Describing mate preference functions and other function-valued traits.

Mate preferences are important causes of sexual selection. They shape the evolution of sexual ornaments and displays, sometimes maintaining genetic diversity and sometimes promoting speciation. Mate preferences can be challenging to study because they are expressed in animal brains and because they are a function of the features of potential mates that are encountered. Describing them requires taking this into account. We present a method for describing and analysing mate preference functions, and introduce a freely available computer program that implements the method. We give an overview of how the program works, and we discuss how it can be used to visualize and quantitatively analyse preference functions. In addition, we provide an informal review of different methods of testing mate preferences, with recommendations for how best to set up experiments on mate preferences. Although the program was written with mate preferences in mind, it can be used to study any function-valued trait, and we hope researchers will take advantage of it across a broad range of traits.

Multivariate female preference tests reveal latent perceptual biases.

The question of why males of many species produce elaborate mating displays has now been largely resolved: females prefer to mate with males that produce such displays. However, the question of why females prefer such displays has been controversial, with an emerging consensus that such displays often provide information to females about the direct fitness benefits that males provide to females and/or the indirect fitness benefits provided to offspring. Alternative explanations, such as production of arbitrarily attractive sons or innate pre-existing female sensory or perceptual bias, have also received support in certain taxa. Here, we describe multivariate female preference functions for male acoustic traits in two chirping species of field crickets with slow pulse rates; our data reveal cryptic female preferences for long trills that have not previously been observed in other chirping species. The trill preferences are evolutionarily pre-existing in the sense that males have not (yet?) exploited them, and they coexist with chirp preferences as alternative stable states within female song preference space. We discuss escape from neuronal adaptation as a possible mechanism underlying such latent preferences.

Conservation of multivariate female preference functions and preference mechanisms in three species of trilling field crickets.

Divergence in mate recognition systems among closely related species is an important contributor to assortative mating and reproductive isolation. Here, we examine divergence in male song traits and female preference functions in three cricket species with songs consisting of long trills. The shape of female preference functions appears to be mostly conserved across species and follows the predictions from a recent model for song recognition. Multivariate preference profiles, combining the pulse and trill parameters, demonstrate selectivity for conspecific pulse rates and high trill duty cycles. The rules for integration across pulse and trill timescales were identical for all three species. Generally, we find greater divergence in male song traits than in associated female preferences. For pulse rate, we find a strong match between divergent male traits and female peak preferences. Preference functions for trill parameters and carrier frequency are similar between species and show less congruence between signal and preference. Differences among traits in the degree of trait-preference (mis)match may reflect the strength of preferences and the potential for linkage disequilibrium, selective constraints and alternative selective pressures, but appear unrelated to selection for mate recognition per se.

Body temperature responses of Pekin ducks (Anas platyrhynchos domesticus) exposed to different pathogens.

Poultry, like mammals and other birds, develop fever when exposed to compounds from gram-negative bacteria. Mammals also develop fever when exposed to the constituents of viruses or gram-positive bacteria, and the fevers stimulated by these different pathogenic classes have discrete characteristics. It is not known whether birds develop fever when infected by viruses or gram-positive bacteria. Therefore, we injected Pekin ducks with muramyl dipeptide, the cell walls of heat-killed Staphylococcus aureus, or the viral mimic polyinosinic:polycytidylic acid and monitored their body temperature (T(b)). For comparative purposes we also injected a group of ducks with lipopolysaccharide, the only known pyrogen in birds. We then compared the T(b) invoked by each injection with the T(b) after an injection of saline. Muramyl dipeptide did not affect T(b). The cell walls of heat-killed S. aureus invoked long-lasting, dose-dependent fevers with relatively low magnitudes. Polyinosinic:polycytidylic acid invoked dose-dependent fevers with high febrile peaks. Fever is a well-known clinical sign of infection in mammals, and the results of this study indicate that the pattern of increase in T(b) could serve as an indicator for diverse pathogenic diseases in birds.

Effect of temperature, oxygen and light on the degradation of ß-carotene, lutein and α-tocopherol in spray-dried spinach juice powder during storage.

The aim of this study was to evaluate the interaction between packaging parameters (transmission of light and oxygen) and storage temperatures (4, 20, 40 °C) on nutrient retention of Spinach (Spinacia oleracea) juice, spray-dried in the absence of an added encapsulant. ß-Carotene was more susceptible to degradation compared with lutein and α-tocopherol. Under our experimental conditions, it was observed that excluding low fluorescent light intensity and air by vacuum packaging at 20 °C did not seem to improve nutrient retention loss over time (p > 0.05). The rate of ß-carotene, lutein and α-tocopherol loss displayed first order reaction kinetic with low activation energy of 0.665, 2.650 and 13.893 kJ/mol for vacuum, and 1.089, 4.923 and 14.142 kJ/mol for non-vacuum, respectively. The reaction kinetics and half-life for ß-carotene, lutein and α-tocopherol at 4 °C and non-vacuumed were 2.2 × 10-2, 1.2 × 10-2, and 0.8 × 10-2 day-1, and 32.08, 58.25 and 85.37 day, respectively.

STY, a tyrosine-phosphorylating enzyme with sequence homology to serine/threonine kinases.

We have cloned a novel kinase (STY) from an embryonal carcinoma cell line. Sequence analysis of the STY cDNA reveals that it shares sequence homology with serine/threonine-type kinases and yet the bacterial expression product of the STY cDNA appears to have serine-, threonine-, and tyrosine-phosphorylating activities. The predicted STY protein is highly basic and contains a putative nuclear localization signal. During differentiation, two new mRNAs were detected in addition to the embryonic transcript.

Proviral inactivation of the Npat gene of Mpv 20 mice results in early embryonic arrest.

The Mpv 20 transgenic mouse strain was created by infection of embryos with a defective retrovirus. When Mpv 20 heterozygous animals were crossed, no homozygous neonatal mice or midgestation embryos were identified. When embryos from heterozygous crosses were cultured in vitro, approximately one quarter arrested as uncompacted eight-cell embryos, indicating that proviral insertion resulted in a recessive lethal defect whose phenotype was manifest very early in development. Molecular cloning of the Mpv 20 insertion site revealed that the provirus had disrupted the Npat gene, a gene of unknown function, resulting in the production of a truncated Npat mRNA. Expression of the closely linked Atm gene was found to be unaffected by the provirus.

Investigation of human pharmacoscintigraphic behavior of two tablets and a capsule formulation of a high dose, poorly water soluble/highly permeable drug (efavirenz).

Human pharmacoscintigraphic behavior of two tablets and a capsule formulation of a high dose, poorly water soluble, highly permeable, micronized drug (efavirenz) was investigated. The tablets and capsule, prepared with samarium oxide and neutron activated to produce radioactive samarium-153, were evaluated for their in vivo disintegration and gastrointestinal (GI) transit in healthy subjects under fasted condition. Scintigraphic images were acquired to coincide with blood sampling times to assess the plasma concentration-time profile in relation to in vivo disintegration and GI transit. The mean gastric emptying times were approximately the same for all three formulations. Although in vivo dosage form disintegration was faster for Tablet A as compared to Tablet B and was similar between Tablet A and the capsule, Tablet A showed a slower rate and extent of drug absorption than Tablet B and the capsule. The results of this study eliminated the initial hypothesis that the difference in in vivo performance between the two tablet formulations is due to a different rate of in vivo disintegration and suggest that for this drug the in vivo dissolution rate of the drug from its disintegrated dosage form was a more important factor affecting the rate and extent of drug absorption.

The Unp proto-oncogene encodes a nuclear protein.

The mouse Unp gene is related to TRE17, a human oncogene, but is not its mouse homolog. Unp is a ubiquitously expressed gene producing two related mRNAs. The protein product of Unp is localized in the nucleus. Expression of Unp from a highly active promoter results in tumorigenic transformation of NIH3T3 cells injected into athymic mice. Unp therefore encodes a novel nuclear oncoprotein.

Association of UNP, a ubiquitin-specific protease, with the pocket proteins pRb, p107 and p130.

The murine Unp gene encodes a widely expressed ubiquitin-specific protease. The predicted sequence of the UNP protein features motifs common to viral oncoproteins through which these proteins interact with the retinoblastoma gene product pRb, as well as the related 'pocket proteins' p107 and p130. We have explored the possibility that UNP interacts with pocket proteins, and report here that such associations can be detected in vitro and in cells. Associations of UNP and pocket proteins are sensitive to site-directed mutations in a manner directly analogous to those documented in viral oncoproteins. We conclude that within cells UNP does physically associate with pRb, and can also associate with p107 and p130.

Unp, a mouse gene related to the tre oncogene.

We have cloned cDNAs from a novel gene designated Unp. Unp cDNAs contain a large open reading frame that would encode a protein of 89 kDa. The predicted protein contains a putative nuclear localization signal, as well as consensus sequences for binding to the retinoblastoma gene product. The latter elements are contained within a region having strong similarity to the human tre oncogene. We have localized the Unp gene to mouse chromosome 9 in a region of homology with human chromosome 3p. This region has been implicated in a number of human malignancies.

Elevated expression of Unph, a proto-oncogene at 3p21.3, in human lung tumors.

The murine Unp proto-oncogene encodes a nuclear ubiquitin protease whose overexpression leads to oncogenic transformation of NIH3T3 cells. We have cloned cDNAs originating from the human homolog of this gene, designated herein as Unph (Unp, human), and have used these cDNAs to map the gene to 3p21.3, a region frequently rearranged in human tumor cells. Unph mRNA levels are consistently elevated in small cell tumors and adenocarcinomas of the lung, suggesting a possible causative role for the gene in the neoplastic process.

Genomic structure of Unp, a murine gene encoding a ubiquitin-specific protease.

The murine Unp gene encodes a ubiquitin-specific protease, a member of a family of enzymes that includes the product of the human tre-2 oncogene. The Unp gene has previously been mapped to chromosome 9. We have cloned in bacteriophage a 50 kilobase region of chromosome 9 containing the Unp gene, and have determined the nucleotide sequence of the gene. The gene has 22 exons, distributed over 47.4 kb. A processed ribosomal S2 pseudogene was identified in the third intron of the Unp gene. Expression of Unp is driven by a GC-rich, 'housekeeping' type promoter.

Modulation of kidney function in conscious Pekin ducks by atrial natriuretic factor.

The influence of avian atrial natriuretic factor (ANF) on renal function was examined in conscious saltwater-acclimated Pekin ducks undergoing a steady state diuresis maintained by iv infused isotonic avian Krebs-Ringer solution at a rate of 1.0 ml/min. Synthetic chicken ANF (chANF) was applied iv at doses of 10, 50, and 90 ng/min.kg BW for 10 min and caused dose-dependent transient increases in urine flow, osmolal excretion, glomerular filtration rate, effective renal plasma flow, and fractional water clearance at decreased urinary osmolality. Using receptor autoradiography, binding sites for [125I]Bolton-Hunter-labeled chANF [( 125I]BH-chANF) were localized in both the reptilian-type glomeruli and the collecting duct system throughout the duck kidney. A RRA for [125I]BH-chANF, established using an enriched kidney membrane fraction, indicated that unlabeled chANF and human ANF competitively displaced [125I]BH-chANF with comparable potencies. ANF-induced modulation of renal salt and water elimination via glomerular and tubular receptor interactions is consistent with the concept that this hormone has a physiological role in avian volume homeostasis.

Plasma atrial natriuretic factor responses to blood volume changes in the Pekin duck.

A RIA was developed for the measurement of immunoreactive atrial natriuretic factor (irANF) in avian plasma and was used to investigate the relationship between vascular volume and plasma irANF concentrations in conscious Pekin ducks. In normally hydrated birds, the mean plasma irANF concentration was 78.5 +/- 6.8 pg/ml. Blood volume expansion by iv infusion of isotonic saline at 1 ml/min elevated irANF concentrations by 132% after 1 h and by 233% after 2 h. Reduction of the vascular volume by nonhypotensive hemorrhage of 10% and then 20% of the total blood volume reduced irANF levels by 16% and 42%, respectively, 5 min after blood removal was completed. Similarly, in birds deprived of water for 24 h and with blood volume reduced by 4.8%, plasma irANF concentrations were 50% lower than in the same euhydrated animals. The observed correlation between plasma irANF concentrations and vascular volume is consistent with the concept that ANF has a physiological role in avian volume homeostasis.

Calcium metabolism and bone mass in female rabbits during skeletal maturation: effects of dietary calcium intake.

This study documents growth and dual-energy X-ray absorptiometry (DXA)-determined peak bone mass profiles in the rabbit model, and tests the hypothesis that rabbits show patterns of bone accretion similar to humans and thus may serve as a viable model for human bone physiology. It is also shown that dietary Ca intake affects peak bone mass and the temporal pattern of its attainment. Groups of weaned animals were administered two nutritionally complete but calcium-altered diets (0.5% or 1.0% Ca). We evaluated growth, bone mass accretion, and Ca metabolism from 20 to 56 weeks of age in both the 1.0% Ca and 0.5% Ca groups of rabbits. For each monthly period, we monitored body mass, naso-tail length, food consumption, and fecal output. In addition, we collected blood and 24 h urine samples for biochemical analyses, and measured bone mass variables of the lumbar spine with DXA. The 1.0% Ca group had a lower apparent fractional absorption of Ca and higher urinary Ca excretion, but retained more Ca than the 0.5% Ca group during the growth phase. Furthermore, the 1.0% Ca group had lower parathyroid hormone (PTH) and bone biochemical marker concentrations throughout the study than the 0.5% Ca group. The lower levels of PTH and bone markers of resorption and formation, may have resulted in a reduction in skeletal remodeling, and this physiological mechanism may have contributed to the 10% increase in peak bone mineral density of the lumbar spine in the 1.0% Ca group of rabbits.

Analysis of ubiquitination in vivo using a transgenic mouse model.

The primary pathway for the proteolytic destruction of cellular proteins is through ubiquitin-mediated targeting to the proteasome. This pathway is pivotal not only in the elimination of damaged or misfolded proteins but also in the temporal, developmental, or signal-mediated destruction of normal cellular substrates. The list of known substrates of the ubiquitin/proteasome pathway is long, but most substrates have been identified in yeast or, more recently, in cultured mammalian cells. It is likely that many mammalian substrates with developmental or disease relevance have yet to be identified because their ubiquitination occurs in tissue or organ systems that cannot be adequately modeled in vitro. We have developed a transgenic mouse model that will allow the isolation and identification of these substrates. The human UbC promoter was used to drive expression of a hexahistidine-tagged version of human ubiquitin in a variety of mouse tissues from early embryonic stages, as assessed by a green fluorescent protein marker. Cleavage of the fusion protein by endogenous enzymes produced epitope-tagged ubiquitin that was detected both in monomeric form and conjugated to cellular proteins. This mouse model should facilitate in the analysis of normal and disease-related ubiquitination events in vivo.

Size-dependent response to conspecific mating calls by male crickets.

Male sexual displays provide females with information that is crucial to their reproductive decisions. That same information is available to eavesdroppers, with potential consequences for both signaller and receiver. We present empirical evidence for size-dependent responses to intersexual communication by conspecific rivals. Acoustic features of a male house cricket's (Acheta domesticus) mating call are positively associated with its size, with females preferring the calls of larger males. In order to investigate whether conspecific males make use of the information content of mating calls, we examined their phonotactic responses to call recordings that differ in attractiveness to females. Males of all sizes exhibited positive phonotaxis, with smaller males showing a clear preference for female-preferred calls. Smaller males were also less likely to seek contact with the speaker playing their chosen call. We discuss possible explanations for this size-dependent male behaviour.

The effect of acetylsalicylic acid on renal function in the Pekin duck.

The acute effects of intravenously administered lysine-acetylsalicylic acid (ASA) on renal function in the Pekin duck have been studied with special reference to possible interactions with the antidiuretic hormone, arginine vasotocin (AVT), in the control of renal water and solute output. ASA produces an immediate increase in urine flow rate which is dose-related in the range 25 to 100 mg kg-1 and is associated with a slight reduction in urine osmolality, but an overall increase in renal osmolal excretion affecting Na+, Cl- and K+ to approximately equal extents. The effects, which are similar in both saltwater and freshwater adapted ducks infused with hyposmotic saline or glucose solution, can also be produced by similar doses of sodium salicylate (SA). The mechanism of action is probably not related to inhibition of prostaglandin synthetases. There is no change in the glomerular filtration rate or peripheral blood pressure following the ASA injection. There is no change in the circulating level of AVT; however, preliminary studies do not exclude the possibility of a partial antagonism of salicylate to AVT at the renal level.

In vitro studies on an antagonist of parathyroid hormone [Nle-8, Nle-18, Tyr-34]bPTH-(3-34)amide.

1 The actions of parathyroid hormone (PTH) are antagonized in vitro by the peptide [Nle-8, Nle-18, Tyr-34]-bPTH-(3-34)amide, an analogue of PTH. In this paper, the actions of the inhibitory peptide were investigated in vivo. 2 Native parathyroid hormone (bPTH-(1-84)), administered i.v. (0.17-1.51 nmol in volume of 0.3 ml) to 7 day old chicks produced hypercalcaemia but administration of the analogue in doses up to 173 nmol was ineffective in this respect. 3 The analogue failed to antagonize the hypercalcaemia produced by bPTH-(1-34) when injected, in 10 fold molar excess, 2 min before or simultaneously with bPTH-(1-34). 4 Normocalcaemia was restored in parathyroidectomized rats by intravenous infusion of bPTH-(1-84) at 32 pmol kg-1 h-1. Addition of the analogue to the infusion fluid in a 200 fold molar excess did not affect the concentrations of calcium and phosphate in the plasma, cyclic adenosine 3',5'-monophosphate (cyclic AMP) in the urine or phosphate clearance but produced a significant (P less than 0.05) rise in urinary calcium clearance. 5 The results suggest that the peptide [Nle-8, Nle-18, Tyr-34]-bPTH-(3-34)amide does not antagonize the actions of PTH in vivo and demonstrate an important dichotomy between in vitro and in vivo biological properties of the PTH analogue.