Photoperiodic regulation of the reproductive axis in male zebra finches, Taeniopygia guttata

In arid central Australia, breeding of zebra finches (Taeniopygia guttata) tends to take place immediately after the aperiodic rainfall-this being irrespective of the time of year. As a result, zebra finches have traditionally been considered nonphotoperiodic. Despite this, there are some published reports of photoperiodic effects on behavior in zebra finches. Therefore, we transferred singly housed male zebra finches from a photoperiod of 14 h light and 10 h dark per day (14L:10D) to either 20L:4D or 8L:16D. Control birds remained exposed to 14L:10D. Plasma LH, testicular volume, and body mass were assessed at the start of the experiment and at intervals for a period of 56 days. Testicular mass was measured at the end of the 56-day period. Plasma LH increased significantly in the 20L:4D group after 14 days, but decreased again by 56 days, presumably an effect of increased gonadal steroid negative feedback. Plasma LH did not change significantly in the other two groups. Testicular volume increased steadily in the 20L:4D group during the treatment period and it was significantly higher than that of the 8L:16D group at the end of the experiment. After 56 days of treatment, combined testicular mass in the 20L:4D group was much greater than that of the 8L:16D group, but not quite statistically different from the 14L:10D group. There was no statistical difference in testicular mass between the 14L and 8L groups. Body mass did not differ between any of the groups at any time in the experiment. Our results are consistent with zebra finches being photoperiodic to some degree, despite their opportunistic breeding strategy. When considered in conjunction with recent reports of photoperiodic responses in tropical avian species, these data suggest that the ability to respond to changing photoperiod is more common among avian species than previously hypothesized.     

Testosterone facilitates some conspecific song discriminations in castrated zebra finches (Taeniopygia guttata).

An experiment was designed to test for the influence of testosterone on song discriminations. We found that testosterone did have an effect, which interacted with practice and the nature of the stimuli. Fourteen adult castrated zebra finches (Taeniopygia guttata) were grouped into seven pairs. In each pair, one bird was implanted with a testosterone-filled silastic tube and the other was implanted with an empty silastic tube. They were then trained on a go/no-go operant task to discriminate between bird songs in six consecutive experiments. The songs to be discriminated were as follows: experiment 1, two canary song segments; experiment 2, the bird's own song and that of the other member of the pair; experiment 3, the same two songs as in experiment 2 but with reversed stimulus-response contingencies; experiment 4, two other zebra finch songs; experiment 5, another two zebra finch songs; and experiment 6, another two canary song segments. There were no reliable learning differences between birds treated with testosterone or with an empty silastic in experiments 1 and 3-6. However, in experiment 2, testosterone-treated birds mastered the discrimination between their own song and the song of the other member of the pair in fewer trials than birds treated with empty silastics. We suggest that a song's ability to control the behavior of male zebra finches is influenced by the nature of the song, prior experience with the training paradigm, and hormone levels.     

Role of gender, season, and familiarity in discrimination of conspecific song by zebra finches (Taeniopygia guttata).

Operant conditioning techniques were used to assess how gender and song familiarity affect song discrimination in adult zebra finches (Taeniopygia guttata). Twenty-five males and females, divided into five cohorts, were trained to discriminate between conspecific songs at different times of year. Males discriminating between their own and another song from their own aviary reached criterion in the fewest number of trials, followed by males discriminating between two songs from their own aviary, then by males discriminating between songs they had not heard before. Females discriminating between two songs from their own aviary required more trials than males to reach criterion, but, unlike in males, song familiarity did not have a significant effect on song discrimination by females. The number of trials required to reach criterion was greater in winter than in summer, suggesting a photoperiodic effect in what has been regarded as a nonphotoperiodic species. Gender, season, and familiarity appear to affect how zebra finches discriminate between conspecific songs.     

Prolactin release and response to vasoactive intestinal peptide in an opportunistic breeder, the zebra finch (Taeniopygia guttata)

Zebra finches in arid regions of Australia are opportunistic breeders that time their breeding cycles to coincide with nonseasonal rainfall. Hormonal profiles associated with reproductive behaviors may differ from those observed in seasonal breeders because these birds need to be reproductively competent on short notice. This study measured plasma prolactin (PRL) levels in nonbreeding and breeding zebra finches and in birds with and without prior reproductive experience. We also investigated the change in plasma PRL following injection with vasoactive intestinal peptide (VIP), the avian PRL-releasing hormone. PRL was lowest in non-paired birds, increased after pair bonds had formed, and was highest in incubating birds. No differences in PRL levels were found between males and females in these biparental care-givers. A single injection of VIP resulted in a rapid increase in plasma PRL in nonbreeding zebra finches, while PRL remained unchanged in incubating birds. When escalating doses of VIP were administered, nonbreeders responded with a maximal response in PRL release, but PRL levels in breeders remained unchanged following even the highest VIP dose. Among nonbreeders, inexperienced birds had significantly lower PRL levels than birds that had successfully reared a clutch, but both groups responded with an equally robust increase in PRL following a VIP challenge. This pattern differs from that observed in most photosensitive species in which only during a breeding cycle do birds secrete significant levels of PRL in response to exogenous VIP. Zebra finches, even when not actively breeding, must maintain competent pituitary lactotrophs that can secrete PRL at maximal rates. This is part of the suite of characters enabling these birds to respond to favorable breeding conditions at any time.     

Sex steroid levels in developing and adult male and female zebra finches (Poephila guttata)

Serum samples from male and female zebra finches ranging in age from 1 day before hatch to 54 days posthatch were assayed for 17 beta-estradiol (E), androgen, testosterone (T), or 5 alpha-dihydrotestosterone (DHT). Additional samples were assayed from intact and gonadectomized adults, gonadectomized adults with intraperitoneal implants of testosterone propionate (TP) or estradiol benzoate (EB), gonadectomized nestlings, and nestlings injected subcutaneously with EB. DHT levels of developing birds did not vary as a function of either sex or age. During development, average androgen and T levels were highest during the nestling period, prior to sexual maturation, and were higher in females than in males. Endogenous androgen levels of most subjects that were sampled repeatedly rose and then declined between 24 and 49 days. TP implants produced higher T levels in adult females than in adult males. Levels of E were higher in both sexes during the hatching period (Days -1 through 0) than during the nestling period (Days 2 through 14). A greater number of males than females had relatively high E levels on Days 12 and 14 and during the second week overall. There was no sex difference in levels of E in adults, and gonadectomized adults had markedly higher E levels than intact adults. Gonadectomized nestlings had the same androgen and E levels as intact nestlings of the same age; EB injected nestlings had elevated E levels. The present results indicate specific endocrine changes that mirror events crucial to sexual differentiation of endocrine and behavioral components of reproduction, and have important implications for models of sexual differentiation in zebra finches.     

Inhibited mitochondrial respiration by amobarbital during cardiac ischaemia improves redox state and reduces matrix Ca2+ overload and ROS release

AIM: Damage to the mitochondrial electron transport chain (ETC) occurs during ischaemia. Blockade of electron flow in the ETC just before ischaemia with the reversible complex I inhibitor amobarbital protects isolated mitochondria against ischaemic damage and preserves oxidative phosphorylation and cytochrome c content. We hypothesized that brief amobarbital perfusion just before ischaemia would improve cardiac recovery and decrease infarct size after ischaemia and reperfusion (IR) by preserving the mitochondrial redox state and reducing mitochondrial superoxide (O(2)(-*)) generation, in turn would decrease mitochondrial Ca(2+) accumulation (mt[Ca(2+)]). METHODS: Guinea pig Langendorff-perfused hearts were treated with Krebs Ringer solution (KR; untreated) or amobarbital (2.5 mM) in KR for 1 min immediately before 30 min of no flow, global ischaemia, followed by reperfusion without additional treatment. Cardiac function, mitochondrial NADH, FAD, mt[Ca(2+)], and O(2)(-*) levels were assessed during the 1 min perfusion period and throughout IR. RESULTS: Amobarbital perfusion alone before ischaemia significantly increased O(2)(-*) levels and NADH, without altering FAD, and decreased mt[Ca(2+)]. During ischaemia, mitochondrial NADH was higher, O(2)(-*) levels were lower, and mt[Ca(2+)] was less elevated in the amobarbital group. On reperfusion O(2)(-*) levels and mt[Ca(2+)] were significantly reduced, NADH-FAD redox state was preserved and cardiac function was markedly improved in the amobarbital group; infarct size was smaller in the amobarbital group compared to the untreated group. CONCLUSION: Temporary blockade of mitochondrial complex I activity by amobarbital protects hearts by reducing production of O(2)(-*) and mtCa(2+) loading during IR injury.     

Effects of embryonic treatment with fadrozole on phenotype of gonads, syrinx, and neural song system in zebra finches.

Previous studies have found that treatment of zebra finch embryos with an aromatase inhibitor on Day 5 or 8 of incubation caused partial sex reversal of gonadal phenotype in females. These females possessed both testicular and ovarian tissue, and the development of the neural circuit for song remained feminine. The present study attempted more complete gonadal reversal by treating zebra finch embryos earlier, on Day 3 of incubation, with Fadrozole (CGS 16949A), an aromatase inhibitor, or with saline. We examined the phenotype of the syrinx (androgen-dependent vocal organ), the gonads, and the telencephallic neural song system in 100-day-old birds. Treated females typically possessed a left ovotestis and a right testis, and significantly larger syringes than control females. The histology and steroid synthetic enzyme activity of the testicular tissue in treated females were quite masculine and similar to that of control males. At the time of sacrifice, the plasma concentrations of testosterone and estradiol for fadrozole-treated females did not differ from those of control females, but dihydrotestosterone was lower in treated females. Despite the large amount of functional testicular tissue and a masculine syrinx, the volumes and soma sizes of song system nuclei (HVC, RA) in treated females remained feminine. These results suggest that testicular secretions masculinize the syrinx, but are not sufficient to masculinize the song system in zebra finches.     

Testing hypotheses of aging in long-lived mice of the genus Peromyscus: association between longevity and mitochondrial stress resistance, ROS detoxification pathways, and DNA repair efficiency

In the present review we discuss the potential use of two long-lived mice of the genus Peromyscus—the white-footed mouse (P. leucopus) and the deer mouse (P. maniculatus) maximum lifespan potential ～8 years for both—to test predictions of theories about aging from the oxidative stress theory, mitochondrial theory and inflammatory theory. Previous studies have shown that P. leucopus cells exhibit superior antioxidant defense mechanisms and lower cellular production of reactive oxygen species (ROS) than do cells of the house mouse, Mus musculus (maximum lifespan ～3.5 years). We present new data showing that mitochondria in P. leucopus cells produce substantially less ROS than mitochondria in M. musculus cells, and that P. leucopus mitochondria exhibit superior stress resistance to those of M. musculus. We also provide evidence that components of the DNA repair system (e.g., pathways involved in repair of DNA damage induced by γ-irradiation) are likely to be more efficient in P. leucopus than in M. musculus. We propose that mitochondrial stress resistance, ROS detoxification pathways and more efficient DNA repair contribute to the previously documented resistance of P. leucopus cells toward oxidative stress-induced apoptosis. The link between these three pathways and species longevity is discussed.     

Changes in suicide rates after reductions in alcohol consumption and problems in Ontario, 1975–1983

In this paper the relationships among suicide rates in Ontario for males, females and the total population, alcohol consumption, and other alcohol-related problems are examined for the period 1963–1983. Suicide rates, like other alcohol-related problems, were strongly correlated with per capita consumption over the total period. However, during the recent period of stabilization in consumption (1975–83), the correlations were markedly reduced and substantial differences in trends for males and females were observed. The contribution to these diverging trends of such factors as increased treatment of alcohol abuse, rising unemployment, and reduced use of tranquillizing drugs is discussed.     

Abnormalities in exercising skeletal muscle in congestive heart failure can be explained in terms of decreased mitochondrial ATP synthesis, reduced metabolic efficiency, and increased glycogenolysis.

OBJECTIVE: To distinguish between the effects of reduced oxidative capacity and reduced metabolic efficiency on skeletal muscle bioenergetics during exercise in patients with congestive heart failure. DESIGN AND PATIENTS: Patients were studied by 31P magnetic resonance spectroscopy during aerobic exercise and recovery, and results compared with controls. RESULTS: In flexor digitorum superficialis muscle (26 patients) there was a 30% decrease in oxidative capacity compared with control (mean (SE) 36 (2) v 51 (4) mM/min) and also a 40% decrease in "effective muscle mass" (5 (1) v 9 (1) arbitrary units), probably at least partly the result of reduced metabolic efficiency. Both contribute to increased phosphocreatine depletion and intracellular acidosis during exercise. However, an increased concentration of ADP (an important mitochondrial regulator) during exercise permitted near-normal rates of oxidative ATP synthesis. Results were similar in gastrocnemius muscle (20 patients), with a 30% decrease in maximum oxidative capacity (29 (4) v 39 (3) mM/min) and a 65% decrease in effective muscle mass (5 (1) v 13 (2) arbitrary units). Exercise training improved maximum oxidative capacity in both muscles, and in gastrocnemius effective muscle mass also. CONCLUSIONS: Skeletal muscle exercise abnormalities in patients with congestive heart failure results more from decreased metabolic efficiency than from the abnormalities in mitochondrial oxidation. Both decreased efficiency and defective mitochondrial oxidation result in an increased activation of glycogen phosphorylase, and may be improved by exercise training.     

The expression of the sex steroid-synthesizing enzymes CYP11A1, 3beta-HSD, CYP17, and CYP19 in gonads and adrenals of adult and developing zebra finches

Songbirds have emerged as important animal models for understanding how sex steroids influence brain and behavior, particularly how they direct the sexually dimorphic development of the neural circuits controlling song and then activate adult song behavior. Presumably, sex steroids synthesized in the gonads are responsible for these actions on brain. However, experiments do not always reveal a direct relationship between gonadal function, circulating sex steroids, and activation and/or organization of song. Thus, it is critical that we understand more about the sites and mechanisms of sex steroid synthesis in this group of birds. Toward this end, we have established the use in zebra finches of chicken cDNA probes to the principal androgen synthetic enzymes, CYP11A1, 3beta-HSD, and CYP17. On Northern blots, these probes recognized bands of the appropriate size and in tissues similar to those seen in chickens. With these probes, and a probe to CYP19 specific to the zebra finch, we used in situ hybridization to examine the cellular expression of these enzymes in gonads and adrenals of adult and developing zebra finches (1 to 20 days posthatching). In adults, we identified significant expression of CYP11A1 and CYP17 in large ovarian follicles, particularly the thecal cell layer and over the testicular interstitial area. 3beta-HSD was expressed by both theca and granulosa and in testicular interstitial and seminiferous tubular cells. In adrenals, CYP11A1 and 3beta-HSD are abundant with lesser amounts of CYP17. Developmentally, we identified high expression of CYP11A1 and 3beta-HSD in the adrenals, CYP17 in both testes and ovaries, and CYP19 in ovaries only. These results suggest that the ovaries but not the testes may secrete estrogen developmentally and the adrenals may contribute precursors for gonadal steroidogensis.     

Cell-surface glycosyltransferases in cultured fibroblasts: increased activity and release during serum stimulation of growth.

Cell-surface galactosyltransferase was studied in suspensions of intact baby hamster kidney fibroblasts with both endogenous and exogenous glycoprotein acceptors. The cell-surface location of galactosyltransferase was demonstrated in experiments with the enzyme modifier alpha-lactalbumin, which does not enter the cell. The addition of alpha-lactalbumin to the assay medium for galactosyltransferase resulted in accumulation of lactose in the medium but not in the cells. There was no detectable hydrolysis of UDP-galactose to free galactose by these cells, nor did a 100-fold molar excess of free galactose inhibit cell-surface galactosyltransferase. There was a marked increase in specific activity of cell-surface exogenous galactosyltransferase in serum-stimulated as compared to resting fibroblasts. Dividing but not resting fibroblasts released galactosyltransferase, but not sialyl- or fucosyltransferase, in soluble form into the tissue culture medium. The release of galactosyltransferase was greater from virally transformed than from nontransformed fibroblasts.     

Coevolution of exceptional longevity, exceptionally high metabolic rates, and mitochondrial DNA-coded proteins in mammals

Mammals' longevity is inversely related to mass-specific basal metabolic rate because the generation of reactive oxygen species constrains lifespan. Longevity increases with body mass because the latter is inversely related to mass-specific basal metabolic rates. In placental mammals the longevity residuals from the power laws that describe longevity as a function of mass-specific basal metabolic rates, or body mass, are positively correlated with the relative rates of evolution of cytochrome b, a generator of reactive oxygen species. Therefore, longevity is more accurately described as a function of both mass-specific basal metabolic rate and the relative rate of cytochrome b evolution. The longevity residuals from the power law that describe longevity as a function of body mass are positively correlated with the relative rate of evolution of most other mtDNA-coded proteins. In taxa with very high rate of cytochrome b evolution exceptional longevity is associated with an increase, rather than the predicted decrease, of basal metabolic rates. These finding are compatible with the hypothesis that, in placental mammals, the accelerated evolution of mtDNA-coded proteins, allowed the extension of lifespan by selecting mutations that reduce the generation of reactive oxygen species, mostly by increasing internal proton leak, that accelerates mitochondrial electron transport.     

HIV testing is associated with increased knowledge and reductions in sexual risk behaviours among men in Cape Town,South Africa

HIV testing benefits those who test positive, allowing them to receive treatment, but the benefits for those who test negative remain controversial. We evaluated the impact of testing on HIV knowledge and sexual risk among men in South Africa. Men were recruited from townships outside Cape Town and completed a survey that assessed testing history, knowledge, and sexual behaviours. Among the 820 participants, 516 (63%) reported being tested (82% tested negative, 6% tested positive, and 12% unknown). Compared to those who had never been tested for HIV, men who tested for HIV were more knowledgeable about HIV transmission, but did not differ on sexual risk behaviour. Knowledge moderated the effect of testing on sexual risk such that men reported fewer sexual partners (incidence rate ratio (IRR) = 0.91, 95% CI = 0.84, 0.98) and fewer unprotected anal sex events (IRR = 0.81, 95% CI = 0.66, 1.00) if they had been tested for HIV and were knowledgeable about HIV transmission. For men testing HIV-negative, knowledge predicted fewer sexual risk behaviours. Previous HIV testing is associated with enhanced knowledge, which moderates sexual risk behaviour among South African men living in Cape Town. Results suggest that HIV testing may increase knowledge and lead to reductions in sexual risk even when results are negative.     

Bile acids induce mitochondrial ROS, which promote activation of receptor tyrosine kinases and signaling pathways in rat hepatocytes

Previous studies have demonstrated in hepatocytes that deoxycholic acid (DCA) promotes inactivation of protein tyrosine phosphatases (PTPases) and activation of ERBB1 and the extracellular-regulated kinase (ERK) 1/2 pathway. The present studies have determined the biochemical mechanism(s) through which these events occur. DCA and taurodeoxycholic acid (TDCA) (100 micromol/L) caused activation of ERBB1, insulin receptor, and the ERK1/2 and AKT pathways in primary rodent hepatocytes. DCA- and TDCA-induced receptor and signaling pathway activations were blocked by the reactive oxygen species (ROS) scavengers N-acetyl cysteine (NAC) and Trolox (TX), as well as by cyclosporin A (CsA) and bongkrekic acid (BKA). DCA activated the ERK1/2 pathway in HuH7 human hepatoma cells that was blocked by the incubation of cells with an ERBB1 inhibitor, NAC, TX, CsA, or BKA. DCA did not activate the ERK1/2 pathway in mitochondria-defective HuH7 Rho 0 cells. In HuH7 cells and primary hepatocytes, DCA enhanced the production of ROS, an effect that was abolished in Rho 0 cells and by prior incubation of cells with CsA or BKA. In hepatocytes and HuH7 cells, DCA inhibited PTPase activity. Incubation of hepatocytes with either CsA or BKA prevented DCA-induced inhibition of PTPase activity. Loss of mitochondrial function in Rho 0 cells also abolished the inhibitory effects of DCA on PTPase activity. In conclusion, DCA and TDCA cause ROS generation in hepatocytes that is dependent on metabolically active mitochondria. The generation of ROS is essential for PTPase inactivation, receptor tyrosine kinase activation, and enhanced signaling down the ERK1/2 and AKT pathways.     

Plasma thrombospondin-1 is increased during acute sickle cell vaso-occlusive events and associated with acute chest syndrome, hydroxyurea therapy, and lower hemolytic rates

Platelets are activated in sickle cell disease (SCD), and particularly during vaso-occlusive episodes (VOE). Thrombospondin-1 (TSP1), a major secretory product of activated platelets, is increased in the circulation in VOE and binds to sickle red blood cells (RBC) promoting vascular adhesion. Thus, we hypothesized that TSP1 may represent a plasma biomarker of disease severity in SCD. We tested the plasma collected from patients in steady state (n = 27) and VOE (n = 14), as well as healthy controls (n = 17) at the University of Pittsburgh Medical Center (UPMC), and from patients in steady state enrolled in the walk-PHaSST clinical trial (n = 483). We found that TSP1 levels were increased in VOE in the UPMC cohort. Among steady-state patients at UPMC, TSP1 values correlated positively with lifetime history of acute chest syndrome (r = 0.72, P < 0.0001) and hemoglobin concentration (r = 0.49, P = 0.01), and negatively with markers of hemolysis, such as LDH (r = -0.50, P = 0.009). Analysis of the walk-PHaSST cohort also showed a positive association between TSP1 levels and hydroxyurea use (r = 0.14, P = 0.003), and confirmed the negative associations with the severity of hemolysis. Our results suggest that TSP1 levels are associated with more VOE, hydroxyurea use and lower rates of hemolysis. High TSP1 concentrations may indicate higher risk of the viscosity/vaso-occlusion phenotype of SCD.     

Elevated plasma norepinephrine concentrations in decompensated cirrhosis. Association with increased secretion rates, normal clearance rates, and suppressibility by central blood volume expansion

Plasma norepinephrine concentrations are elevated in patients with decompensated cirrhosis, and correlate inversely with urinary sodium and water excretion. Increased plasma norepinephrine concentrations may result from a decreased metabolic clearance rate or an increased secretion rate, possibly in response to a decreased "effective arterial blood volume." If the latter hypothesis is correct, plasma norepinephrine might be expected to be suppressed when central blood volume is expanded by head-out water immersion. In the present study, plasma norepinephrine secretion and clearance rates were determined by infusion of tritiated norepinephrine. Norepinephrine secretion rates were elevated in eight cirrhotic patients as compared to control subjects (1.50 +/- 0.25 vs. 0.26 +/- 0.08 micrograms/m2 per min, P less than 0.001), whereas clearance rates were similar (3.13 +/- 0.48 vs. 2.60 +/- 0.28 liters/min, NS). Baseline plasma norepinephrine concentrations were markedly elevated in the cirrhotic patients (830 +/- 136 vs. 185 +/- 12 pg/ml, P less than 0.001). Head-out water immersion significantly suppressed plasma concentrations of both norepinephrine (704 +/- 72 to 475 +/- 70 pg/ml, P less than 0.005) and epinephrine (121 +/- 33 to 57 +/- 10 pg/ml, P less than 0.05) in all seven patients studied. We conclude that the high circulating catecholamine concentrations in cirrhosis are secondary to increased secretion, rather than to decreased metabolic clearance, and are suppressible by central blood volume expansion.