Inositol polyphosphate multikinase mediates extinction of fear memory.

Inositol polyphosphate multikinase (IPMK), the key enzyme for the biosynthesis of higher inositol polyphosphates and phosphatidylinositol 3,4,5-trisphosphate, also acts as a versatile signaling player in regulating tissue growth and metabolism. To elucidate neurobehavioral functions of IPMK, we generated mice in which IPMK was deleted from the excitatory neurons of the postnatal forebrain. These mice showed no deficits in either novel object recognition or spatial memory. IPMK conditional knockout mice formed cued fear memory normally but displayed enhanced fear extinction. Signaling analyses revealed dysregulated expression of neural genes accompanied by selective activation of the mechanistic target of rapamycin (mTOR) regulatory enzyme p85 S6 kinase 1 (S6K1) in the amygdala following fear extinction. The IPMK mutants also manifested facilitated hippocampal long-term potentiation. These findings establish a signaling action of IPMK that mediates fear extinction.

Transcriptome changes associated with fat deposition in the longissimus thoracis of Korean cattle following castration.

The castration of bulls increases the intramuscular fat (IMF) content in skeletal muscle. However, the biological processes of IMF accumulation in skeletal muscle after castration are not completely understood at the molecular level. This study examined the global transcriptomic changes in the longissimus thoracis muscle (LT) of bulls following castration using RNA sequencing (RNA-Seq) and identified new genes or pathways associated with beef quality. Ten bulls and 10 steers castrated at 6 months of age were slaughtered at 26 and 32 months of age respectively. For transcriptome analysis, six LT samples from three bulls and three steers were selected based on age, carcass weight, carcass quantity and beef quality grades. Using RNA-Seq, transcriptomic profiles of the LT were compared between bulls and steers. In all, 640 of the 18,027 genes identified through RNA-Seq were differentially expressed genes (DEGs) between bulls and steers. Pathway analysis of these 640 DEGs showed significant (p < .05) changes in seven Kyoto Encyclopedia of Genes and Genomes pathways, and the most significant terms were complement and coagulation cascade pathways. The transcriptomic expression patterns of 10 genes in the complement and coagulation cascades were validated using all animals through quantitative real-time polymerase chain reaction analysis. In conclusion, transcriptome changes associated with the complement and coagulation cascade pathways provide novel insights into understanding molecular mechanisms responsible for IMF accumulation following castration in beef cattle.

Inositol pyrophosphates inhibit synaptotagmin-dependent exocytosis.

Inositol pyrophosphates such as 5-diphosphoinositol pentakisphosphate (5-IP7) are highly energetic inositol metabolites containing phosphoanhydride bonds. Although inositol pyrophosphates are known to regulate various biological events, including growth, survival, and metabolism, the molecular sites of 5-IP7 action in vesicle trafficking have remained largely elusive. We report here that elevated 5-IP7 levels, caused by overexpression of inositol hexakisphosphate (IP6) kinase 1 (IP6K1), suppressed depolarization-induced neurotransmitter release from PC12 cells. Conversely, IP6K1 depletion decreased intracellular 5-IP7 concentrations, leading to increased neurotransmitter release. Consistently, knockdown of IP6K1 in cultured hippocampal neurons augmented action potential-driven synaptic vesicle exocytosis at synapses. Using a FRET-based in vitro vesicle fusion assay, we found that 5-IP7, but not 1-IP7, exhibited significantly higher inhibitory activity toward synaptic vesicle exocytosis than IP6 Synaptotagmin 1 (Syt1), a Ca(2+) sensor essential for synaptic membrane fusion, was identified as a molecular target of 5-IP7 Notably, 5-IP7 showed a 45-fold higher binding affinity for Syt1 compared with IP6 In addition, 5-IP7-dependent inhibition of synaptic vesicle fusion was abolished by increasing Ca(2+) levels. Thus, 5-IP7 appears to act through Syt1 binding to interfere with the fusogenic activity of Ca(2+) These findings reveal a role of 5-IP7 as a potent inhibitor of Syt1 in controlling the synaptic exocytotic pathway and expand our understanding of the signaling mechanisms of inositol pyrophosphates.

Effects of ambient temperature and rumen-protected fat supplementation on growth performance, rumen fermentation and blood parameters during cold season in Korean cattle steers.

OBJECTIVE: This study was performed to evaluate whether cold ambient temperature and dietary rumen-protected fat (RPF) supplementation affect growth performance, rumen fermentation, and blood parameters in Korean cattle steers. METHODS: Twenty Korean cattle steers (body weight [BW], 550.6±9.14 kg; age, 19.7±0.13 months) were divided into a conventional control diet group (n = 10) and a 0.5% RPF supplementation group (n = 10). Steers were fed a concentrate diet (1.6% BW) and a rice straw diet (1 kg/d) for 16 weeks (January 9 to February 5 [P1], February 6 to March 5 [P2], March 6 to April 3 [P3], and April 4 to May 2 [P4]). RESULTS: The mean and minimum indoor ambient temperatures in P1 (-3.44°C, -9.40°C) were lower (p<0.001) than those in P3 (5.87°C, -1.86°C) and P4 (11.18°C, 4.28°C). The minimum temperature in P1 fell within the moderate cold-stress (CS) category, as previously reported for dairy cattle, and the minimum temperatures of P2 and P3 were within the mild CS category. Neither month nor RPF supplementation affected the average daily gain or gain-to-feed ratio (p>0.05). Ruminal ammonia nitrogen concentrations were higher (p<0.05) in cold winter than spring. Plasma cortisol concentrations were lower (p<0.05) in the coldest month than in the other months. Serum glucose concentrations were generally higher in colder months than in the other months but were unaffected by RPF supplementation. RPF supplementation increased both total cholesterol (p = 0.004) and high-density lipoprotein (HDL) concentrations (p = 0.03). CONCLUSION: Korean cattle may not be significantly affected by moderate CS, considering that the growth performance of cattle remained unchanged, although variations in blood parameters were observed among the studied months. RPF supplementation altered cholesterol and HDL concentrations but did not affect growth performance.

Effects of heat stress and rumen-protected fat supplementation on growth performance, rumen characteristics, and blood parameters in growing Korean cattle steers.

OBJECTIVE: This study was performed to evaluate whether hot temperature and rumen-protected fat (RPF) supplementation affect growth performance, rumen characteristics, and serum metabolites in growing stage of Korean cattle steers. METHODS: Twenty Korean cattle steers (230.4±4.09 kg of body weight [BW], 10.7±0.09 months of age) were divided into a conventional control diet group (n = 10) and a 0.8% RPF supplementation group (n = 10). Steers were fed 1.5% BW of a concentrate diet and 4 kg of tall fescue hay for 16 weeks (July 10 to August 6 [P1], August 7 to September 3 [P2], September 4 to October 1 [P3], October 2 to 30 [P4], of 2015). RESULTS: The mean temperature-humidity index (THI) was higher (p<0.001) in P1 (76.8), P2 (76.3), and P3 (75.9) than in P4 (50.9). The mean THI of P1-3 were within the alert heat stress (HS) category range according to previously reported categories for feedlot cattle, and the mean THI of P4 was under the thermo-neutral range. Neither month nor RPF supplementation affected (p>0.05) average daily gain and gain to feed ratio. Month and RPF supplementation affected concentrations of glucose, albumin, and high-density lipoprotein (HDL); those of albumin and glucose tended to decrease (p<0.10), but HDL concentration increased (p<0.01) by RPF supplementation. Neither month nor RPF affected (p>0.05) ruminal pH, NH3-N, and volatile fatty acid concentrations, whereas the C2:C3 ratio was affected (p<0.05) by month. CONCLUSION: Korean cattle may not have been significantly affected by alert HS during the growing stage. Growth performance was higher during hotter months, although some changes in blood metabolites were observed. The RPF supplementation affected some blood lipids and carbohydrate metabolites but did not affect growth performance.

Genetic, management, and nutritional factors affecting intramuscular fat deposition in beef cattle - A review.

Intramuscular fat (IMF) content in skeletal muscle including the longissimus dorsi muscle (LM), also known as marbling fat, is one of the most important factors determining beef quality in several countries including Korea, Japan, Australia, and the United States. Genetics and breed, management, and nutrition affect IMF deposition. Japanese Black cattle breed has the highest IMF content in the world, and Korean cattle (also called Hanwoo) the second highest. Here, we review results of research on genetic factors (breed and sex differences and heritability) that affect IMF deposition. Cattle management factors are also important for IMF deposition. Castration of bulls increases IMF deposition in most cattle breeds. The effects of several management factors, including weaning age, castration, slaughter weight and age, and environmental conditions on IMF deposition are also reviewed. Nutritional factors, including fat metabolism, digestion and absorption of feed, glucose/starch availability, and vitamin A, D, and C levels are important for IMF deposition. Manipulating IMF deposition through developmental programming via metabolic imprinting is a recently proposed nutritional method to change potential IMF deposition during the fetal and neonatal periods in rodents and domestic animals. Application of fetal nutritional programming to increase IMF deposition of progeny in later life is reviewed. The coordination of several factors affects IMF deposition. Thus, a combination of several strategies may be needed to manipulate IMF deposition, depending on the consumer's beef preference. In particular, stage-specific feeding programs with concentrate-based diets developed by Japan and Korea are described in this article.

Identification of circulating miRNA involved in meat yield of Korean cattle.

Cattle plays an important role in providing essential nutrients through meat production. Thus, we focused on epigenetic factors associated with meat yield. To investigate circulating miRNAs that are involved with meat yield and connect biofluids and longissimus dorsi (LD) muscle in Korean cattle, we performed analyses of the carcass characteristics, miRNA array, qPCR, and bioinformatics. Carcass characteristics relative to the yield grade (YG) showed that the yield index and rib eye area were the highest, whereas the backfat thickness was the lowest for YG A (equal to high YG) cattle among the three YGs. miRNA array sorted the circulating miRNAs that connect biofluids and LD muscle. miRNA qPCR showed that miR-15a (r = 0.84), miR-26b (r = 0.91), and miR-29c (r = 0.92) had positive relationships with biofluids and LD muscle. In YG A cattle, miR-26b was considered to be a circulating miRNA connecting biofluids and LD muscle because the target genes of miR-26b were more involved with myogenesis. Then, miR-26b-targeted genes, DIAPH3 and YOD1, were downregulated in YG A cattle. Our results suggest that miR-15a, miR-26b, and miR-29c are upregulated in biofluids and LD muscle, whereas DIAPH3 and YOD1 are downregulated in the LD muscle of finishing cattle steers.

Muscle-specific deletion of signal transducer and activator of transcription 5 augments lipid accumulation in skeletal muscle and liver of mice in response to high-fat diet.

PURPOSE: Growth hormone (GH) controls liver metabolism through the transcription factor signal transducer and activator of transcription 5 (STAT5). However, it remains to be fully understood to what extent other GH/STAT5 target tissues contribute to lipid and glucose metabolism. This question was now addressed in muscle-specific STAT5 knockout (STAT5 MKO) mice model. METHODS: Changes in lipid and glucose metabolism were investigated at physiological and molecular levels in muscle and liver tissues of STAT5 MKO mice under normal diet or high-fat diet (HFD) conditions. RESULTS: STAT5 MKO mice exhibited an increased intramyocellular lipid (IMCL) accumulation in the quadriceps in HFD group. Decreased lipolytic hormone-sensitive lipase transcript levels may contribute to the increased IMCL accumulation in STAT5 MKO mice. STAT5 MKO induced hepatic lipid accumulation without deregulated STAT5 signaling. The upregulation of lipoprotein lipase and Cd36 mRNA levels, an increased trend of very low-density lipoprotein receptor mRNA levels, and elevated circulating concentrations of free fatty acid, triglyceride, and total cholesterol support the increase in hepatic lipid accumulation. CONCLUSIONS: STAT5 MKO in conjunction with a HFD deregulated both lipid and glucose metabolism in skeletal muscle, and this deregulation induced hepatic fat accumulation via increased circulating glucose, FFA, and TG concentrations. Our study emphasizes that muscle-specific STAT5 signaling is important for balancing lipid and glucose metabolism in peripheral tissues, including muscle and liver and that the deregulation of local STAT5 signaling augments HFD-induced lipid accumulation in both muscle and liver.

Mitochondrial Respiratory Defect Causes Dysfunctional Lactate Turnover via AMP-activated Protein Kinase Activation in Human-induced Pluripotent Stem Cell-derived Hepatocytes.

A defective mitochondrial respiratory chain complex (DMRC) causes various metabolic disorders in humans. However, the pathophysiology of DMRC in the liver remains unclear. To understand DMRC pathophysiology in vitro, DMRC-induced pluripotent stem cells were generated from dermal fibroblasts of a DMRC patient who had a homoplasmic mutation (m.3398T→C) in the mitochondrion-encoded NADH dehydrogenase 1 (MTND1) gene and that differentiated into hepatocytes (DMRC hepatocytes) in vitro. DMRC hepatocytes showed abnormalities in mitochondrial characteristics, the NAD(+)/NADH ratio, the glycogen storage level, the lactate turnover rate, and AMPK activity. Intriguingly, low glycogen storage and transcription of lactate turnover-related genes in DMRC hepatocytes were recovered by inhibition of AMPK activity. Thus, AMPK activation led to metabolic changes in terms of glycogen storage and lactate turnover in DMRC hepatocytes. These data demonstrate for the first time that energy depletion may lead to lactic acidosis in the DMRC patient by reduction of lactate uptake via AMPK in liver.

Effect of knife castration on leukocyte cytokine expression and indicators of stress, pain, and inflammation in Korean cattle bull calves.

OBJECTIVE: This study investigated the effects of surgical castration on behavior, physiological and inflammatory indicators, and leukocyte cytokine mRNA levels in Korean cattle bull calves. METHODS: Nineteen Korean cattle bull calves (average body weight, 254.5 kg; average age, 8.2 months) were divided into two treatment groups: control (n = 9) and castration (n = 10). Surgical castration was performed using Newberry knives and a Henderson castrating tool. Blood was obtained just before castration (0 h) and at 0.5 h, 6 h, 1 d, 3 d, 7 d, and 14 d after castration. Plasma cortisol (PC), saliva cortisol (SC), plasma substance P, and plasma haptoglobin concentrations, and the leucocyte mRNA levels of the interleukin-1-alpha (IL1A), interleukin-1-beta (IL1B), interleukin-1 receptor antagonist (IL1RN), and interleukin-6 (IL6) genes were analyzed. RESULTS: Castration decreased (p<0.01) the average daily gain and gain/feed ratio. Castration reduced the time spent eating (p<0.001) and the eating frequency (p<0.01) and increased (p<0.001) the lying frequency. Castration temporarily increased (p<0.05) circulating PC and SC concentrations at 0.5 h after castration. Castration temporarily increased (p<0.05) plasma substance P concentrations at 1 d after castration. Castration increased (p<0.05) plasma haptoglobin concentrations at 1 and 3 d after castration. Castration increased (p< 0.05) leukocyte mRNA levels of the IL1A, IL1B, IL1RN, and IL6 genes at 6 h after castration. CONCLUSION: Castration temporarily induced stress and expression of leucocyte inflammatory cytokine genes in Korean cattle bull calves.

Deletion of IP6K1 in mice accelerates tumor growth by dysregulating the tumor-immune microenvironment.

A family of inositol hexakisphosphate kinases (IP6Ks) catalyzes the production of inositol pyrophosphate IP7 (5-diphosphoinositolpentakisphosphate) which is known to modulate various biological events such as cell growth. While targeting IP6K1 in various cancer cells has been well reported to control cancer cell motility and invasiveness, the role of host IP6K1 in tumor progression remains unknown. By using a syngeneic MC38 murine mouse colon carcinoma model, here we examined how host IP6K1 in the tumor microenvironment influences tumor growth. In IP6K1 knockout (KO) mice, the growth of MC38 tumor cells was markedly accelerated and host survival was significantly shortened compared with wild-type (WT). Our flow cytometric analysis revealed that tumors grown in IP6K1 KO mice had lower immune suppressive myeloid cells and M1 polarized macrophages. Notably, infiltration of both antigen-presenting dendritic cells and CD8+ cytotoxic T lymphocytes into the tumor tissues was remarkably abrogated in IP6K1 KO condition. These studies suggest that enhanced tumor growth in IP6K1 KO mice resulted from reduced anti-tumor immunity due to disturbed immune cell actions in the tumor microenvironment. In conclusion, we demonstrate that host IP6K1 acts as a tumor suppressor, most likely by fine-tuning diverse tumor-immune cell interactions, which might have implications for improving the host response against cancer progression.

Partial Substitution of Corn Grain in the Diet with Beet Pulp Reveals Increased Ruminal Acetate Proportion and Circulating Insulin Levels in Korean Cattle Steers.

We investigated the effects of the partial substitution of corn grain in the diet with beet pulp on growth performance, ruminal fermentation characteristics, microbial profiles, and blood lipogenic parameters in fattening steers. Twelve Korean cattle steers (body weight, 485 ± 19.32 kg; age, 18.0 ± 0.17 months) were equally divided into corn grain (CG) and beet pulp (BP) groups. Approximately 75% of dry matter of the requirement was offered as a concentrate portion, and the remaining 25% was offered as oat straw. Eighty percent of the concentrate portion was provided by a pelleted basal concentrate, and the remaining 20% with corn grain for the CG group, or 18% beet pulp plus 2.0% rumen-protected fat for the BP group, respectively, by top dressing. The experiment was conducted for 14 weeks, including a 2-week acclimation period. Growth rate was not affected by beet pulp feeding (p = 0.55). The molar proportions of ruminal acetate (p < 0.05) on wk 4, the relative abundances of ruminal cellulolytic bacteria, including Fibrobacter succinogenes (p = 0.01) and Ruminococcus albus (p = 0.04) on wk 12, and serum insulin concentrations (p < 0.05) on wk 12 were higher in the BP group than in the CG group, whereas the molar proportions of propionate (p < 0.05) on wks 8 and 12 and serum nonesterified fatty acids (p < 0.05) on wk 12 were lower in the BP group. Beet pulp could be used as a lipogenic energy source without affecting growth performance during the fattening period of cattle.

Lomitapide, a cholesterol-lowering drug, is an anticancer agent that induces autophagic cell death via inhibiting mTOR.

Autophagy is a biological process that maintains cellular homeostasis and regulates the internal cellular environment. Hyperactivating autophagy to trigger cell death has been a suggested therapeutic strategy for cancer treatment. Mechanistic target of rapamycin (mTOR) is a crucial protein kinase that regulates autophagy; therefore, using a structure-based virtual screen analysis, we identified lomitapide, a cholesterol-lowering drug, as a potential mTOR complex 1 (mTORC1) inhibitor. Our results showed that lomitapide directly inhibits mTORC1 in vitro and induces autophagy-dependent cancer cell death by decreasing mTOR signaling, thereby inhibiting the downstream events associated with increased LC3 conversion in various cancer cells (e.g., HCT116 colorectal cancer cells) and tumor xenografts. Lomitapide also significantly suppresses the growth and viability along with elevated autophagy in patient-derived colorectal cancer organoids. Furthermore, a combination of lomitapide and immune checkpoint blocking antibodies synergistically inhibits tumor growth in murine MC38 or B16-F10 preclinical syngeneic tumor models. These results elucidate the direct, tumor-relevant immune-potentiating benefits of mTORC1 inhibition by lomitapide, which complement the current immune checkpoint blockade. This study highlights the potential repurposing of lomitapide as a new therapeutic option for cancer treatment.

Gut Epithelial Inositol Polyphosphate Multikinase Alleviates Experimental Colitis via Governing Tuft Cell Homeostasis.

BACKGROUND & AIMS: Inositol polyphosphate multikinase (IPMK), an essential enzyme for inositol phosphate metabolism, has been known to mediate major biological events such as growth. Recent studies have identified single-nucleotide polymorphisms in the IPMK gene associated with inflammatory bowel disease predisposition. Therefore, we aimed to investigate the functional significance of IPMK in gut epithelium. METHODS: We generated intestinal epithelial cell (IEC)-specific Ipmk knockout (IPMKΔIEC) mice, and assessed their vulnerability against dextran sulfate sodium-induced experimental colitis. Both bulk and single-cell RNA sequencing were performed to analyze IPMK-deficient colonic epithelial cells and colonic tuft cells. RESULTS: Although IPMKΔIEC mice developed normally and showed no intestinal abnormalities during homeostasis, Ipmk deletion aggravated dextran sulfate sodium-induced colitis, with higher clinical colitis scores, and increased epithelial barrier permeability. Surprisingly, Ipmk deletion led to a significant decrease in the number of tuft cells without influencing other IECs. Single-cell RNA sequencing of mouse colonic tuft cells showed 3 distinct populations of tuft cells, and further showed that a transcriptionally inactive population was expanded markedly in IPMKΔIEC mice, while neuronal-related cells were relatively decreased. CONCLUSIONS: Cholinergic output from tuft cells is known to be critical for the restoration of intestinal architecture upon damage, supporting that tuft cell-defective IPMKΔIEC mice are more prone to colitis. Thus, intestinal epithelial IPMK is a critical regulator of colonic integrity and tissue regeneration by determining tuft cell homeostasis and affecting cholinergic output.

Inositol Polyphosphate Multikinase Signaling: Multifaceted Functions in Health and Disease.

Inositol phosphates are water-soluble intracellular signaling molecules found in eukaryotes from yeasts to mammals, which are synthesized by a complex network of enzymes including inositol phosphate kinases. Among these, inositol polyphosphate multikinase (IPMK) is a promiscuous enzyme with broad substrate specificity, which phosphorylates multiple inositol phosphates, as well as phosphatidylinositol 4,5-bisphosphate. In addition to its catalytic actions, IPMK is known to non-catalytically control major signaling events via direct protein-protein interactions. In this review, we describe the general characteristics of IPMK, highlight its pleiotropic roles in various physiological and pathological conditions, and discuss future challenges in the field of IPMK signaling pathways.

Relationships among carcass traits, auction price, and image analysis traits of marbling characteristics in Korean cattle beef.

Marbling score (MS) is related to beef auction price (AP) in Korea, but how these relate to marbling texture (i.e. number and distribution of marbling particles [MPs]) is not known. We examined relationships among carcass traits, carcass AP, and image analysis marbling texture traits. In experiment 1, carcass data and longissimus thoracis (LT) were obtained from Korean cattle steers reared under similar feeding conditions. MS, quality grade (QG), and AP were related (P < 0.001) to numbers of coarse MPs and fine MPs and fineness index. In experiment 2, LT images photographed at a slaughterhouse were used in regression analysis within individual QG classes (QGs: 1 [middle], 1+, and 1++ [best]). AP was related (P < 0.001) to numbers of coarse MPs and fine MPs and fineness index in both QGs 1+ and 1++ but not in QG 1. Overall, several marbling texture traits were related to AP, but both MS and QG were more strongly related to AP.

Effects of dietary glycerol inclusion on growth performance, carcass and meat quality characteristics, glycogen content, and meat volatile compounds in Korean cattle steers.

OBJECTIVE: We have tested our hypothesis that inclusion of purified glycerol as a replacer of portions of dried distillers grain with solubles (DDGS) would affect growth performance, rumen fermentation and blood parameters, carcass and sensory traits, reducing sugar and glycogen contents, and volatile compound profiles in longissimus thoracis (LT) in Korean cattle steers. METHODS: A total of 20 Korean cattle steers (27.0±0.2 months old; 647±10.5 kg body weight [BW]) were assigned to a conventional control group or a glycerol group (3.17% purified glycerol addition as a replacement for DDGS and molasses). The steers were individually allowed to receive the experimental concentrate at the daily amount of 1.5% of their individual BW and a total 1.0 of kg/d of rice straw twice daily. The feeding trial was conducted for a period of 20 weeks. RESULTS: Glycerol supplementation (GS) increased (p = 0.001) concentrate intake. However, GS did not affect (p>0.05) average daily gain, feed efficiency, and ruminal volatile fatty acid concentrations. GS tended to increase (p≤0.10) serum glucose concentrations at the 16th and 20th weeks. GS decreased (p = 0.001) LT pH. GS did not affect (p>0.05) carcass traits and the chemical or physicochemical compositions, reducing sugar or glycogen contents, sensory traits, and most of volatile compounds in the LT. CONCLUSION: The inclusion of purified glycerol as a replacement for DDGS in the finishing diet did not affect growth performance, rumen fermentation parameters, and carcass quality in Korean cattle. The purified glycerol could be used as a substitute for other energy sources such as DDGS in beef cattle, depending on the price.

Inositol hexakisphosphate kinase-1 is a key mediator of prepulse inhibition and short-term fear memory.

Inositol phosphate metabolism has emerged as one of the key players in synaptic transmission. Previous studies have shown that the deletion of inositol hexakisphosphate kinase 1 (IP6K1), which is responsible for inositol pyrophosphate biosynthesis, alters probability of presynaptic vesicle release and short-term facilitation of glutamatergic synapses in mouse hippocampus. However, the behavioral and cognitive functions regulated by IP6K1 remain largely elusive. In this study, IP6K1-knockout mice exhibited decreased prepulse inhibition with no defects in Y-maze and elevated plus maze tests. Interestingly, IP6K1 knockout led to impaired short-term memory formation in a contextual fear memory retrieval test with no effect on long-term memory. Further, both hippocampal long-term potentiation and long-term depression in IP6K1-knockout mice were similar to those in the wild-type control. Taken together, the findings in this study suggest the physiological roles of IP6K1 and the associated inositol pyrophosphate metabolism in regulating sensorimotor gating as well as short-term memory.

Identification of the Antidepressant Vilazodone as an Inhibitor of Inositol Polyphosphate Multikinase by Structure-Based Drug Repositioning.

Inositol polyphosphate multikinase (IPMK) is required for the biosynthesis of inositol phosphates (IPs) through the phosphorylation of multiple IP metabolites such as IP3 and IP4. The biological significance of IPMK's catalytic actions to regulate cellular signaling events such as growth and metabolism has been studied extensively. However, pharmacological reagents that inhibit IPMK have not yet been identified. We employed a structure-based virtual screening of publicly available U.S. Food and Drug Administration-approved drugs and chemicals that identified the antidepressant, vilazodone, as an IPMK inhibitor. Docking simulations and pharmacophore analyses showed that vilazodone has a higher affinity for the ATP-binding catalytic region of IPMK than ATP and we validated that vilazodone inhibits IPMK's IP kinase activities in vitro . The incubation of vilazodone with NIH3T3-L1 fibroblasts reduced cellular levels of IP5 and other highly phosphorylated IPs without influencing IP4 levels. We further found decreased Akt phosphorylation in vilazodone-treated HCT116 cancer cells. These data clearly indicate selective cellular actions of vilazodone against IPMK-dependent catalytic steps in IP metabolism and Akt activation. Collectively, our data demonstrate vilazodone as a method to inhibit cellular IPMK, providing a valuable pharmacological agent to study and target the biological and pathological processes governed by IPMK.

Inositol Pyrophosphate Metabolism Regulates Presynaptic Vesicle Cycling at Central Synapses.

The coordination of synaptic vesicle exocytosis and endocytosis supports neurotransmitter release from presynaptic terminals. Although inositol pyrophosphates, such as 5-diphosphoinositol pentakisphosphate (5-IP7), are versatile signaling metabolites in many biological events, physiological actions of 5-IP7 on synaptic membrane vesicle trafficking remain unclear. Here, we investigated the role of 5-IP7 in synaptic transmission in hippocampal brain slices from inositol hexakisphosphate kinase 1 (Ip6k1)-knockout mice. We found that presynaptic release probability was significantly increased in Ip6k1-knockout neurons, implying enhanced activity-dependent synaptic vesicle exocytosis. Expression of wild-type but not catalytically inactive IP6K1 in the Ip6k1-knockout hippocampus restored the altered presynaptic release probability. Moreover, Ip6k1-knockout neurons were insensitive to folimycin, a vacuolar ATPase inhibitor, and dynasore, a dynamin inhibitor, suggesting marked impairment in synaptic endocytosis during exocytosis. Our findings collectively establish that IP6K1 and its product, 5-IP7, act as key physiological determinants for inhibition of presynaptic vesicle exocytosis and stimulation of endocytosis at central synapses.