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Fulltext Determination of suitable microspore stage and callus induction from anthers of kenaf (Hibiscus cannabinus L.)

Ibrahim AM, Kayat FB, Hussin ZE, Susanto D, Ariffulah M

ScientificWorldJournal, 2014;2014:284342.
PMID: 24757416 DOI: 10.1155/2014/284342

Kenaf (Hibiscus cannabinus L.) is one of the important species of Hibiscus cultivated for fiber. Availability of homozygous parent lines is prerequisite to the use of the heterosis effect reproducible in hybrid breeding. The production of haploid plants by anther culture followed by chromosome doubling can be achieved in short period compared with inbred lines by conventional method that requires self pollination of parent material. In this research, the effects of the microspore developmental stage, time of flower collection, various pretreatments, different combinations of hormones, and culture condition on anther culture of KB6 variety of Kenaf were studied. Young flower buds with immature anthers at the appropriate stage of microspore development were sterilized and the anthers were carefully dissected from the flower buds and subjected to various pretreatments and different combinations of hormones like NAA, 2,4-D, Kinetin, BAP, and TDZ to induce callus. The best microspore development stage of the flower buds was about 6-8 mm long collected 1-2 weeks after flower initiation. At that stage, the microspores were at the uninucleate stage which was suitable for culture. The best callus induction frequency was 90% in the optimized semisolid MS medium fortified with 3.0 mg/L BAP + 3.0 mg/L NAA.

Matched MeSH terms: Embryonic Development/drug effects; Embryonic Development/physiology*
Determination toxic effects of Hystrix Brachyura Bezoar extracts using cancer cell lines and embryo zebrafish (Danio rerio) models and identification of active principles through GC-MS analysis

Firus Khan AY, Ahmed QU, Nippun TS, Hilles A, Jalal TK, Teh LK, et al.

J Ethnopharmacol, 2020 Nov 15;262:113138.
PMID: 32726681 DOI: 10.1016/j.jep.2020.113138

ETHNOPHARMACOLOGICAL RELEVANCE: Porcupine bezoar (PB) is used as folk medicine for various medical conditions including cancer treatment in Malaysia. However, its toxicity profile has never been thoroughly ascertained to confirm its safe nature as an efficacious traditional medicine in the treatment of cancer as well as other ailments.
AIM OF THE STUDY: This study was aimed to reveal three different PBs' aqueous extracts(viz. PB-A, PB-B, PB-C) chemical constituent's profile using GC-MS analysis, anticancer property on A375, HeLa and MCF7 cancer cells, toxicity profile on zebrafish embryo morphology, EC50, LC50 and teratogenicity index.
MATERIALS AND METHODS: PBs' extracts characterization was performed through GC-MS analysis, in vitro anticancer effect was carried out on A375, HeLa and MCF7 cancer cell lines and finally and toxicity properties on three different PBs aqueous extracts (viz. PB-A, PB-B, PB-C) were determined using zebrafish embryo model.
RESULTS: The GC-MS analysis revealed 10 similar compounds in all PBs' extracts. Dilauryl thiodipropionate was found to be a major compound in all PBs' extracts followed by tetradecanoic acid. An in vitro anticancer study revealed PB extracts exerted median inhibition concentration (IC50) <50 μg/mL, on cancer cells viz. A375, HeLa and MCF7 with no significant toxicity on normal cells viz. NHDF cells. In vivo toxicity of PBs extracts found affecting tail detachment, hatching, craniofacial, brain morphology, soft tissues, edema, spinal, somites, notochord and cardiovascular system (brachycardia, disruption of blood circulation) deformities. The LC50 and EC50 demonstrated PB extracts effect as dose and time dependent with median concentration <150.0 μg/mL. Additionally, teratogenicity index (TI) viz. >1.0 revealed teratogenic property for PB extracts.
CONCLUSIONS: The findings revealed that all three PBs aqueous extracts possessed anticancer activity and exhibited significant toxicological effects on zebrafish embryos with high teratogenicity index. Hence, its use as an anticancer agent requires further investigation and medical attentions to determine its safe dose.

Matched MeSH terms: Embryonic Development/drug effects*; Embryonic Development/physiology
Increasing glucose in KSOMaa basal medium on culture Day 2 improves in vitro development of cloned caprine blastocysts produced via intraspecies and interspecies somatic cell nuclear transfer

Kwong PJ, Abdullah RB, Wan Khadijah WE

Theriogenology, 2012 Sep 1;78(4):921-9.
PMID: 22704387 DOI: 10.1016/j.theriogenology.2012.04.009

This study was conducted to evaluate the efficiency of potassium simplex optimization medium with amino acids (KSOMaa) as a basal culture medium for caprine intraspecies somatic cell nuclear transfer (SCNT) and caprine-bovine interspecies somatic cell nuclear transfer (iSCNT) embryos. The effect of increased glucose as an energy substrate for late stage development of cloned caprine embryos in vitro was also evaluated. Enucleated caprine and bovine in vitro matured oocytes at metaphase II were reconstructed with caprine ear skin fibroblast cells for the SCNT and iSCNT studies. The cloned caprine and parthenogenetic embryos were cultured in either KSOMaa with 0.2 mM glucose for 8 days (Treatment 1) or KSOMaa for 2 days followed by KSOMaa with additional glucose at a final concentration of 2.78 mM for the last 6 days (Treatment 2). There were no significant differences in the cleavage rates of SCNT (80.7%) and iSCNT (78.0%) embryos cultured in KSOMaa medium. Both Treatment 1 and Treatment 2 could support in vitro development of SCNT and iSCNT embryos to the blastocyst stage. However, the blastocyst development rate of SCNT embryos was significantly higher (P < 0.05) in Treatment 2 compared to Treatment 1. Increasing glucose for later stage embryo development (8-cell stage onwards) during in vitro culture (IVC) in Treatment 2 also improved both caprine SCNT and iSCNT embryo development to the hatched blastocyst stage. In conclusion, this study shows that cloned caprine embryos derived from SCNT and iSCNT could develop to the blastocyst stage in KSOMaa medium supplemented with additional glucose (2.78 mM, final concentration) and this medium also supported hatching of caprine cloned blastocysts.

Matched MeSH terms: Embryonic Development/drug effects*; Embryonic Development/physiology
Usual Presentation Has Odds: Unilateral Tibial Hemimelia in One of Dizygotic Twins

Al-Chalabi MMM, Wan Sulaiman WA

Cureus, 2021 Jan 21;13(1):e12834.
PMID: 33633877 DOI: 10.7759/cureus.12834

Tibial hemimelia is a relatively rare congenital tibial longitudinal deficiency (approximately 1 per 1 million live births), unilateral or bilateral, with a relatively intact fibula. Hemimelia results from a disruption of the lower limb developmental field during embryogenesis due to slow or even abort of chondrification process, which results in leg length discrepancy. Affected leg commonly appears short and deformed with knee, ankle, and foot involvement. It may present with a variety of associated anomalies. Surgical treatment varies according to the type and degree of deformity, and reconstructive interventions are still limited. Reported cases of tibial hemimelia are very infrequent, especially tibial hemimelia in twins. Usually, the cases were in single embryo or less frequently in one of the monozygotic twins, but no reported cases regarding tibial hemimelia in one of the dizygotic twins as this article reports.

Matched MeSH terms: Embryonic Development
Fulltext Toxicity assessment of gallic acid loaded graphene oxide (GAGO) nano-formulation in zebrafish (Danio Rerio) embryos

Ahmad Ashraful Hadi Abdul Ghafor, Nurhuda Elias, Suhaili Shams, Faizah Md Yasin, Sarchio, Seri Narti Edayu

Pertanika Journal of Science & Technology, 2020;28(1):311-326.
MyJurnal

Gallic acid (GA) is a phenolic compound found in almost all plants and has been reported to possess powerful health benefits such as anti-oxidant, anti-inflammatory, anti-cancer, and anti-diabetic properties. However, GA suffers a short half-life when administered in vivo. Recent studies have employed graphene oxide (GO), a biocompatible and cost-effective graphene derivative, as a nanocarrier for GA. However, the toxicity effect of this formulated nano-compound has not been fully studied. Thus, the present study aims to evaluate the toxicity and teratogenicity of GA loaded GO (GAGO) against zebrafish embryogenesis to further advance the development of GA as a therapeutic agent. GAGO was exposed to zebrafish embryos (n ≥ 10; 24hr post fertilization (hpf)) at different concentrations (0-500 μg/ml). The development of zebrafish was observed and recorded twice daily for four days. The toxicity of pure GO and GA was also observed at similar concentrations. Distilled water was used as control throughout the experiment. A significantly high mortality rate, delayed hatching rate and low heartbeat were recorded in embryos exposed to GO at concentrations of ≥ 150 μg/ml at 48 hr (p

Matched MeSH terms: Embryonic Development
Citizen science frontiers horseshoe crab population regain at their spawning beach in East Peninsular Malaysia

Mat Zauki NA, Satyanarayana B, Fairuz-Fozi N, Nelson BR, Martin MB, Akbar-John B, et al.

J Environ Manage, 2019 Feb 15;232:1012-1020.
PMID: 33395753 DOI: 10.1016/j.jenvman.2018.12.002

Carcinoscorpius rotundicauda and Tachypleus gigas may co-exist and share common spawning grounds elsewhere but at Balok (East Coast of Peninsular Malaysia), C. rotundicauda is an understudied species. Neglected as research candidate because of inaccessible spawning grounds, smaller size and less commercial value than T. gigas and also, difficulty to attain from the wild has made C. rotundicauda population status remaining unidentified at Balok. This standpoint drove the present attempt because anthropic activities like structure placement and mining are point-source for runoffs that load sediments into Balok River. While erosion-accretion events have altered Balok River width, the shore sediments in Balok Beach were transitioned between medium-fine and fine sand between years 2012 and 2016. Eventually by year 2016, the C. rotundicauda were depositing 5117 eggs in 91 nests from 200 to 1000 m range along this corridor facing South China Sea. From this yield, C. rotundicauda released 2880 eggs in 56 nests during the Southwest monsoon, 1254 eggs in 19 nests during the Northeast monsoon and 983 eggs in 16 nests during the Inter-monsoon seasons. Though female C. rotundicauda opted to lay their eggs in shallow burrows at lower shorelines, the absence of erosion and substantial silt and clay (>20%) deposition facilitates C. rotundicauda embryogenesis with brief periods of temperature and salinity shocks during day-time falling tides. This encourages C. rotundicauda to emerge with increasing abundance and carry out bi-monthly spawning at Balok Beach. In short, shore restoration initiatives like systematic boat docking, proper disposal of nets and waste and, periodic fish-catching operations were effectively led by the Balok fisher citizen scientist. This successful community joint-cooperation proves that citizen-led caretaking of degraded beaches offers marine life protection and are practical for coastal area management especially at areas where other oviparous animals such as turtles and crocodiles are harboured.

Matched MeSH terms: Embryonic Development
Prolonging the interval from ovarian hyperstimulation to laparoscopic ovum pick-up improves oocyte yield, quality, and developmental competence in goats

Abdullah RB, Liow SL, Rahman AN, Chan WK, Wan-Khadijah WE, Ng SC

Theriogenology, 2008 Sep 15;70(5):765-71.
PMID: 18579196 DOI: 10.1016/j.theriogenology.2008.04.052

The objective was to evaluate the effect of the interval between ovarian hyperstimulation and laparoscopic ovum pick-up (LOPU) on quality and developmental competence of goat oocytes before and after in vitro maturation (IVM) and intracytoplasmic sperm injection (ICSI). Estrus was synchronized with an intravaginal insert containing 0.3g progesterone (CIDR) for 10d, combined with a luteolytic treatment of 125 microg cloprostenol 36 h prior to CIDR removal. Ovaries were hyperstimulated with 70 mg FSH and 500 IU hCG given im 36, 60, or 72 h prior to LOPU (n=15, 16, and 7 does, respectively). For these groups, oocyte retrieval rates (mean+/-S.E.M.) were 24.7+/-2.9, 54.5+/-4.7, and 82.8+/-4.6% (P<0.001), and the proportions of cumulus-oocyte complexes (COC) with more than five layers of cumulus cells were 29.7+/-8.3, 37.6+/-6.9, and 37.3+/-7.0% (P<0.001). The proportion of IVM oocytes was highest at 72 h (82.1+/-2.8%; P<0.05), with no significant difference between 36 and 60 h (57.3+/-8.9% and 69.0+/-8.4%). Cleavage rates of ICSI embryos were 4.2+/-4.2, 70.9+/-8.4, and 78.9+/-8.2% with LOPU 36, 60, and 72 h post FSH/hCG (P<0.01), with a lower proportion of Grade-A embryos (P<0.05) following LOPU at 36 h compared to 60 and 72 h (29.7+/-8.3%, 37.6+/-6.9%, and 37.3+/-7.0%). In summary, a prolonged interval from FSH/hCG to LOPU improved oocyte retrieval rate and oocyte quality. Therefore, under the present conditions, LOPU 60 or 72 h after FSH/hCG optimized yields of good-quality oocytes for IVM and embryo production in goats.

Matched MeSH terms: Embryonic Development
Fulltext A rare cause of acute urinary retention in a young man: A median raphe penile cyst

Kamarulzaman MN, Chik A, Ghazali H

Malays Fam Physician, 2021 Mar 25;16(1):114-116.
PMID: 33948149 DOI: 10.51866/cr1016

Background: A penile median raphe cyst is an uncommon congenital lesion that is formed due to failure of the median raphe to close completely during embryo development. The majority of the cysts are asymptomatic and often go unnoticed. Here, we report acute urinary retention as a very rare complication of a penile median raphe cyst.
Case presentation: A 21-year-old single man presented with a sudden onset of acute urinary retention of one days' duration. Urethral catheterization was done at the Emergency Department and this drained 800 ml of urine. On further questioning, he claimed that he had had a painless nodule at the glans penis since childhood. The swelling increased in size in the past week causing discomfort. He denied any history of genitalia trauma or recent sexual intercourse. On examination, there was a bluish lesion over the ventral aspect of the glans penis measuring about 3 x 2 cm. The lesion was later excised and histopathology revealed a median raphe cyst of the penis.
Conclusion: A penile median raphe cyst is a rare lesion. Acute urinary retention caused by this lesion is very rare.

Matched MeSH terms: Embryonic Development
Epitokous metamorphosis, reproductive swimming, and early development of the estuarine polychaete, Neanthes glandicincta Southern, 1921 (Annelida, Nereididae) on the east coast of the Malay Peninsula

Azmi SS, Ibrahim YS, Angsupanich S, Sumpuntarat P, Sato M

Zookeys, 2021;1011:1-24.
PMID: 33551646 DOI: 10.3897/zookeys.1011.59780

The reproductive and developmental characteristics of the nereidid polychaete, Neanthes glandicincta Southern, 1921, commonly recorded in tropical estuaries in the Indo-West Pacific, were examined from Malaysia (the mangrove area of Kuala Ibai, Terengganu) and Thailand (the Lower Songkhla Lagoon) on the east coast of the Malay Peninsula. Epitokous metamorphosis of fully mature males and females and their reproductive swimming behaviour were recorded based on ten Malaysian epitokous specimens, which were collected at night during spring tides in a period of January 2018 to March 2019. Six Thailand epitokes were obtained in February and March 2006 by the laboratory rearing of immature worms. Epitokous metamorphosis is characterised by the enlargement of eyes in both sexes, division of the body into three parts and modification of parapodia with additional lobes in the mid-body of males, and replacement of atokous chaetae in the mid-body by epitokous natatory chaetae, completely in males and incompletely in females. The diameter of coelomic unfertilised eggs in females was 100-140 µm. After fertilisation, each egg formed a jelly layer, inside which embryonic development progressed. Trochophores hatched out of the jelly layer, entering a short free-swimming larval phase followed by demersal life at the early stage of 3-chaetiger nectochaeta one day after fertilisation. Then, the larvae entered benthic life as juveniles, crawling on the bottom, at the late stage of 3-chaetiger nectochaeta two days after fertilisation. The results indicate that N. glandicincta has an annual life cycle, which is usually completed within an estuary with limited larval dispersal ability.

Matched MeSH terms: Embryonic Development
Oral reproductive and developmental toxicity of Lignosus rhinocerotis mycelium in rat

Jhou BY, Liu HH, Yeh SH, Chen CC

J Ethnopharmacol, 2017 Aug 17;208:66-71.
PMID: 28663148 DOI: 10.1016/j.jep.2017.06.029

ETHNOPHARMACOLOGICAL RELEVANCE: Lignosus rhinocerotis (L. rhinocerotis), also known as the tiger milk mushroom, is widely used as traditional medicine and as soup ingredient in Malaysia and Hong Kong. It is edible and is used traditionally for the treatment of fever, cough, asthma, wounds, chronic hepatitis, gastric ulcers and cancers. In view of its safety profile, little information is found in scientific literature. The objective of this study is to investigate developmental toxicity of L. rhinocerotis in pregnant Sprague-Dawley (SD) rats.
MATERIALS AND METHODS: Eighty pregnant SD female rats were used in this study for three treatment groups and a control group, each consisting of 20 pregnant female rats. Three doses of 850mg/kg/day (Low-dose), 1700mg/kg/day (Mid-dose) and 3400mg/kg/day (High-dose) were selected for the study, whereas 10mL/kg distilled water was served as the control. Examinations were conducted on pregnant rats and fetuses respects to mortality, body weight, body weights gains, food consumption and clinical observations. The pregnant females were gross necropsied on G20, followed by maternal and fetus examination, to evaluate the teratogenicity, reproductive and developmental performance of L. rhinocerotis mycelium.
RESULTS: Results showed that no L. rhinocerotis mycelium-related animal death and abnormal clinical sign were noted. No statistical differences were noted in maternal mean body weight and maternal mean body weight gains. Some animals in the high-dose group appeared audible respiration due to dosing accident, it resulted in lower food consumption but not relevant to L. rhinocerotis mycelium treatment. In maternal gross necropsy, no L. rhinocerotis mycelium-related gross lesion was noted. In maternal examination, parameters of gravid uterus weight, implantation number, corpora lutea number, litter size, live or dead fetal number, male or female fetus number, resorption number, fetal sex ratio (M/F), pre-implantation loss and post-implantation loss were all within the normal reference ranges and showed no significant difference when compared to the control group. In fetus examination, including external, visceral and skeletal evaluations, there were no significant changes between any of the L. rhinocerotis mycelium treated groups and the control group.
CONCLUSIONS: Based on the study results, the no-observable-adverse-effect level (NOAEL) for pregnant female rats under the conditions of this study was 3400mg/kg/day.

Matched MeSH terms: Embryonic Development
Fulltext Effect of perivitelline fluid from horseshoe crab on the expression of COL1A1 in dental pulp stem cells

Amanina Fatinah Kamarudin, Najian Ibrahim, Thirumulu Ponnuraj Kannan, Ahmad Aizat Abdul Aziz

Archives of Orofacial Sciences, 2016;11(2):26-30.
MyJurnal

Perivitelline fluid, extracted from the fertilized eggs of horseshoe crabs, has been reported to play a
vital role in supporting embryogenesis as well as cell proliferation. The present study aims to evaluate the effect
of PVF on the expression of COL1A1 in human dental pulp stem cells (DPSCs). The cells were grouped into two;
untreated (control) and treated with a single dose of PVF (0.019 mg/ml). Gene expression was quantified for
COL1A1 on day 1, 3 and 7 using reverse transcriptase PCR. The expression of COL1A1 on day 3 of treated
group with PVF was the highest though there was a decline of COL1A1 expression on day 7. Mann Whitney test
was utilized to determine the significance of COL1A1 expression between treated and untreated groups.
Significant difference in the expression of COL1A1 was observed between the treated and untreated groups on
day 3 though there was no significance in the expression on day 7. The present study indicates that PVF may
have the potential to increase cell proliferation in human DPSCs.

Matched MeSH terms: Embryonic Development
Intracellular ROS scavenging and antioxidant regulation of WL15 from cysteine and glycine-rich protein 2 demonstrated in zebrafish in vivo model

Guru A, Lite C, Freddy AJ, Issac PK, Pasupuleti M, Saraswathi NT, et al.

Dev Comp Immunol, 2021 Jan;114:103863.
PMID: 32918928 DOI: 10.1016/j.dci.2020.103863

Antioxidant peptides are naturally present in food, especially in fishes, and are considered to contain rich source of various bioactive compounds that are structurally heterogeneous. This study aims to identify and characterize the antioxidant property of the WL15 peptide, derived from Cysteine and glycine-rich protein 2 (CSRP2) identified from the transcriptome of a freshwater food fish, Channa striatus. C. striatus is already studied to contain high levels of amino acids and fatty acids, besides traditionally known for its pharmacological benefits in the Southeast Asian region. In our study, in vitro analysis of WL15 peptide exhibited strong free radical scavenging activity in 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS), superoxide anion radical and hydrogen peroxide (H2O2) scavenging assay. Further, to evaluate the cytotoxicity and dose-response, the Human dermal fibroblast (HDF) cells were used. Results showed that the treatment of HDF cells with varying concentrations (10, 20, 30, 40 and 50 μM) of WL15 peptide was not cytotoxic. However, the treatment concentrations showed enhanced antioxidant properties by significantly inhibiting the levels of free radicals. For in vivo assessment, we have used zebrafish larvae for evaluating the developmental toxicity and for determining the antioxidant property of the WL15 peptide. Zebrafish embryos were treated with the WL15 peptide from 4 h of post-fertilization (hpf) to 96 hpf covering the embryo-larval developmental period. At the end of the exposure period, the larvae were exposed to H2O2 (1 mM) for inducing generic oxidative stress. The exposure of WL15 peptide during the embryo-larval period showed no developmental toxicity even in higher concentrations of the peptide. Besides, the WL15 peptide considerably decreased the intracellular reactive oxygen species (ROS) levels induced by H2O2 exposure. WL15 peptide also inhibited the H2O2-induced caspase 3-dependent apoptotic response in zebrafish larvae was observed using the whole-mount immunofluorescence staining. Overall results from our study showed that the pre-treatment of WL15 (50 μM) in the H2O2-exposed zebrafish larvae, attenuated the expression of activated caspase 3 expressions, reduced Malondialdehyde (MDA) levels, and enhanced antioxidant enzymes, including superoxide dismutase (SOD) and catalase (CAT). The gene expression of antioxidant enzymes such as glutathione S-transferase (GST), glutathione peroxide (GPx) and γ-glutamyl cysteine synthetase (GCS) was found to be upregulated. In conclusion, it can be conceived that pre-treatment with WL15 could mitigate H2O2-induced oxidative injury by elevating the activity and expression of antioxidant enzymes, thereby decreasing MDA levels and cellular apoptosis by enhancing the antioxidant response, demonstrated by the in vitro and in vivo experiments.

Matched MeSH terms: Embryonic Development
Fulltext Embryos of the viviparous dermapteran, Arixenia esau develop sequentially in two compartments: terminal ovarian follicles and the uterus

Tworzydlo W, Kisiel E, Bilinski SM

PLoS One, 2013;8(5):e64087.
PMID: 23667700 DOI: 10.1371/journal.pone.0064087

Three main reproductive strategies have been described among insects: most common oviparity, ovoviviparity and viviparity. In the latter strategy, the embryonic development takes place within the body of the mother which provides gas exchange and nutrients for embryos. Here we present the results of histological and EM analyses of the female reproductive system of the viviparous earwig, Arixenia esau, focusing on all the modifications related to the viviparity. We show that in the studied species the embryonic development consists of two "physiological phases" that take place in two clearly disparate compartments, i.e. the terminal ovarian follicle and the uterus. In both compartments the embryos are associated with synthetically active epithelial cells. We suggest that these cells are involved in the nourishment of the embryo. Our results indicate that viviparity in arixeniids is more complex than previously considered. We propose the new term "pseudoplacento-uterotrophic viviparity" for this unique two-phase reproductive strategy.

Matched MeSH terms: Embryonic Development/physiology*
Distinct developmental expression of two elongase family members in zebrafish

Tan SH, Chung HH, Shu-Chien AC

Biochem Biophys Res Commun, 2010 Mar 12;393(3):397-403.
PMID: 20138842 DOI: 10.1016/j.bbrc.2010.01.130

Despite the known importance of long-chained polyunsaturated fatty acids (LC-PUFA) during development, very little is known about their utilization and biosynthesis during embryogenesis. Combining the advantages of the existence of a complete range of enzymes required for LC-PUFA biosynthesis and the well established developmental biology tools in zebrafish, we examined the expression patterns of three LC-PUFA biosynthesis genes, Elovl2-like elongase (elovl2), Elovl5-like elongase (elovl5) and fatty acyl desaturase (fad) in different zebrafish developmental stages. The presence of all three genes in the brain as early as 24 hours post fertilization (hpf) implies LC-PUFA synthesis activity in the embryonic brain. This expression eventually subsides from 72 hpf onwards, coinciding with the initiation of elovl2 and fad expression in the liver and intestine, 2 organs known to be involved in adult fish LC-PUFA biosynthesis. Collectively, these patterns strongly suggest the necessity for localized production of LC-PUFA in the brain during in early stage embryos prior to the maturation of the liver and intestine. Interestingly, we also showed a specific expression of elovl5 in the proximal convoluted tubule (PCT) of the zebrafish pronephros, suggesting a possible new role for LC-PUFA in kidney development and function.

Matched MeSH terms: Embryonic Development/genetics
Fulltext Organotypic slice culture based on in ovo electroporation for chicken embryonic central nervous system

Yang C, Li X, Li S, Chai X, Guan L, Qiao L, et al.

J Cell Mol Med, 2019 03;23(3):1813-1826.
PMID: 30565384 DOI: 10.1111/jcmm.14080

Organotypic slice culture is a living cell research technique which blends features of both in vivo and in vitro techniques. While organotypic brain slice culture techniques have been well established in rodents, there are few reports on the study of organotypic slice culture, especially of the central nervous system (CNS), in chicken embryos. We established a combined in ovo electroporation and organotypic slice culture method to study exogenous genes functions in the CNS during chicken embryo development. We performed in ovo electroporation in the spinal cord or optic tectum prior to slice culture. When embryonic development reached a specific stage, green fluorescent protein (GFP)-positive embryos were selected and fluorescent expression sites were cut under stereo fluorescence microscopy. Selected tissues were embedded in 4% agar. Tissues were sectioned on a vibratory microtome and 300 μm thick sections were mounted on a membrane of millicell cell culture insert. The insert was placed in a 30-mm culture dish and 1 ml of slice culture media was added. We show that during serum-free medium culture, the slice loses its original structure and propensity to be strictly regulated, which are the characteristics of the CNS. However, after adding serum, the histological structure of cultured-tissue slices was able to be well maintained and neuronal axons were significantly longer than that those of serum-free medium cultured-tissue slices. As the structure of a complete single neuron can be observed from a slice culture, this is a suitable way of studying single neuronal dynamics. As such, we present an effective method to study axon formation and migration of single neurons in vitro.

Matched MeSH terms: Embryonic Development*
Fulltext Effect of sonic hedgehog on motor neuron positioning in the spinal cord during chicken embryonic development

Yang C, Li S, Li X, Li H, Li Y, Zhang C, et al.

J Cell Mol Med, 2019 05;23(5):3549-3562.
PMID: 30834718 DOI: 10.1111/jcmm.14254

Sonic hedgehog (SHH) is a vertebrate homologue of the secreted Drosophila protein hedgehog and is expressed by the notochord and floor plate in the developing spinal cord. Sonic hedgehog provides signals relevant for positional information, cell proliferation and possibly cell survival, depending on the time and location of expression. Although the role of SHH in providing positional information in the neural tube has been experimentally proven, the underlying mechanism remains unclear. In this study, in ovo electroporation was employed in the chicken spinal cord during chicken embryo development. Electroporation was conducted at stage 17 (E2.5), after electroporation the embryos were continued incubating to stage 28 (E6) for sampling, tissue fixation with 4% paraformaldehyde and frozen sectioning. Sonic hedgehog and related protein expressions were detected by in situ hybridization and fluorescence immunohistochemistry and the results were analysed after microphotography. Our results indicate that the ectopic expression of SHH leads to ventralization in the spinal cord during chicken embryonic development by inducing abnormalities in the structure of the motor column and motor neuron integration. In addition, ectopic SHH expression inhibits the expression of dorsal transcription factors and commissural axon projections. The correct location of SHH expression is vital to the formation of the motor column. Ectopic expression of SHH in the spinal cord not only affects the positioning of motor neurons, but also induces abnormalities in the structure of the motor column. It leads to ventralization in the spinal cord, resulting in the formation of more ventral neurons forming during neuronal formation.

Matched MeSH terms: Embryonic Development/genetics*
Isolation and characterization of differentially expressed transcripts from the suspension cells of oil palm (Elaeis guineensis Jacq.) in response to different concentration of auxins

Roowi SH, Ho CL, Alwee SS, Abdullah MO, Napis S

Mol Biotechnol, 2010 Sep;46(1):1-19.
PMID: 20390382 DOI: 10.1007/s12033-010-9262-9

Oil palm suspension cultures were initiated by transferring the gel-like friable embryogenic tissue onto liquid medium supplemented with auxins. In this study, transcripts that were differentially expressed in oil palm suspension cells cultured at different auxin concentrations were examined using suppression subtractive hybridization. Total RNA was first isolated from oil palm suspension cells proliferated in liquid medium with different hormone concentrations for 6 months. Four different hormone combinations: T1 (0.1 mg/l 2,4-D and 1.0 mg/l NAA), T2 (0.4 mg/l 2,4-D and 1.0 mg/l NAA), T3 (1.0 mg/l NAA), and T4 (0.4 mg/l 2,4-D) were used for the treatments. The first and second subtractions were performed using samples T1 and T2 in forward and reverse order. The other two subtractions were forward and reverse subtractions of T3 and T4, respectively. Reverse northern analyses showed that 14.13% of these clones were preferentially expressed in T1, 13.70% in T2, 14.75% in T3, and 15.70% in T4. Among the 294 cDNA clones that were sequenced, 61 contigs (assembled from 165 sequences) and 129 singletons were obtained. Among the 61 contigs, 10 contigs consist of sequences from treatment T1, 8 contigs were from treatment T2, 10 contigs were contains sequences of treatment T3 and 13 contigs contains sequences of treatment T4. Northern analyses of five transcripts that were shown to be differentially expressed in the oil palm suspension cells by reverse northern analysis revealed that transcripts 16A1 (a putative lignostilbene-alpha,beta-dioxygenase, EgLSD) and 16H12 (a putative ethylene responsive 6, EgER6) were differentially expressed in oil palm suspension cells treated with different levels of auxin.

Matched MeSH terms: Embryonic Development/drug effects; Embryonic Development/genetics
Fulltext Diverse roles of prostaglandins in blastocyst implantation

Salleh N

ScientificWorldJournal, 2014;2014:968141.
PMID: 24616654 DOI: 10.1155/2014/968141

Prostaglandins (PGs), derivatives of arachidonic acid, play an indispensable role in embryo implantation. PGs have been reported to participate in the increase in vascular permeability, stromal decidualization, blastocyst growth and development, leukocyte recruitment, embryo transport, trophoblast invasion, and extracellular matrix remodeling during implantation. Deranged PGs syntheses and actions will result in implantation failure. This review summarizes up-to-date literatures on the role of PGs in blastocyst implantation which could provide a broad perspective to guide further research in this field.

Matched MeSH terms: Embryonic Development/drug effects
Fulltext Epigenetic modification does not determine the time of POU5F1 transcription activation in cloned bovine embryos

Jafari S, Hosseini SM, Hajian M, Forouzanfar M, Jafarpour F, Abedi P, et al.

J Assist Reprod Genet, 2011 Nov;28(11):1119-27.
PMID: 22020531 DOI: 10.1007/s10815-011-9638-1

To investigate the effect of epigenetic modification on pattern, time and capacity of transcription activation of POU5F1, the key marker of pluripotency, in cloned bovine embryos.

Matched MeSH terms: Embryonic Development/genetics
Time dependent effect of chronic embryonic exposure to ethanol on zebrafish: Morphology, biochemical and anxiety alterations

Ramlan NF, Sata NSAM, Hassan SN, Bakar NA, Ahmad S, Zulkifli SZ, et al.

Behav Brain Res, 2017 08 14;332:40-49.
PMID: 28559182 DOI: 10.1016/j.bbr.2017.05.048

Exposure to ethanol during critical period of development can cause severe impairments in the central nervous system (CNS). This study was conducted to assess the neurotoxic effects of chronic embryonic exposure to ethanol in the zebrafish, taking into consideration the time dependent effect. Two types of exposure regimen were applied in this study. Withdrawal exposure group received daily exposure starting from gastrulation until hatching, while continuous exposure group received daily exposure from gastrulation until behavioural assessment at 6dpf (days post fertilization). Chronic embryonic exposure to ethanol decreased spontaneous tail coiling at 24hpf (hour post fertilization), heart rate at 48hpf and increased mortality rate at 72hpf. The number of apoptotic cells in the embryos treated with ethanol was significantly increased as compared to the control. We also measured the morphological abnormalities and the most prominent effects can be observed in the treated embryos exposed to 1.50% and 2.00%. The treated embryos showed shorter body length, larger egg yolk, smaller eye diameter and heart edema as compared to the control. Larvae received 0.75% continuous ethanol exposure exhibited decreased swimming activity and increased anxiety related behavior, while withdrawal ethanol exposure showed increased swimming activity and decreased anxiety related behavior as compared to the respective control. Biochemical analysis exhibited that ethanol exposure for both exposure regimens altered proteins, lipids, carbohydrates and nucleic acids of the zebrafish larvae. Our results indicated that time dependent effect of ethanol exposure during development could target the biochemical processes thus leading to induction of apoptosis and neurobehavioral deficits in the zebrafish larvae. Thus it raised our concern about the safe limit of alcohol consumption for pregnant mother especially during critical periods of vulnerability for developing nervous system.

Matched MeSH terms: Embryonic Development/drug effects

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