Pattern Of Embryonic Development Research Articles

Genomic and Transcriptional Analysis of the Necroptosis Pathway Elements RIPK and MLKL in Sea Cucumber, Holothuria leucospilota.

Background: Receptor-interacting protein kinases (RIPKs) and mixed-lineage kinase domain-like protein (MLKL) are crucial in regulating innate immune responses and cell death signaling (necroptosis and apoptosis), and are potential candidates for genetic improvement in breeding programs. Knowledge about the RIPK family and MLKL in sea cucumber remains limited. Methods: We searched the genomes of sea cucumber Holothuria leucospilota for genes encoding RIPKs and MLKL, performed phylogenetic tree, motif and functional domain analyses, and examined tissue distribution and embryonic development patterns using qPCR. Results: RIPK5 (Hl-RIPK5), RIPK7 (Hl-RIPK7) and MLKL (Hl-MLKL) were identified in sea cucumber H. leucospilota. Hl-RIPK5 and Hl-RIPK7 were mainly expressed in coelomocytes, suggesting that they play a role in innate immunity, whereas Hl-MLKL exhibited relatively low expression across tissues. During embryonic development, Hl-MLKL was highly expressed from the 2-cell stage to the morula stage, while Hl-RIPK5 and Hl-RIPK7 were primarily expressed after the morula stage, indicating different roles in embryonic development. In primary coelomocytes, Hl-RIPK5 transcriptional activity was significantly depressed by LPS, poly(I:C), or pathogen Vibrio harveyi. Hl-RIPK7 expression levels were unchanged following the same challenges. Hl-MLKL mRNA levels were significantly decreased with poly(I:C) or V. harveyi, but did not change with LPS. Conclusions: These findings provide valuable insights into the evolutionary tree and characterization of RIPK and MLKL genes in sea cucumber, contributing to the broader understanding of the RIPK gene family and MLKL in ancient echinoderms.

The typical developmental trajectory and energy requirements of Shitou goose during the embryonic stage

Low hatchability has been a persistent challenge in the goose industry. Establishing standard atlases and comprehending embryonic development patterns are essential to improving the hatching rates of goose eggs. However, comprehensive descriptions of normal atlases, embryonic development, and energy requirements in geese are lacking. In this study, a total of 120 fertile eggs from well-known large Shitou goose were incubated using 12 nesting purebred female geese. During hatching, both the temperature of the eggshells and the weight of eggs were recorded, and daily photographs of the embryos were captured to monitor their development closely. After hatching, count the number of pores per unit area by choosing eggs without sperm, dead embryos, or abnormal shells. Furthermore, 150 Shitou goose eggs were hatched by automatic incubator, with adjustments made based on observed normal developmental stages that incubated by female geese. The eggs were carefully opened to meticulously document embryonic morphology and create a detailed development map. Measurements were taken of the eye diameter, length of the lower beak, tarsometatarsus bone, and embryo length. Subsequently, an analysis was conducted to assess the calcium, phosphorus, crude protein, and crude fat content to study the energy requirements for embryo development. As a result, the hatch rate of the incubator reached 86.67%, and the cumulative water loss rate increased with embryo age. Notably, normally developing embryos displayed a significantly higher number of pores on the eggshell surface compared to dead embryos (P < 0.05). Additionally, embryonic body length, eyeball diameter, and lower beak length exhibited continuous growth until day 19 of incubation, while tarsometatarsus length increased steadily from days 12 to 31. Liver size measurement began on the 10th day of incubation, while both leg and chest muscles showed continuous growth from the 12th day. For energy demand, the embryo primarily relied on protein sourced from the egg yolk within the first 10 days of development. Afterward, the egg yolk provided both protein and fat for embryonic growth. In summary, this study has generated a comprehensive developmental map for Shitou goose embryos, offering valuable insights into their growth and morphological changes throughout the incubation period. This map can serve as a reference for optimizing machine incubation techniques to enhance goose egg hatching rates and provide fresh perspectives on the development of geese.

From Snapshots to Development: Identifying the Gaps in the Development of Stem Cell-based Embryo Models along the Embryonic Timeline.

In recent years, stem cell‐based models that reconstruct mouse and human embryogenesis have gained significant traction due to their near‐physiological similarity to natural embryos. Embryo models can be generated in large numbers, provide accessibility to a variety of experimental tools such as genetic and chemical manipulation, and confer compatibility with automated readouts, which permits exciting experimental avenues for exploring the genetic and molecular principles of self‐organization, development, and disease. However, the current embryo models recapitulate only snapshots within the continuum of embryonic development, allowing the progression of the embryonic tissues along a specific direction. Hence, to fully exploit the potential of stem cell‐based embryo models, multiple important gaps in the developmental landscape need to be covered. These include recapitulating the lesser‐explored interactions between embryonic and extraembryonic tissues such as the yolk sac, placenta, and the umbilical cord; spatial and temporal organization of tissues; and the anterior patterning of embryonic development. Here, it is detailed how combinations of stem cells and versatile bioengineering technologies can help in addressing these gaps and thereby extend the implications of embryo models in the fields of cell biology, development, and regenerative medicine.

Postpartum Uterine Involution and Embryonic Development Pattern in Chinese Holstein Dairy Cows.

Understanding the postpartum uterine involution pattern and embryonic development could facilitate bovine reproduction management, improve reproductive efficiency, and diagnosis of the reproductive disorder, which would contribute to the success of the dairy business. This study aimed to investigate postpartum uterine involution and embryonic developmental patterns or postconceptional marks of embryonic fetal development in Chinese Holstein dairy cows using B-mode ultrasonography. The results revealed a significant decline in the involution period with an increase of parity and age. The uterine involution period was shorter in multiparous cows when compared with cows with lower parities. Consistently, cows over 4 years old recovered faster than younger cows (2 or 3 years). Besides, the elder cows (over 4 years) had a relatively larger size of resumed cervix uteri and horns. Postpartum uterine involution pattern analysis revealed that the reproductive tract recovered very fast during the first 16 days postpartum for all the parity. Results of postconceptional marks of embryo development revealed a slow increase in diameter of the gravid uterine horn and crown-rump length (CRL) before day 60. In contrast, this increase was dramatic and rapid after the 60th day. We also established two models to estimate gestational age based on gravid uterine horn diameter or CRL. A formula was established to determine the gravid uterine horn size during postconceptional on day 30th–day 90th (r = 0.8714, P < 0.01). In addition, a significant positive correlation between CRL and gestational age (r = 0.98151, P < 0.01) was built. In conclusion, these results illustrated that parity and calving age had significant effects on uterine involution in Chinese Holstein cows. Crown-rump length and gravid uterine horn diameter are both efficient for evaluating the embryo growth. These current findings broaden the understanding of basic reproductive pattern in Chinese Holstein cows and could benefit bovine reproductive management primarily in postpartum and early pregnant cows to reduce the calving interval and avoid periparturient metabolic diseases.

Comparative embryonic development patterns in three deep-water skates from the southwest Atlantic

Embryonic development is a vulnerable and key period during the life cycle of an oviparous elasmobranch. Captivity studies are difficult to carry out on non-coastal species; hence, embryonic development can be analysed from samples collected from the seabed. Here, embryonic development of three shelf and deep-water skates from the southwest Atlantic Ocean, Bathyraja brachyurops, B. macloviana and Amblyraja doellojuradoi was studied. Egg cases containing embryos in different stages were collected from 84 to 1006 m depth in the northern part of the Argentinean continental shelf and continental slope (36°S-41°S), including the Mar del Plata Canyon (38° S). Common development patterns were observed among the three species and also with other skate species previously studied. Anatomical structures and embryo features were similar among species in initial, early and middle stages. Advanced and pre-hatching embryos showed species-specific differences regarding pigmentation and spinulation. This study sheds light on the identification of neonates of shelf and deep-water species, which could be useful to recognize specific nursery areas in the deep ocean. In addition, this embryological comparative study expands the existing database on the different chondrichthyan lineages, thus making a step forward towards understanding of their phylogenetic relationships.

Embryonic and early larval development of two marine angelfish, Centropyge bicolor and Centropyge bispinosa.

Marine angelfish (family: Pomacanthidae) are among the most sought-after fish species in the saltwater aquarium trade. However, there is a lack of information in the literature on their early ontogeny. The objective of this study was to describe the embryonic and early larval development of two dwarf angelfish, the bicolour angelfish, Centropyge bicolor and the coral beauty angelfish, Centropyge bispinosa. The eggs of these two species were obtained from spontaneous spawning of the broodstock fish in captivity and incubated at 26.0 ± 0.2°C throughout the study. Fertilized eggs (n = 15) of both species are transparent, pelagic and spherical; the mean diameters of the eggs were measured at 703.6 ± 7.8 μm for C. bicolor and 627.6 ± 7.8 μm for C. bispinosa. The eggs of both species possessed a narrow perivitelline space, smooth and thin chorion, a homogenous and non-segmented yolk as well as a single oil globule. Overall, the observed embryonic development pattern of C. bicolor and C. bispinosa was very similar, and the main difference was the embryonic pigmentation pattern, which only became evident close to hatching. Larvae of both species started hatching at 13 h 30 min after fertilization, and the larval characteristics of both species also showed high levels of similarities. However, the mouth opening time for C. bicolor was 72 h after hatching (AH) and 96 AH for C. bispinosa. In general, the observed early ontogeny of C. bicolor and C. bispinosa also resembled that of other Centropyge species documented in the literature.

Construction of multicellular aggregate by E-cadherin coated microparticles enhancing the hepatic specific differentiation of mesenchymal stem cells

The differentiation of human mesenchymal stem cells (hMSCs) into hepatocyte-like cells in vitroprovides a promising candidate for cell therapy of liver diseases, and cell aggregates have been proposed to improve the efficiency of expansion and differentiation. Previously, we engineered multicellular aggregates incorporating human E-cadherin fusion protein (hE-cad-Fc)-coated poly(lactic-co-glycolic acid) (PLGA) microparticles (hE-cad-PLGAs), and a significant improvement was obtained in both cellular proliferation of and cytokine secretion by hMSCs. In this study, hepatic differentiation of hMSCs was induced by a biomimetic microenvironment consisting of these engineered aggregates and a cocktail of specific cytokines. The ratio of hE-cad-PLGAs to hMSCs in engineered hMSCs aggregates was optimized to 1:3 for hepatic differentiation. The expressions of hepatic-specific markers were significantly promoted, and cell polarity and activated drug metabolism enzymes were established in MSC/hE-cad-PLGA aggregates compared with MSC and MSC/PLGA aggregates. Moreover, the expressions of stemness and definitive endoderm markers confirmed effectively induced endoderm differentiation in MSC/hE-cad-PLGA aggregates, which was consistent with the pattern of embryonic development. After pre-differentiation for 1 week, the MSC/hE-cad-PLGA aggregates continuously progressed the hepatic phenotype expression in healthy rat peritoneum. Therefore, the biomimetic microenvironment constructed by hE-cad-PLGAs in engineered multicellular aggregates was able to promote the process of endoderm differentiation and the subsequent hepatic differentiation of hMSCs. It would be appropriate for applied research in hepatotoxic drug screening and cell-based treatment of liver diseases. By optimizing with other cytokine cocktail, the engineered multicellular aggregates can be applied to the construction of other endoderm-derived organs. Statement of significanceThe differentiation of mesenchymal stem cells (MSCs) into hepatocyte-like cells in vitroprovides a promising for cell therapy for liver diseases, and cell aggregates have been proposed to improve the expansion and differentiation efficiency. Here, engineered multicellular aggregates were constructed by E-cadherin modified microparticles (hE-cad-PLGAs) construct a biomimetic microenvironment to promote the process of endoderm differentiation and the subsequent hepatic differentiation of hMSCs. Furthermore, after pre-differentiation for 1 week, the MSC/hE-cad-PLGA aggregates continuously progressed the hepatic phenotype expression in healthy rat peritoneum. Therefore, engineered multicellular aggregates with hE-cad-PLGAs would be appropriate for applied research in hepatotoxic drug screening and cell-based treatment of liver diseases, and provide a promising method in the construction of other endoderm-derived organs.

Synthetic circuits reveal how mechanisms of gene regulatory networks constrain evolution

Phenotypic variation is the raw material of adaptive Darwinian evolution. The phenotypic variation found in organismal development is biased towards certain phenotypes, but the molecular mechanisms behind such biases are still poorly understood. Gene regulatory networks have been proposed as one cause of constrained phenotypic variation. However, most pertinent evidence is theoretical rather than experimental. Here, we study evolutionary biases in two synthetic gene regulatory circuits expressed in Escherichia coli that produce a gene expression stripe—a pivotal pattern in embryonic development. The two parental circuits produce the same phenotype, but create it through different regulatory mechanisms. We show that mutations cause distinct novel phenotypes in the two networks and use a combination of experimental measurements, mathematical modelling and DNA sequencing to understand why mutations bring forth only some but not other novel gene expression phenotypes. Our results reveal that the regulatory mechanisms of networks restrict the possible phenotypic variation upon mutation. Consequently, seemingly equivalent networks can indeed be distinct in how they constrain the outcome of further evolution.

DNA damage in human spermatozoa; important contributor to mutagenesis in the offspring.

Nuclear DNA damage in spermatozoa could potentially have a major impact on the fertilizing capacity of these cells, their ability to establish a normal pattern of embryonic development as well as the health and wellbeing of subsequent offspring. Many laboratory techniques have been developed to assess this damage focusing on strand breaks, chromatin stability following exposure to extreme pH conditions and the formation of DNA adducts. Of particular importance may be the dominant role played by oxidative stress in the etiology of defective sperm function and DNA damage. The oxidative base lesions created via this mechanism have the potential to generate genetic and epigenetic mutations in the offspring that could have a profound impact on the latter’s long-term health trajectory. Moreover if oxidative stress is the cause of DNA damage in spermatozoa then there is a potential role for antioxidant therapy in the resolution of this problem, which deserves investigation.

Dead-in-shell positions of near-term ostrich embryos

The patterns of embryonic development in ostriches, especially in the last stage of hatching, are still not well understood. This study examined between 3468 and 3484 dead-in-shell (DIS) eggs with chicks that died between day 35 and day 42 of artificial incubation. Most DIS chicks were positioned correctly with their heads towards the air cell (52.6%). DIS chicks that positioned their heads near the equator of the egg amounted to 46.5%, while a small percentage (0.9%) were positioned with their upper body towards the bottom of the egg. More DIS chicks tended to pip internally near the equator of the egg (37.6%) than DIS chicks that pipped internally through the membranes into the air cell (34.4%). Most DIS chicks had their heads turned in the correct position from left to right (54.4%), though their beaks were mostly positioned towards the air cell (52.9%). The highest percentage of DIS chicks had their feet in the upwards position (52.4%), while 46% had their feet across or below the head. The wings of all DIS chicks were positioned next to the body. Results from the study showed that most of the DIS chicks were roughly in the correct position, but were still unable to hatch. This warrants future research to investigate the reasons that prevent correctly positioned chicks from hatching.

Embryological evidence of a new type of seromucous labial gland in neotropical snail-eating snakes of the genus Sibynomorphus

Snail-eating dipsadine snakes of the genera Sibynomorphus and Dipsas share infralabial glands divided into a distinct lateral portion along the lower lip (il1) and a ventrolateral portion along the mandible (il2). While il1 is constituted by several small and individual glands with their short ducts opening along the margin of the lower lip, il2 is constituted by a single hypertrophied gland with a single duct that opens laterally at the level of the intermandibular raphe. This unique condition seems to be restricted to the goo-eating dipsadine snakes. Here, we describe the embryonic development of the labial glands in the goo-eating dipsadine snakes Sibynomorphus mikanii and S. neuwiedi. Our results indicate that il2 developed independently from il1, being formed through a hypertrophied invagination instead of deriving from several small independent invaginations, as is the case for il1. Additionally, il2 showed a distinct timing of development, starting in younger embryological stages, prior to the development of il1 and other labial glands. The pattern of embryonic development observed for il2 supports the hypothesis that this gland evolved separately from the infralabial glands in both Sibynomorphus and Dipsas, as a novel, independent protein-secretion system associated with their specialized feeding behaviors.

A study of embryonic development in eriophyoid mites (Acariformes, Eriophyoidea) with the use of the fluorochrome DAPI and confocal microscopy.

The embryonic development of four eriophyoid mite species, Cecidophyopsis ribis, Phytoptus avellanae, Oziella liroi and Loboquintus subsquamatus, has been studied with the use of fluorochrome DAPI and confocal microscopy. The first three nuclear divisions occur on the egg periphery (the groups of 2, 4, and 6 nuclei have been recorded), while the biggest part of yolk remains undivided. After four or five nuclear divisions all nuclei are situated only in one sector of the embryo, while other sectors contain only yolk suggesting possible meroblastic cleavage. Later, the formation of superficial blastoderm takes place. A few large yolk cells are situated inside the embryo. Germ band formation initiates as funnel-like cell invagination and leads to formation of a typical stage with four paired prosomal buds (chelicerae, palps, legs I and II). Each palp contains two lobes (anterior and posterior), the adult subcapitulum is presumably a fusion product of the anterior pair of the lobes. Neither rudiments of legs III and IV, traces of opisthosomal segments nor remnants of the prelarval exuvium under the egg shell were detected. Overall, the pattern of embryonic development in eriophyoids re-emphasizes the peculiarity of this ancient group of miniaturized phytoparasitic animals, and invites researches to pursue a deeper investigation of various fundamental aspects of this aberrant group of Acari. Further studies using various fluorescent dyes and transmission electron microscopy are needed to visualize plasma membranes and clarify the pattern of early cleavage of eriophyoids.

Differences in egg nutrient availability, development, and nutrient metabolism of broiler and layer embryos

Selection for production traits of broilers and layers leads to physiological differences, which may already be present during incubation. This study aimed to investigate the influence of strain (broiler vs layer) on egg nutrient availability, embryonic development and nutrient metabolism. A total of 480 eggs with an egg weight range of 62.0 to 64.0 g from Lohmann Brown Lite and Ross 308 breeder flocks of 41 or 42 weeks of age were selected in two batches of 120 eggs per batch per strain. For each batch, 30 eggs per strain were used to determine egg composition, including nutrient and energy content, and 90 eggs per strain were separately incubated in one of two climate respiration chambers at an eggshell temperature of 37.8°C. The results showed that broiler eggs had a higher ratio of yolk: albumen with 2.41 g more yolk and 1.48 g less albumen than layers. The yolk energy content of broiler eggs was 46.32 kJ higher than that of layer eggs, whereas total energy content of broiler eggs was 47.85 kJ higher compared to layer eggs. Yolk-free body mass at incubation day 16 and chick weight and length at hatch were higher in broilers compared to layers. Respiration quotient of broiler embryos was higher than layer embryos during incubation day 8 to incubation day 10. A 0.24 g lower residual yolk at the hatch of broiler embryos than for the layer embryos indicated that broiler embryos used more yolk and had a higher energy utilization and energy deposition in yolk-free body mass. Heat production of broiler embryos was higher than that of layer embryos from incubation day 12 to incubation day 18, but efficiency of converting egg energy used by embryos to form yolk-free body mass was similar. In conclusion, broiler and layer embryos have different embryonic development patterns, which affect energy utilization and embryonic heat production. However, the embryos are equal in efficiency of converting the energy used to yolk-free body mass.

Evolution of viviparous reproduction in Paleozoic and Mesozoic reptiles.

Although viviparity (live-bearing reproduction) is widely distributed among lizards and snakes, it is entirely absent from other extant Reptilia and many extinct forms. However, paleontological evidence reveals that viviparity was present in at least nine nominal groups of pre-Cenozoic reptiles, representing a minimum of six separate evolutionary origins of this reproductive mode. Two viviparous clades (sauropterygians and ichthyopterygians) lasted more than 155 million years, a figure that rivals the duration of mammalian viviparity. Circumstantial evidence indicates that extinct viviparous reptiles had internal fertilization, amniotic fetal membranes, and placentas that sustained developing embryos via provision of respiratory gases, water, calcium, and possibly organic nutrients. Production of offspring via viviparity facilitated the invasion of marine habitats in at least five reptilian lineages. Thus, this pattern of embryonic development and reproduction was central to the ecology and evolution of these ancient animals, much as it is to numerous extant species of vertebrates.

Led by the nose

Hagfish embryos show developmental features that contradict the idea that these jawless fish are the most primitive living vertebrates. The findings also help to trace the evolution of vertebrate cranial structure. See Article p.175 The hagfish is the most primitive extant vertebrate. Its anatomy and development hold clues to understanding the evolution of that most enigmatic of structures, the vertebrate head. Hagfish embryos are notoriously hard to obtain, but a few years ago Shigeru Kuratani and colleagues managed to prepare some, and — for the first time since 1899 — significant work on hagfish embryology began. This paper presents the first detailed report on the craniofacial development of a hagfish species, Eptatretus burgeri. The authors identify an embryonic development pattern shared only by the jawless lampreys and hagfish that may have been primitive for all vertebrates.

Observation on morphological structure of the early development of Theragra chalcogramma

The embryonic development pattern of walleye pollock and the morphological characteristics of fertilized eggs were observed and recorded from October to December in 2008.The egg diameters in six stages were measured and the egg envelopes of seven stages were also studied with scanning electron microscopy.The egg of walleye pollock was pelagic,without oil globules.The diameter was 1.45-1.58 mm,the incubated temperature was(6.3±1.24) ℃ and the salinity was 34 psu.The microscopic observation showed the fertilization tube opened with(4.33±0.31) μm pore diameter;the pore canals of the egg envelope were inconspicuous;the interval of micropyle and density of the pore canals of the egg envelope were(1.46±0.31) μm and 21.1-30.6/100 μm2 respectively.The micropyle with the micropyle canal was completely plugged with material at the 2-cell stage,and the fertilization pore diameter was(5.26±0.29) μm.The egg envelope contained less ruffles and the pore canals of the egg envelope became more prominent and the interval of micropyle was(2.04±0.35) μm.The density of the pore canals of the egg envelope was 14.7-20.3/100 μm2.After the closing phase of blastopore,the fertilization pore of egg presented the open state;few secretions were adhered to its periphery region.The diameter of the fertilization pore,interval of the pore canals and density of the pore canals of the egg envelope were(4.48±0.15) μm;(1.70±0.23) μm;and 23.59-27.53/100 μm2,respectively.Approaching the incubation period,the egg envelope had a rough surface,on which the pore canals were inconspicuous with a(2.56±0.22) μm interval and the density was 12.38-15.97/100 μm2.The diameter of egg,the fertilization pore,as well as the dynamic change of structure of egg envelope,were closely related to the embryonic development of Alaska pollock.The above indexes had great importance in improving the fertility rate of egg,facilitating the normal development and spread of egg on seabed,and preventing both polyspermy and bacterial infection during the egg development process.

Evolution of anterior Hox regulatory elements among chordates

BackgroundThe Hox family of transcription factors has a fundamental role in segmentation pathways and axial patterning of embryonic development and their clustered organization is linked with the regulatory mechanisms governing their coordinated expression along embryonic axes. Among chordates, of particular interest are the Hox paralogous genes in groups 1-4 since their expression is coupled to the control of regional identity in the anterior nervous system, where the highest structural diversity is observed.ResultsTo investigate the degree of conservation in cis-regulatory components that form the basis of Hox expression in the anterior nervous system, we have used assays for transcriptional activity in ascidians and vertebrates to compare and contrast regulatory potential. We identified four regulatory sequences located near the CiHox1, CiHox2 and CiHox4 genes of the ascidian Ciona intestinalis which direct neural specific domains of expression. Using functional assays in Ciona and vertebrate embryos in combination with sequence analyses of enhancer fragments located in similar positions adjacent to Hox paralogy group genes, we compared the activity of these four Ciona cis-elements with a series of neural specific enhancers from the amphioxus Hox1-3 genes and from mouse Hox paralogous groups 1-4.ConclusionsThis analysis revealed that Kreisler and Krox20 dependent enhancers critical in segmental regulation of the hindbrain appear to be specific for the vertebrate lineage. In contrast, neural enhancers that function as Hox response elements through the action of Hox/Pbx binding motifs have been conserved during chordate evolution. The functional assays reveal that these Hox response cis-elements are recognized by the regulatory components of different and extant species. Together, our results indicate that during chordate evolution, cis-elements dependent upon Hox/Pbx regulatory complexes, are responsible for key aspects of segmental Hox expression in neural tissue and appeared with urochordates after cephalochordate divergence.

Genome-Wide Analysis of Sox Genes in Medaka (Oryzias latipes) and Their Expression Pattern in Embryonic Development

Genes of the Sox family encode evolutionarily conserved high-mobility group box containing transcription factors, which play key roles in various events of developmental contexts. In this study, we identified 15 sox genes by searching for the high-mobility group domain in the medaka genome and by polymerase chain reaction using primers designed from the results obtained from homology protein alignment. All medaka sox genes except a novel sox gene, Olsox32, are encoded in 5 groups as follows: 4 sox genes in group B; 3 sox genes in group D and F, respectively; 2 sox genes in group C and E, respectively, while no sox genes were found in groups A, G, H, I, and J. The medaka Olsox32 does not fall within any of the previously defined groups A–J. Here we have assigned it to a new group K. Together with the Sox protein sequences of other species, the phylogenetic relationship was analyzed and compared. Our findings point to recent sox gene loss, duplication and divergence occurring during the evolution of tetrapod and teleost lineages. The expression pattern shows that sox genes play a variety of roles in the early embryonic development of medaka.

PLANNING ON HATCHING TIME OF GROUPER EGGS THROUGH DIFFERENT INCUBATION TEMPERATURES

&lt;p&gt;Groupers were known as a high economically marine commodity and in order to support groupers production, the seed availability was the most important. Eggs are still as limited factor in hatchery production, for this reason the success of eggs transportation is one as base of successful production of seed. Planning on hatching time of eggs through different incubation temperature was an option to solve that problem. This experiment was aimed to find out the optimum temperature for groupers eggs and the minimum temperature to arrange incubation time and to plan the hatching time. Fertilized eggs were incubated into three beaker glasses of 1 liter in volume with the density of ± 250 eggs/liter. The incubation was done under laboratory condition at controlled temperature, i.e. (A) 21-22 ºC, (B) 24-25 ºC, (C) 27-28 ºC and (D) 30-31 ºC. The eggs that used were including orange spotted grouper (Epinephelus coiodes), brown marbled grouper (E. microdon), tiger grouper (E. fuscoguttatus) and humpback grouper (Cromileptes altivelis). Investigated variables were embryonic development pattern, incubation time and hatching rate. The result showed that the eggs incubated in temperature range of 24-31°C had the normal sequence of embryonic development pattern, but in temperature of 21-22°C performed irregular sequence and the embryonic development stopped at blastula or gastrula stage or even the eggs could still develop but the body of hatched larvae were abnormal. In lower temperature incubation, the incubation time was longer and the hatching rate of eggs was lower than those in higher temperature. Therefore the optimum temperature for incubation of orange spotted grouper, marbled grouper, tiger grouper and humpback grouper eggs ranged between 24-31 ºC, while the lowest possible temperature was 24 ºC.&lt;/p&gt;&lt;p&gt;Keywords: incubation temperature, embryonic development pattern, grouper eggs, hatching rate&lt;/p&gt;

PLANNING ON HATCHING TIME OF GROUPER EGGS THROUGH DIFFERENT INCUBATION TEMPERATURES

Groupers were known as a high economically marine commodity and in order to support groupers production, the seed availability was the most important. Eggs are still as limited factor in hatchery production, for this reason the success of eggs transportation is one as base of successful production of seed. Planning on hatching time of eggs through different incubation temperature was an option to solve that problem. This experiment was aimed to find out the optimum temperature for groupers eggs and the minimum temperature to arrange incubation time and to plan the hatching time. Fertilized eggs were incubated into three beaker glasses of 1 liter in volume with the density of ± 250 eggs/liter. The incubation was done under laboratory condition at controlled temperature, i.e. (A) 21-22 ºC, (B) 24-25 ºC, (C) 27-28 ºC and (D) 30-31 ºC. The eggs that used were including orange spotted grouper (Epinephelus coiodes), brown marbled grouper (E. microdon), tiger grouper (E. fuscoguttatus) and humpback grouper (Cromileptes altivelis). Investigated variables were embryonic development pattern, incubation time and hatching rate. The result showed that the eggs incubated in temperature range of 24-31°C had the normal sequence of embryonic development pattern, but in temperature of 21-22°C performed irregular sequence and the embryonic development stopped at blastula or gastrula stage or even the eggs could still develop but the body of hatched larvae were abnormal. In lower temperature incubation, the incubation time was longer and the hatching rate of eggs was lower than those in higher temperature. Therefore the optimum temperature for incubation of orange spotted grouper, marbled grouper, tiger grouper and humpback grouper eggs ranged between 24-31 ºC, while the lowest possible temperature was 24 ºC.Keywords: incubation temperature, embryonic development pattern, grouper eggs, hatching rate