Single Nucleotide Polymorphisms of Candidate Genes Related to Egg Production Traits in Vietnamese Indigenous Chickens

ABSTRACT The objective of this work was to detect single nucleotide polymorphisms (SNPs) in two candidate genes known to relate with reproductive traits in Vietnamese chicken. Two indigenous breeds Ri and Mia chickens were used for genotyping 4 SNPs namely intron 1 of growth hormon gene (GH), exon 5 of prolactin receptor gene (PRLR5), a 24 bp nucleotide sequence insertion-deletion at situation -358 in 5- untranslated region of prolatin gene (PRL24); and at site C2404T of prolactin gene (PRL2402). It was shown that the SNP sites of these candidate genes were identified in two local breeds. Three genotypes of each loci were found (AA, AB and BB for GH; II, ID and DD for 24 bp indel of PRL and CC, CT and TT for PRL2402), except only genotypes of AA and BB of PRLR5 were detected (no heterozygous genotype of AB). The frequencies of the desired genotypes of AA (GH), BB (PRLR5), II (PRL24) and CC (PRL2402) were low in both indigenous Ri and Mia chickens. Based on this study it could be suggested that the study on association between these loci with egg production traits and the selection to increase the desired genotype frequency in poputions are necessary to improve egg production of Ri and Mia chickens.

Development of tetra-primer ARMS-PCR protocol to genotype the prolactin receptor SNP 39136666 and assessment of this SNP in Brazilian locally adapted cattle breeds / [Desenvolvimento de protocolo de tetra-primer ARMS- PCR para genotipagem do SNP 39136666 do receptor da prolactina e avaliação desse SNP em raças bovinas brasileiras localmente adaptadas]

As raças taurinas de origem ibérica Limonero e Carora (Bos primigenius taurus) possuem o fenótipo de pelo curto, liso e com baixa densidade folicular, o que confere a esses animais maior tolerância térmica e melhor produtividade em regiões quentes. Diferentes mutações associadas a esse fenótipo foram descritas no gene do receptor de prolactina PRLR, localizado no cromossomo bovino BTA20. Uma mutação recentemente encontrada é a substituição do nucleotídeo C por T, SNP 39136666 (p. R497*), no exon 11, que gera um códon de parada e, consequentemente, uma menor isoforma desse receptor. Neste trabalho, desenvolveu-se um protocolo rápido e de baixo custo para detecção desse SNP, utilizando-se a técnica de tetra-primer ARMS-PCR. Assim, foi possível detectar essa mutação nas raças brasileiras de origem ibérica localmente adaptadas: Caracu, Crioulo Lageano, Mocho Nacional e Pantaneiro. O alelo T foi mais frequente na raça Caracu (80%), enquanto o alelo C foi mais frequente na raça Crioulo Lageano (84%). Essa simples metodologia pode ser usada para genotipar esse SNP e ajudar na aplicação dessas informações moleculares em programas de melhoramento focados na tolerância térmica em bovinos taurinos e seus mestiços.(AU)

Effects of Vitamin E Supplementation on Serum Hormones and Gene Expression of Anti-season Breeding Xingguo Grey Geese (Anser cygnoides)

The objective of this study was to analyse the impact of dietary vitamin E supplementation on laying performance, serum reproductive hormones concentration and gene expression in ovary and follicles of anti-season breeding goose. A total of 210 anti-season breeding geese were divided into seven treatments with six replications. Each group was supplied with diets containing different vitamin E (DL--tocopherol acetate) contents (0, 10, 20, 40, 80, 160, 320mg/kg). We observed that the egg production and laying rate improved significantly at doses of 10 and 80 mg/kg, while the highest egg weight appeared in the 320 mg/kg group. Meantime, 80 mg/kg of vitamin E supplementation significantly improved the concentration of follicle-stimulating hormone (FSH), luteinizing hormone (LH) and estradiol (E2) in serum (p 0.05). Dietary vitamin E supplementation significantly enhanced mRNA expression of FSHR, LHR and ESR1 at a dose of 80mg/kg, while PRLR increased at doses of 10 and 20mg/kg (p 0.05). It was found that the mRNA expression of Dicer increased at doses of 40 and 80 mg/kg of vitamin E supplementation in the ovary, SWF, LWF and SYF, respectively. Thence, Dietary vitamin E supplementation could improve egg laying performance, plasma reproductive hormones and the mRNA expression of reproductive hormone receptor genes in ovary, as well as the mRNA expression of Dicer in ovary, SWF, LWF and SYF. It was supposed that 80 mg/kg of vitamin E supplementation in dietary was appropriate to improve the fertility of anti-season breeding Xingguo grey goose.(AU)

Effects of Vitamin E Supplementation on Serum Hormones and Gene Expression of Anti-season Breeding Xingguo Grey Geese (Anser cygnoides)

The objective of this study was to analyse the impact of dietary vitamin E supplementation on laying performance, serum reproductive hormones concentration and gene expression in ovary and follicles of anti-season breeding goose. A total of 210 anti-season breeding geese were divided into seven treatments with six replications. Each group was supplied with diets containing different vitamin E (DL--tocopherol acetate) contents (0, 10, 20, 40, 80, 160, 320mg/kg). We observed that the egg production and laying rate improved significantly at doses of 10 and 80 mg/kg, while the highest egg weight appeared in the 320 mg/kg group. Meantime, 80 mg/kg of vitamin E supplementation significantly improved the concentration of follicle-stimulating hormone (FSH), luteinizing hormone (LH) and estradiol (E2) in serum (p 0.05). Dietary vitamin E supplementation significantly enhanced mRNA expression of FSHR, LHR and ESR1 at a dose of 80mg/kg, while PRLR increased at doses of 10 and 20mg/kg (p 0.05). It was found that the mRNA expression of Dicer increased at doses of 40 and 80 mg/kg of vitamin E supplementation in the ovary, SWF, LWF and SYF, respectively. Thence, Dietary vitamin E supplementation could improve egg laying performance, plasma reproductive hormones and the mRNA expression of reproductive hormone receptor genes in ovary, as well as the mRNA expression of Dicer in ovary, SWF, LWF and SYF. It was supposed that 80 mg/kg of vitamin E supplementation in dietary was appropriate to improve the fertility of anti-season breeding Xingguo grey goose.

POLIMORFISMO E EXPRESSÃO DE GENES ENVOLVIDOS COM A MOBILIZAÇÃO DE GORDURA NO PERÍODO DO PERIPARTO EM CABRAS LEITEIRAS

BAGATOLI, Adriana. Polimorfismo e expressão de genes envolvidos com a mobilização de gordura no período do pós-parto em cabras leiteiras. 2015. 68p Tese (Doutorado em Zootecnia). Instituto de Zootecnia, Universidade Federal Rural do Rio de Janeiro, Seropédica, RJ,2015. A apolipoproteína B (APOB), FATP (Slc27 A6), AGPAT6 e PRLR são proteínas distintas associadas ao metabolismo de lipídeos. A principal função da APOB é regular as vias de transporte e redistribuir os triglicerídeos entre os tecidos, por meio de lipoproteínas, como as VLDL. Muitos polimorfismos gênicos já foram descritos, sobretudo em humanos, mas é proposto que mutações neste gene podem estar associadas às variações metabólicas no período do periparto em animais de alta produção. A expressão da proteína completa, a APOB-IOO, ocorre principalmente no fígado e é alterada durante a lactação, e está relacionada com a secreção e transporte de VLDL para os tecidos e acúmulo de gordura nos hepatócitos. A FATP, localizada na membrana das células, capta o complexo de apolipoproteínas e por hidrólise, permite a entrada de AGNE para o meio intracelular. Durante a lactação essa proteína é bastante expressa, principalmente na glândula mamária, para auxiliar a absorção de ácidos graxos a ser incorporado no leite. A proteína AGPAT6 converte ácido lisofosfatídico em ácido fosfatídico, que são intermediários para a formação dos diacilgliceróis, enquanto a PRLR está envolvida diretamente com a regulação do crescimento e desenvolvimento das glândulas mamárias. Neste trabalho o nosso objetivo foi associar os polimorfismos encontrados nestes genes com as características de produção de leite e identificar a relação da expressão dos genes APOB-100 e FATP no tecido hepático entre as raças Saanen e Alpina em diferentes períodos de lactação. Para as análises de polimorfismo dos genes APOB, FATP, AGPAT6 e PRLR foram utilizadas amostras de sangue total de cabras alojadas no setor de caprinocultura da Universidade Federal de Viçosa. O sangue coletado foi utilizado para amplificação dos fragmentos de interesse através da reação de PCR e genotipados por meio da PCR-RFLP, em 90, 85, 118, 105, 247 e 255 amostras relacionadas aos primers Slc27A6-1, Slc27A6-2, Slc27A6-3 (FATP), APOB-l, A GPAT6 e PRLR, respectivamente. Para as análises de expressão gênica da APOB-I00 e FATP foram utilizadas 18 cabras (Capra hircusi das raças Saanen e Alpina em período seco, no início e no pico de lactação. Por meio de biópsia guiada por ultrassom, uma amostra de fígado de cada animal foi coletada e armazenada em nitrogênio líquido para as análises de PCR em tempo real. Por meio de análises estatísticas foi observado que, com exceção da amplificação gerada pelo par de primer Slc27A6-2, em que não houve variação genética aparente, e do gene PRLR, os polimorfismos apresentaram associação com as características de produção de leite, e poderão ser utilizados como marcadores moleculares (P < 0,05). Os genótipos identificados em F ATP foram associados às porcentagens de lactose e proteína no leite, enquanto para a APOB houve associações com as características de produção total de leite, extrato seco, gordura, lactose, proteína e duração da lactação, e para A GPAT6 houve relação com porcentagens de gordura, proteína, lactose e produção de lactose no leite. A expressão dos genes APOB-IOO e FATP foram diferentes entre as raças Saanen e Alpina, sendo que para a Alpina, a expessão da APOB-IOO e FATP foram menores no pico de lactação. Além disso, para a raça Alpina observou-se uma expressão maior no período seco e início da lactação em comparação ao pico para o gene APOB-IOO (P < 0,05).

BAGATOLI, Adriana. Polymorphism and expression of genes involved in fat mobilization in the postpartum period of dairy goats. 2015. 68p Thesis (phD in Animal Science). Institute of Animal Science, Federal Rural University ofRio de Janeiro, Seropédica, RJ,2015. The apolipoprotein B (APOB), FATP (Slc27A6), AGPAT6 and PRLR are distinct proteins associated with lipid metabolismo The main function of APOB is to reguIate the transport pathways and the redístributíon of triglycerides between the tissues through lipoproteins, sueh as VLDL. Several genetic polymorphisms have been described, espeeially in humans, but it is suggested that mutations in this gene may be assoeiated with metabolic changes in the peripartum period in high producing animaIs. The expression of complete protein, APOB-100, occurs mainly in the liver and changes during lactation and is re1ated to the secretion and transport of VLDL to the tissues and associated with the accumulation of fat in hepatocytes. Located in the cell membrane, FATP captures the complex of apoIipoproteins and allows NEF A to entry through the intracellular site by hydrolysis. This protein is fairly expressed during Iactation, especially in the mammary gland to aid the absorption of fatty acids to be incorporated in the milk. The AGPAT6 protein converts lysophosphatidic acid to phosphatidic acid, which are intermediaries for the forrnation of diglycerides, while PRLR is directly involved in the regulation of growth and development ofthe mammary glands. ln this work our aim was to associate the polymorphisms found in genes with milk production traits and identify reIation in the expression of APOB-l 00 genes and FATP in liver tissue between Saanen and Alpine breeds in different periods of lactation. For the analysis of polymorphísms genes APOB, FATP, AGPAT6 and PRLR were used total blood samples from housed goats from the goat section of Federal University of Viçosa. The colleeted blood was used for amplification ofinterest fragments by PCR and genotyped by PCR-RFLP, in 90,85, 118, 105, 247 and 255 samples re1ated with Slc27A6-1, Slc27A6-2, Slc27A6-3 (FATP), APOB-l, AGPAT6 and PRLR primers respectively.To the analysís of gene expression were used 18 goats (Capra hircus) of Alpine and Saanen breeds during dry períod, in the beginning and at the peak of lactation. Through guided biopsy of ultrasound, one of each animal liver sample was collected and stored in liquid nitrogen for real time PCR analysis. By statistical analysis it was found that, except for the amplification generated by the primer pair Slc27A6-2, which has not apparent genetic variation, and PRLR gene, polymorphisms have showed association with milk production traits and can be used as molecular markers (P < 0,05). The genotypes identified in F ATP were associated with percentages of lactose and protein in milk, while for apoB were associated with total milk production characteristics, dry matter, fat, lactose, . protein, and duration of lactation, and AGP AT6 was related with percentages of fat, protein, lactose and lactose production in milk. The expression of ApoB-lOO and FATP genes were different between Saanen and Alpine breeds, whereas in the Alpine breed the expression apoB-100 and FA TP were lower at the peak of lactation. ln addition, for the Alpine breed there was a higher expression in the dry period and early lactation compared to the peak to the gene APOB-l 00 (p < 0,05).