scholarly journals Three-dimensional reconstruction of the rat nephron

2014 ◽  
Vol 306 (6) ◽  
pp. F664-F671 ◽  
Author(s):  
Erik I. Christensen ◽  
Birgitte Grann ◽  
Inger B. Kristoffersen ◽  
Elisabeth Skriver ◽  
Jesper S. Thomsen ◽  
...  
Keyword(s):  
Structural Changes ◽  
Three Dimensional ◽  
Serial Sections ◽  
Mosaic Pattern ◽  
Outer Medulla ◽  
Short Loop ◽  
Collecting Ducts ◽  
Straight Tubules ◽  
Dimensional Reconstruction ◽  
Outer Stripe

This study gives a three-dimensional (3D) structural analysis of rat nephrons and their connections to collecting ducts. Approximately 4,500 2.5-μm-thick serial sections from the renal surface to the papillary tip were obtained from each of 3 kidneys of Wistar rats. Digital images were recorded and aligned into three image stacks and traced from image to image. Short-loop nephrons (SLNs), long-loop nephrons (LLNs), and collecting ducts (CDs) were reconstructed in 3D. We identified a well-defined boundary between the outer stripe and the inner stripe of the outer medulla corresponding to the transition of descending thick limbs to descending thin limbs and between the inner stripe and the inner medulla, i.e., the transition of ascending thin limbs into ascending thick limbs of LLNs. In all nephrons, a mosaic pattern of proximal tubule (PT) cells and descending thin limb (DTL) cells was observed at the transition between the PT and the DTL. The course of the LLNs revealed tortuous proximal “straight” tubules and winding of the DTLs within the outer half of the inner stripe. The localization of loop bends of SLNs in the inner stripe of the outer medulla and the bends of LLNs in the inner medulla reflected the localization of their glomeruli; i.e., the deeper the glomerulus, the deeper the bend. Each CD drained approximately three to six nephrons with a different pattern than previously established in mice. This information will provide a basis for evaluation of structural changes within nephrons as a result of physiological or pharmaceutical intervention.

scholarly journals Mitochondrial Expression of Arginase II in Male and Female Rat Inner Medullary Collecting Ducts

2005 ◽  
Vol 53 (4) ◽  
pp. 533-541 ◽  
Author(s):  
Olivier Levillain ◽  
Sandra Balvay ◽  
Simone Peyrol
Keyword(s):  
Western Blot ◽  
Low Density ◽  
Female Rat ◽  
Outer Medulla ◽  
Male And Female ◽  
Mitochondrial Fractions ◽  
Collecting Ducts ◽  
Straight Tubules ◽  
Outer Stripe ◽  
Arginase Ii

Microdissected rat proximal straight tubules (PST) and inner medullary collecting ducts (IMCD) highly produce urea from L-arginine, supporting the expression of the mitochondrial arginase II. However, IMCD contain a very low density of mitochondria compared with PST. Recently, arginase II has been localized by immunohistochemistry in rat PST but not IMCD. This study was designed to verify whether rat IMCD express arginase II and to identify its subcellular localization. We developed an antibody raised against arginase II that allowed the detection of a band of 38 kDa corresponding to arginase II on immunoblots. In male and female rat kidneys, Western blot analyses revealed that arginase II was highly expressed in the inner medulla (IM), the outer stripe of the outer medulla (osOM), and the deep cortex. Immunocytochemistry demonstrated that arginase II was homogeneously expressed in IMCD. Proteins of the cytosolic and mitochondrial fractions extracted from osOM and IM and analyzed by Western blot showed that 86% of arginase II was associated with mitochondria. The molecular weight of arginase II was similar in the cytosolic and mitochondrial fractions. Immunoelectron microscopy confirmed the presence of arginase II in the mitochondria of IMCD. In conclusion, arginase II is expressed in mitochondria of male and female rat IMCD.

Three-Dimensional Reconstruction Using Stereo Pairs from Serial Thick Sections

1977 ◽  
Vol 35 ◽  
pp. 562-563
Author(s):  
Robert Glaeser ◽  
Thomas Bauer ◽  
David Grano
Keyword(s):  
Three Dimensional ◽  
Dimensional Structure ◽  
Serial Sections ◽  
Thin Sections ◽  
3 Dimensional ◽  
Transmission Electron ◽  
Freeze Dried ◽  
Dimensional Reconstruction ◽  
Stereo Imagery ◽  
Whole Mounts

In transmission electron microscopy, the 3-dimensional structure of an object is usually obtained in one of two ways. For objects which can be included in one specimen, as for example with elements included in freeze- dried whole mounts and examined with a high voltage microscope, stereo pairs can be obtained which exhibit the 3-D structure of the element. For objects which can not be included in one specimen, the 3-D shape is obtained by reconstruction from serial sections. However, without stereo imagery, only detail which remains constant within the thickness of the section can be used in the reconstruction; consequently, the choice is between a low resolution reconstruction using a few thick sections and a better resolution reconstruction using many thin sections, generally a tedious chore. This paper describes an approach to 3-D reconstruction which uses stereo images of serial thick sections to reconstruct an object including detail which changes within the depth of an individual thick section.

Application of stereological analysis of cell volume to isolated myocytes in culture with and without adrenergic innervation

1990 ◽  
Vol 48 (3) ◽  
pp. 416-417
Author(s):  
Jane K. Rosenthal ◽  
Dianne L. Atkins ◽  
William J. Marvin ◽  
Penny A. Krumm
Keyword(s):  
Cardiac Myocytes ◽  
Structural Changes ◽  
Three Dimensional ◽  
Adrenergic Innervation ◽  
Culture Cell ◽  
Serial Sections ◽  
Newborn Rats ◽  
Primary Culture Cell ◽  
Biological Studies ◽  
Cell Volumes

To comprehend structural changes in cardiac myocytes accompanying adrenergic innervation, it is essential that a three dimensional analysis be performed. To date, biological studies which utilize stereological methods have been limited to cells in tissue and in organs. Our laboratory has utilized current stereological techniques for measuring absolute volumes of individual myocytes in primary culture. Cell volumes are calculated for two distinct groups of cells at 96 hours in culture: isolated myocytes and myocytes innervated with adrenergic neurons (Figure 1).Cardiac myocytes are cultured from the ventricular apices of newborn rats. Cells are plated directly onto tissue culture dishes with or without preplated explants from the paravertebral thoracolumbar sympathetic chain. On day four cultures are photographed and marked for one-to-one cell location. Following conventional fixation and embeddment in eponate-12, the cells are relocated and mounted for microtomy. The cells are completely sectioned at 120nm in their parallel orientation to the surface of the dish (Figure 2). Serial sections are collected on formvar coated slotted grids and are recorded in sequence.

Conformational characterization of nucleosome structure by Electron Microscopy

1993 ◽  
Vol 51 ◽  
pp. 194-195
Author(s):  
Gregory J. Czarnota
Keyword(s):  
Electron Microscopy ◽  
Structural Changes ◽  
Physiological State ◽  
Gene Activation ◽  
Three Dimensional ◽  
Macromolecular Complex ◽  
Ultrastructural Studies ◽  
Ionic Environment ◽  
Nucleosome Structure ◽  
Dimensional Reconstruction

Chromatin structure at the fundamental level of the nucleosome is important in vital cellular processes. Recent biochemical and genetic analyses show that nucleosome structure and structural changes are very active participants in gene expression, facilitating or inhibiting transcription and reflecting the physiological state of the cell. Structural states and transitions for this macromolecular complex, composed of DNA wound about a heterotypic octamer of variously modified histone proteins, have been measured by physico-chemical techniques and by enzyme-accessibility and are recognized to occur with various post-translational modifications, gene activation, transformation and with ionic-environment. In spite of studies which indicate various forms of nucleosome structure, all current x-ray and neutron diffraction studies have consistently resulted in only one structure, suggestive of a static conformation. In contrast, two-dimensional electron microscopy studies and three-dimensional reconstruction techniques have yielded different structures. These fundamental differences between EM and other ultrastructural studies have created a long standing quandary, which I have addressed and resolved using spectroscopic electron microscopy and statistical analyses of nucleosome images in a study of nucleosome structure with ionic environment.

scholarly journals THE THREE-DIMENSIONAL RECONSTRUCTION OF THE XY CHROMOSOMAL PAIR IN HUMAN SPERMATOCYTES

1970 ◽  
Vol 45 (1) ◽  
pp. 43-53 ◽  
Author(s):  
A. J. Solari ◽  
Laura L. Tres
Keyword(s):  
Synaptonemal Complex ◽  
Three Dimensional ◽  
Strong Support ◽  
Serial Sections ◽  
Lateral Element ◽  
Spatial Reconstruction ◽  
Chromosomal Pair ◽  
Dimensional Reconstruction ◽  
Short Core ◽  
Apparent Association

The spatial reconstruction of the XY pair of chromosomes from human spermatocytes has been made by the study of serial sections 1000 A in thickness. The sex pair during zygotene-pachytene forms a condensed mass of chromatin that has two filamentous, electron-opaque cores: the long and the short core. During early pachytene both cores have a common ending region, about 0.4–0.8 µ long. This common end is a synaptonemal complex, and each of the cores forms a lateral element of that complex. The cores become more convoluted during middle pachytene forming "ringlike bodies." Nucleoli from spermatocytes have three distinct regions: (a) granular; (b) dense fibrillar; and (c) clear intermediate. Occasional association of the XY pair and the heteropycnotic "basal knobs" results in apparent association of nucleoli and the sex pair in a minority of cells. The evidence presented is interpreted as a strong support of the hypothesis of homologous regions in the human XY pair.

Three-Dimensional Reconstruction From Serial Sections For Medical Applications

1981 ◽  
Author(s):  
Prakairut N. Cook ◽  
Solomon Batnitzky ◽  
Kyo R. Lee ◽  
Errol Levine ◽  
Hilton I. Price ◽  
...  
Keyword(s):  
Three Dimensional ◽  
Medical Applications ◽  
Serial Sections ◽  
Three Dimensional Reconstruction ◽  
Dimensional Reconstruction
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