scholarly journals Three-dimensional genome reorganization during mouse spermatogenesis

2019 ◽  
Author(s):  
Zhengyu Luo ◽  
Xiaorong Wang ◽  
Ruoyu Wang ◽  
Jian Chen ◽  
Yusheng Chen ◽  
...  
Keyword(s):  
Developmental Stages ◽  
Three Dimensional ◽  
Type A ◽  
Pachytene Stage ◽  
Open Chromatin ◽  
Biological Processes ◽  
Primitive Type ◽  
Chromatin Loops ◽  
Genome Reorganization ◽  
Type A Spermatogonia

AbstractThree-dimensional genome organization plays an important role in many biological processes. Yet, how the genome is packaged at the molecular level during mammalian spermatogenesis remains unclear. Here, we performed Hi-C in seven sequential stages during mouse spermatogenesis. We found that topological associating domains (TADs) and chromatin loops underwent highly dynamic reorganization. They displayed clear existence in primitive type A spermatogonia, disappearance at pachytene stage, and reestablishment in spermatozoa. Surprisingly, even in the absence of TADs and chromatin loops at pachytene stage, CTCF remained bound at TAD boundary regions (identified in primitive type A spermatogonia). Additionally, many enhancers and promoters exhibited features of open chromatin and transcription remained active at pachytene stage. A/B compartmentalization and segmentation ratio were conserved in different stages of spermatogenesis in autosomes, although there were A/B compartment switching events correlated with gene activity changes. Intriguingly, A/B compartment structure on the X chromosome disappeared during pacSC, rST and eST stages. Together, our work uncovered a dynamic three-dimensional chromatin organization during mouse spermatogenesis and suggested that transcriptional regulation could be independent of TADs and chromatin loops at specific developmental stages.

scholarly journals Non-random distribution of spermatogonia in rats: evidence of niches in the seminiferous tubules

Reproduction ◽  
2003 ◽  
pp. 669-680 ◽  
Author(s):  
H Chiarini-Garcia ◽  
AM Raymer ◽  
LD Russell
Keyword(s):  
Basal Lamina ◽  
Random Distribution ◽  
Type A ◽  
Seminiferous Epithelium ◽  
Seminiferous Tubules ◽  
Interstitial Tissue ◽  
Primitive Type ◽  
Linear Index ◽  
Type A Spermatogonia

The relationships and distribution of spermatogonia were studied as a function of the stage of the seminiferous epithelium cycle in rats. Primitive spermatogonia in the mouse are located along regions of the basal lamina that face the interstitium. Before studying the distribution of spermatogonia in rats, it was necessary to characterize the various types of spermatogonia, as recently performed for mice. The Strauss' linear index (Li) selectivity method was then used and spermatogonia of the A(single) (A(s)) to A(aligned) (A(al)) lineage were preferentially found to be located in regions opposing the interstitium at stages V, VII and IX of the spermatogenic cycle. Because relatively little tubule-to-tubule contact occurs in rats, the aim of this study was to determine whether tubule-to-tubule contact or tubule proximity (or alternatively, the amount of interstitium) was an important factor in spermatogonial position. In this regard, another method (tubule proximity) was devised to determine spermatogonial position that accounted for the presence of adjacent tubules. This method showed that the position of tubules, rather than tubule contact, was more accurate than the Li method in determining the location of spermatogonia in the rat. The results also showed a non-random distribution of spermatogonia resembling that of the mouse, and that tubule-to-tubule contact is not essential for the positioning of spermatogonia. In conclusion, the results of this study strongly indicate that the most primitive type A spermatogonia (A(s), A(paired) and A(al)) in rats are present in niches located in those areas of the seminiferous tubules that border the interstitial tissue.

scholarly journals Isolation and Characterization of Highly Pure Type A Spermatogonia From Sterlet (Acipenser ruthenus) Using Flow-Cytometric Cell Sorting

2021 ◽  
Vol 9 ◽  
Author(s):  
Xuan Xie ◽  
Tomáš Tichopád ◽  
Galina Kislik ◽  
Lucie Langerová ◽  
Pavel Abaffy ◽  
...  
Keyword(s):  
Germ Cells ◽  
Cell Sorting ◽  
Developmental Stages ◽  
Type A ◽  
Cell Populations ◽  
Forward Scatter ◽  
Flow Cytometric ◽  
Flow Cytometric Cell ◽  
Type A Spermatogonia

Sturgeons are among the most ancient linages of actinopterygians. At present, many sturgeon species are critically endangered. Surrogate production could be used as an affordable and a time-efficient method for endangered sturgeons. Our study established a method for identifying and isolating type A spermatogonia from different developmental stages of testes using flow cytometric cell sorting (FCM). Flow cytometric analysis of a whole testicular cell suspension showed several well-distinguished cell populations formed according to different values of light scatter parameters. FCM of these different cell populations was performed directly on glass slides for further immunocytochemistry to identify germ cells. Results showed that the cell population in gate P1 on a flow cytometry plot (with high forward scatter and high side scatter parameter values) contains the highest amount of type A spermatogonia. The sorted cell populations were characterized by expression profiles of 10 germ cell specific genes. The result confirmed that setting up for the P1 gate could precisely sort type A spermatogonia in all tested testicular developmental stages. The P2 gate, which was with lower forward scatter and side scatter values mostly, contained type B spermatogonia at a later maturing stage. Moreover, expressions of plzf, dnd, boule, and kitr were significantly higher in type A spermatogonia than in later developed germ cells. In addition, plzf was firstly found as a reliable marker to identify type A spermatogonia, which filled the gap of identification of spermatogonial stem cells in sterlet. It is expected to increase the efficiency of germ stem cell culture and transplantation with plzf identification. Our study thus first addressed a phenotypic characterization of a pure type A spermatogonia population in sterlet. FCM strategy can improve the production of sturgeons with surrogate broodstock and further the analysis of the cellular and molecular mechanisms of sturgeon germ cell development.

The behaviour of mitotic nuclei after transplantation to early meiotic ooplasts or mitotic cytoplasts

Zygote ◽  
1997 ◽  
Vol 5 (3) ◽  
pp. 219-227 ◽  
Author(s):  
Michal Kubelka ◽  
Robert M. Moor
Keyword(s):  
High Activity ◽  
Somatic Cells ◽  
Type A ◽  
Chromosome Condensation ◽  
Premature Chromosome Condensation ◽  
Primitive Type ◽  
Dividing Cells ◽  
Pronuclear Formation ◽  
Further Development ◽  
Type A Spermatogonia

SummaryThis study evaluates the ability of the cytoplasm to determine the nature of the division cycle (meiotic or mitotic) in nuclei obtained from mitotically dividing cells. Using mouse oocytes in different stages of development two types of cytoplasm were prepared: firstly, early meiotic ooplasts were obtained by enucleation of non-matured, prophase-stage oocytes; secondly, mitotic cytoplasts were prepared by enucleation and activation of metaphase II (Mll)-stage oocytes. These two types of cytoplasts were then used in fusion experiments, in which mouse primitive type A spermatogonia (prospermatogonia) or mouse fibroblasts were used as a source of donor nuclei. While the fusion of prospermatogonia with mitotic cytoplasts resulted, as expected, in normal premature chromosome condensation (PCC) and subsequent pronuclear formation (58.1%), the majority of hybrids obtained by fusion of prospermatogonia with early meiotic ooplasts (40.3%) displayed unique morphology consisting of two sets of chromosomes organised in two spindle centres connected by microtubules. Each set of chromosomes contained the haploid (1n) number of chromosomes as revealed by chromosome analyses. The same morphology was observed also in 44.2% of hybrids in which the differentiated nuclei of fibroblasts were used as a source of donor mitotic nuclei. In both cases the hybrids were blocked at this stage with high activity of maturation promoting factor (MPF), resistant to any kind of activation and not able to undergo further development. These results suggest that the early meiotic ooplasm was able to induce the initiation of a meiosis-like reducing division in mitotic nuclei originating both from the germline cells and from more differentiated somatic cells.

scholarly journals Spermatogenic cells of the prepuberal mouse: isolation and morphological characterization

1977 ◽  
Vol 74 (1) ◽  
pp. 68-85 ◽  
Author(s):  
AR Bellve ◽  
JC Cavicchia ◽  
CF Millette ◽  
DA O'Brien ◽  
YM Bhatnagar ◽  
...  
Keyword(s):  
Sertoli Cells ◽  
Morphological Characteristics ◽  
Type A ◽  
Cell Types ◽  
Specific Cell ◽  
Spermatogenic Cells ◽  
Type B ◽  
Primitive Type ◽  
Pachytene Spermatocytes ◽  
Type A Spermatogonia

A procedure is described which permits the isolation from the prepuberal mouse testis of highly purified populations of primitive type A spermatogonia, type A spermatogonia, type B spermatogonia, preleptotene primary spermatocytes, leptotene and zygotene primary spermatocytes, pachytene primary spermatocytes and Sertoli cells. The successful isolation of these prepuberal cell types was accomplished by: (a) defining distinctive morphological characteristics of the cells, (b) determining the temporal appearance of spermatogenic cells during prepuberal development, (c) isolating purified seminiferous cords, after dissociation of the testis with collagenase, (d) separating the trypsin-dispersed seminiferous cells by sedimentation velocity at unit gravity, and (e) assessing the identity and purity of the isolated cell types by microscopy. The seminiferous epithelium from day 6 animals contains only primitive type A spermatogonia and Sertoli cells. Type A and type B spermatogonia are present by day 8. At day 10, meiotic prophase is initiated, with the germ cells reaching the early and late pachytene stages by 14 and 18, respectively. Secondary spermatocytes and haploid spermatids appear throughout this developmental period. The purity and optimum day for the recovery of specific cell types are as follows: day 6, Sertoli cells (purity>99 percent) and primitive type A spermatogonia (90 percent); day 8, type A spermatogonia (91 percent) and type B spermatogonia (76 percent); day 18, preleptotene spermatocytes (93 percent), leptotene/zygotene spermatocytes (52 percent), and pachytene spermatocytes (89 percent), leptotene/zygotene spermatocytes (52 percent), and pachytene spermatocytes (89 percent).

Initial Evidence that Testicular Germ Cells from the Prepubertal Rhesus Monkey (Type A Spermatogonia) Undergo Differentiation in Three-Dimensional Culture Systems.

2009 ◽  
Vol 81 (Suppl_1) ◽  
pp. 680-680
Author(s):  
Mahmoud Huleihel ◽  
Brian P. Hermann ◽  
Suresh Ramaswamy ◽  
Tony M. Plant
Keyword(s):  
Rhesus Monkey ◽  
Germ Cells ◽  
Three Dimensional ◽  
Type A ◽  
Culture Systems ◽  
Three Dimensional Culture ◽  
Type A Spermatogonia

Correlated Studies of Cellular Interactions Using TEM, Freeze Etching, and SEM

1974 ◽  
Vol 32 ◽  
pp. 320-321
Author(s):  
J. P. Revel
Keyword(s):  
Developmental Stages ◽  
Three Dimensional ◽  
Cell Movement ◽  
Cellular Interactions ◽  
Serial Sections ◽  
Cell Sheets ◽  
Freeze Etching ◽  
Early Developmental

Movement of individual cells or of cell sheets and complex patterns of folding play a prominent role in the early developmental stages of the embryo. Our understanding of these processes is based on three- dimensional reconstructions laboriously prepared from serial sections, and from autoradiographic and other studies. Many concepts have also evolved from extrapolation of investigations of cell movement carried out in vitro. The scanning electron microscope now allows us to examine some of these events in situ. It is possible to prepare dissections of embryos and even of tissues of adult animals which reveal existing relationships between various structures more readily than used to be possible vithout an SEM.

Electrical Capacitance Volume Tomography (ECVT) for industrial and medical applications-An Overview

2015 ◽  
Vol 11 (1) ◽  
pp. 2897-2908
Author(s):  
Mohammed S.Aljohani
Keyword(s):  
Imaging Technique ◽  
Three Dimensional ◽  
Biological Processes ◽  
Electrical Capacitance ◽  
Image Reconstruction Techniques ◽  
Phase Dynamics ◽  
Non Invasive ◽  
Electrical Capacitance Volume Tomography ◽  
And Performance ◽  
Optimization And Performance

Tomography is a non-invasive, non-intrusive imaging technique allowing the visualization of phase dynamics in industrial and biological processes. This article reviews progress in Electrical Capacitance Volume Tomography (ECVT). ECVT is a direct 3D visualizing technique, unlike three-dimensional imaging, which is based on stacking 2D images to obtain an interpolated 3D image. ECVT has recently matured for real time, non-invasive 3-D monitoring of processes involving materials with strong contrast in dielectric permittivity. In this article, ECVT sensor design, optimization and performance of various sensors seen in literature are summarized. Qualitative Analysis of ECVT image reconstruction techniques has also been presented.

Feeding behaviour and bioerosion: the ecological role of the rock-boring urchin, Echinometra mathaei (de Blainville, 1825), in Okinawa reef flat

2013 ◽  
Vol 1 (1) ◽  
pp. 10
Author(s):  
Noar Muda Satyawan ◽  
Shelly Tutupoho ◽  
Yusli Wardiatno ◽  
Makoto Tsuchiya
Keyword(s):  
Feeding Behaviour ◽  
Reef Flat ◽  
Type A ◽  
Gut Contents ◽  
Biological Processes ◽  
Type B ◽  
Night Time ◽  
Echinometra Mathaei ◽  
Benthic Ecosystems

Erosion rate on corals due to activities of other biota is called bioerosion. The rock-boring urchin, Echinometra mathaei, when it is abundant, plays a significant role in benthic ecosystems, including biological processes like coral erosion. During feeding, E. mathaei erodes calcium carbonate besides grazing on algae living on coral, so it plays an important role in both organic and inorganic carbons in coral reefs. The urchin E. mathaei actively feeds during the night time (nocturnal grazer). Although in Okinawa four types (A-D) of the urchin exist, the research only focused on the types A and B. Type A of E. mathaei produced 0.44951 g feces per day on average while type B produced 0.38030 g feces per day. CaCO3 analysis in feces and gut contents showed bioerosion rate of E. mathaei type A was 0.64492 g/individu/day, and 0.54436 g/individu/day in type B. There were no significant differences in bioerosion impact of E. mathaei type A and B© Laju erosi pada karang yang disebabkan oleh biota, dikenal dengan bioerosi. Bulu babi jenis Echinometra mathaei, ketika melimpah, menjadi sangat berpengaruh terhadap ekosistem bentik termasuk proses biologi seperti erosi karang. Selama aktivitas makan, E. mathaei menggerus kalsium karbonat dalam proporsi yang besar di samping alga yang tumbuh menempel pada karang sehingga memiliki peran penting dalam siklus karbon organik dan anorganik di ekosistem terumbu karang. Bulu babi E. mathaei aktif mencari makan pada malam hari (nocturnal grazer). Meskipun di Okinanawa ada 4 tipe (A-D), pada eksperimen kali ini memfokuskan pada tipe A dan B saja. Tipe A E. mathaei rata-rata memproduksi 0,44951 g feses/hari dan tipe B memproduksi 0,38030 g feses/hari. Berdasarkan analisis CaCO3 yang dilakukan pada feses dan isi lambung, laju bioerosi yang disebabkan oleh E. mathaei tipe A sebesar 0,64492 g/individu/hari sedangkan tipe B sebesar 0,54436 g/individu/hari. Tidak terdapat perbedaan dampak bioerosi yang signifikan antara E. mathaei tipe A dan B©

scholarly journals Structural relation matching: an algorithm to identify structural patterns into RNAs and their interactions

2021 ◽  
Vol 0 (0) ◽  
Author(s):  
Michela Quadrini
Keyword(s):  
Hydrogen Bonding ◽  
Secondary Structure ◽  
Nucleotide Sequence ◽  
Thermus Thermophilus ◽  
Three Dimensional ◽  
Secondary Structures ◽  
Structural Effect ◽  
Biological Processes ◽  
Structural Pattern ◽  
Rna Molecules

Abstract RNA molecules play crucial roles in various biological processes. Their three-dimensional configurations determine the functions and, in turn, influences the interaction with other molecules. RNAs and their interaction structures, the so-called RNA–RNA interactions, can be abstracted in terms of secondary structures, i.e., a list of the nucleotide bases paired by hydrogen bonding within its nucleotide sequence. Each secondary structure, in turn, can be abstracted into cores and shadows. Both are determined by collapsing nucleotides and arcs properly. We formalize all of these abstractions as arc diagrams, whose arcs determine loops. A secondary structure, represented by an arc diagram, is pseudoknot-free if its arc diagram does not present any crossing among arcs otherwise, it is said pseudoknotted. In this study, we face the problem of identifying a given structural pattern into secondary structures or the associated cores or shadow of both RNAs and RNA–RNA interactions, characterized by arbitrary pseudoknots. These abstractions are mapped into a matrix, whose elements represent the relations among loops. Therefore, we face the problem of taking advantage of matrices and submatrices. The algorithms, implemented in Python, work in polynomial time. We test our approach on a set of 16S ribosomal RNAs with inhibitors of Thermus thermophilus, and we quantify the structural effect of the inhibitors.

scholarly journals Membrane fusion studied by colloidal probes

2021 ◽  
Vol 50 (2) ◽  
pp. 223-237 ◽  
Author(s):  
Hannes Witt ◽  
Filip Savić ◽  
Sarah Verbeek ◽  
Jörn Dietz ◽  
Gesa Tarantola ◽  
...  
Keyword(s):  
Membrane Fusion ◽  
Optical Tweezers ◽  
Three Dimensional ◽  
Biological Processes ◽  
Supported Membranes ◽  
Supported Bilayers ◽  
Force Microscopy ◽  
Advantages And Disadvantages ◽  
Atomic Force ◽  
Colloidal Probes

AbstractMembrane-coated colloidal probes combine the benefits of solid-supported membranes with a more complex three-dimensional geometry. This combination makes them a powerful model system that enables the visualization of dynamic biological processes with high throughput and minimal reliance on fluorescent labels. Here, we want to review recent applications of colloidal probes for the study of membrane fusion. After discussing the advantages and disadvantages of some classical vesicle-based fusion assays, we introduce an assay using optical detection of fusion between membrane-coated glass microspheres in a quasi two-dimensional assembly. Then, we discuss free energy considerations of membrane fusion between supported bilayers, and show how colloidal probes can be combined with atomic force microscopy or optical tweezers to access the fusion process with even greater detail.

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