RESUMO
Sex-linked dwarf (SLD) chickens have been widely used in cross breeding of broilers and laying hens. To study the molecular mechanisms underlying growth hormone receptor (GHR) in SLD chickens, the expression profiles of GHR were measured in three growth related tissues (liver, breast, and thigh) in male and female S2 SLD chickens at seven growth stages (1 day, 3 weeks, 7 weeks, 9 weeks, 11 weeks, 13 weeks, and 15 weeks). Growth curves of body weight were fitted using logistic and Gompertz models. The results show that the inflexion week and inflexion weight in male chickens was earlier than in female chickens. Regarding the expression profiles of GHR, there was no significant difference between tissues at hatching. The expression peaked at 7 weeks and dropped by degrees in muscle tissue; hepatic expression increased with age and was positively correlated with body weight. Taken together, these results would provide a basis for further study on the molecular mechanisms underlying GHR regulation in SLD chickens.
Assuntos
Cruzamento , Galinhas/crescimento & desenvolvimento , Galinhas/genética , Regulação da Expressão Gênica no Desenvolvimento , Receptores da Somatotropina/genética , Animais , Feminino , Modelos Logísticos , Masculino , Dinâmica não Linear , Especificidade de Órgãos/genética , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Receptores da Somatotropina/metabolismo , Aumento de Peso/genéticaRESUMO
DNA methylation is an important epigenetic modification in eukaryotes, which plays a significant role in regulating gene expression. When the host is invaded by the influenza virus, gene expression is regulated via changes in DNA methylation levels or patterns, leading to the activation or suppression of relevant signaling pathways or networks, triggering a series of immune responses against viral invasion. Here, we investigated the changes in genomic DNA methylation in the immune organs of chicken infected with H5N1 influenza virus. Genome-wide DNA methylation levels in the spleen, thymus, and bursa of Fabricius of specific pathogen-free (SPF) chicken infected with the Guangdong (G-H5N1) and Anhui (A-H5N1) H5N1 strains, and water (control) were analyzed by fluorescence-labeled methylation-sensitive amplified polymorphism (F-MSAP). The results indicated that total DNA methylation levels did not differ between spleen genomic DNA in chicken treated with different viral strains and the control (P > 0.05). However, the total DNA methylation levels were significantly upregulated in the thymus (P < 0.01) and bursa (P < 0.05) of chicken in the A-H5N1 group compared to those in the G-H5N1 and control groups. These results provide a basis for the screening of avian influenza-resistance genes or methylation markers, analyzing the epigenetic regulation mechanisms of avian influenza, and performing selective breeding for disease resistance.
Assuntos
Metilação de DNA/genética , Resistência à Doença/genética , Virus da Influenza A Subtipo H5N1/patogenicidade , Influenza Aviária/genética , Animais , Bolsa de Fabricius/imunologia , Bolsa de Fabricius/virologia , Galinhas , Metilação de DNA/imunologia , Resistência à Doença/imunologia , Epigênese Genética , Genoma/genética , Virus da Influenza A Subtipo H5N1/imunologia , Influenza Aviária/imunologia , Influenza Aviária/virologia , Transdução de Sinais , Baço/imunologia , Baço/virologia , Timo/imunologia , Timo/virologiaRESUMO
The blue fox, belonging to the family Canidae, is a coat color variant of the native arctic fox (Alopex lagopus). To date, microsatellite loci in blue fox are typically amplified using canine simple sequence repeat primers. In the present study, we constructed an (AC)n enrichment library, and isolated and identified 17 polymorphic microsatellite markers for blue fox. The number of alleles per locus is from two to seven based on 24 examined individuals. The expected and observed heterozygosities were in the range of 0.3112 to 0.8236 and 0.2917 to 0.8750, respectively. The polymorphic information content per locus ranged from 0.2583 to 0.8022. These polymorphic markers can be useful for future population genetic studies of both farmed blue foxes and wild arctic foxes.
Assuntos
Raposas/genética , Repetições de Microssatélites/genética , Polimorfismo Genético , Alelos , Animais , Cães , HumanosRESUMO
An animal model of steroid-induced avascular necrosis of the femoral head (SANFH) was established to investigate the roles of osteocyte apoptosis in this process. Forty five-month-old male and female Japanese white rabbits were randomly divided into groups A (hormone + endotoxin), B (hormone alone), C (endotoxin alone), and D (blank control). Animals were sacrificed two and four weeks following the final treatment (N = 5 for each group at each time point). Bilateral femoral heads were fixed and decalcified, and empty lacunae were counted by hematoxylin staining. At weeks 2 and 4, the empty lacunae percentage was significantly higher in group A than that in groups B, C, or D (P < 0.01), while no significant difference was observed between these latter three. At week 2, all osteocyte apoptosis indexes were within normal ranges in all the groups, which therefore did not significantly differ in this respect (P > 0.05). However, at week 4, the apoptotic index was significantly higher in group A than that in groups B, C, or D (P < 0.01), comparisons between which revealed no such differences. Moreover, a positive correlation was observed between the percentage of empty lacunae and the apoptotic index at week 4 in group A (r = 0.893). We conclude that osteocyte apoptosis plays an important role in SANFH.
Assuntos
Apoptose , Osteócitos/metabolismo , Osteonecrose/metabolismo , Animais , Endotoxinas/toxicidade , Feminino , Fêmur/metabolismo , Fêmur/patologia , Masculino , Osteócitos/patologia , Osteonecrose/etiologia , Osteonecrose/patologia , Coelhos , Esteroides/toxicidadeRESUMO
Amelogenin is a major protein of the developing enamel matrix. There are two amelogenin genes (AMELX and AMELY) located on the X and Y chromosomes, respectively, in dogs. In the present study, we characterized full-length cDNAs and alternative splicing patterns of the AMEL genes in the tooth tissue of a dog by 5'- and 3'-rapid amplification of cDNA ends and AMEL-specific RT-PCR. Sequence analysis revealed that the coding regions of AMELX and AMELY were 579 and 576 bp (accession Nos. KP244310 and KP244311), respectively. The coding sequence of AMELX had 95.1% identity to that of AMELY. The AMEL genes on X and Y chromosomes were both expressed in developing tooth tissue. Eight different alternatively spliced transcripts were identified, five from AMELX and three from AMELY.
Assuntos
Processamento Alternativo , Amelogenina/genética , DNA Complementar , Animais , Sequência de Bases , DNA Complementar/genética , Cães , Dados de Sequência Molecular , Alinhamento de SequênciaRESUMO
Microsatellite markers are widely and evenly distributed, and are highly polymorphic. Rapid and convenient detection through automated analysis means that microsatellite markers are widely used in the construction of plant and animal genetic maps, in quantitative trait loci localization, marker-assisted selection, identification of genetic relationships, and genetic diversity and phylogenetic tree construction. However, few microsatellite markers remain to be isolated. We used streptavidin magnetic beads to affinity-capture and construct a (CA)n microsatellite DNA-enriched library from sika deer. We selected sequences containing more than six repeats to design primers. Clear bands were selected, which were amplified using non-specific primers following PCR amplification to screen polymorphisms in a group of 65 unrelated sika deer. The positive clone rate reached 82.9% by constructing the enriched library, and we then selected positive clones for sequencing. There were 395 sequences with CA repeats, and the CA repeat number was 4-105. We selected sequences containing more than six repeats to design primers, of which 297 pairs were designed. We next selected clear bands and used non-specific primers to amplify following PCR amplification. In total, 245 pairs of primers were screened. We then selected 50 pairs of primers to randomly screen for polymorphisms. We detected 47 polymorphic and 3 monomorphic loci in 65 unrelated sika deer. These newly isolated and characterized microsatellite loci can be used to construct genetic maps and for lineage testing in deer. In addition, they can be used for comparative genomics between Cervidae species.
Assuntos
DNA/genética , DNA/isolamento & purificação , Cervos/genética , Genoma , Repetições de Microssatélites/genética , Animais , Enzimas de Restrição do DNA/metabolismo , Feminino , Biblioteca Gênica , Masculino , Reação em Cadeia da Polimerase , Mapeamento por RestriçãoRESUMO
In this study, expression levels of miRNAs (miRNAs), miR-375 and miR-7, were detected in different tissues of cattle to determine whether adenohypophysis-prefer or exclusively expressed miRNAs, and target genes could be predicted by TargetScan, RNA22, and other software. Target genes related to pituitary function or reproductive traits were identified using a dual-luciferase assay. miR-375 and miR-7 were expressed differently in various tissues. miR-375 and miR-7 showed higher expression in the adenohypophysis, and there was a significant difference compared with expression in other tissues (P < 0.01). The binding sites for miR-7 were the mRNAs of bone morphogenetic protein receptor type II (BMPR2), prostaglandin F2 receptor negative regulator, gonadotropin-releasing hormone receptor, follicle-stimulating hormoneß, somatostatin receptor 1, and interleukin-1ß by bioinformatic analysis; similarly, the mRNAs of BMPR2 and leptin contained binding sites for miR-375, suggesting that these genes are affected by miR-7 or miR-375. Dual-luciferase reporter assays showed that miR-7 regulated prostaglandin F2 receptor negative regulator expression, while miR-375 regulated BMPR2 expression. The mutated plasmid and miRNA mimics were used to co-transfect NIH3T3 cells; luciferase reporter assays showed that the inhibition of luciferase activity in the wild-type cells dramatically decreased from 75 to 26% with a 3-5-nucleotide mismatch mutation into the seed region of miR-7. miR-375 had nearly lost the ability to inhibit luciferase activity, suggesting that GTCTTCC is the site of interaction between miR-7 and the prostaglandin F2 receptor negative regulator sequence and that GAACAAA is the site of interaction between miR-375 and the BMPR2 sequence.
Assuntos
MicroRNAs/genética , Adeno-Hipófise/metabolismo , RNA Mensageiro/genética , Animais , Sequência de Bases , Sítios de Ligação , Bovinos , Expressão Gênica , Perfilação da Expressão Gênica , Regulação da Expressão Gênica , Genes Reporter , MicroRNAs/química , Conformação de Ácido Nucleico , Especificidade de Órgãos/genética , Interferência de RNA , RNA Mensageiro/químicaRESUMO
We investigated the feasibility of interleukin-2 receptor gamma (IL2Rγ) gene based on gene mutation analysis and pre-natal diagnosis of X-linked severe combined immunodeficiency (X-SCID). Blood samples of patients and their parents of X-SCID (family 1) and X-SCID (family 2) were collected. IL2Rγ gene sequences of the 2 families were analyzed using bi-directional direct sequencing by polymerase chain reaction. DNA sequence changes in the IL2Rγ gene exon region and shear zone were also analyzed. We also sequenced the IL2Rγ gene in 100 healthy individuals. Prenatal genetic diagnoses for a high-risk fetus in family 1 were performed by chorionic villus sampling after determining each family's genotypes. The suspect fe-male in family 1 underwent carrier detection. Two novel mutations of IL2Rγ gene were identified, including c.361-363delGAG (p.E121del) in the patient and his mother in family 1, and c.510-511insGAACT (p.W173X) heterozygous mutation in the proband's mother in family 2. These mutations were absent in the 100 controls. Prenatal diagnosis of early pregnancy in the female fetus of family 1 was performed; the fetus was heterozygous, which was confirmed at postnatal follow-up. The suspect female in family 1 showed no mutation in carrier detection. The novel p.E121del and p.W173X mutations in IL2Rγ may have been the primary causes of disease in 2 families with X-SCID. In couples with an X-SCID reproductive history, prenatal gene mutation analysis of IL2Rγ can effectively prevent the birth of children with X-SCID and carrier detection for suspected females.
Assuntos
Amostra da Vilosidade Coriônica/métodos , Análise Mutacional de DNA/métodos , Subunidade gama Comum de Receptores de Interleucina/genética , Doenças por Imunodeficiência Combinada Ligada ao Cromossomo X/genética , Feminino , Doenças Fetais/genética , Humanos , Masculino , Mutação , Linhagem , GravidezRESUMO
The present study aimed to determine the effects of musk ketone on nerve recovery in rats after spinal cord injury. A total of 105 SD female rats were used to establish the rat with dorsal spinal cord injury model (modified Allen's method). The rats weighed from 200 to 250 g and were provided by the Experimental Animal Center of Chongqing Medical University. They were randomly divided into five treatment groups: saline (NS group), methylprednisolone (MP group), and musk ketone groups (MO1, MO2, and MO3 groups). The Swash plate test and BBB behavioral score were used to determine neurological function recovery after spinal cord injury. Hematoxylin-eosin (HE) staining was used to detect general structural changes in spinal cord tissue. The enzyme-linked immunosorbent assay was used for the determination of interleukin 10 (IL-10) in spinal cord tissue. We found that compared with the NS control group, critical angle, BBB score and IL-10 levels in rat spinal cord tissue significantly increased in the MP group and MO groups 7 and 14 days after the operation. HE staining showed that in the NS group, there was hemorrhage, edema, necrosis, axonal demyelination, inflammatory cell infiltration and glial cell response in spinal cord tissue. After 7 days, spinal cord edema and inflammation were reduced and neuronal degeneration and necrosis were not evident in the MP and MO groups. We conclude that musk ketone can reduce secondary damage after spinal cord injury and promote nerve recovery in rats.
Assuntos
Traumatismos da Medula Espinal/tratamento farmacológico , Xilenos/farmacologia , Animais , Modelos Animais de Doenças , Feminino , Masculino , Ratos , Ratos Sprague-Dawley , Medula Espinal/efeitos dos fármacos , Medula Espinal/patologia , Traumatismos da Medula Espinal/fisiopatologiaRESUMO
Congenital cataract is caused by reduced transparency of the lens resulting from metabolic disorders during the fetal period. The disease shows great heterogeneity both clinically and genetically. We identified a 4-generation ethnic Han Chinese family affected by autosomal dominant congenital perinuclear cataract. The patients underwent full clinical and ophthalmologic examinations to rule out any concomitant disorders. Blood samples were collected and genomic DNA was extracted. Potential mutations in the candidate gene alpha A crystallin (CRYAA) were screened. Prenatal diagnosis was then provided for a fetus of the affected proband by chorionic villus sampling. In all patients, DNA sequencing of the CRYAA gene revealed a novel 3-bp deletion mutation in exon 3 (c.246_248delCGC), which led to deletion of codon 117 encoding arginine (p.117delR) in the peptide chain. The same mutation was not found among unaffected and healthy individuals. Bioinformatic analysis revealed that although the c.246_248delCGC is an 'in-frame' mutation, removal of arginine resulted in a significant change in the protein structure. The fetus did not possess this mutation and was confirmed to be healthy at 1-year follow-up. A novel disease-causing mutation, c.246_248delCGC (p.117delR), of the CRYAA gene has been identified in a Chinese family with autosomal-type perinuclear congenital cataracts. This is also the first report of prenatal diagnosis of this type of congenital cataract.