ABSTRACT
Objective To explore the double-labeling method of monitoring the GHRP regulatory function on [Ca2+]i and NO in cardiomyocytes of rats on real time under LSCM.Methods The reformed constant-flow Langendorff system and enzyme-dissociated was used to isolate cardiomyocytes.[Ca2+]i and NO in the cardiomyocytes of SD rats were double-labeled by their molecular probe Rhod-2/AM and DAF-FM/DA,respectively to monitor the regulatory function of GHRP on [Ca2+]i and NO on real time by LSCM.Results Ca2+ signal showed a red fluorescence and NO showed a green fluorescence while the overlapping of the two signals showed a yellow-green fluorescence by this system,and the similar effect presents in both double-labeled state and the single labeled one:GHRP induced a transient[Ca2+]i increase then followed by a plateau phase while there was not significant change in NO signal system after GHRP stimulation under the LSCM in the cardiomyocytes of rats.Conclusions After having established the double-labeling method we monitored the GHRP regulatory function on [Ca2+]i and NO on real time in cardiomyocytes of rats under LSCM causing the [Ca2+]i biphasic increase while no significant change in NO signal system.
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Objective To study the quantitative changes of gap junction (GJ) in the detrusors of unstable bladder(USB) in rats,and to deduce the functional changes of GJ,which mediates intercellular communication,so as to demonstrate one of the mechanisms of USB. Methods Thirteen Wistar rats, which were identified to have USB by manometry of filling bladder after establishment of bladder outlet obstruction (BOO) model, served as study,ie,USB group.Another 10 healthy female Wistar rats served as control group.The content and distribution of connexin (Cx) 43 of the detrusors, which were taken from these rats,were quantitatively analyzed by Western blot and laser confocal microscopy with a double-label immunohistochemical technique. Results Cx43 protein was expressed in both control and USB group, the relative molecular weight was 43000.The mean gray levels of the detrusor protein bands in USB group and control group were 31.066 and 11.701,respectively;the difference of the values between the 2 groups was significant (P
ABSTRACT
Objective To explore the effect of folic acid on neural stem cells(NSCs) proliferation from fetal rats in vitro.Method NSCs were isolated and cultured by microdissection,mechanical blowing and serum-free suspension culture,and identified by immunofluorescent staining using antibody against nestin.BrdU(5’bromo-2’deoxyuridine) was used to mark dividing neural stem cells.Cultured NSCs were divided into four groups:control group,low,high dose group(liquid media with added 4,40 mg/L folic acid),and deficiency group(liquid media with added 0.4 mg/L methotrexate,MTX).Monotetrazolium(MTT) and double-label immunofluorescence technique detected NSCs proliferation under the condition of folic acid.Results In the serum-free suspension medium,neurospheres that consisted of a great number of nestin-positive cells could be obtained.The proliferative ability of NSCs were observed by BrdU labeling methods.MTT assay and double-label immunofluorescence for nestin+BrdU showed that the growth tendency was increased with folate concentration in the medium.Compared with control group,NSCs growth rate of folate group was significantly increased in vitro.Conclusion The culture of NSCs isolated from fetal rats possesses the abilities of proliferation and self-regeneration.Folic acid may stimulate proliferation of NSCs efficiently.