Assoc.Prof. Rangsun Parnpai

Year	Title	Journal
2014	Cytochalasin B efficiency in the cryopreservation of immature bovine oocytes by Cryotop and solid surface vitrification methods Sripunya N., Liang Y., Panyawai K., Srirattana K., Ngernsoungnern A., Ngernsoungnern P., Ketudat-Cairns M., Parnpai R.	Cryobiology
Abstract: The present study was undertaken to compare the efficacies of Cryotop (CT), solid surface vitrification (SSV) methods and cytochalasin B (CB) treatment for the cryopreservation of immature bovine oocytes, in terms of survival, nuclear maturation, and in vitro development. Solution exposed oocytes were in vitro maturated and fertilized. No difference was found in the rates of survival, nuclear maturation and blastocyst among solution exposed groups and fresh control group, except blastocysts rates in oocytes exposed to CB, cryoprotectant (CPA) and fluorescein diacetate (FDA) group (CB-CPA-FDA) (23%) significantly lower than that of control group (32%). CB pretreated ((+)CB) or non-pretreated ((-)CB) COCs were vitrified either by SSV or CT. Among four vitrified groups the nuclear maturation rates (CT(-)CB: 58%, CT(+)CB: 57%, SSV(-)CB: 60%, SSV(+)CB: 63%), cleavage (CT(-)CB: 36%, CT(+)CB: 24%, SSV(-)CB: 34%, SSV(+)CB: 26%) and blastocysts rates (CT(-)CB: 6%, CT(+)CB: 7%, SSV(-)CB: 4%, SSV(+)CB: 6%) did not differ, but the rates of the four vitrified groups were significantly lower than those of non-vitrified group (81%, 71% and 26%, respectively). We thus conclude that CT and SSV perform equally in vitrification of bovine immature oocytes, and CB did not increase the viability, nuclear maturation, or in vitro development of vitrified oocytes. © 2014 Elsevier Inc. Keyword: Bovine; Cryotop; Immature oocytes; Solid surface vitrification; Vitrification Scopus Link: https://www.scopus.com/inward/record.uri?eid=2-s2.0-84922515199&doi=10.1016%2fj.cryobiol.2014.09.001&partnerID=40&md5=ef60f7558ca4fe36445817274919e23a DOI: 10.1016/j.cryobiol.2014.09.001
2014	Effects of trichostatin a on in vitro development and DNA methylation level of the satellite i region of swamp buffalo (Bubalus bubalis) cloned embryos Srirattana K., Ketudat-Cairns M., Nagai T., Kaneda M., Parnpai R.	Journal of Reproduction and Development
Abstract: Trichostatin A (TSA), a histone deacetylase inhibitor, has been widely used to improve the cloning efficiency in several species. This brings our attention to investigation of the effects of TSA on developmental potential of swamp buffalo cloned embryos. Swamp buffalo cloned embryos were produced by electrical pulse fusion of male swamp buffalo fibroblasts with swamp buffalo enucleated oocytes. After fusion, reconstructed oocytes were treated with 0, 25 or 50 nM TSA for 10 h. The results showed that there was no significant difference in the rates of fusion (82–85%), cleavage (79–84%) and development to the 8-cell stage (59–65%) among treatment groups. The highest developmental rates to the morula and blastocyst stages of embryos were found in the 25 nM TSA-treated group (42.7 and 30.1%, respectively). We also analyzed the DNA methylation level in the satellite I region of donor cells and in in vitro fertilized (IVF) and cloned embryos using the bisulfite DNA sequencing method. The results indicated that the DNA methylation levels in cloned embryos were significantly higher than those of IVF embryos but approximately similar to those of donor cells. Moreover, there was no significant difference in the methylation level among TSA-treated and untreated cloned embryos. Thus, TSA treatments at 25 nM for 10 h could enhance the in vitro developmental potential of swamp buffalo cloned embryos, but no beneficial effect on the DNA methylation level was observed. © 2014 by the Society for Reproduction and Development. Keyword: Cloning; DNA methylation; Embryo development; Swamp buffalo; Trichostatin A Scopus Link: https://www.scopus.com/inward/record.uri?eid=2-s2.0-84908213584&doi=10.1262%2fjrd.2013-116&partnerID=40&md5=440725a03c5cbad4ded1723d2d825989 DOI: 10.1262/jrd.2013-116
2014	Ovarian follicular dynamics and hormones throughout the estrous cycle in Thai native (Bos indicus) heifers Chasombat J., Nagai T., Parnpai R., Vongpralub T.	Animal Science Journal
Abstract: Relatively few studies have been reported regarding the reproductive physiology of female Thai native cattle. Therefore, the objective of the present study was to evaluate the follicular dynamics and concentrations of follicle stimulating hormone (FSH), estradiol (E2) and progesterone (P4) during the estrous cycle in Thai native heifers (TNH) and to compare obtained results with those of European and Indian cattle breeds previously reported. For the detection of estrus, ovaries of all 20 heifers were examined twice daily (12 h intervals) by ultrasonography for three consecutive estrous cycles. From data of 60 estrous cycles (n=60 estrous cycles from 20 heifers), it was found that 14 (70%) and 6 heifers (30%) had two (42 estrous cycles collected from 14 heifers) and three follicular waves (18 estrous cycles collected from 6 heifers), respectively. The days when estrus was detected, interovulatory intervals, life-spans of corpus lutea (CL), and days for growing and regression of CLs were shorter in the two follicular waves than those in the three follicular waves (P<0.05). In both two and thre follicular waves, larger maximum diameters and higher growth rates of the dominant follicle (DF) in an ovulatory wave were observed than those of the preceding waves without ovulation (P<0.05). There was a progressive increase in follicular size and FSH and E2 production during follicular growth in each follicular wave. In addition, the FSH and E2 peak concentrations during the ovulatory wave were higher than those of the anovulation waves (P<0.05). Moreover, although the ovarian follicular dynamic patterns in Thai native heifers were similar to those previously reported for European and Indian cattle breeds, the diameter of the largest preovulatory follicle (OF), subordinate follicles (SF) and CLs were smaller than those in European and Indian cattle breeds. In conclusion, when compared with European and some breeds of Indian cattle, the length of interovulatory intervals was shorter, and the sizes of dominant SF and CLs were smaller in Thai native heifers. © 2013 Japanese Society of Animal Science. Keyword: Estrous cycle; Follicular wave pattern; Hormones; Thai indigenous cattle Scopus Link: https://www.scopus.com/inward/record.uri?eid=2-s2.0-84892479273&doi=10.1111%2fasj.12086&partnerID=40&md5=cd3313b6849a1689d83a5bfe059e4e29 DOI: 10.1111/asj.12086
2013	Effects of L-carnitine supplemented in maturation medium on the maturation rate of swamp buffalo oocytes Phongmitr T., Liang Y., Srirattana K., Panyawai K., Sripunya N., Treetampinich C., Parnpai R.	Buffalo Bulletin
Abstract: In vitro maturation (IVM) is a first and crucial step to determine the success of in vitro embryo production. L-carnitine enhances lipid metabolism in cells and exerts antioxidant effects as well. Such properties are known to be beneficial for oocyte maturation. Thus, to investigate the effects of L-carnitine during IVM on maturation rate of swamp buffalo oocytes, oocytes were matured in IVM medium supplemented with 0.3, 0.6 and 1.2 mg/mL of L-carnitine (0.3, 0.6 and 1.2 L-carnitine groups, respectively). Oocytes matured in 0 mg/mL of L-carnitine were treated as a control group. After IVM, oocytes were denuded by hyaluronidase and fixed with ethanol: acetic acid (3:1) for 3 days. After that, the fixed oocytes were stained with 1% (w/v) orcein in acetic acid for 10 min. Oocytes appearing a metaphase plate and one polar body were regarded as metaphase II stage (MII). Supplementation of L-carnitine at 0.3 mg/mL (0.3 L-carnitine group) showed a significantly higher MII rate than that in control group (83/123, 67.5% vs 69/120, 57.5%); however, the rate was not significantly higher than those in 0.6 mg/mL (79/124, 63.7%) and 1.2 mg/mL (79/123, 64.2%) groups. In conclusion, supplementation of L-carnitine during IVM could improve the nuclear maturation rate to MII and 0.3 mg/mL was the optimal concentration. Keyword: L-carnitine; Oocyte maturation; Swamp buffalo Scopus Link: https://www.scopus.com/inward/record.uri?eid=2-s2.0-84897901288&partnerID=40&md5=ede634c180a79bb990daddcf3699ead6 DOI:
2013	Influence of growth factors on survival and development of swamp buffalo early antral follicle cultured in vitro Kaewmungkun K., Srirattana K., Punyawai K., Sripunya N., Liang Y., Sangsritavong S., Parnpai R.	Buffalo Bulletin
Abstract: Buffalo ovary has a large number of follicles but a few follicles are able to grow and ovulate. After collecting oocytes for in vitro maturation, a lot of small follicles still remained in the ovaries. In this study, survival and development of these remaining early antral follicles in the ovaries were investigated after in vitro culture. Growth factors may be essentially required for follicle development. Therefore, the aim of this study was to examine the effects of growth factors, basic fibroblast growth factor (bFGF) and insulin-like growth factor-I (IGF-I), on in vitro culture of swamp buffalo early antral follicles. The follicles isolated from buffalo ovaries were divided into two groups, according to their diameters, Group I: 400-599 μm and Group II: 600-799 μm. The follicles were embedded in collagen gel and cultured in the medium supplemented with either one of the two growth factors or combinations of the two factors for 14 days. The diameters of follicles were measured at 7 and 14 days of culture. Culture medium supplemented with bFGF showed significantly the highest survival rate in all size groups of the follicles (44.8% and 32.7% in groups I and II, respectively), except for bFGF+IGF-I group in Group I. However, when combined with IGF-I, its promoting effect on follicular survival and development was neutralized. Follicles from both Groups I and II cultured in medium supplemented with bFGF for 14 days (also follicles cultured for 7 days in Group I) tended to have increased follicle size compared to other groups; follicles from the bFGF group showed a significantly higher increase in size compared to bFGF+IGF-I group. IGF-I had on effects on follicular survival and development in both Groups I and II. In conclusion, bFGF is a suitable supplement for in vitro culture of the early antral follicles in swamp buffalos in terms of the survival for both Groups I and II and development for Group I. Keyword: Early antral follicle; Growth factor; In vitro cuture; Swamp buffalo Scopus Link: https://www.scopus.com/inward/record.uri?eid=2-s2.0-84897898398&partnerID=40&md5=0f7569f2bd35ca331196a2ee7f8cefd0 DOI:
2013	Vitrification of buffalo oocytes: Current status and perspectives Liang Y., Somfai T., Nagai T., Parnpai R.	Buffalo Bulletin
Abstract: During the past decade vitrification has been acknowledged as a viable alternative to traditional slow-rate freezing in both human and animal embryology. The buffalo is the major milk and meat producing farm animal in many developing countries. Buffalo oocytes obtained from slaughterhouse ovaries and matured in vitro are useful sources. Cryopreservation of buffalo oocytes is very important in preserving this endanger species for future use. This review presents the recent buffalo oocytes vitrification procedures, the principle of vitrification, protocols for buffalo oocytes vitrification and the future of buffalo oocytes vitrification. Keyword: Buffalo; Oocyte; Vitrification Scopus Link: https://www.scopus.com/inward/record.uri?eid=2-s2.0-84888173173&partnerID=40&md5=c5c42f104286b7c3599490187887c546 DOI:
2013	Effect of L-carnitine on maturation, cryo-tolerance and embryo developmental competence of bovine oocytes Phongnimitr T., Liang Y., Srirattana K., Panyawai K., Sripunya N., Treetampinich C., Parnpai R.	Animal Science Journal
Abstract: In this study, the effects of the addition of L-carnitine in in vitro maturation (IVM) medium for bovine oocytes on their nuclear maturation and cryopreservation were investigated; they were matured in IVM medium supplemented with 0.0, 0.3, 0.6 and 1.2mg/mL of L-carnitine (control, 0.3, 0.6 and 1.2 groups, respectively) and some of them were vitrified by Cryotop. Moreover, the effects of L-carnitine during in vitro fertilization (IVF) and in vitro culture (IVC) on the developmental potential and quality of IVF embryos were also examined. A significantly higher maturation rate of oocytes was obtained for 0.3 and 0.6mg/mL groups compared with the control (P<0.05). The blastocyst formation rate in the 0.6 group was significantly improved, whereas the rate in the 1.2 group was significantly decreased when compared with the control group (P<0.05). No significant difference was found in embryo development between the control and the L-carnitine group after oocyte vitrification. Supplementation of IVF and IVC media with L-carnitine had no effect on development to the blastocyst stage of IVM oocytes treated with 0.6mg/mL L-carnitine. In conclusion, the supplementation of L-carnitine during IVM of bovine oocytes improved their nuclear maturation and subsequent embryo development after IVF, but when they were vitrified the improving effects were neutralized. © 2013 Japanese Society of Animal Science. Keyword: Bovine; Embryo development; L-carnitine; Maturation; Vitrification Scopus Link: https://www.scopus.com/inward/record.uri?eid=2-s2.0-84885953685&doi=10.1111%2fasj.12067&partnerID=40&md5=0e74dc559a8eef1b05259a12e449e6ff DOI: 10.1111/asj.12067
2013	Developmental potential of vitrified goat oocytes following somatic cell nuclear transfer and parthenogenetic activation Srirattana K., Sripunya N., Sangmalee A., Imsoonthornruksa S., Liang Y., Ketudat-Cairns M., Parnpai R.	Small Ruminant Research
Abstract: Oocyte cryopreservation is an alternative tool in assisted reproductive technology. The objective of this study was to determine the developmental potential of vitrified goat oocytes after somatic cell nuclear transfer (SCNT). In vitro matured goat oocytes were vitrified by Cryotop method. The survival rate of vitrified oocytes at 1. h after thawing determined by fluorescein diacetate staining was 92.1% which was significantly lower than that of fresh oocytes (99.3%). Live oocytes from both vitrified and fresh groups were used as recipient cytoplasts for SCNT. No significant difference in fusion rate was found between vitrified (97.6%) and fresh (93.2%) oocytes. The cleavage rates of vitrified oocytes in the SCNT and parthenogenetic activation (PA) embryos (29.6% and 27.9%) were significantly lower than those from fresh SCNT and PA oocytes (80.9% and 91.1%). The developmental rates to 8-cell stage of SCNT and PA embryos from vitrified oocytes were also lower than that of fresh oocytes. There was no significant difference among the groups in the development to morula stage (11-27%). However, the blastocyst rate of PA embryos derived from fresh oocytes (12.5%) was significantly higher than the other groups (1.2-3.3%). Although high survival rate of vitrified oocytes was obtained, the cleavage, 8-cell, and blastocyst formation rates of SCNT and PA embryos from vitrified oocytes were still lower than those of fresh oocytes. © 2012 Elsevier B.V. Keyword: Cryotop; Goat oocyte; Somatic cell nuclear transfer; Vitrification Scopus Link: https://www.scopus.com/inward/record.uri?eid=2-s2.0-84876309207&doi=10.1016%2fj.smallrumres.2012.10.011&partnerID=40&md5=513e1b6579a5d31cce9fc40be39205a6 DOI: 10.1016/j.smallrumres.2012.10.011
2013	Superstimulation of follicular growth in Thai native heifers by a single administration of follicle stimulating hormone dissolved in Polyvinylpyrrolidone Chasombat J., Sakhong D., Nagai T., Parnpai R., Vongpralub T.	Journal of Reproduction and Development
Abstract: This study was undertaken to determine whether a single i.m. injection of FSH dissolved in 10 ml of 30% (wt/ vol) polyvinylpyrrolidone (PVP; MW=40,000) to form FSHp would induce follicular growth in Thai native heifers and to determine its optimal dose. In Group 1, heifers (n=4) were given multiple i.m. injections of FSHp every 12 h for 3 days at decreasing doses, for a total of 100 mg (control). In Groups 2, 3 and 4, heifers (n=4 in each group) were given single i.m. injections of FSHp at 50, 100 and 150 mg. All heifers received a single injection of 750 μg PGF2α 48 h after the initiation of exogenous FSH treatment. Ovaries of treated heifers were examined by transrectal ultrasonography every day until they showed estrus. Group 3 showed significantly higher numbers of ovulation follicles, significantly higher growth rates of follicles per day and significantly larger diameters of follicles and corpora lutea than groups 1 and 2 but not Group 4 (P<0.05). Group 4 showed significantly higher numbers of large follicles (≥5 mm in diameter), unovulated follicles and ovulations, a significantly higher growth rate of follicles per day, and significantly larger diameters of follicles and corpora lutea (P<0.05) than those of the other groups. This indicates a state of overstimulation of ovaries in this group. Besides, the plasma levels of FSH in Group 4 were significantly higher (P<0.05) than in the other group and were maintained in the range of 2.2-0.7 ng/ ml over a period of 6 to 66 h after the FSHp injection. Meanwhile, the plasma levels of P4 and E2 did not differ in any of the groups in the period of 0 to 96 h during the superstimulation program. In conclusion, it was demonstrated that a single i.m. injection of 100 mg FSHp was the most effective dose for superstimulation of follicular growth in Thai native heifers under the experimental conditions in this study. ©2013 by the Society for Reproduction and Development. Keyword: Cattle; Follicle stimulating hormone (FSH); Polyvinylpyrrolidone (PVP) Scopus Link: https://www.scopus.com/inward/record.uri?eid=2-s2.0-84876531662&doi=10.1262%2fjrd.2012-119&partnerID=40&md5=a85d8ac40eaf68dc61cab384769c4ee1 DOI: 10.1262/jrd.2012-119
2013	Pathogenic cellular phenotypes are germline transmissible in a transgenic primate model of Huntington's disease Putkhao K., Kocerha J., Cho I.-K., Yang J., Parnpai R., Chan A.W.S.	Stem Cells and Development
Abstract: A transgenic primate model for Huntington's Disease (HD) first reported by our group that (HD monkeys) carry the mutant Huntingtin (HTT) gene with expanded polyglutamine (CAG) repeats and, develop chorea, dystonia, and other involuntary motor deficiencies similar to HD [1]. More recently, we have found that longitudinal magnetic resonance imaging of the HD monkey brain revealed significant atrophy in regions associated with cognitive deficits symptomatic in HD patients, providing the first animal model which replicates clinical phenotypes of diagnosed humans. Here we report germline transmission of the pathogenic mutant HTT in HD monkey by the production of embryos and subsequent derivation of HD monkey embryonic stem cells (rHD-ESCs) using HD monkey sperm. rHD-ESCs inherit mutant HTT and green fluorescent protein (GFP) genes through the gametes of HD monkey. rHD-ESCs express mutant HTT and form intranuclear inclusion, a classical cellular feature of HD. Notably, mosaicism of the pathogenic polyQ region in the sperm as well as derived ESCs were also observed, consistent with intraindividual and intergenerational reports of mosaic CAG repeats [2,3]and CAG expansion in HD patients [4-7]. The confirmation of transgene inheritability and development of pathogenic HD phenotype in derived rHD-ESCs reported in this study is a milestone in the pursuit of a transgenic primate model with inherited mutant HTT for development of novel disease biomarkers and therapeutics. © Mary Ann Liebert, Inc. Keyword: Scopus Link: https://www.scopus.com/inward/record.uri?eid=2-s2.0-84876131051&doi=10.1089%2fscd.2012.0469&partnerID=40&md5=16e16e5c4bb195d5e24c3b5594e117dc DOI: 10.1089/scd.2012.0469