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1.
Sci Rep ; 8(1): 7794, 2018 05 17.
Artículo en Inglés | MEDLINE | ID: mdl-29773831

RESUMEN

DNAJC17 is a heat shock protein (HSP40) family member, identified in mouse as susceptibility gene for congenital hypothyroidism. DNAJC17 knockout mouse embryos die prior to implantation. In humans, germline homozygous mutations in DNAJC17 have been found in syndromic retinal dystrophy patients, while heterozygous mutations represent candidate pathogenic events for myeloproliferative disorders. Despite widespread expression and involvement in human diseases, DNAJC17 function is still poorly understood. Herein, we have investigated its function through high-throughput transcriptomic and proteomic approaches. DNAJC17-depleted cells transcriptome highlighted genes involved in general functional categories, mainly related to gene expression. Conversely, DNAJC17 interactome can be classified in very specific functional networks, with the most enriched one including proteins involved in splicing. Furthermore, several splicing-related interactors, were independently validated by co-immunoprecipitation and in vivo co-localization. Accordingly, co-localization of DNAJC17 with SC35, a marker of nuclear speckles, further supported its interaction with spliceosomal components. Lastly, DNAJC17 up-regulation enhanced splicing efficiency of minigene reporter in live cells, while its knockdown induced perturbations of splicing efficiency at whole genome level, as demonstrated by specific analysis of RNAseq data. In conclusion, our study strongly suggests a role of DNAJC17 in splicing-related processes and provides support to its recognized essential function in early development.


Asunto(s)
Proteínas del Choque Térmico HSP40/metabolismo , Empalme Alternativo , Núcleo Celular/metabolismo , Proteínas del Choque Térmico HSP40/análisis , Proteínas del Choque Térmico HSP40/genética , Células HeLa , Humanos , Mapeo de Interacción de Proteínas , Proteómica , Empalmosomas/metabolismo
2.
Theriogenology ; 65(7): 1265-78, 2006 Apr 15.
Artículo en Inglés | MEDLINE | ID: mdl-16203031

RESUMEN

D-aspartic acid (D-Asp) has been isolated from neuroendocrine tissues of many invertebrates and vertebrates. Recently, it has been demonstrated that this D-amino acid may be converted to N-methyl-D-aspartic acid (NMDA), a neuromodulator associated with sexual activity. In this study, we determined D-Asp and NMDA concentrations in endocrine glands and other tissues in ewes after D-Asp administration and in controls. We also evaluated the effects of d-Asp administration on the reproductive activity of ewes by determining either progesterone concentrations or LH pulses in the presence or absence of estradiol benzoate. The pineal gland showed the highest natural content of D-Asp (1.47+/-0.22 micromol/g tissue), whereas the pituitary gland had the highest capability to store d-Asp, with a peak value (9.7+/-0.81 micromol/g tissue) 6 h after its administration. NMDA increased sharply 12 h following D-Asp administration, reaching values three times higher than the baseline in both the pituitary and brain. D-Asp was quickly adsorbed after subcutaneous administration, with a peak in plasma levels 2 h after administration and a return to baseline values after 6 h. D-Asp administration achieved a significant (P < 0.001) increase in LH values with respect to estradiol or estradiol + D-Asp treatments. d-Asp treatment once or twice a week did not successfully drive acyclic ewes into reproductive activity. In conclusion, the results obtained in this study demonstrated that D-Asp is endogenously present in sheep tissues and electively stored in endocrine glands and brain after its administration. NMDA and LH increase following D-Asp administration suggesting a role of this D-amino acid in the reproductive activity of sheep.


Asunto(s)
Ácido D-Aspártico/administración & dosificación , Ácido D-Aspártico/fisiología , Reproducción/fisiología , Conducta Sexual Animal/fisiología , Ovinos/fisiología , Animales , Encéfalo/metabolismo , Ácido D-Aspártico/análisis , Glándulas Endocrinas/química , Femenino , Lactancia , Hormona Luteinizante/sangre , N-Metilaspartato/análisis , N-Metilaspartato/sangre , Especificidad de Órganos , Glándula Pineal/química , Hipófisis/química , Progesterona/sangre , Reproducción/efectos de los fármacos , Conducta Sexual Animal/efectos de los fármacos
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