Comparative Morphological Study on the Embryonic and Neonatal Development of the Filiform Papillae and Teeth in Mice

정순정 한국치위생과학회 2020 치위생과학회지 Vol.20 No.2

Background: In the early stages of development, teeth and lingual papillae are induced and developed through special and complex epithelial-mesenchymal interactions. Tooth completion indicates the beginning of the weaning phase, and accordingly, many oral tissues and organs are completed, and it is thought that their developmental completion times are related to each other. The purpose of this study was to clarify the embryonic and neonatal development of the filiform papillae and mandibular molar tooth, and discuss the developmental relationship between these organs by comparing the developmental completion times.Methods: Embryos at embryonic day 15 (EM15), 17 (EM17), and 21 (EM21) and mice at neonatal day 1 (NE1), 5 (NE5), 10 (NE10), and 21 (NE21) were used for experimentation. Tissues dissected from embryos and mice were fixed, and processed for histological analysis. Sections from the tissues were stained with hematoxylin and eosin for observation under a light microscope.Results: Based on the histological analysis results, the developmental process of the lingual epithelium covering the dorsal surface of the tongue was classified into three stages: initiation, morphogenesis, and functional. The development of the filiform papillae begins at EM17; undergoes rapid morphological changes in epithelial cells at EM21, PN1 and PN5, and reaches the functional stage at PN10, which is the sucking phase. Tooth development begins at EM13 or 15 and is completed at NE21 through prenatal and postnatal development.Conclusion: The development of the filiform papillae was initiated late and completed quickly through embryonic and neonatal development in comparison with the mandibular molar tooth. The filiform papillae are considered to play an important role in sucking rather than mastication as it is completed in the sucking phase.

Development of a Novel in vitro Maturation System for Pig Oocytes using a Medium with Reduced Sodium Chloride

Joohyeong Lee,Eunsong Lee 한국수정란이식학회 2017 한국수정란이식학회 학술대회 Vol.2017 No.05

In vitro maturation (IVM) systems have become indispensable for the production of large numbers of competent oocytes in domestic species. The quality of in vitro matured oocyte is one of the important factors determining the success of assisted reproductive technologies (ARTs) including intracytoplasmic sperm injection (ICSI), in vitro fertilization (IVF), and somatic cell nuclear transfer (SCNT) in human and livestock. Incomplete cytoplasmic maturation of oocytes can lead not only to a failure of fertilization but also to a developmental arrest after ARTs. Thus, establishment of a stable IVM system to produce a large number of high quality oocytes, especially in domestic animals, is essential for improvement of ARTs efficiency by producing high quality embryos. The morphological characteristics are commonly used to predict the developmental potential of oocytes and embryos. Usually, normal oocytes shrink when exposed to a hypertonic medium, and recover their morphology when returned to an isotonic medium. During this process, oocytes show various morphologic changes, such as shrinkage in spherical (SSP) or irregular shapes (SIR). In the first study, we investigated whether the shrinkage pattern of oocytes that was observed after hyperosmotic treatment could be used as a morphologic characteristic to predict the quality of IVM oocytes in pigs. We found that SSP oocytes showed improved developmental competence after PA and SCNT. This improved embryonic development was most likely because of the more advanced nuclear and cytoplasmic maturation in SSP oocytes compared with SIR oocytes. Pig oocytes shows a wide variation in the size of perivitelline space (PVS) after IVM. Based on this finding, we examined in the second study whether or not there was any correlation between the PVS size of IVM oocytes and their developmental competence after PA and SCNT. Our results demonstrated that in vitro developmental competence to the blastocyst stage positively correlated with the size of the PVS of oocytes after IVM. In addition, we observed that mature oocytes with a larger PVS showed higher levels of intracellular GSH content and transcription factor expression. Furthermore, enlargement of the PVS by culturing in reduced NaCl medium improves the embryonic development after PA and SCNT. In the third study, we investigated the effects of a hypotonic medium with reduced NaCl (61.6 mM) compared with an isotonic medium (108.0 mM NaCl) on oocyte maturation and embryonic development after PA and SCNT. In addition, we attempted to optimize our IVM system using a hypotonic maturation medium by examining the effects of hypotonic medium during various stages of IVM on oocyte maturation and subsequent embryonic development. Our results demonstrated that maturation of pig oocytes in hypotonic medium with reduced NaCl during the last 11 hr of IVM increased the developmental competence of oocytes after PA and SCNT. These beneficial effects was also shown in a commercial medium (a minimum essential medium; aMEM) in which the NaCl concentration was reduced to 61.6 mM. In addition, IVM of oocytes in medium with reduced NaCl increases the proportion of SSP oocytes in pigs. In summary, our results demonstrate that IVM of pig oocytes in a hypotonic medium with low-NaCl is better able to support embryonic development after PA and SCNT, most likely by improving the cytoplasmic maturation via increased intraoocyte GSH content and widened PVS. Based on these results, the newly developed IVM system using a hypotonic medium with reduced NaCl can produce high quality oocytes and be considered a new strategy for improving ARTs efficiency in pigs.

Fluoxetine Treatment during In Vitro Fertilization and Culture Increases Bovine Embryonic Development

Choe, Changyong,Kang, Dawon 韓國受精卵移植學會 2014 한국동물생명공학회지 Vol.29 No.2

K+ channels are involved in the regulation of a variety of physiological functions, including proliferation, apoptosis and differentiation, in mammalian cells. Our previous study demonstrated that the blockage of K+ channels inhibits mouse early embryonic development. This study was designed to identify the effect of K+ channels during bovine embryonic development. K+ channel blockers (tetraethylammonium (TEA), BaCl2, quinine, ruthenium red and fluoxetine) were added to the culture medium during in vitro fertilization (IVF) for 6 h to first identify the short-term effect of these chemicals. Among K+ channel blockers, fluoxetine, which is used as a selective serotonin reuptake inhibitor, significantly increased the blastocyst formation rate by approximately 6% when compared to control. During the in vitro maturation (IVM) of immature oocytes and the in vitro culture (IVC) of embryos, the oocytes and embryos were exposed to fluoxetine for either a short-term (6 h) or a long-term (24 h) to compare the embryonic development in response to exposure time. The 6 h exposure to fluoxetine during IVM did not affect the blastocyst formation rate, but the rate of blastocyst formation was reduced after the 24 h exposure. On the other hand, embryonic development increased approximately 10% in both groups of embryos exposed to fluoxetine for 6 and 24 h during IVC. Taken together, fluoxetine treatment during IVF and IVC, but not IVM, enhances bovine embryonic development. These results suggest that fluoxetine-modulated signals in oocytes and embryos could be an important factor towards enhancing bovine embryonic development.

Fluoxetine Treatment during In Vitro Fertilization and Culture Increases Bovine Embryonic Development

최창용,강다원 사단법인 한국동물생명공학회 2014 한국동물생명공학회지 Vol.29 No.2

K+ channels are involved in the regulation of a variety of physiological functions, including proliferation, apoptosisand differentiation, in mammalian cells. Our previous study demonstrated that the blockage of K+ channels inhibitsmouse early embryonic development. This study was designed to identify the effect of K+ channels during bovineembryonic development. K+ channel blockers (tetraethylammonium (TEA), BaCl2, quinine, ruthenium red andfluoxetine) were added to the culture medium during in vitro fertilization (IVF) for 6 h to first identify the short-termeffect of these chemicals. Among K+ channel blockers, fluoxetine, which is used as a selective serotonin reuptakeinhibitor, significantly increased the blastocyst formation rate by approximately 6% when compared to control. Duringthe in vitro maturation (IVM) of immature oocytes and the in vitro culture (IVC) of embryos, the oocytes and embryoswere exposed to fluoxetine for either a short-term (6 h) or a long-term (24 h) to compare the embryonic developmentin response to exposure time. The 6 h exposure to fluoxetine during IVM did not affect the blastocyst formation rate,but the rate of blastocyst formation was reduced after the 24 h exposure. On the other hand, embryonic developmentincreased approximately 10% in both groups of embryos exposed to fluoxetine for 6 and 24 h during IVC. Takentogether, fluoxetine treatment during IVF and IVC, but not IVM, enhances bovine embryonic development. Theseresults suggest that fluoxetine-modulated signals in oocytes and embryos could be an important factor towardsenhancing bovine embryonic development.

Roles of gangliosides in mouse embryogenesis and embryonic stem cell differentiation

Dong Hoon Kwak,추영국,Byoung Boo Seo,장규태 생화학분자생물학회 2011 Experimental and molecular medicine Vol.43 No.7

Gangliosides have been suggested to play important roles in various functions such as adhesion, cell differentiation,growth control, and signaling. Mouse follicular development, ovulation, and luteinization during the estrous cycle are regulated by several hormones and cell-cell interactions. In addition, spermatogenesis in seminiferous tubules of adult testes is also regulated by several hormones, including follicle-stimulating hormone (FSH) and luteinizing hormone (LH) and cell-cell interactions. The regulation of these processes by hormones and cell-cell interactions provides evidence for the importance of surface membrane components, including gangliosides. During preimplantation embryo development, a mammalian embryo undergoes a series of cleavage divisions whereby a zygote is converted into a blastocyst that is sufficiently competent to be implanted in the maternal uterus and continue its development. Mouse embryonic stem (mES) cells are pluripotent cells derived from mouse embryo, specifically, from the inner cell mass of blastocysts. Differentiated neuronal cells are derived from mES cells through the formation of embryonic bodies (EBs). EBs recapitulate many aspects of lineage-specific differentiation and temporal and spatial gene expression patterns during early embryogenesis. Previous studies on ganglioside expression during mouse embryonic development (including during in vitro fertilization, ovulation, spermatogenesis,and embryogenesis) reported that gangliosides were expressed in both undifferentiated and differentiated (or differentiating) mES cells. In this review,we summarize some of the advances in our understanding of the functional roles of gangliosides during the stages of mouse embryonic development,including ovulation, spermatogenesis, and embryogenesis,focusing on undifferentiated and differentiated mES cells (neuronal cells).

Roles of gangliosides in mouse embryogenesis and embryonic stem cell differentiation

Kwak, Dong-Hoon,Seo, Byoung-Boo,Chang, Kyu-Tae,Choo, Young-Kug Korean Society for Biochemistry and Molecular Bion 2011 Experimental and molecular medicine Vol.43 No.7

Gangliosides have been suggested to play important roles in various functions such as adhesion, cell differentiation, growth control, and signaling. Mouse follicular development, ovulation, and luteinization during the estrous cycle are regulated by several hormones and cell-cell interactions. In addition, spermatogenesis in seminiferous tubules of adult testes is also regulated by several hormones, including follicle- stimulating hormone (FSH) and luteinizing hormone (LH) and cell-cell interactions. The regulation of these processes by hormones and cell-cell interactions provides evidence for the importance of surface membrane components, including gangliosides. During preimplantation embryo development, a mammalian embryo undergoes a series of cleavage divisions whereby a zygote is converted into a blastocyst that is sufficiently competent to be implanted in the maternal uterus and continue its development. Mouse embryonic stem (mES) cells are pluripotent cells derived from mouse embryo, specifically, from the inner cell mass of blastocysts. Differentiated neuronal cells are derived from mES cells through the formation of embryonic bodies (EBs). EBs recapitulate many aspects of lineage-specific differentiation and temporal and spatial gene expression patterns during early embryogenesis. Previous studies on ganglioside expression during mouse embryonic development (including during $in$ $vitro$ fertilization, ovulation, spermatogenesis, and embryogenesis) reported that gangliosides were expressed in both undifferentiated and differentiated (or differentiating) mES cells. In this review, we summarize some of the advances in our understanding of the functional roles of gangliosides during the stages of mouse embryonic development, including ovulation, spermatogenesis, and embryogenesis, focusing on undifferentiated and differentiated mES cells (neuronal cells).

TIP60 contributes to porcine embryonic development by regulating DNA damage response

Guo, Jing,Zhou, Wenjun,Niu, Ying-Jie,Shin, Kyung-Tae,Heo, Young Tae,Kim, Nam-Hyung,Cui, Xiang-Shun Elsevier 2018 Theriogenology Vol.108 No.-

<P><B>Abstract</B></P> <P>The acetyltransferase TIP60 (also known as Kat5) is a member of the MYST family of histone acetyltransferases and was initially identified as a cellular protein. TIP60 acetylates histone and non-histone proteins and is involved in diverse biological processes, including apoptosis, cell cycle, and DNA damage responses. In this study, a specific inhibitor of TIP60 was used to detect the function of TIP60 in porcine parthenogenetic embryos. The results showed that TIP60 inhibition impaired porcine parthenogenetic embryonic development. The mechanism of TIP60 was also determined. We found that the TIP60 inhibition impaired embryonic development by ROS induced DNA damage, as demonstrated by the number of γH2A in the nuclei. TIP60 inhibition triggered DNA damage through the regulation of p53-p21 pathway and TIP60 played a role in DNA repair. TIP60 inhibition decreased the efficiency of DNA repair by regulating 53BP1-dependent repair after DNA damage. Inhibition of TIP60 also increased the adaptive response, autophagy, by modulating LC3. Therefore, TIP60 plays a role in early porcine parthenogenetic embryonic development by regulating DNA damage and repair.</P> <P><B>Highlights</B></P> <P> <UL> <LI> TIP60 inhibition disrupted the embryonic development. </LI> <LI> TIP60 inhibition triggered the DNA damage and incomplete DNA repair due to the excessive generation of ROS. </LI> <LI> TIP60 inhibition induced the apoptosis and autophagy. </LI> </UL> </P>

누에의 알 및 幼蟲의 發育에 따른 體液蛋白質의 變動

장창식,이상몽,이호웅,박상봉 密陽産業大學校 農業技術開發硏究所 1998 農業技術開發硏究所報 Vol.2 No.1

누에배자 및 유충의 발육단계와 체액단백질의 변동과의 상호관계를 Native-PAGE 및 SDS-PAGE 전기영동방법에 의해 비교 검토한 결과 누에알의 체액단백질 전기영동양상은 난령 1-7일, 난령 8-9일, 부화유충단계의 단백질군등 3군으로 구분가능 하였으며 누에유충의 경우는 1-4령, 5령 체액단백질의 2군으로 구분이 가능하였다. 누에알의 체액단백질은 30K이하의 소분자 단백질 보다는 40-200K의 비교적 대분자 단백질에서 발육에 따라 변동이 심하였고, 누에유충 체액단백질은 30K 근처의 소분자 단백질에서 변동이 심한 것으로 관찰되었다. Haemolymph proteins from the silkworm, Bombyx mori, were analysed by Native- and SDS-polyacrylamide gel electrophoresis(PAGE) during embryonic and larval development. From embryonic development, electrophoretic haemolymph protein patterns were grouped into three categories: day 1 ~ day 7, day 8 ~ day 9 and day 10(newly hatched larval stage) egg proteins. From larval development. also the patterns were grouped into two categories; the 1st ~ the 4th instar and the 5th instar larval haemolymph proteins. The electrophoretic patterns of proteins with M.W. of 40-200KD varied during embryonic development, and the patterns of proteins with M.W. of 30KD varied during larval development.

Embryonic Development of Siberian Sturgeon Acipenser baerii under Hatchery Conditions: An Image Guide with Embryological Descriptions

박철홍,이상윤,김동수,남윤권 한국수산과학회 2013 Fisheries and Aquatic Sciences Vol.16 No.1

Normal embryonic development at a constant temperature (18°C) has been described for the Siberian sturgeon Acipenser baerii (Acipenseriformes). Hormone-induced spawning and artificial insemination were performed to prepare embryonic batches for embryologic examination. After insemination, early cleavages of the Siberian sturgeon embryos continued for 7 h post-fertilization (HPF), showing the typical pattern of uneven holoblastic cleavage. Blastulation and gastrulation began at 9 HPF and 19 HPF, respectively. Epiboly formation (2/3 covered) was observed at 25 HPF during gastrulation. Neurulation was initiated with the formation of a slit-like neural groove from the blastopore at 33 HPF. During neurulation, the primary embryonic kidney (pronephros) and s-shaped heart developed. The embryos underwent progressive differentiation, which is typical of Acipenseriform species. A mass hatching was observed at 130 HPF, and the average total length of the hatched prolarvae was 10.5 mm. The hatched prolarvae possessed a typical pigment plug (yolk plug). The results of this study are valuable not only as a reference guide for the artificial propagation of Siberian sturgeon in hatcheries but also as the basis for the derivation of developmental gene expression assays for this species.

Embryonic Development of Siberian Sturgeon Acipenser baerii under Hatchery Conditions: An Image Guide with Embryological Descriptions

Park, Chulhong,Lee, Sang Yoon,Kim, Dong Soo,Nam, Yoon Kwon The Korean Society of Fisheries and Aquatic Scienc 2013 Fisheries and Aquatic Sciences Vol.16 No.1

Normal embryonic development at a constant temperature ($18^{\circ}C$) has been described for the Siberian sturgeon Acipenser baerii (Acipenseriformes). Hormone-induced spawning and artificial insemination were performed to prepare embryonic batches for embryologic examination. After insemination, early cleavages of the Siberian sturgeon embryos continued for 7 h post-fertilization (HPF), showing the typical pattern of uneven holoblastic cleavage. Blastulation and gastrulation began at 9 HPF and 19 HPF, respectively. Epiboly formation (2/3 covered) was observed at 25 HPF during gastrulation. Neurulation was initiated with the formation of a slit-like neural groove from the blastopore at 33 HPF. During neurulation, the primary embryonic kidney (pronephros) and s-shaped heart developed. The embryos underwent progressive differentiation, which is typical of Acipenseriform species. A mass hatching was observed at 130 HPF, and the average total length of the hatched prolarvae was 10.5 mm. The hatched prolarvae possessed a typical pigment plug (yolk plug). The results of this study are valuable not only as a reference guide for the artificial propagation of Siberian sturgeon in hatcheries but also as the basis for the derivation of developmental gene expression assays for this species.