Alteration of nuclear architecture in male germ cells of chromosomally derived subfertile mice

2001 ◽  
Vol 114 (24) ◽  
pp. 4429-4434
Author(s):  
Silvia Garagna ◽  
Maurizio Zuccotti ◽  
Alan Thornhill ◽  
Raul Fernandez-Donoso ◽  
Soledad Berrios ◽  
...  
Keyword(s):  
Sertoli Cells ◽  
Spatial Organization ◽  
Nuclear Architecture ◽  
Spatial Arrangement ◽  
Cell Types ◽  
Condensed State ◽  
Germ Cell Differentiation ◽  
Y Chromosomes ◽  
Dual Colour ◽  
Pachytene Spermatocytes

The mammalian cell nucleus consists of numerous compartments involved in the regular unfolding of processes such as DNA replication and transcription, RNA maturation, protein synthesis and cell division. Knowledge is increasing of the relationships between high-order levels of chromatin organization and its spatial organization, and of how these relationships contribute to the various functions carried out in the nucleus. We have studied the spatial arrangement of mouse telocentric chromosomes 5, 11, 13, 15, 16 and 17, some of their metacentric Robertsonian derivatives, and X and Y chromosomes by whole chromosome painting in male germ (spermatogonia, pachytene spermatocytes and spermatids) and Sertoli cells of homozygous and heterozygous individuals. Using dual-colour fluorescence in situ hybridization we found that these chromosomes occupy specific nuclear territories in each cell type analysed. When chromosomes are present as Robertsonian metacentrics in the heterozygous state, that is, as Robertsonian metacentrics and their homologous telocentrics, differences in their nuclear positions are detectable: heterozygosity regularly produces a change in the nuclear position of one of the two homologous telocentrics in all the cell types studied. In the Robertsonian heterozygotes, the vast majority of the Sertoli cells show the sex chromosomes in a condensed state, whereas they appear decondensed in the Robertsonian homozygotes. As the Robertsonian heterozygosities we studied produce a chromosomally derived impairment of male germ-cell differentiation, we discuss the possibility that changes in chromosome spatial territories may alter some nuclear machinery (e.g., synapsis, differential gene expression) important for the correct unfolding of the meiotic process and for the proper functioning of Sertoli cells.

Frequencies in vibrating wood – Does cell organization matter?

IAWA Journal ◽  
2019 ◽  
Vol 40 (1) ◽  
pp. 124-142
Author(s):  
Oliver Dünisch
Keyword(s):  
Spatial Organization ◽  
Ring Width ◽  
Spatial Arrangement ◽  
Cell Types ◽  
Dominant Frequency ◽  
Fast Fourier Transformation ◽  
Frequency Spectra ◽  
Laser Sensors ◽  
Vibrating Plates ◽  
Dominant Frequencies

ABSTRACT The relationship between the spatial organization of different cell types, of the xylem rays, and of the tree rings and the frequencies in vibrating softwoods and hardwoods was studied under controlled conditions. In total, the frequencies in 1007 standardized vibrating plates from 16 softwoods and 74 hardwoods were analysed using high resolution laser sensors (accuracy ± 0.02 μm, sampling frequency 30 kHz) for vibration measurements. Overlapping frequencies within the frequency spectra were identified by means of Fast Fourier Transformation analysis. With regard to the number of distinct frequencies within the spectra, four different vibration types were identified: type 1–one dominant frequency within the frequency spectra; type 2-two dominant frequencies within the frequency spectra; type 3-three dominant frequencies within the frequency spectra; type 4-no dominant frequencies within the frequency spectra. The presence of distinct frequencies was correlated with a highly organized spatial arrangement of tracheids in softwoods, with a storied arrangement of the xylem rays in hardwoods, and with low variation in tree-ring width in both softwoods and hardwoods. The grid size for repetition in these xylem structures influenced the frequencies of the vibrating wood in absolute numbers. The results indicate that the analysis of the anatomical structure of the wood can contribute to the grading of timber for its vibration characteristics, which is of special interest for the selection of resonance wood for musical instruments.

Immuocytochemical analysis of spatial organization of photoreceptors and amacrine and ganglion cells in the tiger salamander retina

2004 ◽  
Vol 21 (2) ◽  
pp. 157-166 ◽  
Author(s):  
JIAN ZHANG ◽  
ZHUO YANG ◽  
SAMUEL M. WU
Keyword(s):  
Ganglion Cell ◽  
Spatial Organization ◽  
Ganglion Cells ◽  
Electrophysiological Study ◽  
Outer Nuclear Layer ◽  
Amacrine Cells ◽  
Spatial Arrangement ◽  
Cell Types ◽  
Proximal Region ◽  
Dorsal Retina

In the present study, using double- or triple-label immunocytochemistry in conjunction with confocal microscopy, we aimed to examine the population and distribution of photoreceptors, GABAergic and glycinergic amacrine cells, and ganglion cells, which are basic but important parameters for studying the structure–function relationship of the salamander retina. We found that the outer nuclear layer (ONL) contained 82,019 ± 3203 photoreceptors, of which 52% were rods and 48% were cones. The density of photoreceptors peaked at ∼8000 cells/mm2 in the ventral and dropped to ∼4000 cells/mm2 in the dorsal retina. In addition, the rod/cone ratio was less than 1 in the central retina but larger than 1 in the periphery. Moreover, in the proximal region of the inner nuclear layer (INL3), the total number of cells was 50,576 ± 8400. GABAergic and glycinergic amacrine cells made up approximately 78% of all cells in this layer, including 43% GABAergic, 32% glycinergic, and 3% GABA/glycine colocalized amacrine cells. The density of these amacrine cells was ∼6500 cells/mm2 in the ventral and ∼3200 cells/mm2 in the dorsal area. The ratio of GABAergic to glycinergic amacrine cells was larger than 1. Furthermore, in the ganglion cell layer (GCL), among a total of 36,007 ± 2010 cells, ganglion cells accounted for 65.7 ± 1.5% of the total cells, whereas displaced GABAergic and glycinergic amacrine cells comprised about 4% of the cells in this layer. The ganglion cell density was ∼1800 cells/mm2 in the ventral and ∼600 cells/mm2 in the dorsal retina. Our data demonstrate that all three major cell types are not uniformly distributed across the salamander retina. Instead, they exhibit a higher density in the ventral than in the dorsal retina and their spatial arrangement is associated with the retinal topography. These findings provide a basic anatomical reference for the electrophysiological study of this species.

Evidence for the control of testicular interstitial fluid volume in the rat by specific germ cell types

1991 ◽  
Vol 128 (3) ◽  
pp. 359-NP ◽  
Author(s):  
R. M. Sharpe ◽  
J. M. S. Bartlett ◽  
G. Allenby
Keyword(s):  
Germ Cell ◽  
Sertoli Cells ◽  
Interstitial Fluid ◽  
Local Heating ◽  
Cell Types ◽  
Volume Changes ◽  
Interstitial Fluid Volume ◽  
Testicular Weight ◽  
Pachytene Spermatocytes

ABSTRACT Following on from our recent evidence that Sertoli cells may regulate testicular interstitial fluid (IF) volume, this study has assessed whether depletion of specific germ cell types in vivo is associated with changes in recovered IF volume. Germ cell depletion was induced by either a single oral administration of 650 mg methoxyacetic acid (MAA)/kg or exposure of the testes to local heating (43 °C for 30 min). Treatment with MAA induced depletion or loss of most pachytene and later spermatocytes at 1–3 days and, because of maturation depletion, this resulted in the specific depletion of later germ cell types at 7–35 days. Testicular IF volume was unchanged at 1–7 days after MAA treatment but was increased significantly (P < 0·01) at 14 days and was nearly doubled (P< 0·001) at 21 days, before returning to control levels at 28–42 days. Serum LH (and FSH) levels were generally higher in MAA-treated rats, especially at 21 and 28 days, but there was no obvious correlation between LH levels and IF volume changes. Similarly, there was no relationship between IF volume changes and testicular weight or IF levels of testosterone. The increase in IF volume at 14–21 days after MAA treatment coincided with specific depletion of the later elongate spermatids (steps 14–19) and, when these cells reappeared in the testis, IF volume normalized. This possible causal association was studied further in rats exposed to local testicular heating which, within 3 days, caused major depletion of pachytene spermatocytes and early (step 1–8) spermatids. However, testicular IF volume in heat-exposed rats did not change until 14 days, a time at which depletion of the later (step 9–19) spermatids first became evident; IF volume remained increased whilst these germ cells were absent or depleted. The pattern of change in IF volume in heat-exposed rats was not related to LH (or FSH) levels, which were raised at most time-points after heat treatment, nor to testicular weight which was decreased considerably at 3 days and declined progressively thereafter. These data thus provide evidence that specific depletion of the most mature germ cell types (the elongate spermatids) is associated with specific changes in testicular IF volume, presumably via modulation of the secretion of vasoactive factors by the Sertoli cells. These findings also reinforce the growing evidence for the mutual interdependence of all of the cell types in the testis. Journal of Endocrinology (1991) 128, 359–367

scholarly journals A spatial atlas of inhibitory cell types in mouse hippocampus

2018 ◽  
Author(s):  
Xiaoyan Qian ◽  
Kenneth D. Harris ◽  
Thomas Hauling ◽  
Dimitris Nicoloutsopoulos ◽  
Ana B. Muñoz-Manchado ◽  
...  
Keyword(s):  
Spatial Organization ◽  
Memory Function ◽  
Expression Patterns ◽  
Ground Truth ◽  
Spatial Arrangement ◽  
Cell Types ◽  
Cell System ◽  
Inhibitory Neurons ◽  
Hippocampal Area

Understanding the function of a tissue requires knowing the spatial organization of its constituent cell types. In the cerebral cortex, single-cell RNA sequencing (scRNA-seq) has revealed the genome-wide expression patterns that define its many, closely related cell types, but cannot reveal their spatial arrangement. Here we introduce probabilistic cell typing by in situ sequencing (pciSeq), an approach that leverages prior scRNA-seq classification to identify cell types using multiplexed in situ RNA detection. We applied this method to map the inhibitory neurons of hippocampal area CA1, a cell system critical for memory function, for which ground truth is available from extensive prior work identifying the laminar organization of subtly differing cell types. Our method confidently identified 16 interneuron classes, in a spatial arrangement closely matching ground truth. This method will allow identifying the spatial organization of fine cell types across the brain and other tissues.

scholarly journals A single cell atlas reveals unanticipated cell type complexity in Drosophila ovaries

2021 ◽  
Author(s):  
Maija Slaidina ◽  
Selena Gupta ◽  
Ruth Lehmann
Keyword(s):  
Stem Cell ◽  
Single Cell ◽  
Spatial Organization ◽  
Follicle Cell ◽  
Cell Types ◽  
Cell Type ◽  
Organ Function ◽  
Relative Paucity ◽  
Germ Cell Differentiation ◽  
Cell Type Specific

AbstractOrgan function relies on the spatial organization and functional coordination of numerous cell types. The Drosophila ovary is a widely used model system to study the cellular activities underlying organ function, including stem cell regulation, cell signaling and epithelial morphogenesis. However, the relative paucity of cell type specific reagents hinders investigation of molecular functions at the appropriate cellular resolution.Here, we used single cell RNA sequencing to characterize all cell types of the stem cell compartment and early follicles of the Drosophila ovary. We computed transcriptional signatures and identified specific markers for nine states of germ cell differentiation, and 23 somatic cell types and subtypes. We uncovered an unanticipated diversity of escort cells, the somatic cells that directly interact with differentiating germline cysts. Three escort cell subtypes reside in discrete anatomical positions, and express distinct sets of secreted and transmembrane proteins, suggesting that diverse micro-environments support the progressive differentiation of germ cells. Finally, we identified 17 follicle cell subtypes, and characterized their transcriptional profiles. Altogether, we provide a comprehensive resource of gene expression, cell type specific markers, spatial coordinates and functional predictions for 34 ovarian cell types and subtypes.

scholarly journals Nuclear Organization of Mammalian Genomes

1999 ◽  
Vol 146 (6) ◽  
pp. 1211-1226 ◽  
Author(s):  
Nicolas Sadoni ◽  
Sabine Langer ◽  
Christine Fauth ◽  
Giorgio Bernardi ◽  
Thomas Cremer ◽  
...  
Keyword(s):  
Spatial Organization ◽  
Chromosome Structure ◽  
Nuclear Architecture ◽  
Replication Timing ◽  
Cell Types ◽  
Higher Order ◽  
Chromosome Territories ◽  
Mitotic Chromosomes ◽  
Organizational Principle ◽  
Mammalian Genomes

We investigated the nuclear higher order compartmentalization of chromatin according to its replication timing (Ferreira et al. 1997) and the relations of this compartmentalization to chromosome structure and the spatial organization of transcription. Our aim was to provide a comprehensive and integrated view on the relations between chromosome structure and functional nuclear architecture. Using different mammalian cell types, we show that distinct higher order compartments whose DNA displays a specific replication timing are stably maintained during all interphase stages. The organizational principle is clonally inherited. We directly demonstrate the presence of polar chromosome territories that align to build up higher order compartments, as previously suggested (Ferreira et al. 1997). Polar chromosome territories display a specific orientation of early and late replicating subregions that correspond to R- or G/C-bands of mitotic chromosomes. Higher order compartments containing G/C-bands replicating during the second half of the S phase display no transcriptional activity detectable by BrUTP pulse labeling and show no evidence of transcriptional competence. Transcriptionally competent and active chromatin is confined to a coherent compartment within the nuclear interior that comprises early replicating R-band sequences. As a whole, the data provide an integrated view on chromosome structure, nuclear higher order compartmentalization, and their relation to the spatial organization of functional nuclear processes.

scholarly journals BMP4 patterns Smad activity and generates stereotyped cell fate organisation in spinal organoids

2019 ◽  
Author(s):  
Nathalie Duval ◽  
Célia Vaslin ◽  
Tiago Barata ◽  
Stéphane Nédélec ◽  
Vanessa Ribes
Keyword(s):  
Cell Fate ◽  
Bone Morphogenetic Proteins ◽  
Neural Tube ◽  
Spatial Organization ◽  
Time Windows ◽  
Spatial Dynamics ◽  
Spatial Arrangement ◽  
Cell Types ◽  
Dorsal Neural Tube ◽  
Bmp Signalling

AbstractBone Morphogenetic Proteins (BMP) are secreted regulators of cell fate in several developing tissues. In the embryonic spinal cord, they control the emergence of the neural crest, roof plate and distinct subsets of dorsal interneurons. Although a gradient of BMP activity has been proposed to determine cell type identity in vivo, whether this is sufficient for pattern formation in vitro is unclear. Here, we demonstrate that exposure to BMP4 initiates distinct spatial dynamics of BMP signalling within the self-emerging epithelia of both mouse and human pluripotent stem cells derived spinal organoids. The pattern of BMP signalling results in the stereotyped spatial arrangement of dorsal neural tube cell types and concentration, timing and duration of BMP4 exposure modulate these patterns. Moreover, differences in the duration of competence time-windows between mouse and human account for the species specific tempo of neural differentiation. Together the study describes efficient methods to generate patterned subsets of dorsal interneurons in spinal organoids and supports the conclusion that graded BMP activity orchestrates the spatial organization of the dorsal neural tube cellular diversity in mouse and human.

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&gt;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).

scholarly journals Wt1 deficiency causes undifferentiated spermatogonia accumulation and meiotic progression disruption in neonatal mice

Reproduction ◽  
2014 ◽  
Vol 147 (1) ◽  
pp. 45-52 ◽  
Author(s):  
Qiao-Song Zheng ◽  
Xiao-Na Wang ◽  
Qing Wen ◽  
Yan Zhang ◽  
Su-Ren Chen ◽  
...  
Keyword(s):  
Sertoli Cells ◽  
Wilms Tumor ◽  
Post Partum ◽  
Basal Membrane ◽  
Cell Types ◽  
Seminiferous Tubules ◽  
Cell Integrity ◽  
Germ Cell Differentiation ◽  
Multiple Cell

Spermatogenesis is a complex process involving the regulation of multiple cell types. As the only somatic cell type in the seminiferous tubules, Sertoli cells are essential for spermatogenesis throughout the spermatogenic cycle. The Wilms tumor gene, Wt1, is specifically expressed in the Sertoli cells of the mouse testes. In this study, we demonstrated that Wt1 is required for germ cell differentiation in the developing mouse testes. At 10 days post partum, Wt1-deficient testes exhibited clear meiotic arrest and undifferentiated spermatogonia accumulation in the seminiferous tubules. In addition, the expression of claudin11, a marker and indispensable component of Sertoli cell integrity, was impaired in Wt1−/flox; Cre-ERTM testes. This observation was confirmed in in vitro testis cultures. However, the basal membrane of the seminiferous tubules in Wt1-deficient testes was not affected. Based on these findings, we propose that Sertoli cells' status is affected in Wt1-deficient mice, resulting in spermatogenesis failure.

scholarly journals A single-cell atlas reveals unanticipated cell type complexity in Drosophila ovaries

2021 ◽  
pp. gr.274340.120
Author(s):  
Maija Slaidina ◽  
Selena Gupta ◽  
Torsten U. Banisch ◽  
Ruth Lehmann
Keyword(s):  
Stem Cell ◽  
Single Cell ◽  
Spatial Organization ◽  
Follicle Cell ◽  
Cell Types ◽  
Cell Type ◽  
Organ Function ◽  
Relative Paucity ◽  
Germ Cell Differentiation ◽  
Cell Type Specific

Organ function relies on the spatial organization and functional coordination of numerous cell types. The Drosophila ovary is a widely used model system to study the cellular activities underlying organ function, including stem cell regulation, cell signaling and epithelial morphogenesis. However, the relative paucity of cell type-specific reagents hinders investigation of molecular functions at the appropriate cellular resolution. Here, we used single-cell RNA sequencing to characterize all cell types of the stem cell compartment and early follicles of the Drosophila ovary. We computed transcriptional signatures and identified specific markers for nine states of germ cell differentiation, and 23 somatic cell types and subtypes. We uncovered an unanticipated diversity of escort cells, the somatic cells that directly interact with differentiating germline cysts. Three escort cell subtypes reside in discrete anatomical positions, and express distinct sets of secreted and transmembrane proteins, suggesting that diverse micro-environments support the progressive differentiation of germ cells. Finally, we identified 17 follicle cell subtypes, and characterized their transcriptional profiles. Altogether, we provide a comprehensive resource of gene expression, cell type-specific markers, spatial coordinates and functional predictions for 34 ovarian cell types and subtypes.

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