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Miho Matsuda Last modified date:2018.07.04

Associate Professor / Division of Oral Biological Sciences
Department of Dental Science
Faculty of Dental Science


Graduate School


E-Mail
Homepage
http://www.dent.kyushu-u.ac.jp/sosiki/a04/index.html
Phone
092-642-6321
Fax
092-642-6322
Academic Degree
PhD
Country of degree conferring institution (Overseas)
No
Field of Specialization
molecular biology biochemistry cell biology
Total Priod of education and research career in the foreign country
03years00months
Research
Research Interests
  • Function of a novel inositol 1,4,5-trisphosphate binding protein, PRIP, in the regulation of reproductive system
    keyword : hormone, secretion, reproductive system, gonadotropin
    2006.05.
  • Study on the function of PRIP in the regulation of bone metabolism
    keyword : bone metabolism, signal transduction, PRIP, hormone, secretion
    2008.04.
Current and Past Project
  • Study on the involvement of a novel signaling molecule, PRIP in the regulation of bone metabolism.
  • Study on Ca2+ dependent regulation of gene expressions.
  • Study on brain specific regulation of PRIP-1 gene expression.
Academic Activities
Papers
1. Miho Matsuda, Masato Hirata, Phospholipase C-related but catalytically inactive proteins regulate ovarian follicle development, ASBMB/ Journal of Biological Chemistry, 292, 20, 8369-8380, 2017.04, Phospholipase C-related but catalytically inactive proteins
PRIP-1 and -2 are inositol-1,4,5-trisphosphate binding proteins
that are encoded by independent genes. Ablation of the PRIP
genes in mice impairs female fertility, which is manifested
by fewer pregnancies, a decreased number of pups, and the
decreased and increased secretion of gonadal steroids and
gonadotropins, respectively. We investigated the involvement
of the PRIPs in fertility, focusing on the ovaries of Prip-1 and -2
double-knock-out (DKO) mice. Multiple cystic follicles were
observed in DKO ovaries, and a superovulation assay showed a
markedly decreased number of ovulated oocytes. Cumulusoocyte
complexes showed normal expansion, and artificial
gonadotropin stimulation regulated the ovulation-related genes
in a normal fashion, suggesting that the ovulation itself was
probably normal.Ahistological analysis showed atresia in fewer
follicles of the DKO ovaries, particularly in the secondary follicle
stages. The expression of luteinizing hormone receptor
(LHR) was aberrantly higher in developing follicles, and the
phosphorylation of extracellular signal-regulated protein kinase,
a downstream target of LH-LHR signaling, was higher in DKO
granulosa cells. This suggests that the up-regulation of LH-LHR
signaling is the cause of impaired follicle development. The
serum estradiol level was lower but estradiol production was
unchanged in theDKOovaries. These results suggest that PRIPs
are positively involved in the development of follicles via their
regulation of LH-LHR signaling and estradiol secretion. Female
DKO mice had higher serum levels of insulin, testosterone, and
uncarboxylated osteocalcin, which together with reduced fertility,
are reminiscent of polycystic ovary syndrome in humans..
2. Ayako Murakami, Miho Matsuda, Masato Hirata, Phospholipase C-related, but catalytically inactive protein (PRIP) up-regulates osteoclast differentiation via calcium-calcineurin-NFATc1 signaling, Journal of Biological Chemistry, 292, 7994-8006, 2017.03, Phospholipase C-related, but catalytically inactive protein
(PRIP) was previously identified as a novel inositol 1,4,5-trisphosphate-
binding protein with a domain organization similar
to that of phospholipase C- but lacking phospholipase activity.
We recently showed that PRIP gene knock-out (KO) in mice
increases bone formation and concomitantly decreases bone
resorption, resulting in increased bone mineral density and trabecularbonevolume.
However, the role ofPRIPin osteoclastogenesis
has not yet been fully elucidated. Here, we investigated the effects
of PRIP on bone remodeling by investigating dynamic tooth movement
in mice fitted with orthodontic devices. Morphological analysis
indicated that the extent of tooth movement was smaller in
the PRIP-KO mice than in wild-type mice. Histological analysis
revealed fewer osteoclasts on the bone-resorption side in maxillary
bones of PRIP-KO mice, and osteoclast formation assays and flow
cytometry indicated lower osteoclast differentiation in bone marrow
cells isolated from these mice. The expression of genes implicated
inboneresorptionwaslower in differentiatedPRIP-KOcells,
and genes involved in osteoclast differentiation, such as the transcription
factor NFATc1, exhibited lower expression in immature
PRIP-KO cells initiated by M-CSF. Moreover, calcineurin expression
and activity were also lower in the PRIP-KO cells. The
PRIP-KO cells also displayed fewer M-CSF-induced changes in
intracellular Ca2 and exhibited reduced nuclear localization of
NFATc1. Up-regulation of intracellularCa2restored osteoclastogenesis
of the PRIP-KO cells. These results indicate that PRIP deficiency
impairs osteoclast differentiation, particularly at the early
stages, and that PRIP stimulates osteoclast differentiation through
calcium-calcineurin-NFATc1 signaling via regulating intracellular
Ca2..
3. Matsuda, M, Tsutsumi, K., Kanematsu, T., Fukami, K., Terada, Y., Takenawa, T., Nakayama, K.I., Hirata, M., Involvement of phospholipase C-related inactive protein in the mouse reproductive system through the regulation of gonadotropin levels. , Biology of reproduction, 81, 681, 2009.07.
Presentations
1. Miho Matsuda, Ayako Murakami, Masato Hirata, Involvement of phospholipase C-related but catalytically inactive protein in the early stage of osteoclast differentiation, cell biology 2016 ASCB Annual Meeting, 2016.12, Phospholipase C-related but catalytically inactive protein (PRIP) was first identified as an inositol 1,4,5-trisphosphate binding protein. We generated PRIP gene-deficient (PRIP-KO) mice and reported that PRIP-KO mice exhibited the upregulation of the bone mineral density and trabecular bone volume, due to the enhancement of bone formation while decreased bone resorption. In this study, we investigated the possible mechanisms by which PRIP regulates bone resorption. Orthodontic tooth movement was performed using wild type (WT) and PRIP-KO mice equipped with orthodontic devices and analyzed by micro-computed tomography and TRAP staining. In the mutant mice, tooth movement was reduced compared with wild type (WT) mice, and osteoclasts were observed at the lesser amount on pressure side in KO maxillary bones. In vitro osteoclast formation assay and flow cytometric analysis were also performed and revealed the decreased differentiation for osteoclast in bone marrow cells isolated from KO mice. The expression of genes implicated in osteoclast differentiation and bone resorption was decreased in KO mice. Immunoblot analysis using cultured bone marrow cells indicated the lower expression of calcineurin in KO cells, and the phosphatase activity by calcineurin was decreased in KO cells. The expression of nuclear factor of activated T cells, cytoplasmic 1 (Nfatc1), which is an essential factor for osteoclast differentiation and the activity is regulated by calcineurin, was also decreased in KO cells in the early stage of osteoclast differentiation. These results suggest that PRIP positively regulates osteoclast differentiation, especially in the early stage, probably through calcium-calcineurin-NFAT signaling. .
2. Phospholipase C-related catalytically inactive protein (comprising PRIP-1 and -2) was first identified as a novel inositol 1,4,5-trisphosphate binding protein, but the biological functions have remained elusive. We therefore generated PRIP-1 and -2 double knockout (DKO) mice and found their reduced fertility: DKO mice exhibited reduced fertility in the female such as decreased number of pups, longer estrous days, increased secretion of gonadotropins, etc. This time we examined the ability of the ovaries in response to gonadotropins: the total number of the ovulated oocytes were remarkably decreased in DKO female mice, indicating that PRIP plays an important role in follicle maturation and/or ovulation, but not fertilization or implantation. Microarray and quantitative real-time PCR analyses revealed the increased expressions of molecules involved in ovulation including Has2, Ptgs2, PTX3, Tnfaip6 and Areg in both genotype. We examined the ability of COC expansion in both genotypes but little difference was observed. On the other hand, histological analysis revealed more immature follicles and fewer mature follicles or corpus lutea in DKO ovaries. Immunological analysis showed the expression of luteinizing hormone receptors (LHR) at earlier stages of follicle maturation in DKO ovary, and elevated level of ERK phosphorylation in DKO granulosa cells. These results suggest that the appearance of LHR at immature follicle stages leads to suppress follicle maturation by excessive LH signaling in DKO ovary, indicating that PRIP may be involved in positive regulation of ovarian follicle maturation through LH signaling. .
3. Phospholipase C-related catalytically inactive protein (comprising PRIP-1 and -2) was first identified as a novel inositol 1,4,5-trisphosphate binding protein, but the biological functions have remained elusive. We therefore generated PRIP-1 and –2 double knockout (DKO) mice to gain insight into the biological function. DKO mice exhibited reduced fertility in the female such as decreased number of pups, longer estrous days, increased secretion of gonadotropins, etc. We examined the ability of the ovaries in response to gonadotropins: the total number of the ovulated oocytes were remarkably decreased in DKO female mice, indicating that PRIP plays an important role in follicle maturation and/or ovulation, but not fertilization or implantation. Microarray and quantitative real-time PCR analyses revealed the increased expressions of molecules involved in ovulation including Has2, Ptgs2, PTX3, Tnfaip6 and Areg. We examined the ability of COC expansion in both genotypes but little difference was observed. On the other hand, histological analysis of ovaries from both genotypes revealed more immature follicles and fewer mature follicles or corpus lutea in DKO ovaries. Immunofluorescence analysis showed the expression of luteinizing hormone receptors (LHR) at earlier stages of follicle maturation in DKO ovary and more increase of antral follicles. These results are summarized as below: the increased LH secretion from pituitary gland and the appearance of LHR at immature follicle stages lead excessive LH signaling to suppress follicle maturation in DKO females, indicating that PRIP may be involved in positive regulation of ovarian follicle maturation through LH signaling. .
4. Miho Matsuda, Masato Hirata, Involvement of a novel signaling molecule, PRIP in the regulation of reproduction system
, The 22th IUBMB, 37st FEBS Congress, 2012.09, PRIP (PLC-Related, but catalytically Inactive Protein) は、Ins(1,4,5)P3 結合性の分子として見いだされた新規分子であり、種を越えて広く存在するタンパク質である。2つのサブタイプが存在し、PRIP-1は脳に多く発現しているのに対し、PRIP-2は比較的ユビキタスな発現パターンを示す。我々は、PRIP-1及びPRIP-2のダブルノックアウトマウスを作製し、本変異マウスにおいて総出産仔数の減少、血中性腺刺激ホルモン量の増加などの表現型を見いだし、性周期制御にPRIPが強く関与することを明らかにした。排卵数を調べるため、PMSG(妊馬血清性ゴナドトロピン)、HCG(人絨毛生性腺刺激ホルモン)を用いた排卵誘発を行ったところ、変異型で激減していた。このことから、出産仔数の減少は、排卵後の授精や着床における異常によるものではなく、卵成熟を含めた排卵に至るまでの過程における異常によるものであることが示唆された。そこで、まず排卵前後における遺伝子発現の変化を検討するために、卵巣RNAを用いたマイクロアレイ解析を行った。その結果、PMSG投与後48時間でHCGを投与した卵巣において、排卵前の卵胞で発現するHas2 (hyaluronan synthase 2)、Ptgs2 (cyclooxygenase 2)、PTX3 (pentraxin 3)、Tnfαip6 (TNFα-induced protein 6)などの発現量が、変異型で増加していた。また、同時期の卵巣の組織学的解析より、変異型において卵母細胞が残ったまま黄体化したような所見があり、これらのことから排卵時の制御におけるPRIPの関与が示唆された。そこで現在、野生型及び変異型マウスにおいて、排卵直前に起こる急激な変化(卵丘細胞-卵母細胞複合体のexpansionやヒアルロン酸の発現量及び局在等)について解析を進めている。.
5. Phospholipase C-related but catalytically inactive protein (comprising PRIP-1 and -2) was first identified as a novel D-myo-inositol 1,4,5-trisphosphate [Ins(1,4,5)P3] binding protein, but the biological functions have remained elusive. We therefore generated PRIP-1 and –2 double knockout (DKO) mice to gain insight into the biological function. DKO mice showed dysfunction of the reproductive system in the female. We examined the ability of the ovaries in response to gonadotropins: the total number of the ovulated oocytes were significantly decreased in mutant female mice, compared to that seen with wild-type, which was also confirmed by histological analysis of the ovaries after ovulation. These results suggested that PRIP plays an important role in not fertilization or implantation but follicle maturation and/or ovulation. Microarray analysis indicated the increased expressions of ovulation related molecules such as Has2, Ptgs2, PTX3, Tnfαip6 and Ereg. Historogical analysis of ovaries, just before ovulation, showed some abnormal follicles in the DKO. We also examined the ability of COC expansion in both genotypes and the significantly difference was not observed. On the other hand, TUNEL assay was performed to detect apoptotic follicles related to follicle maturation. There were some differences between in WT and DKO ovaries, depending on the follicle stages. Further analyses are underway to reveal the role of PRIP on follicle maturation and ovulation. .
6. Phospholipase C-related but catalytically inactive protein (comprising PRIP-1 and -2) was first identified as a novel D-myo-inositol 1,4,5-trisphosphate [Ins(1,4,5)P3] binding protein, but the biological functions have remained elusive. We therefore generated PRIP-1 and –2 double knockout (DKO) mice to gain insight into the biological function. DKO mice apparently grew normally and became fertile; however, during animal maintenance, we noticed that mutant couples exhibited decreased litter events and litter size, indicating dysfunction of the reproductive system. Cross-mating experiments indicated that the cause appeared to be on the female side. The observation of the estrous cycle in mice by histological analysis of vaginal smears showed that the estrous days were apparently increased in DKO mice. Levels of serum LH and FSH were measured for 5-6 consecutive days, and were significantly higher in the mutant, which was also confirmed by examining the secretion of LH from the explant culture of anterior pituitary glands of wild-type and DKO mice. We also examined the ability of the ovaries in response to gonadotropins: the total number of the ovulated oocytes were significantly decreased in mutant female mice, compared to that seen with wild-type, which was also confirmed by histological analysis of the ovaries after ovulation. These results suggest that PRIP plays an important role in female reproductive system, especially in gonadotropin secretion, ovarian follicle maturation and ovulation..
7. Phospholipase C-related but catalytically inactive protein (comprising PRIP-1 and -2) was first identified as a novel D-myo-inositol 1,4,5-trisphosphate [Ins(1,4,5)P3] binding protein, but the biological functions have remained elusive. We therefore generated PRIP-1 and –2 double knockout (DKO) mice to gain insight into the biological function. DKO mice apparently grew normally and became fertile; however, during animal maintenance, we noticed that mutant couples exhibited decreased litter events and litter size, indicating dysfunction of the reproductive system. Cross-mating experiments indicated that the cause appeared to be on the female side. The observation of the estrous cycle in mice by histological analysis of vaginal smears showed that the estrous days were apparently increased in DKO mice. Levels of serum LH and FSH were measured for 5-6 consecutive days, and were significantly higher in the mutant, which was also confirmed by examining the secretion of LH from the explant culture of anterior pituitary glands of wild-type and DKO mice. We also examined the ability of the ovaries in response to gonadotropins: the total number of the ovulated oocytes were significantly decreased in mutant female mice, compared to that seen with wild-type, which was also confirmed by histological analysis of the ovaries after ovulation. The analysis also showed that the numbers of follicles originally were not significantly different between WT and DKO ovaries. These results suggest that PRIP plays an important role in female reproductive system, especially in gonadotropin secretion and ovarian follicle maturation..
8. PRIP (phospholipase C related, but catalytically inactive protein) was identified as a novel inositol 1,4,5-trisphosphate binding protein, whose domain organization is similar to that of phospholipase C-_1 but is catalytically inactive, comprizing two isoforms, PRIP-1 and PRIP-2. To get insight into the biological functions, we generated the double knockout (DKO) mice of both PRIP-1 and PRIPミ2, followed by the phenotypic analyses. DKO mice grew normally and became fertile. However DKO female exhibited the decreased litter occasion and litter size, indicating the dysfunction of female reproductive system. Then we examined the estrus cycle by cytological analysis of daily vaginal smears, resulting in the irregular cycle in DKO mice. We further noticed that the mutant mice had apparently smaller sized uterus by gross anatomical observation. Basal levels of serum leuteinizing hormone and follicle stimulating hormone were significantly higher in the mutant, the event of which was also confirmed by examining the secretion from the tissue culture of anterior pituitary glands. These results suggest that PRIP is involved in maternal reproduction, through the gonadotropine secretion..
9. PRIP (phospholipase C related, but catalytically inactive protein) was identified as a novel inositol 1,4,5-trisphosphate binding protein, whose domain organization is similar to that of phospholipase C-_1 but is catalytically inactive, comprizing two isoforms, PRIP-1 and PRIP-2. To get insight into the biological functions, we generated the double knockout (DKO) mice of both PRIP-1 and PRIPミ2, followed by the phenotypic analyses. DKO mice grew normally and became fertile. However DKO female exhibited the decreased litter occasion and litter size, indicating the dysfunction of female reproductive system. Then we examined the estrus cycle by cytological analysis of daily vaginal smears, resulting in the irregular cycle in DKO mice. We further noticed that the mutant mice had apparently smaller sized uterus by gross anatomical observation. Basal levels of serum leuteinizing hormone and follicle stimulating hormone were significantly higher in the mutant, the event of which was also confirmed by examining the secretion from the tissue culture of anterior pituitary glands. These results suggest that PRIP is involved in maternal reproduction, through the gonadotropine secretion..
Membership in Academic Society
  • The Molecular Biology Society of Japan
  • The Japanese biochemical Society
Educational
Educational Activities
*Biochemical practical course
*Research exposure
*Lectures of molecular biology
*Early exposure