Facilitation of efficient search of an unbaited radial-arm maze in rats by D1, but not D2, dopamine receptors.

Dopamine (DA) agonists facilitate and antagonists inhibit conditioned preparatory behaviors in rats. We provide added evidence that increased D1 receptor activation facilitates unconditioned preparatory behavior as well, this time in the form of efficient search of an unbaited radial-arm maze. Administration of 0.1, but not 1.0, mg/kg sc SKF81297, a full D1 agonist, increased the number of novel arms chosen in the first eight arms entered. Treatment with 0.1 mg/kg sc D-amphetamine, an indirect DA agonist, also increased search efficiency when given on the first test day but not when given following a test day with a 1.0 mg/kg dose. The 0.1-mg/kg amphetamine-induced facilitation was blocked by coinjection of 0.005 mg/kg SCH23390, a D1 antagonist. Treatment with quinpirole, a D2 agonist, or eticlopride, a D2 antagonist, decreased amount of maze search, but did not affect efficiency. Collectively, our results support the possibility there is a general facilitatory effect of D1 activation on unconditioned preparatory behavior.

Integrating niche-related and general process approaches in the study of learning.

For nearly a century the experimental analysis of learning in animals has been divided into niche-related and general-process approaches, each emphasizing different procedures and conceptual strategies. After considering several current forms of rapprochement, I outline evidence for the integrative hypothesis that niche-related learning provides the basis for results in traditional general-process learning paradigms. Although the full ramifications of this view are not developed here, its advantages include: a clearer relation between laboratory and field results; conceptual and pragmatic guidance in developing new paradigms, and applying old ones to different species and circumstances; clarification of the laws, limits, and anomalies in general-process paradigms; and a more efficient path for inter-relating the study of learning with neurophysiology, genetics, and evolution.

Facilitation of preparatory behavior in an artificial prey paradigm by D1-subfamily dopamine receptor activation.

Dopamine agonists facilitate, and antagonists inhibit, conditioned preparatory behaviors in rats. Similar effects are demonstrated on an unconditioned preparatory behavior: predatory search and contact of a moving artificial prey stimulus. Apomorphine (0.1, 0.2 mg/kg), a direct agonist, had no effect relative to a within-subject injection of saline vehicle but d-amphetamine (0.1 mg/kg), an indirect agonist, increased contact frequency without altering overall motor activation. To determine the relative importance of the D1 and D2 subfamilies of receptors in the amphetamine effect, separate groups of animals received amphetamine co-injected with either SCH23390 (0.01 and 0.005 mg/kg) or eticlopride (0.01 mg/kg), D1 and D2 antagonists, respectively. Whereas the eticlopride-amphetamine group showed no change in contact frequency from baseline, co-injections of either dose of SCH23390 and amphetamine led to near total suppression of contact, as did treatment with SCH23390 (0.005 mg/kg) alone. Treatment with 0.01 mg/kg eticlopride alone increased contact frequency while treatment with a higher dose (0.1 mg/kg) had no effect. Treatment with the D1-subfamily agonist SKF81297 (0.1 mg/kg) increased contact frequency. Collectively, these results support the hypothesis that dopamine mediates unconditioned preparatory behavior and suggest differing roles for the D1 and D2 receptor subfamilies.

Long T methamphetamine schedules produce circadian ensuing drug activity in rats.

Eight female Sprague-Dawley rats were housed in isolated continuous 24-h/day environments under conditions of constant dim light and a rate-limited feeding schedule. Following 2 months of free-running activity, all animals were administered methamphetamine (MA) i.p. (2 mg/kg) once every 31 h for 24 injection cycles. Average wave forms of wheel-running activity showed that animals did not anticipate the 31-h schedule of MA injections, but rather displayed circadian ensuing drug activity (CEDA) between 24 and 28 h following the injections. Post-injection meals failed to meet reliably the threshold necessary to achieve food-engendered anticipatory or ensuing activity. Cosinor analysis showed that the intensity of CEDA was strongly influenced by the relative phase of the free-running rhythm. CEDA was moderately influenced by the size of the post-injection bout of activity. Because injection times rotated daily throughout local time without repeating a time of day, CEDA resulting from a long T schedule of MA administration appeared to be based on one-trial resetting of a circadian-related mechanism by a major drug of abuse.

Incentive downshifts evoke search repertoires in rats.

Negative incentive contrast effects (NCEs) have typically been attributed to frustration or the decremental generalization of learned associations. The purpose of these experiments was to clarify the relation of NCEs to the repertoires of functional search behaviors evoked by incentive downshifts. Rats shifted from 32% to 4% sucrose-solution decreased consummatory responses but increased nose-down locomotion, orientation, location entries, and sampling of alternatives relative to unshifted controls. These changes in behavior were terminated or failed to occur under incentive upshifts. Furthermore, reward downshifts did not produce avoidance of the location of the shifted incentive. Increased search occurred whether or not alternative reward locations were available. Together the evidence suggests that NCEs are related to evoked search modes supporting a repertoire of functional behaviors related to finding food.

The multiply-regulated gabA gene encoding the GABA permease of Aspergillus nidulans: a score of exons.

We describe the cloning, sequence and expression of gabA, encoding the gamma-amino-n-butyrate (GABA) permease of the fungus Aspergillus nidulans. Sequence changes were determined for three up-promoter (gabI ) and six gabA loss-of-function mutations. The predicted protein contains 517 residues and shows 30.3% overall identity with a putative GABA permease of Arabidopsis thaliana, 29.6% identity with the yeast choline transporter and 23.4% identity with the yeast UGA4 GABA permease. Structural predictions favour 11-12 transmembrane domains. Comparison of the genomic and cDNA sequences shows the presence of 19 introns, an unusually large number of introns for, we believe, any fungal gene. In agreement with the wealth of genetic data available, transcript level analyses demonstrate that gabA is subject to carbon catabolite and nitrogen metabolite repression, omega-amino acid induction and regulation in response to ambient pH (being acid-expressed). In agreement with this, we report consensus binding sites 5' to the coding region, six each for CreA and AREA and one for PacC, the transcription factors mediating carbon catabolite and nitrogen metabolite repression and response to ambient pH respectively.

Meal-engendered circadian-ensuing activity in rats.

Large meals scheduled at greater-than-circadian periods (such as T = 31 h) tend to elicit enhanced activity approximately 24 h subsequent to receipt. These studies characterized the process responsible for this meal-engendered "circadian ensuing activity" (meal CEA). Female Sprague-Dawley rats were housed in stations containing a running wheel, pellet dispenser, and lights. Young, middle-aged, or suprachiasmatic-nucleus (SCN)-lesioned rats were given two 1-h meals every 31 or 34 h. Meals were separated by alternating short and long fasts. Most young intact rats engaged in enhanced activity approximately 24 h subsequent to the start of the two-meal series. This circadian ensuing activity underwent large, abrupt daily displacements in response to daily meal delays, was manifested to some degree at all times of day, had an amplitude that was modulated by circadian time of day, was attenuated in middle-aged rats, was evident in SCN-lesioned rats, and oscillated following termination of the feeding schedule. A single experience with food at a novel time of day can "reset" an SCN-independent oscillating process responsible for a circadian activity pattern. CEA has features not readily accommodated by present models of "food-anticipatory activity." The readiness with which the process can be reset implies a keen sensitivity to shifts in the time of food availability but could also produce aberrant behavioral patterns. A T >> 24-h feeding schedule appears to be an ideal procedure with which to study the specific food-related factors responsible for resetting circadian processes and producing a subsequent reallocation of daily activity.

Circadian activity precedes daily methamphetamine injections in the rat.

Scheduled daily injections of methamphetamine (MA) produced locomotor activity that preceded and followed the usual time of injection in rats housed under conditions of constant, moderately dim light and temporally distributed feeding. A circadian basis for pre-injection time activity was supported by its anticipatory timing in the apparent absence of reliable preceding external cues and by its persistence on a test day on which the rats remained undisturbed. Post-injection time locomotor activity also persisted on the test day, occurring from 24 to 29 h after the final MA injection. These results indicate that MA injections engage circadian processes underlying locomotor activity, and they raise the possibility that intake of drugs of abuse by humans may facilitate drug taking or relapse at times of day related to previous drug use.

Increased nuclear traffic chaos in hyphae of Aspergillus nidulans: molecular characterization of apsB and in vivo observation of nuclear behaviour.

Filamentous fungi are model microorganisms for studying nuclear migration in eukaryotic cells. Two genes, apsA and apsB (=anucleate primary sterigmata), were identified in Aspergillus nidulans that affect nuclear distribution in hyphae and specifically block conidiophore development at the metula stage when mutant. Here we describe the cloning, sequencing and molecular analysis of apsB. The gene encodes a 121 kDa coiled-coil, hydrophilic protein that was localized in the cytoplasm. No protein-protein interaction was detected between ApsB and ApsA, a membrane-associated, previously identified protein. An apsB null mutant was characterized by video epifluorescence microscopy using strains that express green fluorescent protein (GFP) in nuclei. With this novel approach, we have discovered a new mutant phenotype and have found that nuclei display an increased chaotic movement in older hyphal compartments that results in clustering and an uneven distribution of these organelles. These results suggest a regulatory role of ApsB in nuclear migration.

The influence of current-visit experience within a prey patch on patch persistence.

A critical component of efficient foraging behavior is leaving a feeding site when prey density in that site declines. If ``real time'' hypotheses are to explain patch-leaving mechanisms, we will need to examine the influence that moment-to-moment experience within a current prey patch has on persistence in that patch. We used linear regression to investigate how current experience of patch quality influenced homing pigeons' patch persistence. We did this by examining the amount of variance in persistence that was accounted for by different measures of within-session patch quality. The importance of several measures of the precise sequence of events in individual sessions were assessed with selected averaging algorithms. Mean inter-capture interval and mean number of inter-capture pecks accounted for a significant amount of the variance in giving-up time in three of four, and four of four birds, respectively. Conversely, only one rate measure in one bird showed an influence on patch persistence. In three birds, recent information had a strong influence on giving-up time. Current-visit experience did influence the patch persistence of the pigeons, but the birds' behavior indicated that different individuals used different measures of patch quality, different mechanisms of assessing those measures, and different patch-exit mechanisms.

In vitro reconstruction of the Aspergillus (= Emericella) nidulans genome.

A physical map of the 31-megabase Aspergillus nidulans genome is reported, in which 94% of 5,134 cosmids are assigned to 49 contiguous segments. The physical map is the result of a two-way ordering process, in which clones and probes were ordered simultaneously on a binary DNA/DNA hybridization matrix. Compression by elimination of redundant clones resulted in a minimal map, which is a chromosome walk. Repetitive DNA is nonrandomly dispersed in the A. nidulans genome, reminiscent of heterochromatic banding patterns of higher eukaryotes. We hypothesize gene clusters may arise by horizontal transfer and spread by transposition to explain the nonrandom pattern of repeats along chromosomes.

Loss of meiosis in Aspergillus.

If strictly mitotic asexual fungi lack recombination, the conventional view predicts that they are recent derivatives from older meiotic lineages. We tested this by inferring phylogenetic relationships among closely related meiotic and strictly mitotic taxa with Aspergillus conidial (mitotic) states. Phylogenies were constructed by using DNA sequences from the mitochondrial small ribosomal subunit, the nuclear ribosomal internal transcribed spacers, and the nuclear 5.8S ribosomal gene. Over 920 bp of sequence was analyzed for each taxon. Phylogenetic analysis of both the mitochondrial and nuclear data sets showed at least four clades that possess both meiotic and strictly mitotic taxa. These results support the hypothesis that strictly mitotic lineages arise frequently from more ancient meiotic lineages with Aspergillus conidial states. Many of the strictly mitotic species examined retained characters that may be vestiges of a meiotic state, including the production of sclerotia, sclerotium-like structures, and hülle cells.

Saccharomyces cerevisiae TEC1 is required for pseudohyphal growth.

Diverse eukaryotic organisms share developmental transcription factors with homologous DNA-binding domains. We showed that the developmental regulator AbaA, a member of the ATTS/TEA (AbaA, TEF-1, TEC1, Scalloped/TEF-1, TEC1, AbaA) class of transcription factors of the filamentous fungus Aspergillus nidulans, induces pseudohyphal development in the yeast Saccharomyces cerevisiae. The S. cerevisiae homologue of AbaA, TEC1p, is required for this morphological transition. We provide evidence that TEC1p functions in co-operation with STE12p to induce pseudohyphal development.

catA, a new Aspergillus nidulans gene encoding a developmentally regulated catalase.

Aspergillus nidulans asexual sporulation (conidiation) is a model system for studying gene regulation and development. The CAN5 cDNA is one of several clones isolated based on transcript induction during conidiation. Here we present the molecular characterization of its corresponding gene, demonstrating that it encodes a developmentally regulated catalase, designated catA. The catA 744-amino-acid-residue polypeptide shows significant identity to other catalases. Its similarity to prokaryotic catalases is greater than to other fungal catalases. catA mRNA is barely detectable in growing mycelia, highly induced during sporulation, and present in isolated spores. However, catA expression is not dependent on the developmental regulatory genes brlA, abaA and wetA. Direct catalase activity determination in native gels revealed the existence of two bands of activity. One of these bands represented the major activity during vegetative growth and was induced during sporulation. The second catalase activity appeared after the induction of sporulation and was the predominant activity in spores. Disruption of catA abolished the major spore catalase without eliminating the vegetative activity, indicating the existence of at least two catalase genes in A. nidulans. catA-disrupted mutants produced spores that were sensitive to H2O2, as compared to wild-type spores. The increase in the activity of the vegetative catalase and the appearance of a second catalase during asexual sporulation is consistent with the occurrence of an oxidative stress during development.

Wild chromosomal variants in Aspergillus nidulans.

Pulsed-field gel electrophoresis and a chromosome-specific cosmid DNA library were used to determine the karyotypes of wild-type Aspergillus nidulans isolates from around the world. Overall, little structural variation was found, with a few major exceptions. One isolate possessed a non-essential B-chromosome of about 1.0 million base pairs (mb). Another isolate had undergone a non-reciprocal translocation of about 1.6 mb of chromosome VI onto chromosome VIII. Other than these chromosomal differences, these isolates appeared phenotypically normal. To analyze its effects on meiosis, the translocation isolate was outcrossed with another wild-type derivative that had a normal electrophoretic karyotype. This cross produced a range of phenotypes, including duplicated progeny that had a barren phenotype similar to that described for Neurospora partial disomics. The duplication was somewhat vegetatively unstable. This is the first association of sterility with chromosomal duplication in A. nidulans.

On the consistency of a physical mapping method to reconstruct a chromosome in vitro.

During recent years considerable effort has been invested in creating physical maps for a variety of organisms as part of the Human Genome Project and in creating various methods for physical mapping. The statistical consistency of a physical mapping method to reconstruct a chromosome, however, has not been investigated. In this paper, we first establish that a model of physical mapping by binary fingerprinting of DNA fragments is identifiable using the key assumption-for a large randomly generated recombinant DNA library, there exists a staircase of DNA fragments across the chromosomal region of interest. Then we briefly introduce epi-convergence theory of variational analysis and transform the physical mapping problem into a constrained stochastic optimization problem. By doing so, we prove epi-convergence of the physical mapping model and epi-convergence of the physical mapping method. Combining the identifiability of our physical mapping model and the epi-convergence of a physical mapping method, finally we establish strong consistency of a physical mapping method.

Two possible determinants of the timing of daily episodes of behavior in rats.

The present experiment examined endogenous ultradian rhythms and regulatory metabolic processes as two potential determinants of daily out-of-nest episodes (ONEs) and feeding episodes (FEs) in rats living in a 24-h environment. Two types of photoperiod were used: a Standard 12:12 h L/D schedule and a Skeleton 12:12 h schedule. During each type of photoperiod rats were exposed to 4 feeding conditions of 7 to 12 days each: (a) a baseline of ad lib food and water; (b) restricted to the diurnal (inactive) portion of the cycle; (c) both food and water restricted to the inactive period; and (d) return to baseline. Time series analyses of nocturnal ONEs in baseline revealed a strong circadian rhythm and weaker ultradian rhythms with periods between 2 and 6 h. Analyses of FEs, though, revealed a general absence of circadian rhythms but strong ultradian rhythms with periods similar to those observed in ONEs. When food and water were restricted to the inactive part of the cycle, ONEs showed no change in frequency, but a decrease in average duration and changes in ultradian periodicities. The results indicated control of daily episodes of behavior in rats by ultradian oscillators that are weaker and more variable in affect than those found in voles, but similar in period.

Decremental carryover effects of sucrose ingestion in the negative anticipatory contrast procedure in rats.

To test for retrospective effects of sucrose ingestion in the anticipatory contrast procedure, 4 experiments examined intake of an initial 0.15% saccharin solution as a function of the unsignaled interspersing of days in which the 2nd solution was 32% sucrose or 0.15% saccharin. In Experiment 1, rats that received alternating saccharin-saccharin days and saccharin-sucrose days drank less saccharin on saccharin-only days, and on both days they drank less saccharin than a control group that received saccharin only. In Experiment 2, rats that received randomized saccharin-saccharin and saccharin-sucrose days drank less saccharin if, and only if, a sucrose day preceded. Experiments 3 and 4 used double and quadruple alternation of saccharin and sucrose days to examine persistence of the effects of a sucrose day. The results highlighted a retrospective carryover effect of sucrose that reduced intake of the initial saccharin solution and apparently was based on sucrose memories persisting over days.

Two meals promote entrainment of rat food-anticipatory and rest-activity rhythms.

Ten female rats were fed early and late in the dark period of a 12-12 h light-dark cycle and then were fed at the same times in constant darkness. In both conditions rats were active prior to mealtimes and manifested no free-running components of activity. When the rats were placed in constant darkness and either were fed early and late in the inactive period, or had free access to food, six of the rats had rest-activity rhythms different from 24.0 h. Though a masking explanation could not be ruled out, two meals during the active period apparently entrained the rest-activity rhythms of these rats. The light-entrainable oscillator appears to integrate information from cycles of both illumination and food availability. Multiple sources of temporal information may promote more stable entrainment of the rest-activity rhythm than the light-dark cycle alone, especially in a burrow dwelling organism, like the rat, that can be exposed to inconsistent light-dark transitions.