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18 décembre 2018 Non Par admin

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have no experimental support, our results indicate the possibility that a t-mediated mate selection system may be more primitive/fundamental than a system based on genetic dissimilarity, because the t-mediated mate selection system works in domestic birds such as japanese quail
taken together, we have demonstrated that female mating preference in the japanese quail is stimulated by male feather coloration in a t-dependent manner and that female birds develop a keen sense for this coloration due to upregulation of retinal red-sensitive opsin under breeding conditions
while we are curious about the mechanism underlying the upregulation of red opsin under ld conditions, further research is required to clarify this point
it is expected that elucidation of this mechanism will open new avenues in avian reproductive physiology, especially in the fields of sexual selection and seasonal breeding activity
five to 15 week-old japanese quail (quail cosmos co
, toyohashi japan) were maintained individually at 20–25 °c under a photoperiod of 14 l:10d (lights on at 05:00) for long-day conditions
for short-day conditions, birds were maintained under a photoperiod of 10 l:14d (lights on at 07:00)
all birds were given ad libitum access to water and a commercial diet (hokkaiya co, aichi, japan)
castration was performed under deep anesthesia by isoflurane inhalation (wako pure chemicals, osaka japan)
the cheek area feathers in males were removed using forceps
the size of the cloacal gland was measured using calipers
blood sampling was performed from the wing-vein and plasma testosterone and estradiol-17β were measured using an elisa kit (cayman, ann arbor, mi)
in the blindfold treatment, we covered the left eyes of females by rubber plunger of 2
5 ml disposable syringe (terumo corp
, tokyo, japan), which was fixed by adhesive in order to shut out the transmitted light
retinas and skin samples of female birds were isolated after cervical dislocation at 2 h after dawn
all experimental procedures for the care and use of animals were carried out in accordance with the approved guidelines of the animal care committees of shizuoka university (approval number: 28–13, 29a-11)
we performed microsatellite analysis to clarify the extent of genetic relatedness among the birds used in this study
dna samples were isolated from blood and used for pcr amplification
primers were prepared for 19 microsatellite loci (guj0005, 11, 13, 14, 17, 21, 23, 26, 28, 40, 41, 44, 49, 54, 55, 65, 69, 71, 84, 85)43
the forward primers were labeled at the 5′ end with 6-fam, 5-hex or ned
multiplex pcr was carried out with two or three primer pairs in a total volume of 12
5 μl (1 μl of genomic dna, 1× pcr buffer, 0
1 μm each of primers and 0
25 u of kod -plus- neo dna polymerase (toyobo, osaka, japan))
the pcr profile consisted of 94 °c for 2 min, followed by 30 cycles of 98 °c for 10 s, 55–62 °c for 30 s and 68 °c for 30 s
annealing temperatures for each reaction were chosen according to a previous report43
5 μl of pcr product was mixed with 0
5 μl of genescan™ 500 liz® size standard (applied biosystems, foster city, ca) and 9 μl of formamide, and denatured at 95 °c for 5 min
this mixture was used to determine the number of repeats in the microsatellites using a 3500 × l genetic analyzer (applied biosystems)
the datasets were then analyzed for allele calling using genemapper 5
genetic relationships were calculated based on the 19 microsatellite genotypes using genepop version 4
estradiol-17β, testosterone and progesterone were obtained from sigma japan (tokyo, japan)
the hormones were dissolved in sesame oil (1 mg/ml) and subcutaneously injected in the back at a dose of 0
1 mg/100 g body weight
the same volume of sesame oil was injected as a vehicle control
hormone injections (in supplementary figs s2 and s7a) were performed twice a week
females were deeply anesthetized by isoflurane inhalation and placed in a stereotaxic instrument (david kopf instruments, tujunga, ca)
a guide cannula (ag-8; eicom, san diego, ca) was implanted stereotaxically into the cerebral ventricle (3
0 mm anterior, 0 mm lateral from the y-point and 7
0 mm deep from the surface of the dura matter) and fixed to the skull with dental cement
an obturator was inserted into the guide cannula until experiments began
3 days after surgery, we injected bovine tsh (t8931, sigma japan) or pbs through a guide cannula and measured the resulting changes in target gene expression
injections were given every 2 days and continued for 2 weeks
we used bovine tsh because avian tsh was not available, and bovine tsh is known to activate the avian tsh receptor46
the testing apparatus used to assess mating preference consisted of a plastic box (60 cm × 30 cm × 30 cm) with two boundaries
each boundary was positioned 15 cm away from the long edge of the box and comprised wire netting (fig
the apparatus was lit from the top by two 20 w lamps
the center area of the apparatus was divided into 3 parts (blocks 1–3), each of which was 10 cm wide
to begin the mating preference test, the stimulus males were each placed on the one side of the focal female, which was placed in a holding box at the center of the apparatus
after 30 s, the holding box was removed, allowing the female to move around the enclosure
the first 1 min was disregarded as acclimation, and preference data were recorded for the next 5 min
we recorded the time that the female spent on each side (block 1 or 3) and assumed that she spent more time near the preferred male
after 5 min, the positions of the stimulus males were reversed (i
the male on one side was placed on the other side and vice versa), and another 5 min recording was performed
in total, 10 min of testing was performed and the time the female spent on each side of the stimulus male was calculated
to investigate if female eyesight is important for mate preference, mating preference tests were performed under lamps covered with a sharp cut filter (sc64; fuji film co
the cheek area feathers of males were removed and placed on a sheet of black paper (3 cm2)
the absolute values of l*, a* and b* were then

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