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Cell fusion during yeast mating requires high levels of a-factor mating pheromone.

During conjugation, two yeast cells fuse to form a single zygote. Cell fusion requires extensive remodeling of the cell wall, both to form a seal between the two cells and to remove the intervening material. The two plasma membranes then fuse to produce a continuous cytoplasm. We report the characterization of two cell fusion defective (Fus-) mutants, fus5 and fus8, isolated previously in our laboratory. Fluorescence and electron microscopy demonstrated that the fus5 and fus8 mutant zygotes were defective for cell wall remodeling/removal but not plasma membrane fusion. Strikingly, fus5 and fus8 were a specific; both mutations caused the mutant phenotype when present in the MATa parent but not in the MAT alpha parent. Consistent with an a-specific defect, the fus5 and fus8 mutants produced less a-factor than the isogenic wild-type strain. FUS5 and FUS8 were determined to be allelic to AXL1 and RAM1, respectively, two genes known to be required for biogenesis of a-factor. Several experiments demonstrated that the partial defect in a-factor production resulted in the Fus- phenotype. First, overexpression of a-factor in the fus mutants suppressed the Fus- defect. Second, matings to an MAT alpha partner supersensitive to mating pheromone (sst2 delta) suppressed the Fus- defect in trans. Finally, the gene encoding a-factor, MFA1, was placed under the control of a repressible promoter; reduced levels of wild-type a-factor caused an identical cell fusion defect during mating. We conclude that high levels of pheromone are required as one component of the signal for prezygotes to initiate cell fusion.

Alleles↗

Delayed selfing as an optimal mating strategy in preferentially outcrossing species: theoretical analysis of the optimal age at first reproduction in relation to mate availability.

The evolution of mating systems and that of life history have usually been modeled separately. However, they may be to some extent coupled in natural situations because they rely on the same phenotypic traits. Here, we focus on one of these traits, the age at first reproduction, in a species able to self- and cross-fertilize. When inbreeding depression is strong, self-fertile species preferentially cross-fertilize. However, outcrossing is not always possible when the availability of sexual partners is limited. The optimal reproductive strategy in this case would be to wait for a sexual partner for a certain period of time (the waiting time) and then switch to selfing if no mates have been encountered (reproductive assurance strategy). We predict the evolution of an optimal waiting time depending on the efficiency of resource reallocation to late fecundity, on the inbreeding depression, and on the instantaneous probability of encountering a partner versus dying. As a consequence of reduced mate availability, intermediate selfing rates can be generated in preferentially outcrossing populations, but they are lowered by the existence of a waiting time. Our model may thus explain low selfing rates observed in natural populations of many self-fertile, preferentially outcrossing plants or animal species.

Age Factors↗

Aedes triseriatus and La Crosse virus: similar venereal infection rates in females given the first bloodmeal immediately before mating or several days after mating.

Venereal infection rates of Aedes triseriatus females mated to males transovarially infected with La Crosse virus were determined in 6 cage-mating trials. In trials 1-4, venereal infection rates averaged 46% and 45% in F2 females bloodfed 6-8 hr before and 7 days after exposure, respectively, to transovarially-infected males. These rates were similar to rates previously reported only in mosquitoes receiving a bloodmeal 6-8 days prior to mating. Lower rates (24%-31%) were obtained using F4 and F7 generation mosquitoes in trials 5B and 6. In most trials, oral and transovarial transmission rates by venereally infected females were less than 25%. In trial 5B, however, the transovarial transmission rates reached 60% and 94% in the second and third ovarian cycles, respectively, with filial infection rates of 46% and 65%, respectively. The oral transmission rate in this trial reached 38% after 32 days. LAC virus was not detected in first ovarian cycle progeny. It is concluded that higher venereal infection rates must be found and/or first ovarian cycle progeny shown to become infected, before venereal transmission can be considered to make more than a modest contribution to offsetting the erosion of virus prevalence during TO transmission.

Aedes↗

Expression of MFA1 and STE6 is sufficient for mating type-independent secretion of yeast a-factor, but not mating competence.

The yeast a-factor mating peptide and its transporter Ste6 are normally expressed only in MATa haploid cells. The a-factor is initially produced as a 36- or 38-residue peptide precursor and must undergo extensive post-translational processing to produce an active 12 amino-acid lipopeptide. To better understand the steps required for Ste6-dependent a-factor transport, we have reconstituted a-factor synthesis and transport in MATalpha haploids and MATa/alpha diploids. Ste6 and a-factor were stably expressed in MATalpha and MATa/alpha cells and the ectopically expressed a-factor was correctly processed. In addition, Ste6 was able to transport a-factor from all cell types, indicating that once expressed no other MATa-specific functions are required. However, despite significant levels of a-factor secretion, MATalpha cells are unable to support efficient mating.

ATP-Binding Cassette Transporters↗

Interspecific actions of alpha mating pheromones on the a mating-type cells of three Saccharomyces yeasts.

The alpha mating pheromones synthesized in three Saccharomyces yeasts (S. cerevisiae, S. kluyveri, and S. exiguus) displayed interspecific actions on the a cells of all three species despite the fact that the amino acid sequences of all three alpha pheromones are different. Mating between species, however, did not occur. The interspecific alpha pheromone--a cell reaction was not necessarily more effective than the interspecific one.

Amino Acid Sequence↗

Mating reaction in Saccharomyces cerevisiae. X. Agglutinability-inactivating factor: a factor which destroys sexual agglutinability of a mating-type cells.

From cells of Saccharomyces cerevisiae a factor has been extracted that destroys the agglutinability of a mating-type cells specifically. It was found in the cell extracts of diploid and tetraploid strains as well as haploid strains of a and alpha mating types. It is heat-labile and the molecular weight is about 50,000. It is adsorbed by neither a cells nor alpha cells. Its biological activity is dependent on the incubation temperature and the pH, and is completely inhibited by phenylmethylsulfonyl fluoride, a potent inhibitor of the serine proteases. All the results described in this paper indicate that this factor is a proteolytic enzyme.

Adsorption↗

Flagellar adhesion between mating type plus and mating type minus gametes activates a flagellar protein-tyrosine kinase during fertilization in Chlamydomonas.

When Chlamydomonas gametes of opposite mating type are mixed together, flagellar adhesion through sex-specific adhesion molecules triggers a transient elevation of intracellular cAMP, leading to gamete activation in preparation for cell-cell fusion and zygote formation. Here, we have identified a protein-tyrosine kinase (PTK) activity that is stimulated by flagellar adhesion. We determined that the protein-tyrosine kinase inhibitor genistein inhibited fertilization, and that fertilization was rescued by dibutyryl cAMP, indicating that the genistein-sensitive step was upstream of the increase in cAMP. Incubation with ATP of flagella isolated from non-adhering and adhering gametes followed by SDS-PAGE and immunoblotting with anti-phosphotyrosine antibodies showed that adhesion activated a flagellar PTK that phosphorylated a 105-kDa flagellar protein. Assays using an exogenous protein-tyrosine kinase substrate confirmed that the activated PTK could be detected only in flagella isolated from adhering gametes. Our results indicate that stimulation of the PTK is a very early event during fertilization. Activation of the PTK was blocked when gametes underwent flagellar adhesion in the presence of the protein kinase inhibitor staurosporine, but not in the presence of the cyclic nucleotide-dependent protein kinase inhibitor, H8, which (unlike staurosporine) does not block the increases in cAMP. In addition, incubation of gametes of a single mating type in dibutyryl cAMP failed to activate the PTK. Finally, flagella adhesion between plus and minus fla10-1 gametes, which have a temperature-sensitive lesion in the microtubule motor protein kinesin-II, failed to activate the PTK at elevated temperatures. Our results show that kinesin-II is essential for coupling flagellar adhesion to activation of a flagellar PTK and cAMP generation during fertilization in Chlamydomonas.

Animals↗

The evolution of mate choice and mating biases.

We review the current status of three well-established models (direct benefits, indirect benefits and sensory drive) and one newcomer (antagonistic chase-away) of the evolution of mate choice and the biases that are expressed during choice. We highlight the differences and commonalities in the underlying genetics and evolutionary dynamics of these models. We then argue that progress in understanding the evolution of mate choice is currently hampered by spurious distinctions among models and a misguided tendency to test the processes underlying each model as mutually exclusive alternatives. Finally, we suggest potentially fruitful directions for future theoretical and empirical research.

Animals↗

Multiple mating and sequential mate choice in guppies: females trade up.

The trade-up hypothesis outlines a behavioural strategy that females could use to maximize the genetic benefits to their offspring. The hypothesis proposes that females should be more willing to accept a mate when the new male encountered is a superior genetic source to previous mates. We provide a direct test of the trade-up hypothesis using guppies (Poecilia reticulata), and evaluate both behavioural and paternity data. Virgin female guppies were presented sequentially with two males of varying attractiveness, and their responsiveness to each male was quantified. Male attractiveness (ornamentation) was scored as the amount of orange coloration on their body. Females were generally less responsive to second-encountered males, yet responsiveness to second males was an increasing function of male ornamentation. These attractive second males also sired a greater proportion of the offspring. There was an overall tendency for last-male advantage in paternity, and this advantage was most exaggerated when the second male was more ornamented than the first. Finally, we found that our estimate of relative sperm number did not account for any significant variation in paternity. Our results suggest that female guppies may use pre-copulatory mechanisms to maximize the genetic quality of their offspring.

Animals↗

DEFENSE OF MATE AND MATING CHAMBER IN A WOOD ROACH.

Studies of the eastern wood roach, Cryptocercus punctulatus, reveal that, under laboratory conditions, the mating chambers in rotten wood are inhabited by mated pairs, the male of which will usually defend the chamber successfully against intruding males by a form of fighting. In every staged contest in which the intruder won the fight, it also won the resident female. Females, as well as unmated males singly inhabiting a chamber, could not be induced to defend against an intruder of either sex.

Aggression↗

Simulation of the mating advantage in mating of rare drosophila males.

In several species of Drosophila, certain males mate more frequently when they are less abundant than they do when they are in the majority. The mating advantage may be artificially induced by the use of a "double chamber" technique, even when there is no difference in the actual frequencies of the competing males.

Animals↗

a-Factor from Saccharomyces cerevisiae: partial characterization of a mating hormone produced by cells of mating type a.

Conjugation between haploid cells of Saccharomyces cerevisiae is mediated through the action of diffusible mating hormones, two of which have been designated as a-factor and alpha-factor. Partially purified fractions exhibiting a-factor activity have been obtained from culture filtrates of a cells by ultrafiltration, ion-exchange chromatography, and gel filtration. The a-factor preparations specifically caused both G1 arrest and morphological alterations in cells of alpha-mating type, whereas a cells, a/alpha diploids, and nonmating alpha mutants were not affected. The a-factor activity was found in the culture filtrates of all a strains tested, but not in filtrates of alpha or a/alpha cell cultures. The hormone is sensitive to various proteases, showing that it is associated with a peptide or protein. Gel filtration studies suggest an apparent molecular weight greater than 600,000; however, this result may be due to aggregation with carbohydrate present in the preparations. Although the biological activities of a-factor are analogous to those described previously for alpha-factor, the chemical properties of these two hormones appear to be quite different.

Cell Division↗

Serving capacity of crossbred yearling beef bulls. I. Single-sire mating behavior and fertility during average and heavy mating loads at pasture.

Eighty crossbred, virgin, yearling beef bulls were subjected to three serving capacity (SC) tests. Ten low SC (LSC) and 10 high SC (HSC) bulls similar in weight (avg wt = 452 kg), testicular size and seminal traits but differing in average services were selected. Objectives were to evaluate the mating behavior and fertility of LSC and HSC bulls at pasture when exposed to an average and a heavy mating load and to determine the effect of sexual activity on body weight and testicular characteristics. One LSC and one HSC bull per block were exposed single-sire to 25 naturally cyclic (N) cows for 3 d and to 9 estrus-synchronized (S) cows for 1 d in a randomized complete block design consisting of 10, 4-d blocks. Bulls were fitted with pedometers to record distance travelled. Behavioral data were collected by periodic visual observation. During the N cow treatment, LSC bulls had fewer (P less than .05) services per cow, total services and a higher mount to service ratio than HSC bulls. During the S cow treatment, LSC bulls showed increased sexual activity, though they achieved fewer (P less than .09) services per cow. Pregnancy rates for LSC and HSC bulls did not differ (P greater than .20) and across SC groups averaged 53.6% for N cows and 31.9% for S cows. Distance travelled also was similar (P greater than .20) between SC groups for both treatments. Sexual activity during pasture exposure had no effect (P greater than .20) on testicular characteristics but resulted in an average decline in body weight of 25.7 kg across SC groups during the 4-d period. Post-exposure SC tests found that LSC bulls had increased (P less than .05) services, decreased mounts and mount to service ratio compared to pre-exposure measurements, but LSC were still lower (P = .06) than HSC bulls for average services. These results suggest that LSC virgin, yearling bulls should be offered sexual experience and retested before their inherent SC can be determined.

Animals↗

Effects of adjustment for mate's yield, mate's sire's merit, and days open in sire evaluation.

First lactations were from the North Carolina Institutional Breeding project sine 1955 for cows and their dams where Modified Contemporary Comparisons were available for sires of both females. Each sire of cows was required to have a minimum of 20 progeny in the data. A total of 1771 cow-dam pairs by 33 sires in 7 herds and 217 herd-year-seasons passed all edits. Models including combinations of Predicted Difference milk for sire and maternal grandsire, dam's milk yield deviated from herd-year-season mean, and dam's age at cow's birth were used to predict cow's milk yield deviated from herd-year-season mean. Adjustment of yields of cow and dam for days open also was examined. Predicted Difference milk of her sire was the most important variable for predicting cow's yield; next was dam's yield. Predicted Difference of maternal grandsire and dam's age at cow's birth were not significant. Adjustment for days open reduced the percent of variation of cow's yield explained by sire's Predicted Difference. Sire's contributions to yield also were obtained by least squares and mixed model procedures in the various models with Predicted Difference milk of sire omitted. Maximum change in ranks among the 33 sires was four places, and maximum change of differences among sires was 129 kg when mate's yield and mate's sire's Predicted Difference were included in the model. Dam's yield was more useful than maternal grandsire's merit in predicting cow's yield. Differences among sires changed as much as 185 kg when cow's yield was adjusted for days open.

Animals↗

A mating-type-specific sterility gene map1 is required for transcription of a mating-type gene mat1-Pi in the fission yeast Schizosaccharomyces pombe.

Transcriptional activation of the mating-type gene mat1-Pi essential for meiosis in the fission yeast Schizosaccharomyces pombe was studied. A homozygous diploid harboring the mutation at the h+-specific sterility gene, map1, was arrested before premeiotic DNA synthesis in the nitrogen-free sporulation medium. Transcription of mat1-Pi was totally absent in the map1 mutant. The mei2 gene encoding a positive regulator for meiosis was normally transcribed in the map1 mutant, suggesting that the map1 function was specific to mat1-Pi.

Blotting, Northern↗

Mating stimulation required for mating-induced estrous abbreviation in female rats: effects of repeated testing.

Mating stimulation, particularly vaginal-cervical stimulation, causes estrous abbreviation in female rats. In most previous studies, female rats were repeatedly tested for sexual behavior until estrous termination occurred. Thus, it was not clear whether sensory stimulation (e.g., flank stimulation, olfactory cues) received during the repeated testing procedure contributed to estrous abbreviation. In Experiment 1, we determined the effect of premating to two or four ejaculations on the rate of estrous termination when a repeated testing procedure was used. We compared ovariectomized, hormone-primed, female rats receiving (1) four ejaculations, (2) two ejaculations, or (3) no premating. Females premated to either two or four ejaculations showed significantly lower levels of sexual receptivity 12 h later than did nonpremated females. These results confirm that premating induces estrous abbreviation when a repeated testing procedure is used. In Experiment 2, we determined whether the repeated testing procedure was necessary for estrous abbreviation. Ovariectomized, hormone-primed female rats were premated to two ejaculations or not premated. The rats were then tested for sexual behavior repeatedly or only once. Females that were premated and repeatedly tested for sexual behavior showed a statistically significant decrease in sexual receptivity compared to females that were not premated; however, the level of sexual receptivity in premated females did not differ from that in non-premated females when they were tested only once. The results suggest that heat duration is the result of a complex interplay between those factors that promote the expression of sexual receptivity and those that inhibit it.

Animals↗

Efficiency of single-sire mating programs with beef bulls mated to estrus synchronized females.

Efficiency in reference to pregnancy rates of breeding beef bulls with estrus synchronized cows and heifers was tested. Most bulls (104 of 112) were given a breeding soundness examination and two 10-min libido/serving capacity tests. Females received either Syncro-Mate-B (SMB) or two injections of Prostaglandin F(2)alpha (PGF) to synchronize estrus. They were assigned to single-sire breeding groups with bull-to-female ratios ranging from 1:7 to 1:51. Control groups consisted of untreated females maintained in single-sire breeding pastures with ratios from 1:24 to 1:37. Continuous observations of sexual activity were made for 30 h (SMB) and 48 h (PGF). After the 120-h posttreatment breeding period, females were placed in breeding pastures. During the synchronized breeding period the percentage of pregnant cattle of total treated was 43.5 +/- 1.7% compared (P < 0.01) with 58.9 +/- 3.3% for the control group after 23 d of breeding. At end of 28-d (treated) and 46-d (control) period, the percentage of pregnant females was 75.0 +/- 2.4 and 79.6 +/- 4.7, respectively (P > 0.05). In SMB trials, the percentages of females exhibiting estrus, those serviced at estrus and those pregnant following service during the synchronized breeding period were 90.8 +/- 1.5, 73.3 +/- 4.5 and 56.4 +/- 5.6%, respectively. In PGF trials, the means for these same factors were 78.3 +/- 2.4, 70.4 +/- 5.9 and 56.1 +/- 6.5%, respectively.

Journal Article↗

Behavior of triatomines (Hemiptera: Reduviidae) vectors of Chagas' disease. II. Influence of feeding, lighting and time of day on the number of matings, mating speed and duration of copulation of Panstrongylus megistus (Burm, 1835) under laboratory conditions.

To determine the influence of feeding, lighting and time of day on the copulating behavior of Panstrongylus megistus, 480 insect pairs were divided into four groups of 120 each and tested in the following respective situations: without food deprivation (F.D.), with five days of F.D., with ten days of F.D., and with 20 days of F.D. The tests were performed between 9:00 a.m. to 12:00 a.m. and 7:00 p.m. to 10:00 p.m., with light (700-1400 lux) and in the dark (1.4-2.8 lux) and behavior was recorded by the time sampling technique. Mating speed (MS) and duration of copulation (DC) were also calculated for each situation. The maximum frequency of copulation was observed after five days of F.D., at night, in the dark (n = 16), and the minimum was observed for recently-fed pairs, at night, with light (n = 4). Males approached females more often than females approached males. MS was lowest in pairs with twenty days of F.D., at night, with light (mean = 23.0 +/- 16.0 minutes), and highest in recently-fed pairs, during the day, with light (mean = 2.9 +/- 2.5 minutes). DC was shortest in recently-fed insects, during the day, in the dark (mean = 23.5 +/- 6.7 minutes), and longest in recently-fed animals, at night, in the dark (mean = 38.3 +/- 6.9 minutes).

Animals↗