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G H Jones

Publications and source records attributed to G H Jones.

At least 37 records · Page 2Linked to original sources

Comparative analysis of crossover exchanges and chiasmata in Allium cepa x fistulosum after genomic in situ hybridization (GISH).

Genomic in situ hybridization (GISH) successfully differentiated homoeologous genomes in the inter-specific hybrid Allium cepa x fistulosum, thus allowing the detection of reciprocal crossover events as label exchanges in separating anaphase I chromosomes. Three of the eight chromosome pairs were positively identified by fluorescence in situ hybridization (FISH) to rDNA sequences. There was a general similarity of the GISH-based label exchange frequencies and metaphase I chiasma frequencies, but with a 20% deficit of chiasmata. Reasons for this apparent deficit are discussed. The locations of chiasmata and label exchanges are in broad agreement.

Allium↗

Cytological characterization of four meiotic mutants of Arabidopsis isolated from T-DNA-transformed lines.

A secondary screen of the Feldmann collection of T-DNA transformed Arabidopsis lines identified several meiotic mutants. We used a spreading technique combined with DAPI staining in a detailed cytogenetic analysis of meiotic chromosome behaviour in four of these mutants, all of which are putatively T-DNA tagged and therefore candidates for molecular and functional analysis of the mutated genes. Two of them are defined as 'synaptic' mutants, showing greatly reduced association of homologous chromosomes at metaphase I: one is asynaptic, showing failure of synapsis during prophase I, whereas the other is desynaptic and is characterized by normal but non-maintained synapsis. Another mutant is defective in meiotic cell cycle control and undergoes a third meiotic division, resembling a second division but without an additional round of chromosome duplication. A further mutant shows meiosis-limited chromosome disruption, resulting in extensive chromosome fragmentation combined with other defects. All four mutants experience very irregular chromosome distribution during the meiotic divisions, resulting in abnormal numbers and/or sizes of microspores, with resulting reduced fertility.

Arabidopsis↗

Chiasma interference is blind to centromeres.

It has long been assumed, since the early works of Mather, that the centromere plays a central role in chiasma position determination, so much so that in all sequential models chiasma determination was supposed to start or finish at this point. More specifically, it has been assumed that the centromere acts as a barrier to the transmission of interference, so that a chiasma in the vicinity of a centromere would not affect the probability of chiasma formation across at this point. Some statistical analyses seemed to ratify this supposition. However, a reassessment of the literature led us to the conclusion that the statistical analyses that were not flawed were consistent in showing that interference may act across the centromere. Using large sets of chiasma data from the grasshoppers Leptysma argentina and Chorthippus brunneus and applying statistical approaches that involved either the calculation of coincidence or correlating the distances between the centromere and the nearest chiasma in either arm, it is concluded: 1. that interference acts across the centromere; 2. that the action of interference is not changed by the presence of an intervening centromere.

Animals↗

Sigma-E is required for the production of the antibiotic actinomycin in Streptomyces antibioticus.

The phsA gene encodes phenoxazinone synthase (PHS), which catalyses the penultimate step in the pathway for actinomycin biosynthesis in Streptomyces antibioticus. The phsA promoter strikingly resembles a putative Streptomyces sigma E cognate promoter, and purified E sigma E holoenzyme transcribed the phsA promoter in vitro. However, the phsA promoter was still active in an S. antibioticus sigE null mutant and the level of PHS activity was unaffected. Despite this, disruption of sigE blocked actinomycin production completely. The loss of actinomycin production correlated with a 10-fold decrease in the activity of actinomycin synthetase I, the enzyme which catalyses the activation of the precursor of the actinomycin chromophore.

Amino Acid Sequence↗

A light microscopic atlas of meiosis in Arabidopsis thaliana.

An atlas of meiosis in Arabidopsis thaliana, encompassing all stages from preleptotene to telophase II and early microspore formation, is presented in detail for the first time. The photomicrographs and descriptions are based on staining with the DNA fluorochrome 4',6-diamidino-2-phenylindole (DAPI) combined with a spreading procedure, or haematoxylin-iron alum (HIA) staining. Despite previous reservations about the practicality of cytogenetic meiotic analysis in Arabidopsis due to its small genome size, good-quality and clearly analysable preparations of all meiotic stages were obtained. This atlas of normal, wild-type meiosis is considered an essential prerequisite to informed analyses of meiotic mutants. Furthermore, extended prophase I chromosomes, particularly at the pachytene stage, offer considerable potential for producing a detailed cytogenetic map (karyotype) of Arabidopsis chromosomes with the additional prospect of high-resolution physical mapping based on fluorescence in situ hybridization (FISH) of defined DNA probes to these extended chromosomes.

Anaphase↗

Guanosine pentaphosphate synthetase from Streptomyces antibioticus is also a polynucleotide phosphorylase.

The gene for the enzyme guanosine pentaphosphate synthetase I (GPSI) from Streptomyces antibioticus has been cloned and sequenced. The cloned gene functioned as a template in the streptomycete coupled transcription-translation system and directed the synthesis of a protein with the properties expected for GPSI. Sequencing of the cloned gene identified an open reading frame of 740 amino acids whose amino terminal sequence corresponded to the N terminus of purified GPSI. The GPSI protein sequence was found to possess significant homology to polynucleotide phosphorylase from Escherichia coli. Indeed, like E. coli polynucleotide phosphorylase, purified GPSI was shown to catalyze the polymerization of ADP and the phosphorolysis of poly(A). However, the E. coli enzyme was unable to catalyze the synthesis of guanosine pentaphosphate under conditions in which GPSI was highly active in that reaction. Overexpression of the cloned gpsI gene in E. coli led to an increase in both polynucleotide phosphorylase and guanosine pentaphosphate synthetase activities in the cloning host. The polynucleotide phosphorylase activities of GPSI and of the E. coli enzyme were strongly inhibited by dCDP, but the pppGpp synthetase activity of GPSI was not inhibited and indeed was slightly stimulated by dCDP. These results strongly support the identity of GPSI as a bifunctional enzyme capable of both pppGpp synthesis and polynucleotide phosphorylase activities.

Amino Acid Sequence↗

Do chiasmata disappear? An examination of whether closely spaced chiasmata are liable to reduction or loss.

The questions of whether closely spaced crossovers could be misidentified as single chiasmata or could cancel out each other and whether exchange could occur without chiasma formation were examined in Locusta migratoria. Monochiasmate bivalents that showed differential sister chromatid staining following bromodeoxyuridine incorporation were screened for their patterns of label distribution. Half of the chiasmata were associated with an exchange between dark and lightly stained chromatids, as expected if recombination involved any two non-sister chromatids chosen at random. Two variant types of label distribution were also seen: approximately 10% of all monochiasmate bivalents had an anomalous distribution of dark and light chromatids around the chiasma, and in three of the 1365 bivalents screened a second type of anomalous pattern was observed for the first time, in which all four chromatids had a label exchange at the chiasma. The observed incidence of the latter was considerably less than expected if they originated through a four-strand double crossover with closely positioned exchanges. Analysis of label distribution patterns in monochiasmate bivalents did not produce evidence of the other configurations expected if two closely spaced exchanges could be misidentified as single chiasmata or could cancel one another out so as not to form a chiasma. We conclude that analysis of chiasma frequencies and distributions offers an accurate means of assessing recombination in organisms with favourable cytology.

Animals↗

Meiosis in the leek (Allium porrum L.) revisited. I. Prophase I pairing.

Meiotic chromosome pairing of the tetraploid species Allium porrum, the cultivated leek, was analysed by electron microscopy of 83 surface-spread nuclei in the late zygotene to early diplotene interval of prophase I, from four different varieties. Prophase I quadrivalent frequency, at 71%, marginally but significantly exceeds the two-thirds expected on a simple random end-pairing model, suggesting that more than two autonomous pairing sites occur, in some tetrasomes at least. This pattern of synaptic behaviour is consistent with an autotetraploid status, but comparison with other tetraploids, including other Allium species, indicates that Allium porrum may be a weak segmental allopolyploid displaying limited preferential homologous pairing. The incidence of pairing partner switches (PPSs) in prophase I quadrivalents is relatively low; 90% of all analysed quadrivalents had only one or two PPSs. The positional distribution of PPSs along quadrivalents was distinctly uneven with peaks in mid-chromosome arms and reduced frequencies around centromeres and near the ends. The four different varieties of leek analysed were remarkably similar in their meiotic behaviour despite their diverse breeding history, but individual plants within varieties displayed substantial variation in quadrivalent and PPS frequencies.

Allium↗

Double dissociation between the effects of muscarinic antagonists and benzodiazepine receptor agonists on the acquisition and retention of passive avoidance.

Both muscarinic antagonists, such as scopolamine, and benzodiazepine receptor (BZR) agonists, such as diazepam, produce a reliable impairment in the performance of one trial passive avoidance. Such deficits are frequently interpreted as drug-induced amnesia. However, these deficits could also result from a learning impairment. The present experiments compared the effects of two BZR agonists, lorazepam (0, 0.125, 0.25, and 0.375 mg/kg, IP) and diazepam (0, 0.78, 1.56, and 3.13 mg/kg, IP) with the effects of two muscarinic antagonists, scopolamine (0, 0.6, 0.8 and 1.0 mg/kg, SC) and atropine (0, 15, 30 and 60 mg/kg, IP) on a multiple trial passive avoidance task. In this procedure, the rats were trained with a 5-min inter-trial interval until a learning criterion was achieved. Retention was assessed 24 h later. This enabled the effects of the drugs on the acquisition and the retention of a passive avoidance response to be dissociated. Both atropine and scopolamine produced a marked impairment in the acquisition of the passive avoidance response, but did not impair retention. In contrast, diazepam and lorazepam did not alter the acquisition of a passive avoidance response, but did produce a dose-dependent impairment of retention. These results therefore demonstrate a double dissociation between the effects of muscarinic antagonists and BZR agonists on the acquisition and retention of passive avoidance.

Animals↗

Effects of benzodiazepine receptor partial inverse agonists in the elevated plus maze test of anxiety in the rat.

The present series of experiments examined the effects of five benzodiazepine receptor (BZR) partial inverse agonists on the behaviour of rats on an elevated plus maze. The drugs were tested in a standard plus maze with 3-cm walls added to the open arms, as this has been shown to increase the sensitivity of the plus maze to anxiogenic-like drugs effects (Jones and Cole 1995). The drugs tested were FG 7142 (0-100 mg/kg), beta-CCE (0-30 mg/kg), ZK 132,556 (0-100 mg/kg), ZK 90 886 (0-30 mg/kg) and Ro 15-4513 (0-30 mg/kg). In addition, to allow a comparison with previous studies, the effects of three reference substances, DMCM (0-2.5 mg/kg), pentylenetetrazol (PTZ; 0-30 mg/kg) and yohimbine (0-5 mg/kg), were also examined. These three reference compounds produced a dose-dependent reduction in the duration of open arm exploration and the total number of open arm entries, indicative of anxiogenic-like effects. DMCM produced significant effects at the doses of 1.25 and 2.5 mg/kg, PTZ at 30 mg/kg, and yohimbine at 5 mg/kg. The BZR partial inverse agonist FG 7142 (10, 30 and 100 mg/kg) also reduced the duration of open arm exploration and the total number of arm entries. The minimally effective dose resulted in a receptor occupancy of approximately 80%. Ro 15-4513 also produced anxiogenic-like effects, but only at a dose (30 mg/kg) that resulted in a receptor occupancy of approximately 95%.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Nucleotide sequence, transcriptional analysis, and glucose regulation of the phenoxazinone synthase gene (phsA) from Streptomyces antibioticus.

The nucleotide sequence of a 2.3-kb SphI fragment containing the structural gene (phsA) for phenoxazinone synthase (PHS) of Streptomyces antibioticus was determined. The sequence was found to contain an open reading frame (ORF) with a G+C content of 71.5% oriented in the direction of transcription that was confirmed by primer extension. The ORF encodes a protein with an M(r) of 70,223 consisting of 642 amino acids and is preceded by a potential ribosome-binding site. The codon usage pattern is in agreement with the general pattern for streptomycete genes, with a 92.5 mol% G+C content in the third position. The N-terminal sequence of the mature PHS subunit corresponds exactly to that predicted from the nucleotide sequence. Neither ATG nor GTG initiator codons were identified for the protein. However, a TTG codon was located near the amino terminus of the mature protein and is a good candidate for the initiator codon. The transcriptional start point of phsA was located 36 bp upstream of the start codon by primer extension. The -10 region of the putative promoter showed some similarity to the consensus sequence for the major class of prokaryotic promoters, but the -35 region was less similar. Comparison of the primary amino acid sequence of PHS of S. antibioticus with other amino acid sequences indicated that PHS is a blue copper protein with copper binding domains in the N-terminal and C-terminal regions of the polypeptide chain. A BsrBI fragment containing the promoter region of phsA and a portion of the ORF was shown to promote xylE expression when cloned in the streptomycete promoter probe vector pIJ2843. This phsA promoter-dependent xylE expression could be repressed by glucose in S. antibioticus when the organism was grown on glucose or galactose plus glucose. Thus, the cloned promoter region appears to contain the sequences responsible for catabolite repression of PHS production.

Amino Acid Sequence↗

Individual differences in schedule-induced and conditioned behaviors.

Previous experiments have shown that subjects which exhibit a high locomotor response to novelty (HR) also show a greater locomotor response to psychomotor stimulants than subjects which have a low locomotor response to a novel environment (LR). The current experiments were designed to examine in more detail the behavioral differences between HR and LR rats in non-drug paradigms. In the first experiment HR rats acquired schedule-induced polydipsia (SIP) more readily than LR rats. Panel pressing to gain access to the food pellets, however, was greater in LR rats compared to HR rats, especially after stable levels of SIP had been attained. In the second experiment one group of rats were fed daily after a 30-min period in photocell-cages (food conditioning; FC) while a control group was fed in the home-cage (non-conditioned; NC). FC subjects developed heightened locomotor activity in anticipation of feeding in the initial 30 min in the test-cage compared to NC rats. This anticipatory locomotor activity developed more rapidly and to a greater level in HR rats than in LR rats. The concentrations of dopamine, dihydroxyphenylacetic acid, homovanillic acid, serotonin, 5-hydroxyindoleacetic acid, and norepinephrine were determined at the completion of behavioral testing in both the food conditioned and non-conditioned rats. The food conditioned experiment showed that variations in both the dopaminergic and serotoninergic systems may underlie individual differences in behavioral responsiveness. However, no clear pattern of neurochemical differences emerged. The current set of experiments have demonstrated differences between HR and LR rats in non-drug related paradigms and that HR rats appear to show a greater motivational excitement induced by periodic food delivery than LR rats.

Animals↗

Comparison of several benzodiazepine receptor ligands in two models of anxiolytic activity in the mouse: an analysis based on fractional receptor occupancies.

This study compared the effects of the beta-carboline anxiolytic, abecarnil, with other benzodiazepine receptor (BZR) ligands, including the full agonists diazepam and alprazolam, and the partial agonists ZK 95962 and bretazenil (Ro 16-6028), and alpidem, in the mouse four-plate test and plus-maze. The efficacy and potency of each compound was related to the fraction of BZR occupied by the drug. Abecarnil was efficacious in both tests and showed anxiolytic effects comparable with alprazolam and diazepam. In the four-plate test, abecarnil, bretazenil, and ZK 95962 had selective effects on releasing exploratory locomotor activity suppressed by footshock (punished crossings). None of these compounds significantly altered non-punished crossings. In contrast, diazepam and alprazolam increased both unpunished and punished crossings at low to medium doses (receptor occupancies of approximately 20-60%). The number of punished and unpunished crossings fell to control levels or below at higher, more sedative doses (approximately 80% receptor occupancy). Alpidem had very weak anxiolytic-like effects in this test and markedly reduced unpunished crossings at relatively low receptor occupancies (> 15%). In the plus-maze, abecarnil increased the time spent in the open arms and the percentage open arm entries to an extent equal to that observed following diazepam or alprazolam administration. Bretazenil and ZK 95962 had weak effects on the measures of anxiolytic activity in this test. Alpidem also had little anxiolytic-like activity in the plus-maze but markedly reduced the total number of arm entries.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

5-HT1A receptor agonists improve the performance of normal and scopolamine-impaired rats in an operant delayed matching to position task.

A series of experiments examined the effects of 5-HT1A ligands alone and in combination with the muscarinic antagonist scopolamine on short term working memory in the rat. The behavioural paradigm was a discrete trial, operant delayed matching to position task, with delays of 0, 5, 15 and 30 s. The 5-HT1A ligands tested were the full agonist, 8-OH DPAT (0, 0.1, 0.3 and 1 mg/kg), the partial agonist, ipsapirone (0, 1, 3 and 10 mg/kg), and the purported antagonist, NAN 190 (0, 1, 2, and 4 mg/kg). 1-PP (0, 0.1, 0.3, 1 mg/kg), the major metabolite of ipsapirone, was also tested. The lowest dose of 8-OH DPAT significantly improved matching accuracy at the longest delay, whereas the highest dose impaired matching accuracy and increased the latency to respond. Ipsapirone also significantly improved the accuracy of performance at a dose of 3 mg/kg, but the doses of 1 and 10 mg/kg did not significantly affect performance. NAN-190, at the highest dose tested (4 mg/kg), impaired matching accuracy, whereas the two lower doses did not significantly affect performance. The highest dose also increased the latency to respond. 1-PP had no effect on performance. Scopolamine HBr (0.14 mg/kg) caused a delay dependent impairment in matching accuracy, and had no effect on missed trials or the latency to respond. Low doses of 8-OH DPAT (0.1 and 0.3 mg/kg) significantly attenuated the scopolamine induced accuracy impairment, whereas 1 mg/kg 8-OH DPAT potentiated the impairment. Ipsapirone (3 mg/kg) also significantly improved the performance of scopolamine impaired rats.(ABSTRACT TRUNCATED AT 250 WORDS)

8-Hydroxy-2-(di-n-propylamino)tetralin↗

Effects of cocaine microinjections into the nucleus accumbens and medial prefrontal cortex on schedule-induced behaviour: comparison with systemic cocaine administration.

The effects of cocaine HCl infusions into either the nucleus accumbens (NACC) or medial prefrontal cortex (PFC) were compared on the performance of schedule-induced polydipsia (SIP) and related behaviours. Food-deprived rats were exposed to a fixed-time 60-s schedule of food delivery in daily 30-min sessions until stable levels of behaviour were obtained (14 days). Rats were then bilaterally infused with cocaine into either the NACC or PFC via chronically indwelling guide cannulae. Each subject received a sequence of five cocaine infusions (0, 12.5, 25, 50, 100 micrograms) according to a Latin Square design. For comparison, following these intracranial infusions each rat received a sequence of five IP injections of cocaine (0, 2.5, 5, 10, 20 mg/kg) also in a counterbalanced order. NACC and PFC infusions of cocaine and IP cocaine dose-dependently reduced SIP. Cocaine infusions into the NACC, but not the PFC, increased locomotor activity but the characteristic temporal profile of locomotor activity during SIP was retained. IP cocaine also increased locomotor activity in a dose-dependent manner, but the temporal profile of activity was flattened following 20 mg/kg cocaine. NACC and PFC infusions of cocaine had little effect on the total number of panel presses to gain access to the food pellets, but did slightly decrease the high rates of responding immediately prior to the pellet delivery. IP cocaine increased the total number of panel presses at the higher doses, mainly by increasing the low rates of responding. The effects of cocaine infusions into the PFC were behaviourally the most selective, as they reduced SIP without having substantial effects either on locomotor activity or panel pressing.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Assessment of the relative contribution of peripheral and central components in cocaine place conditioning.

A balanced place conditioning paradigm was used to assess the contribution of peripheral and central factors mediating place conditioning induced by cocaine HCl. The first experiment was conducted to examine changes in locomotor activity and extracellular dopamine (DA) concentrations in the nucleus accumbens (NACC) following intraperitoneal (IP) injections of cocaine HCl (15 mg/kg) or cocaine methiodide (19.6 mg/kg). IP cocaine HCl significantly increased locomotor activity and extracellular NACC DA, whereas IP cocaine methiodide failed to increase either locomotor activity or extracellular DA in the NACC. In the second experiment, IP cocaine HCl (15 mg/kg) induced a significant conditioned place preference; however, neither IP procaine HCl (25 or 50 mg/kg) nor IP cocaine methiodide (4.9, 9.8, or 19.6 mg/kg) induced preferences for the drug-paired compartment. In the third experiment, intracerebroventricular (ICV) infusions of cocaine HCl (25 micrograms/2 microliters) or cocaine methiodide (1 or 5 micrograms/2 microliters) induced significant place conditioning for the drug-paired compartment. These results suggest place conditioning induced by cocaine HCl is mediated centrally and that the local anaesthetic properties alone do not contribute to this effect to any significant degree.

Animals↗