scholarly journals Serotonin in pre-implantation mouse embryos is localized to the mitochondria and can modulate mitochondrial potential

Reproduction ◽  
2008 ◽  
Vol 135 (5) ◽  
pp. 657-669 ◽  
Author(s):  
Basudha Basu ◽  
Radha Desai ◽  
J Balaji ◽  
Raghothama Chaerkady ◽  
V Sriram ◽  
...  
Keyword(s):  
Fluorescence Microscopy ◽  
Serotonin Transporter ◽  
Tryptophan Hydroxylase ◽  
Developmental Stages ◽  
Mouse Embryos ◽  
Mitochondrial Potential ◽  
Early Embryos

Serotonin is reported to be present in early embryos of many species and plays an important role in early patterning. Since it is a fluorophore, it can be directly visualized using fluorescence microscopy. Here, we use three-photon microscopy to image serotonin in live pre-implantation mouse embryos. We find that it is present as puncta averaging 1.3 square microns and in concentrations as high as 442 mM. The observed serotonin puncta were found to co-localize with mitochondria. Live embryos pre-incubated with serotonin showed a higher mitochondrial potential, indicating that it can modulate mitochondrial potential. Pre-implantation mouse embryos were also examined at various developmental stages for the presence of transcripts of the peripheral and neuronal forms of tryptophan hydroxylase (Tph1andTph2respectively) and the classical serotonin transporter (Slc6a4). Transcripts ofTph2were seen in oocytes and in two-cell stages, whereas transcripts ofTph1were not detected at any stage. Transcripts of the transporter,Slc6a4, were present in all pre-implantation stages investigated. These results suggest that serotonin in embryos can arise from a combination of synthesis and uptake from the surrounding milieu.

Equilibrium vitrification of mouse embryos at various developmental stages

2012 ◽  
Vol 79 (11) ◽  
pp. 785-794 ◽  
Author(s):  
Bo Jin ◽  
Keiji Mochida ◽  
Atsuo Ogura ◽  
Chihiro Koshimoto ◽  
Kazutsugu Matsukawa ◽  
...  
Keyword(s):  
Developmental Stages ◽  
Mouse Embryos

The histogenetic capacity of tissues in the caudal end of the embryonic axis of the mouse

Development ◽  
1984 ◽  
Vol 82 (1) ◽  
pp. 253-266
Author(s):  
P. P. L. Tam
Keyword(s):  
Developmental Stages ◽  
Primitive Streak ◽  
Mouse Embryos ◽  
Embryonic Axis ◽  
Tail Bud ◽  
Germ Layers ◽  
Kidney Capsule ◽  
Caudal Region ◽  
Caudal Portion ◽  
Adult Body

The caudal end of the embryonic axis consists of the primitive streak and the tail bud. Small fragments of this caudal tissue were transplanted from mouse embryos of various developmental stages to the kidney capsule in order to test their histogenetic capacity. The variety of mature tissues obtained from these small fragments was similar to that obtained by grafting a larger caudal portion of the embryo. Initially, the grafted tissue broke up into loose masses of embryonic mesenchyme and this was later re-organized into more compact tissues and into cysts that were lined with various types of epithelia. After 14 days in the ectopic site, grafted tissues coming from embryos of the primitive-streak, the early-somite and the forelimb-bud stages differentiated into structures that has presumably originated from the three embryonic germ layers. Many of these structures were related to the caudal region of the adult body, such as the mid- and hindgut segments and urogenital derivatives. The histogenetic capacity for endodermal tissues and urogenital organs was lost when the grafted tissue consisted entirely of the tail bud of the hindlimb-bud-stage embryos. The behaviour of the caudal tissues suggested that (1) the primordia for the various parts of embryonic body were derived from a small progenitor population in the primitive streak and the tail bud, and (2) the histogenetic capacity of this population changed during development.

scholarly journals PML Regulates the Epidermal Differentiation Complex and Skin Morphogenesis during Mouse Embryogenesis

Genes ◽  
2020 ◽  
Vol 11 (10) ◽  
pp. 1130
Author(s):  
Anna Połeć ◽  
Alexander D. Rowe ◽  
Pernille Blicher ◽  
Rajikala Suganthan ◽  
Magnar Bjørås ◽  
...  
Keyword(s):  
Gene Expression ◽  
Developmental Stages ◽  
Expression Patterns ◽  
Growth Control ◽  
Wild Type Mouse ◽  
Global Gene Expression ◽  
Mouse Embryos ◽  
Epidermal Differentiation ◽  
Envelope Gene ◽  
Epidermal Differentiation Complex

The promyelocytic leukemia (PML) protein is an essential component of nuclear compartments called PML bodies. This protein participates in several cellular processes, including growth control, senescence, apoptosis, and differentiation. Previous studies have suggested that PML regulates gene expression at a subset of loci through a function in chromatin remodeling. Here we have studied global gene expression patterns in mouse embryonic skin derived from Pml depleted and wild type mouse embryos. Differential gene expression analysis at different developmental stages revealed a key role of PML in regulating genes involved in epidermal stratification. In particular, we observed dysregulation of the late cornified envelope gene cluster, which is a sub-region of the epidermal differentiation complex. In agreement with these data, PML body numbers are elevated in basal keratinocytes during embryogenesis, and we observed reduced epidermal thickness and defective hair follicle development in PML depleted mouse embryos.

Focal adhesion kinase PTK2 autophosphorylation is not required for the activation of sodium–hydrogen exchange by decreased cell volume in the preimplantation mouse embryo

Zygote ◽  
2019 ◽  
Vol 27 (3) ◽  
pp. 173-179
Author(s):  
Jane C. Fenelon ◽  
Baozeng Xu ◽  
Jay M. Baltz
Keyword(s):  
Focal Adhesion Kinase ◽  
Cell Volume ◽  
Focal Adhesion ◽  
Mouse Embryo ◽  
Hydrogen Exchange ◽  
Cell Types ◽  
Mouse Embryos ◽  
Cell Stage ◽  
Early Embryos ◽  
Early Mouse

SummaryRecovery from decreased cell volume is accomplished by a regulated increase of intracellular osmolarity. The acute response is activation of inorganic ion transport into the cell, the main effector of which is the Na+/H+ exchanger NHE1. NHE1 is rapidly activated by a cell volume decrease in early embryos, but how this occurs is incompletely understood. Elucidating cell volume-regulatory mechanisms in early embryos is important, as it has been shown that their dysregulation results in preimplantation developmental arrest. The kinase JAK2 has a role in volume-mediated NHE1 activation in at least some cells, including 2-cell stage mouse embryos. However, while 2-cell embryos show partial inhibition of NHE1 when JAK2 activity is blocked, NHE1 activation in 1-cell embryos is JAK2-independent, implying a requirement for additional signalling mechanisms. As focal adhesion kinase (FAK aka PTK2) becomes phosphorylated and activated in some cell types in response to decreased cell volume, we sought to determine whether it was involved in NHE1 activation in the early mouse embryo. FAK activity requires initial autophosphorylation of a tyrosine residue, Y397. However, FAK Y397 phosphorylation levels were not increased in either 1- or 2-cell embryos after cell volume was decreased. Furthermore, the selective FAK inhibitor PF-562271 did not affect NHE1 activation at concentrations that essentially eliminated Y397 phosphorylation. Thus, autophosphorylation of FAK Y397 does not appear to be required for NHE1 activation induced by a decrease in cell volume in early mouse embryos.

The distribution of motor proteins in the muscles and flame cells of the Schistosoma mansoni miracidium and primary sporocyst

Parasitology ◽  
2006 ◽  
Vol 133 (3) ◽  
pp. 321-329 ◽  
Author(s):  
D. BAHIA ◽  
L. G. A. AVELAR ◽  
F. VIGOROSI ◽  
D. CIOLI ◽  
G. C. OLIVEIRA ◽  
...  
Keyword(s):  
Fluorescence Microscopy ◽  
Schistosoma Mansoni ◽  
Specific Antibody ◽  
Developmental Stages ◽  
Longitudinal Muscle ◽  
Tubular Structures ◽  
Filamentous Actin ◽  
Optical Sectioning ◽  
Flame Cell ◽  
Confocal Fluorescence

Schistosoma mansoni eggs, miracidia and primary sporocysts were labelled with phalloidin-rhodamine to visualize filamentous actin structures. Analysis of these forms by confocal fluorescence microscopy revealed the presence of previously well-defined circular and longitudinal muscle layers. Besides these muscular layers that sustain and provide motility to these parasite forms, we found in these 3 consecutive developmental stages of the parasite previously unidentified actin-rich tubular structures. In the 3 forms, 4 actin-rich tubules could be observed by optical sectioning underneath the well-developed muscle layers. The tubules appear in pairs, transversal to the length of the parasite, and located towards the extremities. By using an anti-flame cell specific antibody we confirmed that the tubules co-localize with flame cells and also determined that the tubule core is filled with microtubules. The additional presence of myosin in these tubules strongly suggests that they are contractile structures.

scholarly journals Microarray analysis of gene expression during early development: a cautionary overview

Reproduction ◽  
2010 ◽  
Vol 140 (6) ◽  
pp. 787-801 ◽  
Author(s):  
Claude Robert
Keyword(s):  
Gene Expression ◽  
Early Development ◽  
Microarray Analysis ◽  
Developmental Stages ◽  
Sample Handling ◽  
Early Embryos ◽  
High Throughput Method ◽  
Full Benefit ◽  
Key Steps ◽  
Early Developmental

The rise of the ‘omics’ technologies started nearly a decade ago and, among them, transcriptomics has been used successfully to contrast gene expression in mammalian oocytes and early embryos. The scarcity of biological material that early developmental stages provide is the prime reason why the field of transcriptomics is becoming more and more popular with reproductive biologists. The potential to amplify scarce mRNA samples and generate the necessary amounts of starting material enables the relative measurement of RNA abundance of thousands of candidates simultaneously. So far, microarrays have been the most commonly used high-throughput method in this field. Microarray platforms can be found in a wide variety of formats, from cDNA collections to long or short oligo probe sets. These platforms generate large amounts of data that require the integration of comparative RNA abundance values in the physiological context of early development for their full benefit to be appreciated. Unfortunately, significant discrepancies between datasets suggest that direct comparison between studies is difficult and often not possible. We have investigated the sample-handling steps leading to the generation of microarray data produced from prehatching embryo samples and have identified key steps that significantly impact the downstream results. This review provides a discussion on the best methods for the preparation of samples from early embryos for microarray analysis and focuses on the challenges that impede dataset comparisons from different platforms and the reasons why methodological benchmarking performed using somatic cells may not apply to the atypical nature of prehatching development.

scholarly journals An improved deyolking procedure facilitates the proteomics analysis of early developmental stages of zebrafish embryos

2019 ◽  
Author(s):  
Purushothaman Kathiresan ◽  
Christopher Presslauer ◽  
Prem Prakash Das ◽  
Lim Teck Kwang ◽  
Steinar Dae Johansen ◽  
...  
Keyword(s):  
Developmental Stages ◽  
Cell Cycle Control ◽  
Fold Increase ◽  
Shotgun Proteomics ◽  
Egg Yolk ◽  
Vertebrate Development ◽  
Proteomics Analysis ◽  
Early Embryos ◽  
Mitochondrial Functions ◽  
Innovative Tool

Abstract Background: Zebrafish is a well-recognised organism for investigating vertebrate development and human diseases. However, the data on zebrafish proteome are scarce, particularly during embryogenesis. This is mostly due to the overwhelming abundance of egg yolk proteins, which tend to mask the detectable presence of less abundant proteins. Results: In this study, we developed an efficient procedure to reduce the amount of yolk in zebrafish early embryos to improve the LC-MS-based shotgun proteomics analysis. We demonstrated that the deyolking procedure resulted in a greater number of proteins being identified. This protocol resulted in approximately two-fold increase in the number of proteins identified in deyolked samples at cleavage stages, and the number of identified proteins increased greatly by 3-4 times compared to non-deyolked samples in both oblong and bud stages. Gene Ontology and KEGG analysis revealed a high number of functional proteins differentially accumulated in the deyolked versus non-deyolked samples. The most prominent enrichments after the deyolking procedure included processes, functions and components related to cellular organization, cell cycle, control of replication and translation, and mitochondrial functions. Conclusion: This deyolking procedure improves both qualitative and quantitative proteome analyses and provides an innovative tool in molecular embryogenesis of polylecithal animals, such as fish, amphibians, reptiles, or birds.

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