ABSTRACT
We recently found that extracellular administration of nicotine indirectly excited hypothalamic paraventricular nucleus (PVN) corticotropin-releasing hormone (CRH) mRNA-expressing neurons. In this study, we studied the effect of nicotine on PVN oxytocin (OT) mRNA-expressing neuron in vitro in rats, by whole-cell patch-clamp recording technique, immunohistochemistry methods and single-cell reverse-transcription multiplex polymerase chain reaction (SC-RT-mPCR) methods Our results showed that 79.3% (73/92) of the 92 PVN putative magnocellular neurons co-expressed GAPDH mRNA and OT mRNA. Under current-clamp recording conditions, local micro application of nicotine (1-300µM) induced a decrease in spontaneous firing rate accompanied with a hyperpolarization of membrane potential in 76.7% (56/73) of PVN OT mRNA-expressing magnocellular neurons. The nicotine induced inhibition in spontaneous activity of PVN OT mRNA-expressing magnocellular neurons was dose-dependent. The half-inhibitory concentration (IC50) is 2.9µM. The nicotine induced hyperpolarization of PVN OT mRNA-expressing magnocellular neurons was sensitive to GABAA receptor antagonist, SR95531 (10µM) and tetrodotoxin (TTX, 1µM). In addition, local micro application of nicotine induced a significant increase in frequency of spontaneous inhibitory postsynaptic potentials (sIPSPs), but without changes in the sEPSPs amplitude of the OT-mRNA expressing neurons. Biocytin staining confirmed that the nicotine-sensitive OT-mRNA expressing neurons were the PVN magnocellular neurons. These results demonstrated that nicotine enhances the GABAergic inhibition, resulting in a decrease in spontaneous firing rate of the PVN OT-mRNA expressing neurons. These findings suggested that nicotine modulated PVN OT secretion via enhancement of both presynaptic action potential drive and quantal GABA release.
Subject(s)
Neurons/drug effects , Nicotine/pharmacology , Oxytocin/metabolism , Paraventricular Hypothalamic Nucleus/drug effects , RNA, Messenger/metabolism , gamma-Aminobutyric Acid/metabolism , Action Potentials/drug effects , Animals , Excitatory Postsynaptic Potentials/drug effects , GABA-A Receptor Antagonists/pharmacology , Inhibitory Postsynaptic Potentials/drug effects , Male , Neurons/metabolism , Oxytocin/genetics , Paraventricular Hypothalamic Nucleus/metabolism , Pyridazines/pharmacology , Rats, Wistar , Tetrodotoxin/pharmacologyABSTRACT
OBJECTIVE: To evaluate the application of scrotal ultrasonography in the diagnosis of the causes of azoospermia. METHODS: We performed scrotal ultrasonography for 96 patients with azoospermia, and analyzed the ultrasonographic images in comparison with the results of the final diagnosis. RESULTS: Based on the characteristics of the ultrasonographic images, 35 of the cases were diagnosed as obstructive azoospermia and the other 61 as non-obstructive azoospermia, with an accuracy rate of 97.0%. The causes and sites of the lesions were manifested in 44.4% of the obstructive and 46.7% of the non-obstructive cases, with accuracy rates of 100% and 75.0%, respectively. As for the rest of the cases, scrotal ultrasonography helped narrow the scope of further examinations by giving directional indications. CONCLUSION: Scrotal ultrasonography is an essential means for the diagnosis of the causes of azoospermia, or for obtaining directional indications of its causes.