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5-Aminolevulinic acid improves chicken sperm motility
Taniguchi Shin,Zhu Zhendong,Matsuzaki Mei,Tsudzuki Masaoki,Maeda Teruo 아세아·태평양축산학회 2021 Animal Bioscience Vol.34 No.12
Objective: This study investigated the effects of 5-aminolevulinic acid (5-ALA) on the motility parameters, mitochondrial membrane depolarization, and ATP levels in chicken sperm. Methods: The pooled semen from Barred Plymouth Rock males was used. In the first experiment, the semen was diluted 4-times with phosphate-buffered saline (PBS (-)) containing various concentrations (0, 0.01, 0.05, and 0.1 mM) of 5-ALA, and then the sperm motility parameters after incubation were evaluated by computer-assisted sperm analysis (CASA). In the second experiment, the semen was diluted 4-times with PBS (-) containing 0.05 mM 5-ALA, and then sperm mitochondrial membrane depolarization and ATP levels after 1.5 h of incubation were analyzed with the MitoPT® JC-1 Assay and ATP Assay kits, respectively. In the third experiment, the semen was removed from the seminal plasma and resuspended with the mediums of PBS (-), PBS (-) supplemented with CaCl2 and MgCl2 (PBS (+)) + 5-ALA, PBS (+) + caffeine, and PBS (+) + caffeine + 5-ALA. Then, the sperm motility parameters after incubation were evaluated by CASA. In the last experiment, the semen was treated with the mediums of PBS (-), PBS (-) + 5-ALA, 5.7% glucose, 5.7% glucose + 5-ALA after removing the seminal plasma, and then the sperm motility parameters were evaluated by CASA. Results: The addition of 0.05 mM 5-ALA significantly increased the chicken sperm motility, progressive motility, linearity, average path velocity, curvilinear velocity, straight-line velocity, and the wobble. The sperm mitochondrial membrane depolarization was also increased by the 5-ALA treatment. The 5-ALA treatment decreased the sperm ATP levels. Both the caffeine treatment and glucose treatment decreased the sperm motility during incubation period. Conclusion: 5-ALA might increase sperm mitochondrial membrane depolarization to utilize the ATP for enhancing sperm movement. Objective: This study investigated the effects of 5-aminolevulinic acid (5-ALA) on the motility parameters, mitochondrial membrane depolarization, and ATP levels in chicken sperm.Methods: The pooled semen from Barred Plymouth Rock males was used. In the first experiment, the semen was diluted 4-times with phosphate-buffered saline (PBS (-)) containing various concentrations (0, 0.01, 0.05, and 0.1 mM) of 5-ALA, and then the sperm motility parameters after incubation were evaluated by computer-assisted sperm analysis (CASA). In the second experiment, the semen was diluted 4-times with PBS (-) containing 0.05 mM 5-ALA, and then sperm mitochondrial membrane depolarization and ATP levels after 1.5 h of incubation were analyzed with the MitoPT<sup>®</sup> JC-1 Assay and ATP Assay kits, respectively. In the third experiment, the semen was removed from the seminal plasma and resuspended with the mediums of PBS (-), PBS (-) supplemented with CaCl2 and MgCl2 (PBS (+)) + 5-ALA, PBS (+) + caffeine, and PBS (+) + caffeine + 5-ALA. Then, the sperm motility parameters after incubation were evaluated by CASA. In the last experiment, the semen was treated with the mediums of PBS (-), PBS (-) + 5-ALA, 5.7% glucose, 5.7% glucose + 5-ALA after removing the seminal plasma, and then the sperm motility parameters were evaluated by CASA.Results: The addition of 0.05 mM 5-ALA significantly increased the chicken sperm motility, progressive motility, linearity, average path velocity, curvilinear velocity, straight-line velocity, and the wobble. The sperm mitochondrial membrane depolarization was also increased by the 5-ALA treatment. The 5-ALA treatment decreased the sperm ATP levels. Both the caffeine treatment and glucose treatment decreased the sperm motility during incubation period.Conclusion: 5-ALA might increase sperm mitochondrial membrane depolarization to utilize the ATP for enhancing sperm movement.
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Osman, S.A.-M.,Sekino, M.,Nishibori, M.,Yamamoto, Y.,Tsudzuki, M. Asian Australasian Association of Animal Productio 2005 Animal Bioscience Vol.18 No.6
Blood samples were collected from eight native Japanese breeds of chickens (Miyadi-dori, Ohiki, Onaga-dori, Shoukoku, Tosa-Jidori, Tosa-Kukin, Toutenkou and Uzurao) and two foreign breeds of chickens (White Leghorn and Rhode Island Red) to examine the genetic variability and relationships among the breeds by using a microsatellite DNA technique. Except for the Shoukoku breed, the other Japanese chicken breeds all originate from Kochi Prefecture. Ohiki, Onaga-dori, Tosa-Jidori, Toutenkou and Uzurao are fancy fowl, and Miyadi-dori and Tosa-Kukin are utility fowl. Among the fancy fowl, Ohiki, Onaga-dori, and Toutenkou males have thick and long feathers in the saddle and tail. Genetic variabilities of the 20 microsatellites examined, varied depending on the breed: the mean number of alleles per locus ranged from 2.05 (Miyadi-dori) to 3.90 (Rhode Island Red); proportion of polymorphic loci ranged from 0.75 (Miyadi-dori) to 1.00 (Rhode Island Red, Shoukoku and Uzurao); and mean expected heterozygosity ranged from 0.330 (Miyadi-dori) to 0.607 (Rhode Island Red). Unique microsatellite alleles were detected in each breed. Using the neighbour-joining method, phylogenetic trees were constructed based on the genetic distances of D$_{A}$ and D$_{ST}$. Among the breeds originating from Kochi Prefecture, fancy and utility breeds belonged to different clusters. Among the fancy breeds, those having thick and long feathers in the tail and saddle showed a close genetic relationship to the Shoukoku breed, which also has thick and long feathers in the tail and saddle.
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Osman, S.A.M.,Sekino, M.,Nishihata, A.,Kobayashi, Y.,Takenaka, W.,Kinoshita, K.,Kuwayama, T.,Nishibori, M.,Yamamoto, Y.,Tsudzuki, M. Asian Australasian Association of Animal Productio 2006 Animal Bioscience Vol.19 No.10
This is the first study in which genetic variability and relationships of a large number of Japanese chicken breeds were revealed along with those of several foreign breeds by using microsatellite DNA polymorphisms. Twenty-eight breeds (34 populations) of native Japanese chickens and seven foreign breeds or varieties were analyzed. The mean number of alleles per locus, the proportion of the polymorphic loci, and the expected average heterozygosity ranged from 1.75 to 4.70, from 0.55 to 1.00, and from 0.21 to 0.67, respectively. Microsatellite alleles being unique to a particular population were detected in some populations. The $D_A$ genetic distance between populations was obtained from allele frequency for every pair of the populations to construct a neighbor-joining tree. According to the phylogenetic tree, excluding a few exceptions, native Japanese chicken breeds and foreign breeds were clearly separated from each other. Furthermore, the tree topology divided native Japanese chickens into four main classes, which was almost in accordance with the classification based on body morphology; that is, (1) Cochin type, (2) Malay type, (3) layer type, and (4) intermediate type between Malay and layer types. This is the first finding for native Japanese chickens.
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