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PubMed1994

[Effect of xylazine on acetylcholine content of the hippocampal CA1 region in urethane anesthetized rats].

By using acetylcholine selective microelectrode technique, a biphasic effect of xylazine on acetylcholine content of the hippocampal CA1 region in urethane anesthetized rats was observed as evidenced by the fact that small doses of xylazine (2.0 and 6.0 mg.kg-1) significantly increased, while higher dose (10.0 mg.kg-1) decreased the acetylcholine content. Though idazoxan (0.6 mg.kg-1) significantly antagonized the effect of xylazine when used in combination, the acetylcholine content in the hippocampal CA1 region of urethane anesthetized rats was still much lower than that of the control group. In rats whose nucleus locus coeruleus was chemically lesioned, xylazine (2.0 and 6.0 mg.kg-1) significantly decreased the acetylcholine content, and idazoxan (0.6 mg.kg-1), which increased the acetylcholine content in the hippocampal CA1 region when used alone, completely antagonized the effect of xylazine. These results suggest that the biphasic effect of xylazine and the lower than normal acetylcholine content observed when idazoxan was used to antagonize xylazine induced changes in acetylcholine content of the hippocampal CA1 region in urethane anesthetized rats might be related to the effects of xylazine and idazoxan on the central noradrenergic neurotransmitter system.

Acetylcholine↗
PubMed1963

Homograft tolerance in mice: use of urethan and sublethal irradiation.

Adult mice [CBA/J (H-2(k))], which received either a single sublethal dose of x-radiation (500 rad) or urethan plus 500 rad, were given intravenous injections of C3H/HeJ (H-2(k)) spleen or bone marrow cells (18 to 42 x 10(6) cells per mouse) or both, for 3 days. C3H/HeJ tail-skin homografts were retained (over 130 days) by these mice, whereas BALB/cJ (H-2(d)) homografts all were rejected within 33 days. Similarly irradiated or urethan-treated controls (or controls treated with a combination of both), which did not receive C3H cells, rejected both homografts. Specific homograft tolerance is induced in adult mice by this procedure.

Allografts↗
PubMed1996

Urethane-protected alpha-amino acid N-carboxyanhydrides and peptide synthesis.

The preparation, physical properties and analytical data are reported for seventy urethane-protected (Boc, Cbz, FMOC) amino acid N-carboxyanhydrides (UNCAs). Most of the UNCAs are crystalline and the X-ray diffraction patterns for several of these are described. UNCAs are stable to routine laboratory manipulations and can be stored for extended periods of time (1-2 years at below 0 degrees C). Most are completely stable to the conditions commonly employed for peptide synthesis. The correct choice of base is key for the successful introduction of urethane protecting groups into NCAs. N-Methylmorpholine is used for the introduction of FMOC, Cbz or Boc from the chloroformates, and pyridine is used for the introduction of the Boc group from Boc anhydride. UNCAs represent a unique class of preactivated, isolable and stable amino acid derivatives that generate no side products or co-products, other than CO2, during condensation reactions. The application of UNCAs in peptide synthesis in both solid phase and in solution is reviewed in detail.

Amino Acid Sequence↗
PubMed1994

Effects of perforant path procaine on hippocampal type 2 rhythmical slow-wave activity (theta) in the urethane-anesthetized rat.

Previous research has suggested that the entorhinal cortex plays a major role in the production of type 1 rhythmical slow-wave activity (RSA) recorded in the hippocampus of the freely moving preparation. In the present experiment we investigated the contribution of the entorhinal cortex to the type 2 fields recorded under urethane anesthesia. Rats had stimulating electrodes and cannulae filled with procaine positioned in the perforant pathway of one or both hemispheres. Recording electrodes were positioned in the dorsal hippocampus of each hemisphere to record perforant path and commissural/associational evoked potentials and RSA fields. Following unilateral procaine blockade, a decrease in RSA amplitude was observed in the stratum oriens and fissure regions of both hemispheres. Concomitant with this change in RSA, there was a loss of perforant path evoked responses, although commissural/associational control potentials remained unaltered. A greater reduction in RSA amplitude was observed following bilateral procaine microinfusion. RSA phase reversal also occurred more dorsally in microelectrode depth profiles conducted through the hippocampus during perforant path inactivation. In current source density analyses performed under baseline conditions, large rhythmic sinks were observed in stratum oriens, in stratum radiatum, and in strata adjacent to the hippocampal fissure. A rhythmic source was often observed in stratum pyramidale. Following perforant path inactivation decreases in the magnitude of the phasic sinks located near the fissure and stratum radiatum were observed. In contrast to the reduction in RSA amplitude observed in the stratum oriens region, the sink in this region and the source in stratum pyramidale remained relatively unaltered. These results demonstrate that the entorhinal region contributes to the production of RSA observed under urethane anesthesia. Furthermore, the CSD and amplitude changes following perforant path inactivation suggest that a substantial portion of RSA recorded in stratum oriens may result from ventrally located RSA dipoles.

Anesthesia↗
PubMed1989

Activation of the cellular Harvey ras gene in mouse skin tumors initiated with urethane.

Mouse skin tumors, benign papillomas, and squamous cell carcinomas (SCCs) were initiated by a single topical application of urethane followed by repeated promotion with 12-O-tetradecanoylphorbol-13-acetate (TPA). Using the NIH 3T3 focus forming assay, dominant transforming activity was detected in DNA isolated from SCC samples. Rearranged and amplified copies of the c-Ha-ras gene were detected in NIH 3T3 transformant cell lines, indicating that an activated Ha-ras gene had been transferred to the NIH 3T3 recipient cells. Analysis of p21ras from the transformant cell lines suggested that the activating ras mutation was present in codon 61. Ultimately, the Ha-ras gene was shown to be activated by a specific A----T transversion at the second position of codon 61. This mutation was detected in both benign papillomas and SCCs, suggesting the activation occurred early in tumor development. The results demonstrate a highly consistent activation of the Ha-ras oncogene by a specific point mutation, suggesting a functional role for an activated ras gene in the initiation of mouse skin tumors by urethane.

Animals↗
PubMed2003

Effects of a selective metabotropic glutamate receptor agonist on the micturition reflex pathway in urethane-anesthetized rats.

AIMS: To determine a possible role of metabotropic glutamate receptors in the spinobulbospinal micturition reflex pathway in the rat. MATERIALS AND METHODS: A selective metabotropic glutamate receptor agonist, trans-(+/-)-1-amino1,3-cyclopentanedicarboxylic acid (trans-ACPD) was administered to the lumbosacral spinal cord via an intrathecal catheter in urethane anesthetized rats. Amplitude of reflex bladder contractions evoked by bladder distension under isovolumetric condition as well as amplitude of bladder contractions elicited by electrical stimulation of the pontine micturition center (PMC) were examined before and after administration of trans-ACPD. The effect of trans-ACPD on the urethral activity during isovolumetric bladder contractions was also examined by monitoring urethral perfusion pressure and electromyography of the external urethral sphincter (EUS-EMG). RESULTS: Trans-ACPD (3-10 microg) completely inhibited reflex bladder contractions evoked by bladder distension and the duration of inhibition was dose dependent (3 microg: 11.4 +/- 2.8 min, 5 microg: 13.2 +/- 1.3 min, 10 microg: 36.2 +/- 2.4 min). The mean amplitude of bladder contractions evoked by electrical stimulation of the PMC was reduced to 12.6 +/- 2.3% of control by 10 microg of trans-ACPD. In addition, bursting activity of EUS-EMG and corresponding high frequency oscillations of urethral pressure during isovolumetric bladder contractions were completely abolished by 10 microg of trans-ACPD. CONCLUSIONS: These results indicate that intrathecal administration of a selective metabotropic glutamate receptor agonist to the lumbosacral spinal cord has an inhibitory effect on the spinobulbospinal micturition reflex pathway in urethane-anesthetized rats. This pharmacological action is attributed at least to the inhibitory effect on the descending pathway from the PMC to the lumbosacral spinal cord.

Anesthesia, Intravenous↗
PubMed1977

Digital anomalies of mouse limbs induced by treatment with urethan in vitro.

The effects of urethan on ICR/JCL mouse limbs were examined in forelimbs of day-11 embryos cultured in concentrations of 0, 50, and 100 mug/ml for one to four days, and in forelimbs of day-12 embryos and in fore- and hind limbs of day-13 embryos in 0, 10, 50, and 100 mug/ml for one to four days. Urethan did not cause digital malformations on day-11 limbs but induced syndactyly in day-12 forelimbs and polydactyly in day-12 forelimbs and day-13-fore- and hind limbs. The polydactyly may have been due to effects on mesenchymal cells producing localized cell proliferation, whereas syndactyly may have been due to effects on precartilage cells, resulting in excess migration of mesenchymal cells toward the digits, leaving the connective tissue at the interdigital space to develop inadequately.

Abnormalities, Drug-Induced↗
PubMed1997

Urban air pollution enhances the formation of urethane-induced lung tumors in mice.

This paper investigates the association between air pollution and lung neoplasia in an animal model. The experimental exposures were done in two locations with different air pollution profiles: a polluted area (downtown São Paulo) and a "clean" environment (Atibaia). Swiss mice were employed and urethane (3 g/kg) was used as carcinogenic substance. Two experiments were performed: Experiment I was designed to verify whether air pollution acts as initiator and/or promoter of lung cancer, using 300 mice; Experiment II employed 250 animals and aimed to verify if the effects of air pollution on the development of lung tumors was dose dependent. A significant effect of air pollution in augmenting lung carcinogenecity induced by urethane was observed. This effect was shown to be dose-dependent and reproducible on two different occasions. In addition, morphometric studies revealed that pollution may influence tumor phenotype. These results support the hypothesis that air pollution plays a significant role in the development of lung tumors.

Air Pollutants↗
PubMed1999

Preconception urethane or chromium(III) treatment of male mice: multiple neoplastic and non-neoplastic changes in offspring.

Increase in neoplasia in offspring after preconception exposure of parents presents puzzling features such as high frequency of effects and lack of Mendelian inheritance. The present study examined the hypothesis that preconception carcinogenesis involves an increase in the rate of occurrence of neoplasms with a spontaneous incidence. Male NIH Swiss mice (12 per group) were exposed 2 weeks before mating (once, ip) to urethane (1.5 g/kg) or chromium(III) chloride (1 mmol/kg). Offspring (48-78/sex/group) were examined for all grossly apparent changes when moribund or at natural death, followed by histopathological diagnosis and statistical analysis. Significant exposure-related changes occurred in multiple organs. Ten to 20 percent of offspring showed changes related to paternal exposure, including at least one sired by most treated males. Pheochromocytomas occurred in both male and female offspring after both treatments, with none in controls. These neoplasms are rare in mice and suggest endocrine dysfunction as a component of preconception carcinogenesis. This was supported by increases in thyroid follicular cell and Harderian gland tumors, ovarian cysts, and uterine abnormalities. Lung tumors were increased in female offspring only. Effects seen in offspring only after paternal urethane exposure were an increase in preneoplasia/neoplasia in the glandular stomach (males) and in females, increased lymphoma but decreased incidence of histiocytic sarcoma. Increases in incidence of male reproductive gland tumors and of renal non-neoplastic lesions occurred only after chromium exposure. Thus, preconception exposure of fathers to toxicants had a significant impact on both neoplastic and non-neoplastic changes in almost all tissues in which these lesions often occur naturally during the aging process.

Adenoma↗
PubMed1977

Inhibition of choline incorporation into brain lipids in rats by urethane, a proposed mechanism of depression of the central nervous system.

Concentrations and specific radioactivities of choline, acetylcholine, phosphorylcholine, lipid choline, and sn-glycero-3-phosphorylcholine after i.v. injection of methyl-14C-choline were measured in the brain of untreated controls and of rats anesthesized with urethane. The specific activity was found to be decreased during deep anesthesia by 40% in acetylcholine, 20-30% in phosphorylcholine, 50-75% in lipid choline, and 30-40% in sn-glycero-3-phosphoryl-choline. No significant change was detected in the specific activity of choline. The brain concentration of acetylcholine was increased by 40%, the concentration of sn-glycero-3-phosphorylcholine, however, was diminished by 10% during anesthesia. No change was found in the concentration of the other choline containing compounds investigated. Measuring choline incorporation into 4 subcellular fractions of brain tissue specific activities were found to be decreased by the same percentage, although 2 fractions (nuclei and microsomes) were higher labelled than the 2 other fractions (crude mitochondria with synaptosomes and lysosomes). A correlation between the biochemical and the functional alterations is supported by the dose-effect relationships on both parameters. It is suggested that urethane reduces turnover of lipids and by that mechanism inhibits the exocytotic release of the transmitter from presynaptic nerve endings.

Animals↗
PubMed1977

Compared effects of N-hydroxyurethan, urethan and hydroxyurea on DNA synthesis. In vivo and in vitro studies.

The effect of N-hydroxyurethan (HUR) on DNA synthesis has been tested both in vivo on various tissues and in vitro on concanavalin A (ConA)-stimulated rat thymocytes and compared with the action of urethan and hydroxyurea. HUR suppresses scheduled DNA synthesis, except that of non-stimulated spleen cells in vitro. The inhibition is efficient and rapid and takes place immediately if the drug is administered at the peak of the S-phase. UR inhibits DNA synthesis in vitro only at much higher doses and with different time course. It is effective or slightly effective if it is administered at the peak of the S-phase. A conversion of urethan into HUR the latter depressing DNA synthesis could partly explain the differences observed. No toxicity was found after treatment with drugs at the concentration employed. Finally, the relationships between drug doses and cell responses have been particularly observed in vivo.

Animals↗
PubMed1984

Hippocampal excitability related to the phase of theta rhythm in urethanized rats.

Urethanized rats were studied to determine if excitability in the hippocampus is related to the phase of the theta rhythm. The responses recorded from area CA1 were clearly modulated at the theta frequency. Those recorded from the dentate region were not. The most excitable phase for CA1 differed significantly from that previously reported for walking rats. These findings suggested that the mechanisms for theta production differ for normal and urethanized rats.

Animals↗
PubMed1986

Laminar distribution of hippocampal rhythmic slow activity (RSA) in the behaving rat: current-source density analysis, effects of urethane and atropine.

This study investigated the laminar distribution of rhythmic slow wave activity (RSA) in the dorsal hippocampus of the rat during running. Depth analyses of field EEG were performed by stepping the recording electrode in 82.5 micron increments and sampling RSA at each depth. One-dimensional current-source density (CSD) was calculated from the RSA profiles to enhance spatial resolution of current sources and sinks. Laminar analysis of power, coherence, and phase of RSA with respect to a stationary electrode in the stratum oriens of CA1 was performed with spectral methods. RSA waves in the CA1-dentate axis had power maxima at about the hippocampal fissures, hilus, outer molecular layer of the endal leaf of dentate gyrus and stratum oriens of CA1, in that order. A gradual shift of phase occurred in stratum radiatum of CA1. Large phase-shifts were found in both the endal and ectal leaves of the fascia dentata. A null zone and associated sudden phase-reversal of RSA were observed in stratum lucidum of CA3. Multiunit activity showed phase-locked modulation with RSA in the granule cell layer of the dentate gyrus and pyramidal cell layer of CA1, CA3, and subiculum. CSD analysis in the CA1-dentate axis revealed multiple source-sink pairs. The sinks and sources showed cyclic changes with RSA, and were attributed to the rhythmic, but time-shifted, activity of hippocampal afferents from the septum and entorhinal cortex. The gradual phase-shift in CA1, and the configurational changes of RSA waves with depth, are explained by the summation of extracellular currents produced by time-delayed sink-source pairs (RSA dipoles). When the cholinergic septohippocampal path was blocked by atropine a null zone in the middle of stratum radiatum of CA1 occurred and the phase-shift of RSA became steeper. Under urethane anesthesia a null zone was present in the inner stratum radiatum associated with a sudden phase-reversal of RSA. Urethane reduced the power of RSA in the hilus and decreased the firing rate of the granule cells. It is suggested that field RSA is produced by several rhythmical dipoles along the somadendritic surface of pyramidal cells and granule cells and the spatiotemporal relations of the individual dipoles determine the actually observed extracellular RSA.

Anesthesia, General↗
PubMed1987

Hippocampal rhythmical slow activity following ibotenic acid lesions of the septal region. I. Relations to behavior and effects of atropine and urethane.

The effects of intraseptal injections of various concentrations of ibotenic acid on hippocampal electrical activity were studied in freely moving and urethane-anesthetized rats. Ibotenic acid selectively abolished the atropine-sensitive form of hippocampal rhythmical slow activity (RSA) normally seen during urethane anesthesia. Large amplitude irregular activity (LIA) and RSA in the waking state were somewhat depressed as well. Despite this, clear RSA persisted in the waking state in association with locomotion or struggling (Type 1 behavior). As in normal rats, such RSA was resistant to systemic administration of atropine. Analysis of brain sections stained with gallocyanin or for acetylcholinesterase showed that ibotenic acid produced cell loss in the dorsal lateral septal nucleus and the septohippocampal nucleus. Cells in the medial septal and diagonal band nuclei were resistant to ibotenic acid. The results suggest that intrinsic septal circuitry is critically involved in the generation of the atropine-sensitive (presumably cholinergic) form of RSA. The mechanisms by which LIA and the two forms of RSA are generated in the hippocampus is discussed.

Animals↗
PubMed1989

Detection of an atropine-resistant component of the hippocampal theta rhythm in urethane-anesthetized rats.

An important pharmacological feature of the hippocampal theta rhythm in urethane-anesthetized animals is its apparent sensitivity to antimuscarinic drugs. This sensitivity may be partly due to a masking of the theta frequency by increases in both higher and lower frequency EEG components that are unrelated to any residual theta rhythm. The discovery of atropine-resistant, rhythmic medial septal neurons has provided a physiological trigger for averaging EEG and unit activity after large atropine doses. Such averaging has permitted the detection of an atropine-resistant component of the hippocampal theta rhythm in urethane-anesthetized rats. The postatropine theta activity recorded from both CAl (superficial to the pyramidal cell layer) and dentate (near the hippocampal fissure) in 15 rats was typically reduced in amplitude, but the recordings from the two locations maintained their phase relations to the septal units and to each other. The presence of this residual theta component after doses as large as 100 mg/kg indicates that it cannot be mediated by muscarinic cholinergic receptors. The coupling of the signal to the atropine-resistant septal cells strengthens our previous suggestion that these septo-hippocampal neurons are not cholinergic, and are therefore probably GABAergic.

Anesthesia↗
PubMed1990

Spontaneous single cell discharge in rat somatosensory cortical slices and its relationship to discharge in the urethane-anaesthetized rat.

A slice preparation of rat somatosensory cortex has been used to observe the nature of spontaneous single cell discharge in layer V. On the basis of their discharge pattern, cells recorded using extracellular microelectrodes fell in to one of two categories: 70% of cells discharged single action potentials separated by relatively constant time intervals, while the remainder discharged in bursts of 2-7 action potentials. The firing patterns and the range of discharge frequencies observed were similar to those recorded in the same region and layer of the cortex in urethane-anaesthetized rats. However, unlike cells recorded in vivo, the majority of cells in slices never changed their discharge pattern. This suggests that while these layer V cells in cortical slices have the basic ability to discharge spontaneously, the potential for switching between discharge patterns requires circuitry absent from this isolated preparation. Urethane, over the dose range 6-24 mM, caused a dose-dependent decrease in the discharge frequency of cells with a regular firing pattern but did not affect their basic pattern of discharge.

Anesthesia, General↗
PubMed1992

Vasopressin perfusion within the medial amygdaloid nucleus attenuates prostaglandin fever in the urethane-anaesthetized rat.

The antipyretic effect of arginine vasopressin (AVP) introduced into the ventral septal area (VSA) by push-pull perfusion was investigated in the urethane-anaesthetized rat. In addition, experiments were carried out to determine whether AVP could suppress fever when similarly perfused within the medial amygdaloid nucleus (meA). During push-pull perfusion of artificial cerebrospinal fluid within the VSA or meA, PGE1 injected intracerebroventricularly evoked fevers with respective magnitudes of 1.3 +/- 0.2 degrees C and 1.4 +/- 0.3 degrees C above baseline. Perfusion of AVP (6.5 micrograms/ml) within the VSA had significantly reduced the magnitude of PGE1 fever to 0.3 +/- 0.3 degrees C above baseline, while having no significant effect on afebrile colonic temperature. Perfusion of AVP (6.5 micrograms/ml) within the meA had significantly attenuated the magnitude of PGE1 fever to 0.7 +/- 0.2 degrees C above baseline, while having no significant effect on afebrile colonic temperature. These results support further the utility of the urethane-anaesthetized rat model for future investigations of the central control of fever and antipyresis. In addition, these data are consistent with the hypothesis that AVP may act within the meA as an endogenous antipyretic.

Alprostadil↗
PubMed1992

The origin of rhythmic fast subthreshold depolarizations in thalamic relay cells of rats under urethane anaesthesia.

Intracellular recordings were performed in relay neurons of the dorsal thalamus in rats under urethane anaesthesia. In 77 out of 127 neurons of the ventro-posterolateral and ventral lateral nuclei, but not in neurons of the ventro-posteromedial and posterior nuclei, a highly rhythmic pattern of subthreshold depolarizations was present at rest. The average frequency of these rhythmic depolarizations in ventro-posterolateral cells was 23.36 +/- 11.48 Hz (range: 6-60 Hz); in ventral lateral relay cells higher frequencies were observed (65.86 +/- 17.42 Hz; range: 17-95 Hz). The rhythmic subthreshold events were identified as excitatory postsynaptic potentials generated by the regular firing of prethalamic afferents located in dorsal column and deep cerebellar nuclei. Indeed, in cells of the ventro-posterolateral nucleus these spontaneous potentials had a waveform similar to that of synaptic potentials triggered by somatosensory stimulation. They increased in amplitude with membrane hyperpolarization and their rhythmic occurrence was not affected by the injection of large inward currents. Moreover, they persisted after capsular transection, but they could no more be recorded in ventro-posterolateral cells after lesion of dorsal column nuclei. Finally, it was found that prethalamic afferents within the deep cerebellar nuclei discharged spontaneously in a rhythmic manner within the same frequency band as that of the rhythmic synaptic potentials recorded in ventral lateral cells. On the basis of these results, it is concluded that the rhythmic subthreshold depolarizations observed in thalamic neurons of animals under urethane anaesthesia are not generated intrinsically but that they represent excitatory postsynaptic potentials of prethalamic origin.(ABSTRACT TRUNCATED AT 250 WORDS)

Anesthesia↗