A major flood caused by a typhoon did not affect the population genetic structure of a river mayfly metapopulation.

Floods affect the population structure of organisms that inhabit streams. In recent decades, the scale of floods has become larger due to climate change. Under these circumstances, on 12 October 2019, the largest typhoon in the history of observation in Japan struck the Japanese Archipelago. This typhoon caused heavy rainfall in various places, and the Chikuma-Shinano River System (Japan's largest) suffered great damage. Eight years before the large-scale disturbance in the river system, the population structure of the mayfly Isonychia japonica was studied in detail using quantitative sampling (population numbers and biomass) and by sequencing the mtDNA cytochrome c oxidase subunit I. To understand the impact of the flood on the population and genetic structures, we repeated the same research approximately 1 year after the flood. Direct comparison of sites before and after the flood revealed no significant changes between pre- and post-flood population genetic structure. This indicates high in situ resistance and/or resilience recovery of the populations to the disturbance. We hypothesize that this high resistance/resilience to flood disturbance is a result of strong selection for such traits in the rivers of the Japanese Archipelago, which are short, steep, flow rapidly and violently, and are strongly affected by floods.

Therapeutic effects of eperisone on pulmonary fibrosis via preferential suppression of fibroblast activity.

Although the exact pathogenesis of idiopathic pulmonary fibrosis (IPF) is still unknown, the transdifferentiation of fibroblasts into myofibroblasts and the production of extracellular matrix components such as collagen, triggered by alveolar epithelial cell injury, are important mechanisms of IPF development. In the lungs of IPF patients, apoptosis is less likely to be induced in fibroblasts than in alveolar epithelial cells, and this process is involved in the pathogenesis of IPF. We used a library containing approved drugs to screen for drugs that preferentially reduce cell viability in LL29 cells (lung fibroblasts from an IPF patient) compared with A549 cells (human alveolar epithelial cell line). After screening, we selected eperisone, a central muscle relaxant used in clinical practice. Eperisone showed little toxicity in A549 cells and preferentially reduced the percentage of viable LL29 cells, while pirfenidone and nintedanib did not have this effect. Eperisone also significantly inhibited transforming growth factor-ß1-dependent transdifferentiation of LL29 cells into myofibroblasts. In an in vivo study using ICR mice, eperisone inhibited bleomycin (BLM)-induced pulmonary fibrosis, respiratory dysfunction, and fibroblast activation. In contrast, pirfenidone and nintedanib were less effective than eperisone in inhibiting BLM-induced pulmonary fibrosis under this experimental condition. Finally, we showed that eperisone did not induce adverse effects in the liver and gastrointestinal tract in the BLM-induced pulmonary fibrosis model. Considering these results, we propose that eperisone may be safer and more therapeutically beneficial for IPF patients than current therapies.

Repeated restraint stress modifies fatty acid and amino acid metabolism in the mouse skin.

In modern society, stress caused by relationships and emotions is one of the greatest social problems. Similar to humans, domestic and captive animals live under various stresses. Several stresses have been associated with skin disorders, such as atopic dermatitis, but there is a lack of reliable and objective indicators for the characterization of this association. This study aimed to define the changes in fatty acid composition and amino acid concentration in the skin following repeated restraint stress in ICR mice. Mice subjected to 30 min of daily restraint stress for 8 days showed changes in the composition of saturated fatty acids, such as an increase in palmitic acid content, which are the substrates of Δ-9 desaturase. Conversely, unsaturated fatty acids decreased with stress treatment, which appeared to be a result of these fatty acids being the substrate of Δ-6 desaturase. Changes in fatty acid composition after stress treatment may be one of the factors that cause skin inflammation. The water-retention capacity may have been lowered by stress treatment because histidine and leucine, which are natural moisturizing factors, were significantly decreased. The collagen content in the skin gradually decreased after repeated stress treatment. Our results indicate that repeated restraint stress may impact skin health through changes in both the fatty acid composition and amino acid concentration in mice.

Produced ß-hydroxybutyrate after ß-hydroxy-ß-methylbutyrate (HMB) administration may contribute HMB function in mice.

ß-Hydroxy-ß-methylbutyrate (HMB) is an intermediate in the metabolism of the branched-chain amino acid leucine. HMB has several demonstrated effects on skeletal muscle function, some of which are contradictory. In addition, the effect of exogenous HMB intake on the levels of intermediate metabolites is not known. Therefore, we investigated changes in HMB metabolites after oral HMB administration in mice. First, ICR mice were treated with either distilled water or HMB (0.215 g/10 mL/kg). Sampling was performed at 0, 1, 6, 12, and 24 h after administration. Next, ICR mice were given distilled water or HMB (0.215 g/10 mL/kg/d) for 10 d. Mice given HMB shown a significant increase in liver ß-methylcrotonyl-CoA and increased ß-hydroxybutyrate in plasma and the gastrocnemius muscle 1 h after HMB administration. Mice administered HMB for 10 d showed significantly decreased food intake and body weight; however, the relative weight of the gastrocnemius muscle was significantly increased. These results may be attributed to an increase in ß-hydroxybutyrate resulting from exogenous HMB, since ß-hydroxybutyrate inhibits food intake and suppresses skeletal muscle catabolism. In conclusion, ß-hydroxybutyrate, a metabolite of HMB, was found to play an important role in the function of HMB.

Cortisol induces follicle regression, while FSH prevents cortisol-induced follicle regression in pigs.

During follicular development, a few dominant follicles develop to large antral dominant follicles, whereas the remaining follicles undergo atretic degeneration. Because vascularization on the follicular surface is a morphological feature of dominant follicles, we previously classified these follicles as vascularized follicles (VFs) and non-VFs (NVFs). In NVFs, progesterone producing genes were expressed similarly to that in VFs; however, the progesterone concentration in follicular fluid was low in large NVFs. Therefore, we estimated that progesterone is converted to cortisol, which induces the loss of follicular functions. In this study, we comparative analyzed the expression of genes for progesterone converting enzymes (Cytochrome (CYP)11B1, CYP21A2, Hydroxysteroid (HSD)11B2) and cortisol receptor (NR3C1) in VF and NVF granulosa cells. In NVFs, expression of cortisol producing genes (CYP11B1 and CYP21A2) was higher than in VFs. Expression of the gene for the cortisol metabolizing enzyme HSD11B2 in NVFs was significantly lower than in VFs. In NVFs, accompanied by increasing cortisol concentration in follicular fluid, apoptosis of granulosa and cumulus cells was observed. Cultivation with FSH and metyrapone (a CYP11B1 inhibitor) of NVF cumulus-oocyte complexes inhibited apoptosis of cumulus cells and induced cumulus cell proliferation and oocyte maturation. Cortisol-induced CYP11B1 and CYP21A2 expression, whereas FSH-induced HSD11B2 mRNA expression in VF granulosa cells in the presence of cortisol. Furthermore, an addition of 18ß-glycyrrhetinic acid (18-GA; a HSD17B2 inhibitor) to cortisol and FSH-containing medium increased apoptosis of VF granulosa cells. These results suggested that cortisol is a stimulatory factor that induces follicular atresia; furthermore, inhibition of cortisol production by FSH might increase the number of healthy preovulatory follicles in pigs.

Serum Progesterone Levels with The Use of Four Different Types of Vaginal Progesterone in Frozen-Thawed Embryo Transfer Cycles and Related Pregnancy Outcomes.

BACKGROUND: Luteal phase support (LPS) is essential for hormone replacement therapy (HRT) for frozen-thawed embryo transfer (FET). However, the optimal dose and serum progesterone (P4) levels required for pregnancy are controversial. We attempted to determine the association between pregnancy outcomes and serum P4 levels administered via vaginal suppository for HRT-FET cycles on embryo transfer day. MATERIALS AND METHODS: This was a secondary analysis of the dataset from the EXCULL trial, which prospectively investigated pregnancy outcomes of four different P4 vaginal suppositories (Lutinus, Utrogestan, Luteum, and Crinone) for HRT-FET. It was conducted at a private fertility clinic between December 2016 to December 2017. During this trial, 235 cycles were divided into four groups based on serum P4 values (quartile [Q] 1 group: <7.8 ng/mL; Q2 group: 7.8-10.8 ng/mL; Q3 group: 10.8-13.7 ng/mL; Q4 group: >13.7 ng/mL). We investigated clinical pregnancy rate (CPR), positive fetal heart rate (FHR), live birth rate (LBR), and miscarriage rate (MR) for each group. A logistic regression analysis was performed using age, body mass index (BMI), and transferred embryos as covariates. RESULTS: Serum P4 values (ng/mL) of each drug were as follows: Lutinus, 13.3 ± 4.9; Utrogestan, 9.3 ± 3.3; Luteum, 13.6 ± 4.2; and Crinone, 8.7 ± 3.2 (mean ± standard deviation, P<0.001).The percentages of Utrogestan and Crinone were higher in the Q1 group, while the percentages of Lutinus and Luteum were higher in the Q4 group. Nonetheless, there were no statistical differences between the Q1 and Q4 groups in CPR, FHR, LBR, and MR. CONCLUSION: When vaginal P4 was used for FET, although serum P4 levels on transfer day differed based on the drug that was administered, no relationship was observed between serum progesteronelevels and pregnancy outcomes (Registration number: UMIN000032997).

Central pontine myelinolysis during treatment of hyperglycemic hyperosmolar syndrome: a case report.

BACKGROUND: Central pontine myelinolysis (CPM) is a non-inflammatory demyelinating lesion of the pons. CPM and extrapontine demyelination (EPM) are together termed osmotic demyelination syndrome (ODS), a known and serious complication of acute correction of hyponatremia. Conversely, hyperglycemic hyperosmolarity syndrome (HHS) develops in patients with type 2 diabetes who still have some insulin secretory ability due to infection, non-compliance with treatment, drugs, and coexisting diseases, and is often accompanied by ketosis. HHS represents a life-threatening endocrine emergency (mortality rate, 10-50%) associated with marked hyperglycemia and severe dehydration. HHS may develop ODS, and some cases have been associated with hypernatremia. CASE PRESENTATION: The patient was an 87-year-old woman with hyperglycemia, dehydration, malnutrition, and potential thrombus formation during long-term bed rest. HHS was suspected to have developed due to progression of hyperglycemia and dehydration caused by pneumonia. Furthermore, ketoacidosis developed from ketosis and prerenal renal failure associated with circulating hypovolemia shock, which was also associated with disseminated intravascular coagulation. Treatment was started with continuous intravenous injection of fast-acting insulin and low-sodium replacement fluid. In addition, ceftriaxone sodium hydrate, heparin sodium, thrombomodulin α, human serum albumin, and dopamine hydrochloride were administered. Blood glucose, serum sodium, serum osmolality, and general condition (including vital, infection/inflammatory findings, and disseminated intravascular coagulation) improved promptly, but improvements in disturbance of consciousness were poor. Diffusion-weighted imaging of the brain 72 h after starting treatment showed no obvious abnormalities, but high-intensity signals in the midline of the pons became apparent 30 days later, leading to definitive diagnosis of CPM. CONCLUSIONS: Fluctuation of osmotic pressure by treatment from hyperosmolarity due to hyperglycemia and hypernatremia in the presence of risk factors such as malnutrition, severe illness, and metabolic disorders may be a cause of CPM onset. When treating HHS with risk factors, the possibility of progression to ODS needs to be kept in mind.

The Association Between Alcohol-Flavoured Non-Alcoholic Beverages and Alcohol Use in Japanese Adolescents.

AIMS: There are no legal regulations in Japan governing minors' consumption of alcohol-flavoured non-alcoholic beverages (AFNAB); therefore, we examined if their consumption could lead to increased alcohol use among adolescents in Japan. METHODS: This cross-sectional study used a nonclinical, nationally representative sample of 38,494 junior (19,662 boys) and 61,556 senior (31,925 boys) high school students recruited in 2012. We measured AFNAB consumption rates and the order that adolescents first consumed AFNAB and alcohol. RESULTS: The AFNAB consumption was strongly associated with alcohol use in high school students. Among all age groups, alcohol was more commonly consumed before AFNAB for both males and females. CONCLUSIONS: Consumption of AFNAB is more prevalent among minors than alcohol consumption and it has a strong association with alcohol consumption. However, concerns that AFNAB use would lead to increased alcohol use were not supported because AFNAB consumption usually started after adolescents began consuming alcohol. SHORT SUMMARY: Consumption of AFNAB is more prevalent among high school students than alcohol consumption and it has a strong association with alcohol consumption. However, concerns that AFNAB use would lead to increased alcohol use were not supported because AFNAB consumption usually started after adolescents began consuming alcohol.

The Novel Pig In Vitro Maturation System to Improve Developmental Competence of Oocytes Derived from Atretic Non-Vascularized Follicle.

The in vitro maturation (IVM) technique is beneficial for producing animal offspring, but the blastocyst rate is low after IVM. In this technique, cumulus-oocyte complexes (COCs) are collected from medium size follicles. The follicles are ultimately selected as large dominant follicles or atretic follicles; therefore it is possible that the COCs collected using IVM are contaminated by follicles that will develop into large follicles and induce atresia. In the dominant follicles, estradiol-17beta and progesterone induce the differentiation of follicular somatic cells which exhibit the ability to respond to ovulation during follicular development. Thus, we hypothesized that changes in the hormonal condition of healthy follicles are essential for oocyte maturation during IVM. In this study, we performed a comparative analysis of the steroid hormone concentrations in non-vascularized follicles (NVFs) and vascularized follicles (VFs). The estradiol-17beta concentration increased in medium VFs, whereas the level was low in NVFs of the same size. The progesterone concentration increased with large follicular size in VFs, but the level remained low in follicles of any size among NVFs. To improve the oocyte quality derived from NVFs, NVF COCs were cultured with FSH alone or FSH under theVF hormonal conditions. Cultivation under the VF hormonal conditions dramatically improved the proliferation and survival of cumulus cells, meiotic maturation of oocytes, cumulus expansion, and blastocyst rate following in vitro fertilization. Thus, the cultivation of NVF COCs under VF hormonal conditions improves the developmental potential to the blastocyst stage by NVF oocytes.

Anger and Impulsivity Among Japanese Adolescents: A Nationwide Representative Survey.

OBJECTIVE: This study aimed to clarify the prevalence of anger and impulsivity and its associated factors through a nationwide survey of junior and senior high school adolescent students in Japan. METHODS: A self-administered questionnaire covering (1) personal data, (2) lifestyle, (3) mental health status, and (4) feelings of anger and impulsivity was distributed to junior and senior high school students in Japan. Among the total of 10,955 junior high schools and 5,115 senior high schools nationwide, 130 and 110 were randomly selected, respectively. Of those, 92 junior and 80 senior high schools participated in the survey. The survey period was from December 2008 to the end of January 2009. A total of 95,680 questionnaires were collected. After excluding invalid responses, the remaining 94,777 responses (response rate: 62.3%) were analyzed. RESULTS: From the questions regarding anger and impulsivity, 8.7% (95% CI, 8.5%-8.9%) and 7.5% (95% CI, 7.3%-7.7%) of the participants were considered to have experienced intense anger and impulsivity, respectively. Logistic regression analysis indicated that the odds ratios for experiencing intense feelings of anger were significantly higher (all P values < .05) among students who smoked, consumed alcohol, skipped breakfast, did not wish to go to university, had short sleep duration, had decreased positive feelings, had increased depressive feelings, or used mobile phones for longer hours. The odds ratios for experiencing intense impulsivity were significantly higher among students who smoked, consumed alcohol, skipped breakfast, did not participate in club activities, had short sleep duration, had decreased positive feelings, had increased depressive feelings, or used mobile phones for longer hours. CONCLUSIONS: The results suggest that healthy lifestyle habits, good sleep habits, and improved mental health are important for preventing intense feelings of anger and impulsivity among adolescents.

Internet use and problematic Internet use among adolescents in Japan: A nationwide representative survey.

INTRODUCTION: Japan is assumed to have serious health and social problems due to Internet overuse, but little is known about the actual conditions. This study was conducted to investigate the prevalence of problematic Internet use (PIU) and associated Internet use in adolescents in Japan. METHODS: A nationwide survey of adolescent Internet use was conducted in 2012 and 2013. The participants were 100,050 students from randomly selected junior and senior high schools nationwide who gave valid responses to a self-reported questionnaire. The questionnaire included questions on Internet use and the Japanese version of the Young's Diagnostic Questionnaire (YDQ) as well as other questions related to lifestyle habits. Internet users were classified by gender according to three categories on the basis of their YDQ scores: adaptive use, maladaptive use, and PIU. RESULTS: The estimated prevalence of PIU was 6.2% in males, 9.8% in females, and 7.9% in total; it closely correlated with female gender, school grades, and number of Internet hours. The following common and gender-specific applications that conferred a risk of PIU were identified: downloading (both genders), online gaming (males), social networking services, blogs, and bulletin boards (females). CONCLUSIONS: A cross-sectional survey using YDQ of a large number of high school students yielded a PIU prevalence of 7.9% in Japan. This study showed that problems associated with Internet overuse have already become serious; therefore, planning and implementation of prevention and control measures is urgently required.

The oncogenic role of GASC1 in chemically induced mouse skin cancer.

Gene amplified in squamous cell carcinoma (SCC) 1 (GASC1), also known as KDM4C/JMJD2C, encodes a histone demethylase that specifically demethylates lysine residues (H3K9, H3K36, and H1.4K26) and plays a crucial role in the regulation of gene expression as well as in heterochromatin formation. GASC1 is located at human chromosome 9p23-24, where frequent genomic amplification is observed in human esophageal cancer, and its aberrant expression is detected in a variety of human cancers, such as breast, colon, and prostate. Therefore, it is highly likely that GASC1 contributes to the genesis and/or development of cancer. However, there is a lack of direct evidence of GASC1 having an oncogenic function. In this study, we aimed to clarify the role of GASC1 in the skin SCC carcinogenesis. For this purpose, we generated Gasc1-heterozygous mice (Gasc1+/-) with reduced expression of Gasc1. On the basis of our results, Gasc1+/- mice displayed a significantly lower incidence and multiplicity of both benign and malignant tumors induced by the two-stage skin carcinogenesis protocol than wild-type mice. In addition, the volume of carcinoma was significantly lower in Gasc1+/- mice. Consistent with these observations, knocking down of Gasc1 resulted in reduced cell viability of SCC cells in vitro. Our findings clearly demonstrated that GASC1 has an oncogenic role in skin carcinogenesis.

Development of Augmentative and Alternative Communication Assessment Tools for Patients with ALS.

ALS patients usually use augmentative and alternative communication tools to communicate with other people, but the assessment tools, including the selection of an input switch, are very difficult to operate. In this study, we developed a novel device to measure the physical ability of patients to operate the input switch with a push lever. The study focused on the amount of pushing and the power required to operate the input switch, and the effectiveness was verified.

Irritable bowel syndrome among Japanese adolescents: A nationally representative survey.

BACKGROUND AND AIM: No nationally representative survey of irritable bowel syndrome (IBS) among adolescents has ever been performed in Japan. In the present study, we aimed to clarify the prevalence of IBS among Japanese adolescents and the factors associated with it. METHODS: The items related to the diagnostic criteria for IBS based on the Rome III Diagnostic Criteria for Functional Gastrointestinal Disorders were included in a cross-sectional nationwide survey of "alcohol consumption and smoking habits among junior and senior high school students." The participating schools were sampled from among all junior and senior high schools in Japan using the cluster-sampling method, and self-administered questionnaires were sent to the selected schools by mail. Among 99 416 questionnaires that were collected, data from 98 411 valid responses were analyzed. RESULTS: The results showed that the prevalence of IBS was 18.6%. Although no sex difference was observed in the overall prevalence of IBS, the prevalence of diarrhea-predominant IBS was higher among boys than among girls, and the prevalence of constipation-predominant IBS was higher among girls than among boys. The prevalence of IBS increased with progression of the school grade, and there were the significant relationships between IBS and sleep-phase delay and insomnia symptoms. IBS was also significantly associated with poor mental health status. CONCLUSION: These results indicate that IBS is common among junior and senior high school students, and associated with lifestyle and mental health.

Identification of a novel E-box binding pyrrole-imidazole polyamide inhibiting MYC-driven cell proliferation.

The MYC transcription factor plays a crucial role in the regulation of cell cycle progression, apoptosis, angiogenesis, and cellular transformation. Due to its oncogenic activities and overexpression in a majority of human cancers, it is an interesting target for novel drug therapies. MYC binding to the E-box (5'-CACGTGT-3') sequence at gene promoters contributes to more than 4000 MYC-dependent transcripts. Owing to its importance in MYC regulation, we designed a novel sequence-specific DNA-binding pyrrole-imidazole (PI) polyamide, Myc-5, that recognizes the E-box consensus sequence. Bioinformatics analysis revealed that the Myc-5 binding sequence appeared in 5'- MYC binding E-box sequences at the eIF4G1, CCND1, and CDK4 gene promoters. Furthermore, ChIP coupled with detection by quantitative PCR indicated that Myc-5 has the ability to inhibit MYC binding at the target gene promoters and thus cause downregulation at the mRNA level and protein expression of its target genes in human Burkitt's lymphoma model cell line, P493.6, carrying an inducible MYC repression system and the K562 (human chronic myelogenous leukemia) cell line. Single i.v. injection of Myc-5 at 7.5 mg/kg dose caused significant tumor growth inhibition in a MYC-dependent tumor xenograft model without evidence of toxicity. We report here a compelling rationale for the identification of a PI polyamide that inhibits a part of E-box-mediated MYC downstream gene expression and is a model for showing that phenotype-associated MYC downstream gene targets consequently inhibit MYC-dependent tumor growth.

Factors affecting unhappiness at school among Japanese adolescents: an epidemiological study.

BACKGROUND: Unhappiness at school is one of the main reasons for truancy among adolescents. In order to assess this problem more thoroughly in the context of Japanese adolescents, the present study examined the associations between feelings of unhappiness at school and lifestyle habits, school life realities, and mental health status. METHOD: This study was designed as a cross-sectional survey. A self-administered questionnaire was provided to students enrolled in randomly selected junior and senior high schools throughout Japan. We calculated the percentages of both junior and senior high school students who felt unhappy at school based on factors related to school life, lifestyle habits, and mental health status. Multiple logistic regression analyses were performed in order to examine the associations between those factors and students' feelings of unhappiness at school. RESULTS: A total of 98,867 valid responses were analysed, 7.9% (Boys: 8.4%, Girls: 7.4%) of which came from students who responded that they felt unhappy at school. For both junior and senior high school students, the percentages of those who felt unhappy at school were significantly higher among those who had not yet decided on their future life course, who did not participate in extracurricular activities, did not eat breakfast every day, went to bed late, had used tobacco or alcohol in the previous 30 days, and had poor mental health compared with others. The results of multiple logistic regression analyses indicated that the adjusted odds ratios for feeling unhappy at school with regard to the above-mentioned factors were significantly high for both junior and senior high school students. CONCLUSIONS: The present results suggest that school employees and administrators must provide health guidance to students, considering that irregular lifestyle habits, lower school engagement, smoking, drinking alcohol, and poor mental health status are all associated with maladaptation to school among adolescents.

[The newest epidemiology trend of insomnia].

Sleep disturbance such as insomnia is one of common complaint among adults in developed countries. Insomnia induces sleepiness and drowsiness, resulting in a reduction of working efficiency in the daytime. Drivers in the transportation system and machine operators could cause serious industrial accidents if they have sleep disturbances. Recent studies reported that approximately 20% of Japanese population had sleep disturbances. Here we provide a general account of sleep disturbance about Japanese population.

Protein kinase C (PKC) increases TACE/ADAM17 enzyme activity in porcine ovarian somatic cells, which is essential for granulosa cell luteinization and oocyte maturation.

During in vitro maturation of porcine cumulus cell-oocyte complexes and in vitro luteinization of porcine granulosa cells, FSH induces the expression of the protease TNFα-converting enzyme/A disintegrin and metalloproteinase domain 17 (TACE/ADAM17) and the epidermal growth factor (EGF)-like factors, which activate the EGF receptor (EGFR)-MAPK3/1 pathway in both cumulus and granulosa cells. FSH is known to activate not only protein kinase A and p38MAPK pathways in both cell types but also activates protein kinase C (PKC). Because PKC-induced association of cellular-Sarcoma (c-Src) and TACE/ADAM17 is required for TACE/ADAM17 enzyme activation in some cancer cells, we hypothesized that PKC and c-Src impact TACE/ADAM17-mediated activation of EGFR signaling pathway in porcine granulosa and cumulus cells. When granulosa cells or cumulus cell-oocyte complexes were cultured with FSH, PKC activity and c-Src phosphorylation increased and were associated with increased TACE/ADAM17 enzyme activity. The PKC inhibitor calphostin C (CalC) and the c-Src inhibitor (4 amino 5 (4 chlorophenyl) 7 (t butyl)pyrazolo[3,4 d]pyrimidine [PP2]) suppressed TACE/ADAM17 enzyme activity, whereas these inhibitors did not affect Tace/Adam17 mRNA expression. Immunoprecipitation analysis showed that FSH mediated the association of c-Src with TACE/ADAM17 via a PKC-dependent mechanism. Either CalC or PP2 suppressed EGFR downstream signaling pathway (MAPK3/1) in these ovarian cell types and reduced cumulus expansion, meiotic maturation of oocytes, and progesterone production. The negative effects were overcome by the addition of amphiregulin. Collectively, these results indicate that activation of TACE/ADAM17 via a PKC-induced c-Src-dependent manner mediates proteolytic activation of the EGF-like factors that are involved in the induction of granulosa cell differentiation, cumulus expansion, and meiotic maturation of porcine oocytes in vitro.