An effective double gene knock-in strategy using small-molecule L755507 in the medaka fish (Oryzias latipes).

Homology-directed repair (HDR)-mediated genome editing has become a powerful method for altering chromosomal sequences in a seamless and accurate manner. However, the low efficiency of HDR in most cells hinders the establishment of desired strains harboring accurately modified genomes. To enhance HDR-mediated knock-in events, we explored two approaches, namely low-temperature incubation and chemical compound administration using medaka embryos after microinjection. We validated the performance of each method by calculating the knock-in efficiencies according to the expression area of fluorescent protein in the embryos. The in vivo assay indicated that the reduction in temperature did not promote HDR events, whereas among the nine compounds screened, the small molecule L755507 could enhance the HDR-mediated targeted integration of reporter cassettes. Additionally, the L755507-based approach allowed for the simultaneous integration of two different DNA fragments into the two targeted loci, that is, double knock-in. Our established knock-in system combining L755507, donor plasmids, and the CRISPR/Cas9 nickase system can reduce the workload for genetically modified strain generation, thus accelerating studies on the molecular mechanisms of biological phenomena.

[Metastatic Brain Tumor from Urachal Cancer:A Case Report].

Urachal cancers usually do not metastasize until the late stages, and local invasion and recurrence are observed with high frequency. We report a rare case of brain metastasis from urachal cancer in a 67-year-old female. She presented with abnormal behavior and right hemiparesis that started one month previously. MRI revealed a 55-mm lesion in the left frontal lobe, 5-mm in the right parietal lobe, and 2-mm lesion in the left cerebellum. The tumors showed ring enhancement on gadolinium-enhanced T1WI and were suspected to be metastatic brain tumors. Craniotomy was performed to excise the biggest tumor;the histological diagnosis was a poorly differentiated adenocarcinoma. As a result of the search for the primary tumor, the urachal carcinoma and multiple lung metastases were found. The patient chose best supportive care after diagnosis and died due to progression of multiple systemic metastases five months after surgery.

Single-laboratory validation for the determination of caffeic acid and seven caffeoylquinic acids in sweet potato leaves.

A single-laboratory validation study was conducted on an HPLC method for the detection and quantification of caffeic acid (CA) and seven species of caffeoylquinic acids (CQAs) in lyophilized sweet potato leaves. The procedure for extraction of the analytes from the matrix and the HPLC conditions for the efficient separation of CA and CQAs were optimized. In the proposed method, a relative response factor to one of the CQAs (5-CQA) was used to quantify the others. The method performed well in terms of precision when carried out on five different days and demonstrated Horwitz ratio (HorRat) scores ranging from 0.5 to 1.0 for all analytes, which were well within the limits of performance acceptability. Accuracy testing at three levels showed an overall recovery of 94% when duplicated on five different days. Moreover, a stability study revealed that all analytes in both standard solution and sample extract were stable for 28 days.

[Anti TNF-α (infliximab) treatment for intravenous immunoglobulin (IVIG) resistance patients with acute Kawasaki disease the effects of anticytokine therapy].

Among the patients with acute Kawasaki disease treated with intravenous immunoglobulin (IVIG), 10-20 % demonstrate resistance or incomplete effects. Cardiac complication such as the coronary arterial aneurysm is frequent in these patients. For patients with IVIG-resistance, we have surveyed the efficacy and safety of anti-cytokine therapy with use of infliximab (Remicade), chimera type anti TNF-α agent, for children. After May, 2005, Remicade has been used in >500 pediatric patients in whom IVIG and intravenous methylprednisolone pulse therapy did not show significant effects. The efficacy and safety of Remicade on patients with IVIG-resistant Kawasaki disease has been observed but 10~20 % of patients was Remicade-resistant. Re-treatment with IVIG or steroids was also effective. The efficacy of Remicade for reducing the fever duration, CRP, WBC counts was promising, but reduction of the incidence of coronary aneurysm was not confirmed. Randomized clinical trial will be needed.

Mechanical Thrombectomy for Patients with Occlusions in Both the Anterior Cerebral Artery and Middle Cerebral Artery: Case Series and Review of the Literature.

Objective: Most large-vessel occlusions (LVOs) amenable to acute recanalization occur in the internal carotid or middle cerebral artery. However, few LVOs with a multivessel disease can be difficult to treat. This study aimed to determine the outcomes of mechanical thrombectomy in patients with both anterior and middle cerebral artery occlusions. Methods: We retrospectively collected data for patients who had undergone mechanical thrombectomy since January 2016 at Fukushima Medical University and its affiliated institutions (10 institutions). Patients with occluded vessels in the anterior and middle cerebral arteries were selected, and patient background, treatment course, and outcomes were reviewed. Results: A total of 341 mechanical thrombectomies were performed during the study period. Seven patients had occlusions involving both anterior and middle cerebral arteries. In these seven patients, the median time from onset to imaging, imaging to puncture, and puncture to recanalization was 106, 60, and 74 min, respectively. Only one patient (14%) had a modified Rankin Scale of 0-2 at 90 days. Conclusion: Comorbid anterior cerebral artery occlusion may worsen the outcome of patients with middle cerebral artery occlusion.

Targeted deletion of ecto-5'-nucleotidase results in retention of inosine monophosphate content in postmortem muscle of medaka (Oryzias latipes).

Inosine monophosphate (IMP) is an important indicator of meat freshness and contributes to its umami taste. An attractive strategy for enhancing umami is to suppress the IMP-degrading activity and increase the IMP content in the skeletal muscle through genome editing technology using the CRISPR-Cas9 system. However, the molecular mechanisms underlying IMP degradation remain unclear. We cloned two ecto-5'-nucleotidase genes, designated as ecto-5'-nucleotidase-a (nt5ea) and ecto-5'-nucleotidase-b (nt5eb), from medaka (Oryzias latipes), a vertebrate model organism. Expression analysis using embryos showed that nt5ea or nt5eb overexpression remarkably upregulated IMP degradation, and that the IMP-degrading activity was higher in Nt5ea than in Nt5eb. Furthermore, we established frame-shifted or large deletion (lacking nt5ea or nt5eb locus) mutant strains and assayed the effects of gene disruptions on the amount of IMP in skeletal muscle. The nt5ea-deficient medaka showed considerable higher levels of IMP at 48 h postmortem than did the wild-type fish. The nt5eb mutants also exhibited higher IMP contents than that in the wild types, but the increase was less than that in the nt5ea mutants. Our results demonstrated that nt5e is an important regulator of IMP levels in skeletal muscle and that its loss of function was effective in maintaining IMP content.

Hyporheic zone, a blind spot: Discovery of the larva of Kisaura (Trichoptera: Philopotamidae) from Japan.

Four genera of Philopotamidae (Trichoptera) are known from Japan: Chimara, Dolophilodes, Kisaura, and Wormaldia. Among them, adults of the genus Kisaura are known in the Oriental and East Palearctic region; but until now, larvae of this genus have not been observed. In this study, molecular evidence is presented to support the relationship between adults and larvae of Kisaura minakawai Arefina 2005 from hyporheic zones of Japanese rivers. Morphological features of the larva of K. minakawai, which include very small eyes and somewhat dorsoventrally flattened (depressed) head capsules, compared with those of other philopotamid species, are also suggested to be adapted to the hyporheic zone. Detailed morphology of K. minakawai larval characters are provided.

Initial symptoms and diagnostic delay in children with brain tumors at a single institution in Japan.

BACKGROUND: A prolonged interval between onset of symptoms and diagnosis of childhood brain tumor is associated with worse neurological outcomes. The objectives of this study are to determine factors contributing to diagnostic delay and to find an interventional focus for further reduction in the interval between symptom onset and diagnosis in Japan. METHODS: We retrospectively analyzed 154 patients younger than 18 years with newly diagnosed brain tumors who visited our institution from January 2002 to March 2013. RESULTS: The median age at diagnosis was 6.2 years and the median total diagnostic interval (TDI) was 30 days. Patients with low-grade tumors and cerebral midline tumor location had significantly long TDI. Durations between the first medical consultation and diagnosis (diagnostic interval, DI) were exceedingly longer for patients with visual, hearing, or smelling abnormalities as the first symptom (median, 303 days). TDI and DI of patients who visited ophthalmologists or otolaryngologist for the first medical consultation were significantly longer. Among these patients, longer DI was associated with worse visual outcome. CONCLUSION: Raising awareness of brain tumor diagnosis among ophthalmologists and otolaryngologists may reduce diagnostic delay and may improve the neurological impairment of children with brain tumors in Japan.

Needs Assessment for Supports to Promote Pediatric Clinical Research Using an Online Survey of the Japanese Children's Hospitals Association.

INTRODUCTION: Infrastructure and the capacity to conduct clinical research in pediatrics have not been fully established in Japan. To elucidate the physicians' perspectives on clinical research, level of experience, existing barriers, and requests for support, we conducted a survey at 34 children's hospitals in Japan. METHODS: In January 2016, an online survey with 13 questions was sent to approximately 2000 physicians working in 34 pediatric hospitals belonging to the Japanese Association of Children's Hospitals and Related Institutions. RESULTS: Of the 360 respondents, 318 (88.3%) had presentations at academic conferences, and 261 (72.5%) had publications in academic journals, in the previous year. The most common study designs of clinical research conducted were case reports and case series. The most requested supports were for statistical analysis, followed by study design, grant application, and English-language editing. Younger physicians were more likely to prefer educational lectures (p < 0.001), whereas experienced physicians were more likely to request support for conducting statistical analysis (p = 0.002). Whereas physicians who had ever led a clinical trial requested support for the development of study protocol (p = .013), those without this experience preferred support for literature review (p = .002) and consultation services for study design (p = .027). CONCLUSIONS: The requests for supports were different, depending on the physicians' years after graduation and experience level in clinical research. In order to enhance clinical research in pediatrics, it is essential to provide appropriate types and levels of educational and support programs.

Sphenoparietal sinus transposition technique: optimization of the surgical corridor with preservation of prominent bridging veins between the brain and the cranial base during aneurysm clipping via the pterional approach.

OBJECTIVE: The sylvian bridging veins between the brain and the dura on the inner surface of the sphenoid wing can restrict brain retraction for widening of the lateral retrocarotid space during clipping surgery for internal carotid artery (ICA)-posterior communicating artery (PCoA) and basilar apex (BX) aneurysms. In such cases, the authors perform extradural anterior clinoidectomy with peeling of the temporal dura propria from the periosteal dura and inner cavernous membrane around the superior orbital fissure, with the incision of the dura mater stretching from the base of the temporal side to just before the distal dural ring of the ICA (termed by the authors as the sphenoparietal sinus transposition [SPST] technique). This technique displaces the bridging segment of the sylvian vein posteriorly and enables widening of the surgical space without venous injury. In this study, the authors observed the operative nuances and investigated the usefulness of this technique. METHODS: The authors retrospectively reviewed the medical charts of 66 consecutive patients with ICA-PCoA and BX aneurysms between January 2016 and July 2018. This technique was performed in 8 patients (5 patients with PCoA aneurysms and 3 with BX aneurysms) in whom the bridging segments of the sylvian veins between the brain and the skull base restricted brain retraction for widening of the surgical space. The surface areas of the lateral retrocarotid space and the aneurysm were measured at the most visible working angle before and after the SPST technique was performed. RESULTS: With the use of the SPST technique, an adequate surgical space for aneurysm clipping was obtained with preservation of the bridging veins in all patients. The mean surface areas of the lateral retrocarotid space (p = 0.002) and aneurysm (p = 0.001) were significantly increased from 18.3 ± 18.8 and 2.8 ± 2.5 cm2 before to 64.2 ± 21.1 and 20.9 ± 20.6 cm2, respectively, after the SPST technique was performed. CONCLUSIONS: The SPST technique enables displacement of the bridging segments of the sylvian veins without venous injury and enables widening of the surgical space around the lateral retrocarotid area.

Prediction of the difficulty of proximal vascular control using 3D-CTA for the surgical clipping of internal carotid artery-posterior communicating artery aneurysms.

OBJECTIVE: During surgical clipping of internal carotid artery (ICA)-posterior communicating artery (PCoA) aneurysms, proximal vascular control (PVC) is difficult to achieve in some cases because of variations in the anatomy of this type of aneurysm and its parent arteries. The authors investigated morphometric features that may be predictive for the necessity of anterior clinoidectomy (ACL) or cervical ICA exposure for PVC. METHODS: The authors retrospectively reviewed 65 patients with an ICA-PCoA aneurysm treated with clipping during the previous 3 years. The factors considered for assessing the difficulty of attaining PVC included the following: the maximum diameter of the aneurysm; the distance between the tip of the anterior clinoid process (ACP) and the proximal aneurysmal neck; the presence of calcification at the ophthalmic segment of the ICA; and the angles between the communicating segment of the ICA and the ophthalmic segment of the ICA and a line perpendicular to the cranial base, which reflect the tortuosity of the ICA. These parameters were measured based on preoperative CTA results. RESULTS: In a total of 21 patients (32.3%), PVC was difficult to perform with the usual pterional approach. In 6 patients, temporary artery occlusions (TAOs) were difficult to achieve because of severe atherosclerotic wall changes in the ophthalmic segment of the ICA. For 15 patients, the ACPs overhanging the ophthalmic segment of the ICA obstructed the ability to secure a space for TAO. In the 21 patients with PVC difficulty, ACL alone, cervical ICA exposure alone, and both ACL and cervical ICA exposure were conducted in 6, 8, and 7 patients, respectively. Multivariate analysis with binary logistic regression revealed that the maximum diameter of the aneurysm (p = 0.041), the distance between the proximal neck of the aneurysm and the ACP tip (p = 0.002), and calcification of the ICA ophthalmic segment (p = 0.001) were significant predictive factors for difficulties with PVC. A receiver operating characteristic curve analysis revealed that a distance between the proximal aneurysmal neck and the ACP tip of ≤ 5.4 mm was the best cutoff value for predicting the difficulty of attaining PVC (area under the curve 0.800, sensitivity 80.0%, specificity 80.0%). CONCLUSIONS: A short distance between the proximal aneurysmal neck and the ACP tip and the presence of calcification at the ophthalmic segment of the ICA on preoperative CTA are helpful for predicting the difficulty of achieving PVC.

Inhibitory effects of the antiepileptic drug ethosuximide on G protein-activated inwardly rectifying K+ channels.

Antiepileptic drugs protect against seizures by modulating neuronal excitability. Ethosuximide is selectively used for the treatment of absence epilepsy, and has also been shown to have the potential for treating several other neuropsychiatric disorders in addition to several antiepileptic drugs. Although ethosuximide inhibits T-type Ca(2+), noninactivating Na(+), and Ca(2+)-activated K(+) channels, the molecular mechanisms underlying the effects of ethosuximide have not yet been sufficiently clarified. G protein-activated inwardly rectifying K(+) channels (GIRK, or Kir3) play an important role in regulating neuronal excitability, heart rate and platelet aggregation. In the present study, the effects of various antiepileptic drugs on GIRK channels were examined first by using the Xenopus oocyte expression assay. Ethosuximide at clinically relevant concentrations inhibited GIRK channels expressed in Xenopus oocytes. The inhibition was concentration-dependent, but voltage-independent, and time-independent during each voltage pulse. However, the other antiepileptic drugs tested: phenytoin, valproic acid, carbamazepine, phenobarbital, gabapentin, topiramate and zonisamide, had no significant effects on GIRK channels even at toxic concentrations. In contrast, Kir1.1 and Kir2.1 channels were insensitive to all of the drugs tested. Ethosuximide also attenuated ethanol-induced GIRK currents. These inhibitory effects of ethosuximide were not observed when ethosuximide was applied intracellularly. In granule cells of cerebellar slices, ethosuximide inhibited GTPgammaS-activated GIRK currents. Moreover, ADP- and epinephrine-induced platelet aggregation was inhibited by ethosuximide, but not by charybdotoxin, a platelet Ca(2+)-activated K(+) channel blocker. These results suggest that the inhibitory effects of ethosuximide on GIRK channels may affect some of brain, heart and platelet functions.

Exclusive Cerebellar and Leptomeningeal Metastases from Early Gastric Cancer 14 Months after Proximal Gastrectomy: An Autopsy Case Report.

We report a rare autopsy case in which the patient received gastrectomy after an endoscopic diagnosis of early gastric cancer, and had deteriorated due to exclusive metastatic cerebellar tumors identified 14 months after surgery. A 65-year-old male was diagnosed as having a 0-IIc-type early gastric cancer on the posterior wall of the upper stomach by gastrointestinal endoscopy in search of a cause of epigastralgia, and thus received proximal gastrectomy and pyloroplasty. Although the tumor was in the early stages and limited within the mucosal layer, adjuvant chemotherapy was started by using S-1 80 mg daily due to evidence of metastasis into lymph node #3 at the lesser curvature. Evidence of both recurrence and metastases was not detected by CT scans of the chest, abdomen, and pelvis, and the chemotherapy was completed 12 months after surgery. However, the patient was admitted to hospital 14 months postoperatively due to dizziness and gait disturbance. Cranial MRI (Magnetic Resonance Imaging) revealed multiple tumors in the bilateral cerebellar hemispheres with additional leptomeningeal involvement. The patient died 2 weeks after admission. An autopsy revealed metastatic cerebellar tumors and leptomeningeal lesions from the early gastric cancer, and obstructive hydrocephalus due to metastatic cerebellar tumors. To our knowledge, this case is the first report of metastasis exclusive to the cerebellum and leptomeninges from early gastric cancer limited to the mucosal layer.

Complete regression of xenografted human carcinomas by a paclitaxel-carboxymethyl dextran conjugate (AZ10992).

Clinically available taxanes, such as paclitaxel and docetaxel, represent one of the most promising classes of anticancer agents, despite their toxicity. To improve their pharmacological profiles, AZ10992 was synthesized based on the concept that a rational design of a polymer-drug conjugate would increase the efficacy of the parent drug. This prodrug is a paclitaxel-carboxymethyl dextran conjugate (molecular weight 150,000 g/mol) via a gly-gly-phe-gly linker. The in vivo antitumor study using AZ10992 against colon26 carcinoma cells, resistant to paclitaxel, supported this concept. Additionally, the comparative efficacy studies of AZ10992 and paclitaxel using a panel of human tumor xenografts in nude mice showed the advantages of drug-polymer conjugation. The maximum tolerated dose of AZ10992 was more than twice as high as the MTD of paclitaxel. A repeated intravenous administration of AZ10992 at 30 mg/kg/day (five injections for 4-days) showed complete regression of MX-1 mammary carcinoma xenografts. Also, HT-29 colorectal tumor xenografts, which are highly refractory to paclitaxel, showed complete regression after AZ10992 administered at 30 mg/kg/day (seven injections for 4-days). Pharmacokinetic studies showed that there were significant increases in the amount and the exposure time of total paclitaxel in the tumors after intravenous administration of AZ10992, which explains the enhanced efficacy of AZ10992.

Inhibition by cocaine of G protein-activated inwardly rectifying K+ channels expressed in Xenopus oocytes.

Cocaine, a commonly abused psychostimulant, interacts with not only transporters for dopamine, serotonin and norepinephrine but also several receptors and channels. However, the molecular mechanisms underlying the various effects of cocaine remain to be clarified. Using the Xenopus oocyte expression assay, we investigated the effects of cocaine on G protein-activated inwardly rectifying K+ (GIRK) channels, which regulate neuronal excitability and the heart rate. In oocytes injected with mRNAs for GIRK1/GIRK2, GIRK2 or GIRK1/GIRK4 subunits, cocaine reversibly reduced basal GIRK inward currents. The inhibition by cocaine at the toxic levels was concentration-dependent, but voltage-independent and time-independent during each voltage pulse. However, methylphenidate, methamphetamine and 3,4-methylenedioxymethamphetamine (MDMA) at their toxic concentrations had little effect on the channels. Additionally, Kir1.1 and Kir2.1 channels were insensitive to all of the drugs. The inhibition by cocaine, which exists mainly in a protonated form at pH 7.4, was not affected by extracellular pH 9, at which the proportion of the uncharged form increases, suggesting the inhibition by both forms with similar effectiveness, and at physiological pH the effect being predominantly due to the protonated cocaine. Our results suggest that inhibition of GIRK channels by cocaine may contribute to some of its toxic effects.

Polycystic ovary syndrome with asynchronous bilateral adnexal torsion in a natural cycle.

Cases involving polycystic ovaries (PCOs) with adnexal torsion in a natural cycle without ovulation induction are rare, and no reports of such cases have described asynchronous bilateral adnexal torsion. Here, we report a case of PCO syndrome (PCOS) with asynchronous bilateral adnexal torsion in a natural cycle. The patient was a 37-year-old woman with a history of 2 gravidas 1 para. Her primary complaint was left lower abdominal pain. Ultrasonography and MRI identified multiple uterine myomas occupying the pelvis and the left ovary, with oedematous swelling that had moved ventrally to the uterus. She was diagnosed with adnexal torsion and underwent emergency laparoscopic adnexectomy. Nine months after surgery, she experienced right lower abdominal pain. Ultrasonography revealed suspected right adnexal torsion and she underwent emergency surgery. The right ovary was twisted 540° counterclockwise and swollen to 7 cm in size, with partial polycystic changes. She was histopathologically diagnosed with a PCO, and the final diagnosis, which also considered the endocrine test results, was PCOS. In PCOS, adnexal torsion may occur if the swollen ovary moves because of a hysteromyoma or other cause. Accordingly, torsion should be considered during the follow-up of patients with PCOS.

Inhibition of G protein-activated inwardly rectifying K+ channels by ifenprodil.

G protein-activated inwardly rectifying K+ channels (GIRK, also known as Kir3) are regulated by various G-protein-coupled receptors. Activation of GIRK channels plays an important role in reducing neuronal excitability in most brain regions and the heart rate. Ifenprodil, which is a clinically used cerebral vasodilator, interacts with several receptors, such as alpha1 adrenergic, N-methyl-D-aspartate, serotonin and sigma receptors. However, the molecular mechanisms underlying the various clinically related effects of ifenprodil remain to be clarified. Here, we examined the effects of ifenprodil on GIRK channels by using Xenopus oocyte expression assays. In oocytes injected with mRNAs for GIRK1/GIRK2, GIRK2 or GIRK1/GIRK4 subunits, ifenprodil reversibly reduced inward currents through the basal GIRK activity. The inhibition was concentration-dependent, but voltage- and time-independent, suggesting that ifenprodil may not act as an open channel blocker of the channels. In contrast, Kir1.1 and Kir2.1 channels in other Kir channel subfamilies were insensitive to ifenprodil. Furthermore, GIRK current responses activated by the cloned kappa-opioid receptor were similarly inhibited by ifenprodil. The inhibitory effects of ifenprodil were not observed when ifenprodil was applied intracellularly, and were not affected by extracellular pH, which changed the proportion of the uncharged to protonated ifenprodil, suggesting its action from the extracellular side. The GIRK currents induced by ethanol were also attenuated in the presence of ifenprodil. Our results suggest that direct inhibition of GIRK channels by ifenprodil, at submicromolar concentrations or more, may contribute to some of its therapeutic effects and adverse side effects.

Gene expression in the brain from fluoxetine-injected mouse using DNA microarray.

Previously we have examined the effects of phencyclidine and clozapine upon the gene expression in the mouse brain. Recently, fluoxetine (Prozac) has been introduced for the therapeutic purpose as an antidepressant drug. Miledi et al. reported blockage of mouse muscle and neuronal nicotinic acetylcholine receptor by various concentrations of fluoxetine. Furthermore, Kobayashi et al. discovered that fluoxetine inhibits G protein activated inwardly rectifying G protein activated K(+) (GIRK) channels using Xenopus oocyte expression assay. From these experiments, we considered that it might be interesting to study the effects of fluoxetine on the gene expression in the mouse brain. After we have injected fluoxetine once a day into mouse for 20 days, we sacrificed mouse by decapitation and extracted RNA from mouse cerebral cortex. We used DNA microarray method for examining the gene expression in the brain. We found the downregulation of many spot signals in the fluoxetine-treated mouse, for example cholecystockinin and prostaglandin D2 synthase.

G protein-activated inwardly rectifying K+ channel inhibition and rescue of weaver mouse motor functions by antidepressants.

Antidepressants, including tricyclic antidepressants (TCAs) and selective serotonin reuptake inhibitors (SSRIs), have been widely used for the treatment of not only depression but also other psychiatric disorders, although the molecular mechanisms of the drug effects have not yet been sufficiently revealed. Here, we investigated the in vivo effects of these antidepressants on G protein-activated inwardly rectifying K+ (GIRK) channels, which are important for regulating the excitability of various cells, by using weaver (wv) mice, which have mutant GIRK channels and show abnormal neuronal cell death and motor disturbances. First, we found that a widely used SSRI fluoxetine (also known as Prozac) effectively inhibited wv GIRK2 channels like wild-type GIRK channels, expressed in Xenopus oocytes. Next, we found that weaver motor disturbances were remarkably alleviated by chronic treatment with fluoxetine or desipramine. Furthermore, the chronic fluoxetine treatment substantially suppressed the abnormal neuronal cell death in the weaver mouse cerebellum and pontine nuclei. These results suggest that continuous inhibition of wv GIRK2 channels by a group of antidepressants caused substantial suppression of the neuronal cell death and resulted in improvement of motor abilities in weaver mice. These results provide evidence for in vivo GIRK channel inhibition by a group of antidepressants.