RESUMO
Polymorphonuclear neutrophil (PMN) leukocytes are primary phagocytic cells of the bovine mammary gland and a first line of defense against invading pathogens during bovine mastitis infection. Cluster of differentiation 14 (CD14) is mainly expressed in macrophages and neutrophils and acts as a co-receptor that binds bacterial lipopolysaccharide (LPS) and recruits PMNs to CD14-LPS complexes in mammary epithelial cells. In this study, we identified a novel splice variant in PMNs, named CD14-SV, characterized by a deleted region from c.143-579 nt compared to the CD14 reference mRNA sequence. Moreover, a single nucleotide polymorphism (c.523 A>G) in exon 2 of CD14 was identified and found to modify the secondary structure and hydrophilicity of the CD14 protein. Association analysis also showed that the milk somatic cell score, an indicator of mastitis, of cows with the GG genotype was lower than that of cows with the AA and AG genotypes. Our findings suggest that the expression of CD14 in bovine blood PMNs is regulated by alternative splicing, and that CD14-SV is a candidate functional marker that may influence mastitis-resistance in dairy cows.
Assuntos
Bovinos/genética , Receptores de Lipopolissacarídeos/genética , Mastite Bovina/genética , Neutrófilos/metabolismo , RNA Mensageiro/genética , Processamento Alternativo , Animais , Feminino , Variação Genética , Receptores de Lipopolissacarídeos/metabolismo , Masculino , Mastite Bovina/sangue , Polimorfismo de Nucleotídeo Único , Isoformas de Proteínas/genética , Isoformas de Proteínas/metabolismo , RNA Mensageiro/metabolismoRESUMO
The aim of this study was to evaluate the correlation between chromosomal polymorphisms and male infertility. The patients were diagnosed with azoospermia or oligospermiaby a semen analysis. Chromosomal analysis was performed on peripheral blood lymphocytes obtained from the patients, with standard G-banding and C-banding. Y chromosome microdeletions were detected by multiplex polymerase chain reaction (PCR) amplification. The parents of 35 polymorphic probands were also subjected to chromosomal analysis, and their detailed reproductive histories were surveyed. The frequency of autosomal polymorphisms did not differ significantly among the infertile patients and fertile control individuals. The frequency of the Yqh-variant increased with the decrease in sperm count; this appeared at a significantly higher frequency in the azoospermia group (57.2 vs 24.3 vs 0%). The results of PCR amplification indicated that 32.14% of the patients with Yqh ± had microdeletions in the Y chromosome. The parents of the probands with the same chromosomal polymorphisms as the probands (among the 35 recalled families) did not show any adverse reproductive history. We observed no significant correlations between autosomal polymorphisms and male infertility. However, we observed a significant increase in the frequency of Yqh- in the azoospermic patients. This may be attributed to Y chromosome microdeletions, although the association between Y chromosome microdeletions and Y chromosome variants remains to be elucidated.