Regulation of Dopamine Release from Interplexiform Cell Processes in the Outer Plexiform Layer of the Carp Retina

The gamma-aminobutyric acid (GABA) antagonists bicuculline and picrotoxin stimulate a four- to fivefold increase in endogenous dopamine release from isolated intact carp retina. The release evoked by these agents is Ca2+ dependent, a finding suggesting a vesicular release. Using light microscopic autoradiography, we have localized the sites of dopamine release to the dopaminergic interplexiform cell processes of the outer plexiform layer, which synapse onto horizontal cells. Our findings support previous suggestions that the dopaminergic interplexiform cells receive GABAergic inhibitory input and that the effects of GABA antagonists on horizontal cells are mediated by dopamine release from the interplexiform cells.     

Circannual rhythms of ground squirrels: a test of the frequency demultiplication hypothesis

The frequency demultiplication hypothesis (FDH) posits that circannual rhythms are generated from circadian cycles by frequency transformation to the lower-frequency rhythm. To test the FDH, we determined the periods of the circannual body mass and estrous cycles of golden-mantled ground squirrels with circadian locomotor activity rhythms entrained to 23-, 24-, or 25-hr days (T-cycles). Circannual period length did not differ among squirrels entrained to the different T-cycles; intergroup ranges were 298-314 days and 303-312 days, respectively, for body mass and estrus. These results are not consistent with the FDH and suggest instead that separate mechanisms generate circadian and circannual rhythms. In ground squirrels the circannual system influences circadian organization, but a reciprocal influence of circadian on circannual rhythms has yet to be demonstrated.     

Seasonal variations in plasma luteinizing hormone levels of gonadectomized male ground squirrels (Spermophilus lateralis)

Plasma luteinizing hormone (LH) levels were determined at monthly intervals in intact and orchidectomized ground squirrels maintained in a constant 14L:10D photoperiod at a temperature of 23 +/- 2 degrees C. Animals were orchidectomized or sham operated at different ages and at different stages of the annual reproductive cycle. LH levels were elevated in orchidectomized as compared to intact males within 1 month, and often within 48 h, after orchidectomies performed either before or after the normal breeding season. LH levels tended to remain chronically elevated in long-term orchidectomized males. Although negative feedback regulation of LH by the testes was evident at most stages of the reproductive cycle, effects of orchidectomy on LH were most pronounced during the nonbreeding season when plasma LH titers of intact males were at their annual nadirs, and less pronounced or absent during the breeding season when levels were normally elevated in intact males. For most of the year male gonadal hormones exert negative feedback on LH release. We hypothesize that under the influence of a circannual timing mechanism, sensitivity to steroid feedback is reduced as the breeding season approaches and LH secretion increases despite the attendant rise in plasma androgen levels. These results contrast with data for female squirrels in which there is a seasonally restricted and characteristically brief elevation of plasma LH after ovariectomy; in females, but not in males, onset and termination of LH secretion appear to be regulated by a circannual clock that functions independently of gonadal secretions.     

Turnover and metabolism of chlorogenic Acid in xanthium leaves and potato tubers

The active turnover of chlorogenic acid (3-caffeoylquinic acid(3)), a major phenolic component of Xanthium leaves and potato tuber disks, has been demonstrated in these tissues. Pulse-labelling experiments with radioactive l-phenylalanine and trans-cinnamic acid as well as direct feeding experiments with chlorogenic acid-(14)C labelled in the caffeoyl moiety have been employed in the turnover studies. The rate of turnover is calculated to be on the order of 50 to 100 mmumoles per hour per gram fresh weight of tissue.In Xanthium leaves chlorogenic acid is in part converted to an isochlorogenic acid identified by silica gel chromatography as 3,5-dicaffeoylquinic acid. Radioactivity of the caffeoyl moiety of chlorogenic acid is also incorporated into lignin-like insoluble polymers in the leaf. Turnover of chlorogenic acid in tuber tissue is largely accounted for by the incorporation of the caffeoyl moiety into insoluble polymers in the tissue.The significance of chlorogenic acid turnover is discussed in relation to the perception of the photoperiodic stimulus by leaves and to the possible role of chlorogenic acid in lignin synthesis.     

Control of intragroup aggression by a captive male orangutan

An adult male orangutan, housed with two adult females, intervened in female-female aggressive interactions at least seven times over a 4.5-year period. Five of these seven episodes involved multiple interventions rather than a single act. One series of interventions was directly observed during an ongoing study of spatial relationships, and six other episodes were described in the zoo records. The intervention that was observed directly began with one female displacing the other, after which the male located himself between the two females. He remained nearer the displaced female than he did to the other female. These seven episodes demonstrate the male's mediation and regulation of intragroup aggression. Whereas the opportunities for the control of aggression are limited in the wild, the conditions of captivity likely allowed for the expression of this behavior pattern.     

Delayed courtship in the fiddler crab Uca musica terpsichores

Courting male fiddler crabs of the species Uca musica terpsichores congregate in the upper central portion of the colony, while receptive females leave their burrows located at the colony's periphery and wander among the communally displaying males prior to choosing a mate. I observed that courting males in a newly-established population were significantly smaller than courting males in large high-density colonies. This observation led to a series of translocation experiments designed to ascertain whether high population density influences the size (=age) at which males begin to court. Smaller courting males from a low-density population failed to court after being placed among larger courting males in a high-density population. The reciprocal translocation revealed that smaller noncourting males from the high-density population would start courting shortly after being placed in a low-density population. Smaller males placed in the high-density population were subsequently observed significantly further away from where they were initially placed than were larger males similarly translocated. The results suggest that smaller males delay courtship activities once they are forced, via encounters with larger males, to the periphery of the colony. I believe that both intrasexual selection (competition from larger males) and intersexual selection (female choice of large males) are responsible for the delay in male courtship activities.     

Azaserine: further evidence for DNA damage in Escherichia coli

Azaserine causes DNA damage in stationary-phase cells. In our investigation of this damage, we used strains of Escherichia coli differing in repair capabilities to study azaserine-induced DNA damage, detected as DNA strand breaks by sucrose gradient sedimentation techniques. Reduced sedimentation in alkaline and neutral sucrose gradients indicated the presence of both alkali-labile sites and in situ strand breaks. Azaserine induced DNA single-strand breaks (SSBs) abundantly in all but the recA strain, in which SSBs were greatly reduced. Treatment of purified DNA with azaserine from bacteriophages T4 and PM2 produced no detectable SSBs. Several other studies also failed to detect DNA damage induced directly by azaserine. Increased levels of beta-galactosidase were induced in an E. coli strain possessing a rec::lac fusion, providing further evidence for azaserine induction of the recA gene product. In addition, azaserine induced adaptation against killing but not against mutagenesis in wild-type E. coli strain.     

Enhanced peripheral chemoreflex function in conscious rabbits with pacing-induced heart failure.

The present study aimed to determine whether peripheral and/or central chemoreflex function is altered in chronic heart failure (CHF) and whether altered chemoreflex function contributes to sympathetic activation in CHF. A rabbit model of pacing-induced CHF was employed. The development of CHF (3-4 wk of pacing) was characterized by an enlarged heart, an attenuated contractility, and an elevated central venous pressure. Renal sympathetic nerve activity (RSNA) and minute volume (MV) of ventilation in response to stimulation of peripheral chemoreceptors by isocapnic/hypoxic gases were measured in the conscious state. It was found that the baseline RSNA at normoxia was higher in CHF rabbits than in sham rabbits (35. 00 +/- 4.03 vs. 20.75 +/- 2.87% of maximum, P < 0.05). Moreover, the magnitudes of changes in RSNA and MV in response to stimulation of the peripheral chemoreceptors and the slopes of RSNA-arterial PO2 and MV-arterial PO2 curves were greater in CHF than in sham rabbits. Inhibition of the peripheral chemoreceptors by inhalation of 100% O2 decreased RSNA in CHF but not in sham rabbits. The central chemoreflex function, as evaluated by the responses of RSNA and MV to hyperoxic/hypercapnic gases, was not different between sham and CHF rabbits. These data suggest that an enhancement of the peripheral chemoreflex occurs in the rabbit model of pacing-induced CHF and that the enhanced peripheral chemoreflex function contributes to the sympathetic activation in the CHF state.     

Apoptosis and Morphology in Mouse Embryos by Confocal Laser Scanning Microscopy

Confocal laser scanning microscopy combined with a vital stain was used to study apoptosis in organogenesis-stage mouse embryos. Apoptosis has previously been visualized in whole embryos using the vital dyes acridine orange, Nile blue sulfate, and neutral red. In the present study, mouse embryos were harvested on Gestation Day 9 and stained with the vital lysosomal dye LysoTracker Red. Following incubation in the stain, embryos were fixed overnight in 4% paraformaldehyde, dehydrated in a graded methanol series, and cleared in benzyl alcohol/benzyl benzoate. The resulting embryo is almost transparent and retains specific LysoTracker Red staining. To achieve optical sectioning through embryos, it was necessary to use low-power objectives. With this procedure, the entire embryo can be optically sectioned and reconstructed in three dimensions to reveal areas of dye staining. Our results demonstrate specific regions undergoing programmed cell death in normal development and increased LysoTracker staining in embryos exposed to hydroxyurea. This procedure allows for the optical imaging of whole Day 9 ( approximately 22 somites) embryos that were greater than 700 microm thick in the z axis and can be applied to studies involving neural tube formation or other aspects of organogenesis.