Role of histone deacetylase activity in the developing lateral line neuromast of zebrafish larvae

Histone deacetylases are involved in many biological processes and have roles in regulating cell behaviors such as cell cycle entry, cell proliferation and apoptosis. However, the effect of histone deacetylases on the development of hair cells (HCs) has not been fully elucidated. In this study, we examined the influence of histone deacetylases on the early development of neuromasts in the lateral line of zebrafish. Hair cell development was evaluated by fluorescent immunostaining in the absence or presence of histone deacetylase inhibitors. Our results suggested that pharmacological inhibition of histone deacetylases with inhibitors, including trichostatin A, valproic acid and MS-275, reduced the numbers of both HCs and supporting cells in neuromasts. We also found that the treatment of zebrafish larvae with inhibitors caused accumulation of histone acetylation and suppressed proliferation of neuromast cells. Real-time PCR results showed that the expression of both p21 and p27 mRNA was increased following trichostatin A treatment and the increase in p53 mRNA was modest under the same conditions. However, the expression of p53 mRNA was significantly increased by treatment with a high concentration of trichostatin A. A high concentration of trichostatin A also led to increased cell death in neuromasts as detected in a TUNEL assay. Moreover, the nuclei of most of these pyknotic cells were immunohistochemically positive for cleaved caspase-3. These results suggest that histone deacetylase activity is involved in lateral line development in the zebrafish and might have a role in neuromast formation by altering cell proliferation through the expression of cell cycle regulatory proteins.

Streptozotocin-induced mechanical hypernociception is not dependent on hyperglycemia

Since streptozotocin (STZ)-induced diabetes is a widely used model of painful diabetic neuropathy, the aim of the present study was to design a rational protocol to investigate whether the development of mechanical hypernociception induced by STZ depends exclusively on hyperglycemia. Male Wistar rats (180-200 g; N = 6-7 per group) received a single intravenous injection of STZ at three different doses (10, 20, or 40 mg/kg). Only the higher dose (40 mg/kg) induced a significant increase in blood glucose levels, glucose tolerance and deficiency in weight gain. However, all STZ-treated rats (hyperglycemic or not) developed persistent (for at least 20 days) and indistinguishable bilateral mechanical hypernociception that was not prevented by daily insulin treatment (2 IU twice a day, sc). Systemic morphine (2 mg/kg) but not local (intraplantar) morphine treatment (8 µg/paw) significantly inhibited the mechanical hypernociception induced by STZ (10 or 40 mg/kg). In addition, intraplantar injection of STZ at doses that did not cause hyperglycemia (30, 100 or 300 µg/paw) induced ipsilateral mechanical hypernociception for at least 8 h that was inhibited by local and systemic morphine treatment (8 µg/paw or 2 mg/kg, respectively), but not by dexamethasone (1 mg/kg, sc). The results of this study demonstrate that systemic administration of STZ induces mechanical hypernociception that does not depend on hyperglycemia and intraplantar STZ induces mechanical sensitization of primary sensory neurons responsive to local morphine treatment.

The Reflex Effects on the Respiratory Regulation of the CO2 at the Different Flow Rate and Concentration

PURPOSE: The purpose of this study was to investigate the activation of the respiratory centers during insufflation of the larynx with CO2 at different flow rates and concentrations. MATERIALS AND METHODS: The experiments were carried out in spontaneous air breathing rabbits, anesthetized with thiopental sodium (25mg kg(-1) i.v.). The larynx was separated from the oropharyngeal cavity and the trachea. The tidal volume (VT) and respiratory frequency (f min(-1)) were recorded from the lower tracheal cannula. The respiratory minute volume (VE) was calculated, the action potentials from the right phrenic nerve were recorded and the inspiratory (TI) and expiratory (TE) periods and the mean inspiratory flow rate (VT/TI) were calculated. The larynx was insufflated at flow rates of 500mL min(-1) and 750mL min(-1), with 7 and 12% CO2-Air by means of a respiratory pump. RESULTS: Insufflation of the larynx, with both gas mixtures, decreased the f and VT significantly. The TI and TE were found to increase significantly due to the decreasing in f. There was a significant decrease in VT/TI ratio. Following bilateral midcervical vagotomy, on the passing of both gas mixtures, significant decreases were observed in the VT, and the responses of f, TI and TE were abolished. After cutting the superior laryngeal nerve, the responses of the VT to both gas mixtures were abolished. CONCLUSION: In conclusion, the results of this study purpose that the stimulation of the laryngeal mechanoreceptors by the effect of hyper- capnia decreases the activation of the respiratory center

Mecanorreceptores cardíacos / Cardiac mechanoreceptors

Utilizou-se a noradrenalina na dose padräo de 2 ug/Kg, medindo-se o fluxo na artéria femoral antes e após o estímulo. A injeçäo foi feita em diversos níveis da aorta, intracardíaca e na veia axilar. Para estudar a dependência do fenômeno com as vias nervosas, estimularam-se cäes com os nervos femoral e ciático seccionados, com desnervaçäo dos seios carotídeos e aórtico e com os vagos seccionados bilateralmente. Foi possível observar que: 1 - O aumento da atividade cardíaca que se acompanha de aumento da contratilidade é estímulo eficaz para desencadear o aumento do fluxo na artéria femoral. O aumento da contratilidade cardíaca e a diminuiçäo da resistência periférica säo proporcionais à dose de noradrenalina injetada (dentro dos limites de 1 a 3 ug). 2- No cäo com uma pata posterior submetida à secçäo dos nervos femoral ciático, a injeçäo de noradrenalina causa diminuiçäo da resistência vascular na pata normal, simultaneamente com o aumento da resistência na pata desnervada. Essa observaçäo demonstra ser esse fenômeno um reflexo. 3- A secçäo dos seios carotídeos e aórticos quase näo influencia essa resposta reflexa. 4- A injeçäo de noradrenalina na artéria femoral e em diversas posiçöes da aorta näo causa diminuiçäo da resistência periférica. A injeçäo endovenosa no bulbo aórtico e intracardíaca (ventrículo esquerdo) sempre leva à vasodilataçäo na regiäo estudada. 5- A secçäo vagal deprime acentuadamente a resposta, demonstrando ser essa a principal via nervosa responsável pelo reflexo