exogenous NE is increased significantly over the few days post-treatment. During this period the photophores continuously emitted a steady low-level glow. Electronmicroscopic studies of such photophores revealed progressive destruction of the nerve endings. Photophore luminescent sensitivity to NE subsequently became sub-normal, and at this stage electron microscopy revealed an increasingly larger number of damaged photocytes, supportive cells and, in one case, lens cells. From these results it is suggested that 6-OHDA initially impairs neuro-photocyte transmission by destroying catecholaminergic nerve endings. In turn, the transmitter reuptake mechanism is also impaired, thus accounting for development of supersensitive responses to exogenous NE. Subnormal luminescent responses to NE appear as a result of loss of photocyte competence due to structural deterioration. The latter are interpreted as the consequence of removal of trophic factors supplied by the photophore adrenergic innervation.Suppression of luminescent response to both electrical stimulation and exogenous NE in photophores treated with higher doses of 6-OHDA, may be due to a direct effect of this drug on the receptor sites of the photocytes.     

Mimecan is involved in aortic hypertrophy induced by sinoaortic denervation in rats

This study was designed to investigate whether mimecan was involved in aortic hypertrophy induced by sinoaortic denervation in rats. 8 weeks after sinoaortic denervation, when compared to sham-operated rats, sinoaortic denervated rats exhibited aortic hypertrophy and down-regulation of mimecan. Through classic univariate correlation analysis, it was found that mimecan mRNA was negatively related to extent of aortic hypertrophy. Treatment of primary cultured vascular smooth muscle cells with the Ang II (1 μM), which was found locally increased in the aortae of sinoaortic denervated rats, resulted in a reduction of mimecan expression. In vitro, knockdown of mimecan in vascular smooth muscle cells promoted cell proliferation induced by 15% of fetal bovine serum or Ang II (1 μM). We concluded that down-regulation of mimecan was involved in aortic hypertrophy induced by sinoaortic denervation in rats.     

Selective sympathetic denervation induced by 6-hydroxy-dopamine in the small intestine.

The structure of the nerve fibres in the intact and neurally isolated intestine of the cat was studied after 6-OHDA treatment by fluorescence and electron microscopy. A gradual disappearance of green fluorescence in the monoaminergic fibers was observed. Degeneration of nerve fibres containing 30--60 nm agranular and 30--60 nm densecore (granular) vesicles could be seen in all layers of the small intestine. Sporadically, degeneration of the presynaptic elements occurred also in the synapses of myenteric ganglia. A large number of degenerating fibres could be observed in close relation to blood vessels. These were thought to be monoaminergic because 6-OHDA selectively affected their terminals. In the chronically isolated small intestine degenerated nerve processes could not be observed following 6-OHDA treatment. It is concluded that the intrinsic nerve elements do not contain monoamines. Accordingly, the previously observed yellow fluorescence of the nerve elements of the isolated small intestine might be due to some other kind of neurotransmitter, possibly tryptamine.     

Carboxy-terminus recruitment induced by substrate binding in eukaryotic fructose bis-phosphate aldolases

The crystal structures of Leishmania mexicana fructose-1,6-bis(phosphate) aldolase in complex with substrate and competitive inhibitor, mannitol-1,6-bis(phosphate), were solved to 2.2 A resolution. Crystallographic analysis revealed a Schiff base intermediate trapped in the native structure complexed with substrate while the inhibitor was trapped in a conformation mimicking the carbinolamine intermediate. Binding modes corroborated previous structures reported for rabbit muscle aldolase. Amino acid substitution of Gly-312 to Ala, adjacent to the P1-phosphate binding site and unique to trypanosomatids, did not perturb ligand binding in the active site. Ligand attachment ordered amino acid residues 359-367 of the C-terminal region (353-373) that was disordered beyond Asp-358 in the unbound structure, revealing a novel recruitment mechanism of this region by aldolases. C-Terminal peptide ordering is triggered by P1-phosphate binding that induces conformational changes whereby C-terminal Leu-364 contacts P1-phosphate binding residue Arg-313. C-Terminal region capture synergizes additional interactions with subunit surface residues, not perturbed by P1-phosphate binding, and stabilizes C-terminal attachment. Amino acid residues that participate in the capturing interaction are conserved among class I aldolases, indicating a general recruitment mechanism whereby C-terminal capture facilitates active site interactions in subsequent catalytic steps. Recruitment accelerates the enzymatic reaction by using binding energy to reduce configurational entropy during catalysis thereby localizing the conserved C-terminus tyrosine, which mediates proton transfer, proximal to the active site enamine.     

Role for PKC-epsilon in neuronal death induced by oxidative stress

We investigated which isoforms of PKCs can be modulated and what their roles are during l-buthionine-S,R-sulfoximine (BSO)-induced neuronal death. We observed the isoform specific translocation of PKC-epsilon from the soluble fraction to the particulate in cortical neurons treated with 10 mM BSO. The translocation of PKC-epsilon by BSO was blocked by antioxidant trolox, suggesting the PKC-epsilon as a downstream of reactive oxygen species (ROS) elevated by BSO. Trolox inhibited the ROS elevation and the neuronal death in BSO-treated cortical cells. The BSO-induced neuronal death was remarkably inhibited by both the pharmacological inhibition of PKC-epsilon with epsilonV1-2 and the functional blockade for PKC-epsilon through overexpression of PKC-epsilon V1 region, suggesting the detrimental role of PKC-epsilon. These results suggest that PKC-epsilon is the major PKC isoform involved in the pathways triggered by ROS, leading to neuronal death in BSO-treated cortical neurons.     

Gender differences in behavioural changes induced by latent toxoplasmosis

There is growing evidence that the protozoan Toxoplasma gondii modifies behaviour of its intermediate hosts, including humans, where it globally infects about 20–60% of the population. Although it is considered asymptomatic in its latent stage, it was previously found to have remarkable and gender different effects on the personality factors A (warmth), G (rule consciousness), L (vigilance, mistrust) and Q3 (self-control, self-image) from Cattell’s 16PF Questionnaire. We performed a double blind experiment testing 72 and 142 uninfected men and women, respectively, and 20 and 29 infected men and women, respectively, in order to verify these gender differences using behavioural experiments. Our composite behavioural variables Self-Control and Clothes Tidiness (analogue to the 16PF factors G – conscientiousness and Q3 – self-control) showed a significant effect of the toxoplasmosis–gender interaction with infected men scoring significantly lower than uninfected men and a trend in the opposite direction in women. The effect of the toxoplasmosis–gender interaction on our composite behavioural variable Relationships (analogue to factor A – warmth) approached significance; infected men scored significantly lower than uninfected men whereas there was no difference in women. In the composite behavioural variable Mistrust (analogue to factor L), the pattern was affected by environment (rural versus urban). Possible interpretations of the gender differences are discussed.     

Nucleolar lesions induced by ribonuclease in living cultured cells

The nucleolar lesions provoked by the action of ribonuclease (RNase) on living chick embryo fibroblasts were studied by means of microcinematographic analysis and at the ultrastructural level using oxidized diaminobenzidine as a differentially contrasting stain for nucleic acids. This study has shown that the induction of nucleolar dispersion by RNase was only the beginning of a series of discrete steps. The following sequences are described: dispersion of the nucleolus into fragments, their reassembly, and the emission of spherules which appear of chromatin origin. At that step nucleoli are typically segregated. The alteration of the nucleolar associated chromatin seemed to be primordial in these processes. Moreover, the large mass of heterochromatin intimately associated with the nucleolus and which has been considered to be a part of the nucleolar organizer region apparently plays a chief part in the reassembly of the nucleolar fragments into a segregated nucleolus. Ribonuclease is compared to other drugs known to act on nucleolar DNA.     

Sex differences in oxidative stress induced by benzene in rats

Role of sex differences on oxidative stress induced by benzene has been studied in liver, kidney and lungs of rat. It was observed that benzene administration enhanced lipid peroxidation in liver, kidney and lungs of rat, nevertheless, significant variations were recorded in male and female rats. Decrease of GSH and CYTP(450)2E1 was higher in female rats than male rats except lungs. The results suggest that oxidative stress induced by benzene is higher in female rats.     

Effect of ventilation on sodium excretion in the anesthetized dog with unilateral renal denervation

We studied if the effect of mechanical ventilation induced to keep arterial blood gas values within normal physiological limits has any influence on renal sodium excretion in anesthetized dogs (n = 17) subjected to acute unilateral renal denervation. Compared to the control and the postcontrol periods, ventilation elevated arterial pO2 from 86 +/- 5 to 96 +/- 5 mmHg and blood pH from 7.37 +/- 0.02 to 7.41 +/- 0.01 while arterial pCO2 was decreased from 38 +/- 2 to 33 +/- 1 mmHg (p less than 0.05 in all cases). Compared to the innervated kidney urine flow, urinary sodium and potassium excretion from the denervated kidney were markedly elevated both during spontaneous respiration and during mechanical ventilation but GFR and cPAH were similar on the two sides. Ventilation decreased sodium excretion by the denervated kidney from 314 +/- 26 to 252 +/- 31 mumols/min/100 g k. w. (p less than 0.05). No other excretory changes were noted either in the innervated or in the denervated kidneys. Difference in sodium excretion between innervated and denervated kidneys was decreased from 209 +/- 19 to 126 +/- 20 mumole/min/100 g k. w. (p less than 0.001), due to the ventilation induced diminution of sodium excretion from the denervated kidney. It is concluded that mechanical ventilation of anesthetized dogs modifies sodium excretion, and this phenomenon can be demonstrated only in the denervated kidney.     

Preneoplastic lesions in rodent kidney induced spontaneously or by non-genotoxic agents: predictive nature and comparison to lesions induced by genotoxic carcinogens

The current literature on non-genotoxic renal carcinogens and the associated neoplastic and preneoplastic lesions has been reviewed in order to determine their occurrence and predictive nature with regard to tumor formation. In addition the mechanisms involved in the genesis of renal tumors are discussed. A more generalized classification of preneoplastic and neoplastic renal lesions was introduced, based on studies conducted with genotoxic and non-genotoxic renal carcinogens. Reports on preneoplastic lesions were found in the literature for control animals as well as animals treated with non-genotoxic carcinogens. Due to the paucity of data regarding preneoplastic lesions in control animals and animals treated with non-genotoxic carcinogens, new data were also generated by rereading kidney slides of control animals of a randomly selected NTP study and kidney slides of male rats treated with the highest dose of ochratoxin A, one of the most potent non-genotoxic renal carcinogens known. The control slides and the slides from the ochratoxin A study indicated that the cytologic and morphologic types of preneoplastic lesions characteristically observed in bioassays using genotoxic carcinogens are also present in control animals and animals treated with non-genotoxic carcinogens. The incidence of preneoplastic lesions was low in control animals and higher in animals treated with non-genotoxic carcinogens. The diverse classifications used in the literature did not allow a direct comparison of lesions and corresponding incidences with those of the newly generated data. However, three major tendencies were observed: (a) whenever a high incidence of preneoplastic lesions was reported, renal neoplasms were also found, (b) the larger the size and the further a lesion had progressed, the higher was the probability of tumor formation, and (c) not all preneoplastic lesions are irreversible, but reversibility seemed to decrease with increasing lesion size and progression. It must be emphasized that the data available for these conclusions are limited. This is not due to the lack of adequate numbers of bioassays with non-genotoxic carcinogens, but rather to the lack of consistent reporting of data. A generalized and more widely used classification which incorporates early lesions would certainly improve the current data base on renal lesions and provide future improvements in the predictive nature of these lesions.     

CHOP is involved in neuronal apoptosis induced by neurotrophic factor deprivation

Neurotrophic factors are essential for the survival of neurons. We found that the endoplasmic reticulum (ER) stress-C/EBP homologues protein (CHOP) pathway to be activated during neurotrophic factor deprivation-induced apoptosis in PC12 neuronal cells and in primary cultured neurons, and this apoptosis was suppressed in the neurons from chop(-/-) mice. In addition, we found that CHOP is expressed in the subventricular zone (SVZ) and striatum of the young adult mouse brain. The number of apoptotic cells in the SVZ decreased in chop(-/-) mice. These results indicate that the ER stress-CHOP pathway plays a role in neuronal apoptosis during the development of the brain.     

Extrinsic denervation elevates neuronal aromatic l-amino acid decarboxylase immunoreactivity in rat small intestine

Summary In order to clarify further the neural control of digestive tract function, we have compared the neuronal localization of tyrosine hydroxylase (TH) and aromatic amino acid decarboxylase (AADC) in rat small intestine. Immunoreactivity for TH was found in numerous varicose axons associated with neurons of the enteric plexuses and in axons within the circular muscular coat and the mucosal villi. Axons with AADC immunoreactivity had a similar distribution, but were sparser in the enteric plexuses and musculature than those containing TH. Chronic extrinsic denervation of a segment of intestine removed all TH-positive nerves from that region. By contrast, the intensity of AADC immunoreactivity was enhanced and more AADC-positive axons were visible than in adjacent intact areas of intestine. The AADC-positive axons appear to represent the intrinsic amine-handling neurons rather than intrinsic tryptaminergic neurons or extrinsic dopaminergic neurons, and the effect on AADC activity of removing the extrinsic nerve supply suggests that this normally exerts some restraining influence on the metabolism of the amine-handling population.