RESUMEN
<p><b>OBJECTIVE</b>To investigate the effect of Hath1 (human atonal homolog 1) overexpression on greater epithelial ridge (GER) cells from postnatal rat cochlea in vitro.</p><p><b>METHODS</b>GER cells were isolated by using a combinatorial approach of enzymatic digestion and mechanical separation from P1 rat cochlear. The GER cell cultures were infected by adenovirus containing Hath1 and enhanced green fluorescent protein (ad-Hath1-EGFP), while transfecting EGFP(ad-EGFP) was as controls. Immunostaining were performed at different time points after adenovirus infection.</p><p><b>RESULTS</b>Some of the infected GER cells became myosin VIIa-positive following ad-Hath1-EGFP infection. The earliest time point to see induction of hair cell differentiation (hair cell marker expression) by ad-Hathl was 5 days post-infection. In contrast, infection of the GER sheet cultures with ad-EGFP control virus did not show any myosin VIIa-positive cells at 3-12 days post-infection in all cultures examined.</p><p><b>CONCLUSIONS</b>GER cells may potentially serve as hair cell progenitors and they are capable of differentiating hair cell-like cells when forced to express Hath1.</p>
Asunto(s)
Animales , Ratas , Animales Recién Nacidos , Factores de Transcripción con Motivo Hélice-Asa-Hélice Básico , Genética , Diferenciación Celular , Células Cultivadas , Cóclea , Biología Celular , Células Epiteliales , Biología Celular , Células Ciliadas Auditivas , Biología Celular , Ratas Sprague-Dawley , TransfecciónRESUMEN
<p><b>OBJECTIVE</b>To evaluate the relationship between labeling index (LI) Ki-67, proliferating cell nuclear antigen (PCNA) and transforming growth factor-beta1 (TGF-beta1) with the clinical behavior of acoustic neuroma.</p><p><b>METHODS</b>Expression of Ki-67, PCNA and TGF-beta1 was detected by immunohistochemistry in 53 specimens of acoustic neuromas. The relationship among tumor proliferation, histological representation, size of tumor, clinical proliferation index of tumor and tumor proliferation activity were analyzed.</p><p><b>RESULTS</b>In all 53 cases, the positive rate of Ki-67 was 77.4% (41/53) but the positive rate of PCNA was 84.9% (45/53). There was significant difference between the proliferate index, clinic growth rate and course of disease (t = 2.14, t = 2.70; P < 0.05). The positive rate of TGF-beta1 was 83.0% (44/53). The correlation of TGF-beta1 with LI (Ki-67) was significant difference (r = 0.36, P < 0.05). Cystic degeneration often occurred in large-size tumor (Z = 4.44, P < 0.05). There was no significant relationship between the expression of LI (Ki-67), LI (PCNA) and TGF-beta1 and the course of disease as well as between the cystic degeneration and the non-cystic degeneration. Although clinic growth rate of cystic degeneration was bigger than that of non-cystic degeneration, there was not statistically significant.</p><p><b>CONCLUSIONS</b>Ki-67 and PCNA are reflected proliferation activities of tumor cells in acoustic neuromas. Cell proliferation-labeling index LI (PCNA) was related with clinical growth rates. TGF-beta1 might participate in the biological behavior of acoustic neuroma. Cystic degeneration was one of special pattern of acoustic neuroma, however, tumor enlargement might due to the volume of the cystic but unrelated to fast proliferation of parenchyma cell.</p>
Asunto(s)
Adolescente , Adulto , Anciano , Femenino , Humanos , Masculino , Persona de Mediana Edad , Adulto Joven , Proliferación Celular , Antígeno Ki-67 , Metabolismo , Neuroma Acústico , Diagnóstico , Metabolismo , Patología , Antígeno Nuclear de Célula en Proliferación , Metabolismo , Estudios Retrospectivos , Factor de Crecimiento Transformador beta1 , Metabolismo , Nervio VestibulococlearRESUMEN
<p><b>OBJECTIVE</b>To establish in vitro culture systems of greater epithelial ridge (GER) cells from rat cochlear and to investigate the characterization, growth pattern and ultrastructure of GER cells.</p><p><b>METHODS</b>Using a combinatorial approach of enzymatic digestion and mechanical separation to allow isolation and culture of GER cells from P1 rat cochleae. The dissociated GER cells were cultured in serum-free or 10% fetal bovine serum DMEM respectively. BrdU, phalloidin, ZO1, calretinin and myosin VIIa immunostaining and scanning electron microscope observation were performed in GER cell cultures.</p><p><b>RESULTS</b>The dissociated GER cell cultures showed positive to ZO1, phalloidin and BrdU staining, but negative to myosin VIIa and calretinin. They assumed a polygonal morphology which was similar to epithelial cells and grew in islands-like patches in medium containing 10% fetal bovine serum while forming spheres in serum-free medium. The GER cells presented significant ability to proliferate in both conditions. Scanning electron microscope showed that there was microvillus and centre bodies but not hair cell specific stereociliary bundles on the surface of GER cultures.</p><p><b>CONCLUSIONS</b>The GER cell cultures showed significant ability to proliferate and grew in islands-like patches in medium containing 10% fetal bovine serum while forming spheres in serum-free medium. The dissociated GER cells expressed epithelial cell specific marker but not marker of hair cells.</p>
Asunto(s)
Animales , Ratas , Animales Recién Nacidos , Técnicas de Cultivo de Célula , Células Cultivadas , Cóclea , Biología Celular , Células Epiteliales , Biología Celular , Células Ciliadas Auditivas , Biología Celular , Ratas Sprague-DawleyRESUMEN
<p><b>OBJECTIVE</b>To detect the expression of Math1, Hes1 and Hes5 in greater epithelial ridge (GER) cells of rat cochlear and explore their influence on hair cell differentiation.</p><p><b>METHODS</b>Postnatal day 0 (P0), day 1 (P1) , day 3 (P3) day 4 (P4) and day 5 (P5) rat cochlear were dissected respectively and then pure GER cells were separated by a combinatorial approach of attachment and mechanical separation. The total RNA of GER cells was extracted by Trizol one step method and the expression of Math1, Hes1 and Hes5 in GER cells was detected with reverse transcription polymerase chain reaction.</p><p><b>RESULTS</b>Math1 was expressed in P0 - P5 rat GER cells and Hes1 was expressed only in PO - P3 rat GER cells, while there was no expression of Hes5 in P0 - P5 rat GER cells.</p><p><b>CONCLUSIONS</b>Probably only when the expression of Math1 reaches a certain level can it induce GER cells to differentiate into hair cells. Meanwhile this process might controlled by Hes1 to some extent.</p>
Asunto(s)
Animales , Ratas , Factores de Transcripción con Motivo Hélice-Asa-Hélice Básico , Genética , Metabolismo , Diferenciación Celular , Cóclea , Biología Celular , Metabolismo , Células Epiteliales , Metabolismo , Regulación del Desarrollo de la Expresión Génica , Células Ciliadas Auditivas , Biología Celular , Metabolismo , Proteínas de Homeodominio , Genética , Metabolismo , Ratas Sprague-Dawley , Factor de Transcripción HES-1RESUMEN
<p><b>OBJECTIVE</b>To investigate the effect of noise on the antioxidant enzymes of cochleae.</p><p><b>METHODS</b>16 male pigmented guinea pigs (250 - 300 g) were randomly divided into 2 groups, control group and noise group. Each group had 8 animals. The animals in noise group were performed auditory evoked brainstem responses (ABR) recording before and after exposure to a continuous noise (4 kHz, octave band, 100 dB, SPL) 8 h/d for 3 consecutive days. Immediately at the end of the third day's noise exposure after ABR recording, guinea pigs were decapitated. Both the right and the left cochlea with the bony capsule removed were homogenized, and the supernatants were prepared for assays. Reactive oxygen species (ROS), superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSH-Px) were measured.</p><p><b>RESULTS</b>ROS level of the noise group [(281.2 +/- 3.5) U/mg pro] was significantly higher than that of the control group [(273.0 +/- 3.2) U/mg pro, P < 0.05] and SOD, CAT and GSH-Px activities of the noise group [(206.5 +/- 5.1) NU/mg pro, (47.0 +/- 9.0) U/g pro, (14.1 +/- 2.5) U/mg pro respectively] were significantly lower than that of the control group [(221.8 +/- 4.8) NU/mg pro, (60.8 +/- 9.9) U/g pro, (21.1 +/- 3.1) U/mg pro respectively, P < 0.05].</p><p><b>CONCLUSION</b>Noise may damage the defensive system of antioxidant enzymes in cochlea.</p>