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Accumulation kinetics of cotton late embryogenesis-abundant mRNAs and storage protein mRNAs: coordinate regulation during embryogenesis and the role of abscisic acid.

The accumulation of total RNA transcripts of 18 late embryo-abundant (Lea) gene families, each encoding two closely related Lea mRNAs, was measured in cotyledon total RNA during embryogenesis and germination of Gossypium hirsutum L. by RNA dot hybridization. Transcript abundance of the three storage protein families was also followed. The Lea mRNAs belong to only two related groups of commonly regulated mRNAs. The transcript level of each of the 6 members of Class I has two transient maxima during early maturation and a maximum level at 3 days prior to desiccation. The transcript level of each of the 12 members of Class II increases abruptly in late maturation with a maximum concentration at 3 days before desiccation (Class IIA) or at desiccation (Class IIB). Several patterns of early accumulation also exist within Class II, some overlapping with those of storage protein or Lea Class I mRNAs. The concentrations of Lea mRNAs increase at least 10- to 1700-fold during embryogenesis and decline 15- to 220-fold during the first day of germination. Earlier studies indicated that most Lea mRNAs, but no storage protein mRNAs, are induced in excised embryos exposed to abscisic acid (ABA). Free (+)ABA was measured during embryogenesis using a monoclonal antibody-based ELISA. The ABA concentration shows maxima correlating with the maxima in early Class I Lea mRNA abundance. However, ABA declines during the accumulation of Class I and Class II Lea mRNAs in late embryogenesis. Consequently, ABA could be an endogenous regulator of Class I Lea mRNAs during early maturation but is not the primary regulator of Lea mRNAs during late maturation. In contrast, cotyledon water potential exhibits a decrease that correlates with the late induction of Lea mRNAs. These findings are consistent with the hypothesis that low water potential is the endogenous regulator of some ABA-inducible mRNAs. The cessation of vascular flow to the embryo may also be involved in the disappearance of storage protein mRNAs and the late induction of Lea mRNAs.

Abscisic Acid

Integrated metabolomics, transcriptional, and physicochemical analysis reveals key metabolites and genes associated with somatic embryogenesis in Phyllostachys pubescens.

Phyllostachys pubescens (Moso bamboo) is a significant perennial crop species that provides valuable nutritional and industrial uses, as well as carbon sequestration. Due to its remarkable growth rate, bamboo offers an ideal system for studying organogenesis, particularly in monocots. Somatic embryogenesis (SE) serves as a useful technique for crop breeding and improvement. SE in moso bamboo (Phyllostachys pubescens) remains challenging due to limited knowledge of its transcriptional and metabolomic reprogramming. To address this, we optimized callus initiation (MS + 18.1 µM 2,4-D + 8.5 µM picloram), callus proliferation (MS + 12.5 µM 2,4-D + 8.5 µM picloram), and somatic embryogenesis (MS + 1.1 µM 2,4-D + 3.3 µM metatopolin), using nodal segments as explants. UHPLC-Q-TOF-MS-based metabolite profiling revealed distinct biochemical trajectories across developmental stages of P. pubescens. NEC (non-embryogenic callus) was enriched in flavonoids, alkaloids, and saponins, while in-vitro shoots showed flavonoids and glycosides enrichment, and ex-vitro shoots showed high accumulation of glycosides and terpenoids. In contrast, EC (embryogenic callus) showed elevated levels of fatty acid derivatives (α-ESA, 26-Methyl Nigranoate), phytoalexins (Wyerone acid), sesquiterpene (Alpha-santalal, Beta-guaiene), flavonoid glycosides, and plant hormones (Cis-Zeatin, Gibberellin A45), indicating a metabolically active state supporting somatic embryogenesis. Similarly, genes and transcription factors controlling cell differentiation and embryogenesis were upregulated during SE. This study provides a comprehensive resource to facilitate future genomic and genetic investigations aimed at deciphering the molecular basis of organogenesis and advancing research on somatic embryogenesis in bamboo.

Plant Somatic Embryogenesis Techniques

Independent expression of cardiac type I and II cyclic AMP-dependent protein kinase during murine embryogenesis and postnatal development.

The amount of total cyclic AMP-dependent protein kinase and of the protein kinase isozymes present in mouse heart changes during development. During embryogenesis, the total cardiac protein kinase activity increases most markedly during the 6 days prior to birth. A maximum kinase level is achieved in the 7 day-old neonate, and then activity progressively declines to an adult level approximating that of the mid-embryo. The type II kinase exhibits a moderate increase during late embryogenesis which declines by the time of birth. The type I isozyme increases throughout embryogenesis and the first neonatal week to a maximum specific activity five-fold higher than the mid-embryogenesis level. The isozyme level then falls to an adult activity similar to the mid-embryonic. These changes in isozyme profile are reflected in a changing type I to type II kinase ratio of 1.1 at 13--14 days embryogenesis, 2.4 at birth, 3.0 in the 7 day-old neonates, and 1 in the adult heart. Thus, the two protein kinase isozymes change in association with the developmental process in an independent fashion.

Animals

[Tumors in animals irradiated during embryogenesis].

In experiments on 465 male and female white non-lineal rats irradiated in their embryonal (7th day--240 rats), organogenesis (14th day--105 rats) and foetal periods of embryogenesis (19th day--120 rats) with 60Co--rays in a dose of 100 r, its actual power 168 r/m, and on 73 control rats the authors have studied the frequency and terms of tumors appearance. A group comprising 219 experimental rats and 53 control animals, died during a period of 200--600 days following their birth, was analysed. Irradiation at different stages of embryogenesis would provide a statistically significant increase in the frequency of tumors and shorten the terms of their appearance. In control animals tumors arose in 16,9% (in 9 of 53 animals). Embryonal and foetal periods of embryogenesis proved to be mostly susceptible as evidenced by the yield of radiation tumors. In animals exposed to irradiation in their embryonal period tumors arose in 64,5% (in 69 of 107 rats), in the foetal period--in 62,8% (in 44 of 70 rats), and during the period of organogenesis--in 42,8% (in 18 of 42 rats). Rats exposed to irradiation in the embryonal and foetal period of embryogenesis developed tumors to the 120--180th day of the experiment, while in the organogenesis period--to the 400th day. Maximum tumor appearance was observed in the last third of the lifetime. Male rats irradiated in the period of embryogenesis developed tumors of the mammary gland and bones, females- ovarian and bone tumors, which were absent in control animals.

Animals

Proteome Unravels Mechanism Differences in Embryogenesis Between Honey Bee Drone and Worker (Apis mellifera L.).

The physiological and social behaviors differ widely between honeybee workers and drones. All the organ rudiments of adult bees are formed during the embryonic stage. The initial molecular bases at the proteomic level for both embryonic developments have been identified, but a comprehensive understanding of the significant events involved in embryonic establishment remains elusive. To elucidate the molecular regulatory mechanisms underlying tissue differentiation during the embryogenesis of drones and workers, we implemented a state-of-the-art approach that combines in-hive inspection and targeted sampling (at nine embryogenesis stages) with high-throughput proteomics technology to investigate the developmental differences. In-hive inspection of hatching timing revealed an average developmental gap of approximately 3.6 h between the two embryos. Furthermore, proteomic analyses indicate that drone and worker embryos adopt distinct developmental strategies. Notably, proteins involved in fatty acid metabolism and key biological pathways related to organ formation-such as the Hedgehog and Wnt signaling pathways-are activated earlier in drones, suggesting that tissue development begins sooner in drone embryos than in workers. Additionally, the upregulation of cytoskeletal proteins and antioxidants in drone embryos likely supports their larger cell size and higher metabolic stress, reflecting distinct molecular characteristics of male development. Ribosomal proteins essential for biosynthetic support remain consistently expressed throughout the late stages in male embryos, indicating that drone embryogenesis lasts longer than that of workers. This work provides novel insights into the molecular foundations of honeybee embryogenesis and lays both theoretical and practical groundwork for future research into the mechanisms driving embryonic development.

Animals

The expression of beta 1 and beta 3 tubulin genes of Drosophila melanogaster is spatially regulated during embryogenesis.

In Drosophila beta tubulins are encoded by a small gene family and the four members of this family are differentially expressed. mRNAs transcribed from two of these genes, namely the beta 1 and beta 3 tubulin genes, are abundant during embryogenesis. While the beta 1 tubulin gene is constitutively expressed during development, beta 3 mRNA is restricted to two distinct phases: mid embryogenesis and metamorphosis. The transcription initiation sites are identical in both these stages and comparison of presumptive promoter regions reveals no extensive homologies between the genes. In situ localization shows beta 1 tubulin mRNA to be maternally expressed in the nurse cells of the egg chambers and evenly distributed during early embryogenesis. In contrast, during later stages of embryogenesis beta 1 tubulin transcripts are predominantly expressed in neural derivatives. The beta 3 tubulin gene expression is also spatially regulated, beta 3 mRNA being restricted to the mesoderm.

Animals

An analysis of dopa decarboxylase expression during embryogenesis in Drosophila melanogaster.

Dopa decarboxylase (DDC) activity appears near the end of embryogenesis in Drosophila. High titers of 20-OH-ecdysone on the other hand are found at midembryogenesis. Several explanations for this lag were investigated, since this hormone has been shown to induce a rapid increase in DDC activity at pupariation (G. P. Kraminsky, W. C. Clark, M. A. Estelle, R. D. Gietz, B. A. Sage, J. D. O'Connor, and R. B. Hodgetts, 1980, Proc. Natl. Acad. Sci. USA 77, 4175-4179). Using immunological and genetical criteria, it was shown that the same structural gene encodes DDC in embryos, mature larvae, and young adults. This rules out the existence of a distinct embryonic DDC gene unresponsive to 20-OH-ecdysone. Second, no evidence was found to support the hypothesis that a delay in the translation of DDC transcripts, produced in response to the elevated titer of 20-OH-ecdysone at midembryogenesis, caused the lag. Northern analysis of the RNA molecules homologous to cloned genomic sequences revealed that DDC transcripts were present at two different times during embryogenesis. A transcript was found in both ovaries and 0- to 2-hr embryos. However, this species disappeared by 4 hr and DDC transcript levels remained low until late in embryogenesis, when a significant increase occurred. This increase was presumably responsible for the appearance of the enzyme at this time. The northern blotting revealed nine DDC transcript species were present during embryogenesis and hybridization to intron-specific probes indicated that five of these contained at least part of one (or both) of the two introns. Three putative mature mRNA species were identified by their small size, relative abundance, apparent lack of intron sequence, and their presence on polysomes. The two mature species found during the late stages were postulated to differ in the length of their poly(A)+ tails. The third mature species was found only in ovaries and very young embryos and may well be of maternal origin. Data are examined in light of the possibility that this species is derived from a precursor initiated at a novel promotor.

Animals

Biochemical effects of juvenile hormone III on the tick, Argas (Persicargas) arboreus (Acari: Argasidae), during embryogenesis.

The biochemical effects of juvenile hormone III (JH III) on developing embryos from treated female Argas (Persicargas) arboreus Kaiser, Hoogstraal and Kohls were examined. Exogenous JH III resulted in a decrease in total proteins (P less than 0.001) only during the first 2 d of embryogenesis. There was no significant difference (P greater than 0.05) between RNA and DNA content in eggs from control and JH III-treated females. No significant difference (P greater than 0.05) was observed between control and JH III eggs in their lipid or phospholipid contents throughout embryogenesis. A total of 14-17 protein bands and 6-8 glycoprotein bands were separated by electrophoresis during embryogenesis of A. arboreus with some differences in mobility ratio between bands from control and JH III eggs. Differences in activity and isozyme patterns of malic acid, lactic acid, glucose 6-phosphate dehydrogenase, acid phosphatase, and alkaline phosphatase were not observed during embryogenesis of control and JH III-treated A. arboreus. Differences were observed in esterase activity.

Animals

Relationship between Merkel cells and nerve endings during embryogenesis in the mouse epidermis.

Close relationships between Merkel cells (MC) and nerve endings (NE) exist in the adult mouse. Because MC may serve as targets for the ingrowth of NE during embryogenesis, the purpose of the present study was to analyze the relationship between MC and NE during embryogenesis. Frozen tissue from whisker pads and backs of NMRI mouse embryos (12-17 d gestational age) were studied by double-labeling indirect immunofluorescence (IIF) with a cytokeratin monoclonal antibody that recognizes MC and with a neurofilament anti-serum. Such an approach allowed the analysis of a large number of MC (up to 5000), thus yielding quantitative data. At day 12 of gestational age, no MC were observed by IIF. From day 13 to 17, the number of MC, as well as their association with NE, progressively increased. On day 13, only 57% of whisker pad MC were NE associated, whereas by day 17, 95% were NE associated. These results were confirmed by electron microscopic (EM) observations. On the back, the same chronologic relationship between MC and NE was observed, but was later in the course of embryogenesis. There was also a time- and zone-dependent increase in MC association with NE in the epidermal zones studied (isthmic, parafollicular, interfollicular). These observations 1) establish the time course of MC and NE contacts during embryogenesis in the mouse epidermis, 2) show that MC are present in the epidermis and appendages before NE reach the epithelium, and 3) support the hypothesis that MC could act as targets for the growing NE.

Animals

Ontogenic appearance of MHC class I (B-F) antigens during chicken embryogenesis.

Expression of chicken MHC class I (B-F) antigens during ontogeny was determined by binding of anticlass I antibody and appearance of B-F transcripts by Northern blotting in chicken organs during embryogenesis until 2 weeks after hatching. MHC class I transcripts first become detectable in day 6.5 of embryogenesis. B-F cell-surface expression first becomes detectable in hemopoietic organs by day 10-12 of embryogenesis and somewhat later in nonhemopoietic organs. Flow cytometry analysis of hemopoietic cells throughout embryogenesis revealed B-Fhi and B-Flo cell populations. The percentage of B-F+ cells in spleen and bone marrow decreased around hatching, which could reflect either cell flows in these organs during this period or the sensitivity of hemopoietic cells to hatching stress.

Animals

Heat shock and thermotolerance in plant and animal embryogenesis.

Although the strategies of early embryogenesis differ greatly among multicellular eukaryotes, there are certain parallels in structure, form, and function that cross even kingdom lines: the extreme heat sensitivity of zygotes and very early embryos, followed by the acquisition of thermotolerance during subsequent development, is one such parallel. The heat sensitivity may be so extreme that even moderate increases in temperature result in lethality (generally associated with the earliest phases of embryogenesis), or the effects may be less severe, resulting in defects in development but not in lethality. Mechanistically, and molecularly, these two forms of thermosensitivity appear to have different origins. On the one hand, outright lethality appears to result from an inability to induce heat shock genes and proteins; on the other hand, heat-induced developmental defects appear to result from an alteration in expression of non-heat shock genes and from a delay in the overall developmental program that generally accompanies the cell's response to heat shock. This review is focused on the developmental regulation of the heat shock response during early embryogenesis and on the impact of this regulation on the development of both animal and plant embryos. The two basic issues that we address here are (i) the expression of heat shock genes in the absence of heat shock during embryogenesis and (ii) the expression (or lack of expression) of heat shock genes after deliberate exposure of the embryos to heat shock and the consequences of this expression on its subsequent survival and development.

Animals

Changes in nucleus, nucleolus and cell size accompanying somatic embryogenesis of Theobroma cacao L. II. Relation between basic protein content and size of nucleus, nucleolus and cell.

Embryo formation from callus of Theobroma cacao L. was associated with the changes in relationship between nuclear, nucleolar and cell sizes and the content of basic proteins (FG-FCF-stained). Together with the increase in nuclear size of callus and proembryo cells the increase in the amount of nuclear basic proteins was found. In the callus cells the increase in nucleolar protein content exceeded that in nucleolus size, which led to the rise in basic protein concentration in the nucleolus. However, in the early stage of embryogenesis the increase in protein content was not so marked as that in callus, which indicated that embryogenesis involved a decrease in concentration of nucleolar basic proteins. Differences between callus and proembryo cells were also observed in the concentration of cytoplasmic proteins. The increase in size of callus cells was the same as the increasing amount of cytoplasmic proteins. In proembryos a significant increase in cell size was accompanied by only slight changes in cytoplasmic proteins. The stimulation of embryogenesis by 2,4-D resulted in an increase of nuclear concentration of basic proteins in proembryos. The intensification of embryogenesis involved the decrease of the concentration of nucleolar proteins together with the increase in concentration of basic cytoplasmic proteins.

Cacao

The contents of proteins, carbohydrates, lipids and DNA during the embryogenesis of Drosophila.

The concentration of proteins, sugars, lipids and DNA has been determined during the embryogenesis of Drosophila. The protein content decreases after fertilization, being in the late embryo only 60% the value of the oocyte. The total sugar increases about 2.5-fold, from 3.5 h on until the end of embryogenesis. The lipids increase with a sharp peak at about 4 h and decrease during the rest of embryogenesis. DNA increases exponentially from the beginning of embryogenesis.

Animals

Cardiac embryogenesis: a three-dimensional approach.

In order to aid students in the understanding of cardiac embryogenesis, seven three-dimensional fiberglass models depicting stages eleven to eighteen of cardiac embryogenesis, and one model of a mature heart were constructed. The details of these models, and a review of cardiac embryogenesis is presented. The results we have had with these models in the teaching of cardiac embryogenesis to medical students, cardiac technologists, and nurses have been encouraging, and this approach to teaching is highly recommended.

Heart

Acquisition of insulin-dependent protein tyrosine kinase activity during Drosophila embryogenesis.

We have previously reported (Petruzzelli, L., Herrera, R., Garcia, R., and Rosen, O. M. (1985) Cancer Cells 3, 115-121) that adult Drosophila melanogaster contain a specific, high-affinity insulin-binding protein. Insulin-dependent protein tyrosine kinase activity has now been identified in Drosophila. Activity first appears at 6-12 h of embryogenesis, increases during the 12-18-h period and falls to low levels in the adult. 125I-insulin was cross-linked specifically and with high affinity to a protein (Mr = 135,000) throughout embryogenesis and in the adult. However, during the 6-12- and 12-18-h periods of embryogenesis when insulin-dependent protein tyrosine kinase activity is expressed, another protein (Mr = 100,000) becomes cross-linked to 125I-insulin. Crosslinking to both proteins was competitively inhibited by the addition of 100 nM insulin. We conclude that the insulin-binding and insulin-dependent protein tyrosine kinase activities of the mammalian insulin receptor are conserved in Drosophila. However, the insulin-dependent protein tyrosine kinase activity of the receptor is detected only during specific times in embryogenesis.

Amino Acids