Ca2+ mobilization by nicotine through synaptic activation in rat parotid acini

Nicotine has been reported to increase the intracellular Ca2+ concentration ([Ca2+]i) in sublingual acini due to neurotransmitter release from nerve terminals associated with the cell preparation (1). However, it is unclear whether or not the same reaction exists in parotid cells. Therefore, we investigated effect of nicotine on Ca2+ mobilization in digested parotid acini from rats. After removing the parotid gland from Wistar rats, the tissues were minced and digested with collagenase. Then, the intracellular Ca2+ indicator fura-2 was added to the preparation, and the change in [Ca2+]i was monitored using fluorescent microscope. In many but not all parotid acini, K+ stimulation induced transient increases in [Ca2+]i. The K+-induced Ca2+ response in parotid acini was completely blocked by Cd2+-containing solution. These results suggest that the parotid cell preparation has nerve terminals. In all high-K+-sensitive parotid acini, over 3μM of nicotine increased [Ca2+]i, and the response was blocked by a Cd2+-containing solution and nicotinic receptor antagonists. All high-K+-insensitive acinar cells were resistant to the effect of nicotine on Ca2+ mobilization. These results suggest that nicotine induces increases in [Ca2+]i in parotid acini due to neurotransmitter release from associated nerve terminals

Effects of pilocarpine and cevimeline on Ca2+ mobilization in rat parotid acini and ducts

Previous reports suggested that there is no significant difference in the binding affinity of pilocarpine and cevimeline on muscarinic receptors (1). However, in vivo studies from our laboratory suggested that pilocarpine-induced salivation from the parotid gland is greater than that induced by cevimeline in rats (2, 3). Therefore, in the present study, sialogogue-induced intracellular Ca2+ mobilization was investigated in isolated parotid gland cells in vitro. Pilocarpine and cevimeline increased the intracellular Ca2+ concentration of parotid gland acinar and duct cells over 1 μM in a dose-dependent manner. Pilocarpine-induced responses were higher than cevimeline-induced responses. Ca2+ responses to both agents were completely blocked by 1 μM 4-DAMP, an M3 muscarinic receptor antagonist. In the absence of extracellular Ca2+, both sialogogues induced transient Ca2+ increase in parotid gland acinar cells. These results suggest that the sialogogues stimulate some common routes via the Ca2+ signaling in parotid gland acinar cells. We also report a significant difference of Ca2+ responses in concentration between pilocarpine and cevimeline in parotid gland acinar cells. The different Ca2+ responses between the sialogogues would explain the different saliva volumes from the parotid gland between them that we have observed in previous in vivo studies (2, 3)

Ca2+ mobilization by nicotine through synaptic activation in rat parotid acini

Nicotine has been reported to increase the intracellular Ca2+ concentration ([Ca2+]i) in sublingual acini due to neurotransmitter release from nerve terminals associated with the cell preparation (1). However, it is unclear whether or not the same reaction exists in parotid cells. Therefore, we investigated effect of nicotine on Ca2+ mobilization in digested parotid acini from rats. After removing the parotid gland from Wistar rats, the tissues were minced and digested with collagenase. Then, the intracellular Ca2+ indicator fura-2 was added to the preparation, and the change in [Ca2+]i was monitored using fluorescent microscope. In many but not all parotid acini, K+ stimulation induced transient increases in [Ca2+]i. The K+-induced Ca2+ response in parotid acini was completely blocked by Cd2+-containing solution. These results suggest that the parotid cell preparation has nerve terminals. In all high-K+-sensitive parotid acini, over 3μM of nicotine increased [Ca2+]i, and the response was blocked by a Cd2+-containing solution and nicotinic receptor antagonists. All high-K+-insensitive acinar cells were resistant to the effect of nicotine on Ca2+ mobilization. These results suggest that nicotine induces increases in [Ca2+]i in parotid acini due to neurotransmitter release from associated nerve terminals

Assessment of intraoral mucosal pain induced by the application of capsaicin

Objective To develop an objective method for assessing nociceptive behaviour in an animal model of capsaicin-induced intraoral pain. Changes in nociceptive responses were also examined after injury to the inferior alveolar nerve (IAN). Design Nociceptive responses evoked by the intraoral application of various doses of capsaicin were analyzed in lightly anaesthetized rats. The number of c-Fos protein-like immunoreactive (Fos-LI) neurons in the medullary dorsal horn (MDH) induced by the intraoral application of capsaicin was measured. Behavioural and c-Fos responses were also examined 14 days after injury to the IAN. Results Larger doses of intraoral capsaicin (1, 10 and 100 μg) induced vigorous licking behaviour and c-Fos response in the MDH in a reproducible manner. The magnitudes of both behavioural activity and the c-Fos response from the 10 and 100 μg doses of capsaicin were significantly greater than that by the 1 μg dose. Injury to the IAN exaggerated the behavioural and c-Fos responses evoked by intraoral capsaicin. Conclusions The intraoral application of capsaicin is a valid and reliable method for studying intraoral pain and hyperalgesia following nerve injury

SINC-seq: correlation of transient gene expressions between nucleus and cytoplasm reflects single-cell physiology

We report a microfluidic system that physically separates nuclear RNA (nucRNA) and cytoplasmic RNA (cytRNA) from a single cell and enables single-cell integrated nucRNA and cytRNA-sequencing (SINC-seq). SINC-seq constructs two individual RNA-seq libraries, nucRNA and cytRNA, per cell, quantifies gene expression in the subcellular compartments, and combines them to create novel single-cell RNA-seq data. Leveraging SINC-seq, we discover distinct natures of correlation among cytRNA and nucRNA that reflect the transient physiological state of single cells. These data provide unique insights into the regulatory network of messenger RNA from the nucleus toward the cytoplasm at the single-cell level

The Satb1 Protein Directs Hematopoietic Stem Cell Differentiation toward Lymphoid Lineages

SummaryHow hematopoietic stem cells (HSCs) produce particular lineages is insufficiently understood. We searched for key factors that direct HSC to lymphopoiesis. Comparing gene expression profiles for HSCs and early lymphoid progenitors revealed that Satb1, a global chromatin regulator, was markedly induced with lymphoid lineage specification. HSCs from Satb1-deficient mice were defective in lymphopoietic activity in culture and failed to reconstitute T lymphopoiesis in wild-type recipients. Furthermore, Satb1 transduction of HSCs and embryonic stem cells robustly promoted their differentiation toward lymphocytes. Whereas genes that encode Ikaros, E2A, and Notch1 were unaffected, many genes involved in lineage decisions were regulated by Satb1. Satb1 expression was reduced in aged HSCs with compromised lymphopoietic potential, but forced Satb1 expression partly restored that potential. Thus, Satb1 governs the initiating process central to the replenishing of lymphoid lineages. Such activity in lymphoid cell generation may be of clinical importance and useful to overcome immunosenescence

Association between the examination rate of treatment-resistant schizophrenia and the clozapine prescription rate in a nationwide dissemination and implementation study

Background: The decision to initiate clozapine treatment should be made on an individual basis and may be closely related to the early detection of treatment-resistant schizophrenia (TRS), although there is evidence that the early use of clozapine results in a better response to treatment. Therefore, we investigated the relationship between the examination rate of TRS and the prescription rate of clozapine. Methods: After attending a 1-day educational program on schizophrenia based on the "Guidelines for the Pharmacological Treatment of Schizophrenia," we asked the participating facilities to submit records of whether or not TRS was evaluated for each patient. We calculated the clozapine prescription rate from the schizophrenic patients prescribed clozapine and all of the schizophrenic patients. Forty-nine facilities in 2017 were included in the study. Results: There were dichotomous distributions in the examination rate of TRS and a non-normal distribution in the prescription rate of clozapine. There was a significant correlation between the prescription rate of clozapine and the examination rate of TRS (r s = 0.531, P = 1.032 × 10−4). A significant difference was found in the prescription rate of clozapine between the three groups of facilities according to the examination rate of TRS. Conclusion: As a preliminary problem for the use of clozapine, in Japan, the examination rate of TRS varies, and there are many facilities that typically do not consider the possibility of TRS; this trend leads to a low rate of clozapine use. Clearly, further clinician training is needed for the early detection and appropriate management of TRS that includes an explanation of TRS and how to introduce clozapine therapy to patients and their families

Clozapine and Antipsychotic Monotherapy

Background: Although clozapine is effective for treatment-resistant schizophrenia (TRS), the rate of clozapine prescription is still low. Whereas antipsychotic monotherapy is recommended in clinical practice guidelines, the rate of antipsychotic polypharmacy is still high. There is little evidence on whether a clozapine prescription influences changes in the rate of monotherapy and polypharmacy, including antipsychotics and other psychotropics. We therefore hypothesized that the rate of antipsychotic monotherapy in patients with TRS who were prescribed clozapine would be higher than that in patients with schizophrenia who were not prescribed clozapine. Methods: We assessed 8306 patients with schizophrenia nationwide from 178 institutions in Japan from 2016 to 2019. We analyzed the psychotropic prescription data at discharge in patients diagnosed with TRS and with no description of TRS (ND-TRS) based on the diagnosis listed in the discharge summary. Results: The rate of antipsychotic monotherapy in the TRS with clozapine group (91.3%) was significantly higher than that in the TRS without clozapine group (45.9%; P < 2.0 × 10−16) and the ND-TRS without clozapine group (54.7%; P < 2.0 × 10−16). The rate of antipsychotic monotherapy without any other concomitant psychotropics in the TRS with clozapine group (26.5%) was significantly higher than that in the TRS without clozapine group (12.6%; P = 1.1 × 10−6) and the ND-TRS without clozapine group (17.0%; P = 5.9 × 10−6). Conclusions: Clozapine prescription could be associated with a high rate of antipsychotic monotherapy. Patients will benefit from the correct diagnosis of TRS and thus from proper clozapine prescription

EGUIDE project and treatment guidelines

Background Clinical practice guidelines for schizophrenia and major depressive disorder have been published. However, these have not had sufficient penetration in clinical settings. We developed the Effectiveness of Guidelines for Dissemination and Education in Psychiatric Treatment (EGUIDE) project as a dissemination and education programme for psychiatrists. Aims The aim of this study is to assess the effectiveness of the EGUIDE project on the subjective clinical behaviour of psychiatrists in accordance with clinical practice guidelines before and 1 and 2 years after participation in the programmes. Method A total of 607 psychiatrists participated in this study during October 2016 and March 2019. They attended both 1-day educational programmes based on the clinical practice guidelines for schizophrenia and major depressive disorder, and answered web questionnaires about their clinical behaviours before and 1 and 2 years after attending the programmes. We evaluated the changes in clinical behaviours in accordance with the clinical practice guidelines between before and 2 years after the programme. Results All of the scores for clinical behaviours in accordance with clinical practice guidelines were significantly improved after 1 and 2 years compared with before attending the programmes. There were no significant changes in any of the scores between 1 and 2 years after attending. Conclusions All clinical behaviours in accordance with clinical practice guidelines improved after attending the EGUIDE programme, and were maintained for at least 2 years. The EGUIDE project could contribute to improved guideline-based clinical behaviour among psychiatrists

EGUIDE project and treatment guidelines

Aim: Although treatment guidelines for pharmacological therapy for schizophrenia and major depressive disorder have been issued by the Japanese Societies of Neuropsychopharmacology and Mood Disorders, these guidelines have not been well applied by psychiatrists throughout the nation. To address this issue, we developed the ‘Effectiveness of Guidelines for Dissemination and Education in Psychiatric Treatment (EGUIDE)’ integrated education programs for psychiatrists to disseminate the clinical guidelines. Additionally, we conducted a systematic efficacy evaluation of the programs. Methods: Four hundred thirteen out of 461 psychiatrists attended two 1‐day educational programs based on the treatment guidelines for schizophrenia and major depressive disorder from October 2016 to March 2018. We measured the participants’ clinical knowledge of the treatment guidelines using self‐completed questionnaires administered before and after the program to assess the effectiveness of the programs for improving knowledge. We also examined the relation between the participants’ demographics and their clinical knowledge scores. Results: The clinical knowledge scores for both guidelines were significantly improved after the program. There was no correlation between clinical knowledge and participant demographics for the program on schizophrenia; however, a weak positive correlation was found between clinical knowledge and the years of professional experience for the program on major depressive disorder. Conclusion: Our results provide evidence that educational programs on the clinical practices recommended in guidelines for schizophrenia and major depressive disorder might effectively improve participants’ clinical knowledge of the guidelines. These data are encouraging to facilitate the standardization of clinical practices for psychiatric disorders