scholarly journals Permselectivity of blood follicle barriers in mouse ovaries of the mifepristone-induced polycystic ovary model revealed by in vivo cryotechnique

Reproduction ◽  
2008 ◽  
Vol 136 (5) ◽  
pp. 599-610 ◽  
Author(s):  
Hong Zhou ◽  
Nobuhiko Ohno ◽  
Nobuo Terada ◽  
Sei Saitoh ◽  
Ichiro Naito ◽  
...  
Keyword(s):  
Blood Flow ◽  
Blood Vessels ◽  
Histopathological Examination ◽  
Polycystic Ovary ◽  
Basement Membranes ◽  
In Vivo Cryotechnique ◽  
Preparation Methods ◽  
Perfusion Fixation ◽  
Pco Syndrome

Despite the potential association of polycystic ovary (PCO) syndrome with hemodynamic changes, follicular microenvironment and the involvement of blood follicle barriers (BFB), a histopathological examination has been hampered by artifacts caused by conventional preparation methods. In this study, mouse ovaries of a mifepristone-induced PCO model were morphologically and immunohistochemically examined byin vivocryotechnique (IVCT), which prevents those technical artifacts. Ovarian specimens of PCO model mice were prepared by IVCT or the conventional perfusion fixation after s.c. injection of mifepristone. Their histology and immunolocalization of plasma proteins, including albumin (molecular mass, 69 kDa), immunoglobulin G (IgG, 150 kDa), inter-α-trypsin inhibitor (ITI, 220 kDa), fibrinogen (340 kDa), and IgM (900 kDa), were examined. In the PCO model, enlarged blood vessels with abundant blood flow were observed in addition to cystic follicles with degenerative membrana granulosa. The immunolocalization of albumin and IgM in the PCO model were similar to those in normal mice. Albumin immunolocalized in the blood vessels, interstitium or follicles, and IgM was mostly restricted within the blood vessels. In contrast, immunolocalization of IgG, ITI, and fibrinogen changed in the PCO model. Both IgG and ITI were clearly blocked by follicular basement membranes, and hardly observed in the membrana granulosa, though fibrinogen was mostly observed within blood vessels. These findings suggest that increased blood flow and enhanced selectivity of molecular permeation through the BFB are prominent features in the PCO ovaries, and changes in hemodynamic conditions and permselectivity of BFB are involved in the pathogenesis and pathophysiology of PCO syndrome.

Ultrastructural Analyses of Living Animal Organs Prepared by “in vivo Cryotechnique” for Electron Microscopy

2001 ◽  
Vol 7 (S2) ◽  
pp. 730-731
Author(s):  
S. Ohno ◽  
I. Takayama ◽  
N. Terada ◽  
Y. Fujii ◽  
H. Ueda ◽  
...  
Keyword(s):  
Blood Circulation ◽  
Chemical Fixation ◽  
Considerable Time ◽  
In Vivo Cryotechnique ◽  
Preparation Methods ◽  
Morphological Studies ◽  
Perfusion Fixation ◽  
Excised Tissues ◽  
Animal Organs

It is well known that hemodynamic factors, such as blood pressure and flow, exert an important influence on native morphology of various animal organs. However, information about the significance of ultrastructures which are revealed by conventional preparation methods has been limited, because routine chemical fixation of cells and tissues takes considerable time, during which their morphology could easily change. Therefore, morphological studies with routine immersion or perfusion fixation have not revealed real ultrastructures of functioning animal organs with normal blood circulation.2 The ultimate goal of morphological study is that all features to be examined should reflect the physiological meaning under investigation. For that purpose, the preservation of cells and tissues in functioning organs is necessary for studies to define their ultrastructures. Most cryotechniques have been based on the use of prior excised tissues. Such small specimens are commonly frozen within several seconds following the excision of the tissues.

scholarly journals Involvement of follicular basement membrane and vascular endothelium in blood–follicle barrier formation of mice revealed by ‘in vivo cryotechnique’

Reproduction ◽  
2007 ◽  
Vol 134 (2) ◽  
pp. 307-317 ◽  
Author(s):  
Hong Zhou ◽  
Nobuhiko Ohno ◽  
Nobuo Terada ◽  
Sei Saitoh ◽  
Yasuhisa Fujii ◽  
...  
Keyword(s):  
Basement Membrane ◽  
Blood Vessels ◽  
Vascular Endothelium ◽  
Serum Proteins ◽  
Vascular Endothelial Cells ◽  
Follicular Development ◽  
Basement Membranes ◽  
In Vivo Cryotechnique ◽  
Quick Freezing

The molecular sieve with size- and charge selectivity in ovarian follicles, the so-called blood–follicle barrier (BFB), was examined during follicular development under physiological conditions to reveal ovarian structures responsible for the BFB by using our ‘in vivocryotechnique’ (IVCT). Mouse ovary specimens were prepared with different methods including IVCT, immersion, or perfusion chemical fixation and quick-freezing following resection or perfusion. Their paraffin sections or cryosections were stained with hematoxylin–eosin or immunostained for serum proteins with different molecular weights: albumin, immunoglobulin (Ig) G1 heavy chain, inter-α-trypsin inhibitor (IαI), fibrinogen, and IgM. Their immunoreactivity was better preserved with IVCT. The immunostaining for albumin was clearly observed in blood vessels, interstitium, and developing follicles, but that of IgG1, IαI, or fibrinogen was significantly decreased inside the follicles. IgM was immunohistochemically decreased throughout the interstitium outside blood vessels. The immunoreactivities of IgG1 and IgM, as compared with albumin, were clearly changed along follicular basement membranes and around vascular endothelial cells respectively. These findings indicate that BFB functions throughout follicular development, and the follicular basement membrane and the vascular endothelium could play some significant roles in the permselectivity for such soluble proteins with intermediate and high molecular weight respectively.

scholarly journals Amylin: Localization, Effects on Cerebral Arteries and on Local Cerebral Blood Flow in the Cat

2001 ◽  
Vol 1 ◽  
pp. 168-180 ◽  
Author(s):  
Lars Edvinsson ◽  
Peter J. Goadsby ◽  
Rolf Uddman
Keyword(s):  
Blood Flow ◽  
Cerebral Blood Flow ◽  
Blood Vessels ◽  
Cerebral Arteries ◽  
In Vivo Studies ◽  
Cerebral Blood Vessels ◽  
Local Cerebral Blood Flow ◽  
Doppler Flowmetry

Amylin and adrenomedullin are two peptides structurally related to calcitonin gene-related peptide (CGRP). We studied the occurrence of amylin in trigeminal ganglia and cerebral blood vessels of the cat with immunocytochemistry and evaluated the role of amylin and adrenomedullin in the cerebral circulation by in vitro and in vivo pharmacology. Immunocytochemistry revealed that numerous nerve cell bodies in the trigeminal ganglion contained CGRP immunoreactivity (-ir); some of these also expressed amylin-ir but none adrenomedullin-ir. There were numerous nerve fibres surrounding cerebral blood vessels that contained CGRP-ir. Occasional fibres contained amylin-ir while we observed no adrenomedullin-ir in the vessel walls. With RT-PCR and Real-Time�PCR we revealed the presence of mRNA for calcitonin receptor-like receptor (CLRL) and receptor-activity-modifying proteins (RAMPs) in cat cerebral arteries. In vitro studies revealed that amylin, adrenomedullin, and CGRP relaxed ring segments of the cat middle cerebral artery. CGRP and amylin caused concentration-dependent relaxations at low concentrations of PGF2a-precontracted segment (with or without endothelium) whereas only at high concentration did adrenomedullin cause relaxation. CGRP8-37 blocked the CGRP and amylin induced relaxations in a parallel fashion. In vivo studies of amylin, adrenomedullin, and CGRP showed a brisk reproducible increase in local cerebral blood flow as examined using laser Doppler flowmetry applied to the cerebral cortex of the a-chloralose�anesthetized cat. The responses to amylin and CGRP were blocked by CGRP8-37. The studies suggest that there is a functional sub-set of amylin-containing trigeminal neurons which probably act via CGRP receptors.

Edema, Fainting, and Strokes

2021 ◽  
pp. 216-240
Author(s):  
Graham Mitchell
Keyword(s):  
Blood Pressure ◽  
Blood Flow ◽  
Blood Vessels ◽  
High Blood Pressure ◽  
Fluid Pressure ◽  
Basement Membranes ◽  
Tissue Pressure ◽  
Efficient Mechanisms ◽  
The Brain ◽  
Mediated Reduction

High blood pressure in humans is often associated with heart failure, edema, strokes, and episodes of fainting. Giraffes never show these. Edema, the abnormal collection of fluid in the lower legs, is prevented in giraffes by a combination of thick basement membranes of capillary blood vessels, which probably reduce their permeability to proteins, a very high tissue pressure that resists flow of fluid out of capillaries, and efficient mechanisms for returning blood to the heart. Fainting occurs when blood flow (and thus oxygen and glucose supply) to the brain is reduced. When a giraffe lifts its head after drinking water there is a sudden reduction of blood flow to the head, and fainting should result. Fainting is avoided because the blood flow that remains is diverted completely to the brain by a unique arrangement of blood vessels and nerves, and by structures that maintain the perfusion pressure of the blood flowing through the brain. Strokes can be caused by rupture of small blood vessels in the brain when they are exposed to high blood pressure of the kind reached in the head of a giraffe when it drinks surface water. Rupture of brain blood vessels is prevented in giraffes by mechanisms that reduce pressure. The posture adopted while drinking, baroreceptor-mediated reduction in cardiac output, the effects of the carotid rete, diversion of blood away from the brain, an increase in cerebrospinal fluid pressure, and passive and active constriction of blood vessels, all contribute.

1506 In vivo Three-Dimensional Mapping of Mesenteric Blood Vessels and Blood Flow Using GPU

2013 ◽  
Vol 2013.50 (0) ◽  
pp. 150601-150602
Author(s):  
Hikari ICHISHIMA ◽  
Yudai NAKAYAMA ◽  
Hiroki ISHIDA ◽  
Shunsuke AKIGUCHI ◽  
Tadashi HACHIGA ◽  
...  
Keyword(s):  
Blood Flow ◽  
Blood Vessels ◽  
Three Dimensional ◽  
Three Dimensional Mapping ◽  
Dimensional Mapping

scholarly journals Morphological study by an 'in vivo cryotechnique' of the shape of erythrocytes circulating in large blood vessels

1998 ◽  
Vol 193 (1) ◽  
pp. 73-79 ◽  
Author(s):  
MEI XUE ◽  
YASUKO KATO ◽  
NOBUO TERADA ◽  
YASUHISA FUJII ◽  
TAKESHI BABA ◽  
...  
Keyword(s):  
Blood Vessels ◽  
Morphological Study ◽  
In Vivo Cryotechnique

Abstract P342: Histopathological Correlates of MRI-Visible Perivascular Spaces in Cerebral Amyloid Angiopathy

Stroke ◽  
2021 ◽  
Vol 52 (Suppl_1) ◽  
Author(s):  
Valentina Perosa ◽  
Leon P Munting ◽  
Whitney Freeze ◽  
Ashley A Scherlek ◽  
Anand Viswanathan ◽  
...  
Keyword(s):  
Blood Vessels ◽  
Cerebral Amyloid Angiopathy ◽  
Ex Vivo ◽  
Histopathological Examination ◽  
Amyloid Β ◽  
Amyloid Angiopathy ◽  
Perivascular Spaces ◽  
Cerebral Amyloid

Perivascular spaces (PVS) are fluid-filled spaces surrounding cerebral blood vessels. MRI-visible, supposedly enlarged, PVS in the centrum semiovale (CSO) have been associated with cerebral amyloid angiopathy (CAA). PVS enlargement may be due to perivascular clearance impairments, potentially caused by increased amyloid-β (Aβ) accumulation in the walls of vessels in the overlying cortex. We test this hypothesis, using MRI-guided histopathological examination of PVS in CAA autopsy cases. The cohort included 19 CAA (74.1±8.2y, 7F) and 5 non-CAA control cases (88.0±4.9y, 3F). Formalin-fixed hemispheres were scanned on a 3T MRI scanner, including a 500μm T2-weighted sequence. PVS enlargement was assessed in the CSO on in vivo and ex vivo MRI. In addition, local score of PVS enlargement was assessed in four pre-defined juxtacortical areas (Fig.A), using a semiquantitative score and on the corresponding histological sections (Fig.B). Severity of leptomeningeal and cortical CAA were assessed on adjacent Aβ-stained sections, using a semiquantitative scale.PVS enlargement was more severe in CAA cases compared to controls, both on in vivo and ex vivo MRI (p<0.05). PVS enlargement on ex vivo MRI positively correlated with the severity of PVS enlargement on the corresponding histopathological samples (Fig.C). Within CAA cases, the degree of PVS enlargement on ex vivo MRI was positively associated with leptomeningeal CAA severity (n=52 samples, ρ=0.35, p=0.011), but not cortical CAA severity (n=52 samples, ρ=0.10, p=0.472). These preliminary findings confirm that the degree of MRI-visible PVS in juxtacortical brain areas reflects enlargement on histopathology. Moreover, they suggest that PVS enlargement in cases with CAA corresponds to increased CAA severity in the overlying leptomeningeal vessels, possibly as a result of impaired perivascular clearance. Future directions include characterization of individual blood vessels associated with PVS enlargement.

Scanning Electron Microscopic Study of Flowing Erythrocytes in Hepatic Sinusoids as Revealed by “in Vivo Cryotechnique”

1997 ◽  
Vol 3 (S2) ◽  
pp. 239-240
Author(s):  
N. Terada ◽  
Y. Fujii ◽  
Y. Kato ◽  
H. Ueda ◽  
T. Baba ◽  
...  
Keyword(s):  
Body Weight ◽  
Blood Vessels ◽  
Morphological Changes ◽  
Surface Membrane ◽  
Heart Beat ◽  
Ultrastructural Changes ◽  
Sodium Pentobarbital ◽  
Electron Microscopic ◽  
In Vivo Cryotechnique

The flowing behavior of individual erythrocytes in blood vessels is usually determined by their deformability, which is controlled mainly by the nature of their interior constituents and the flexibility of their surface membrane. Moreover, the physical behavior of erythrocytes passing through capillaries has been examined in vivo by light microscopy. However, little has been known about ultrastructural changes of such erythrocyte shapes flowing in blood vessels in vivo. Recently, a new technique was developed for freezing cells and tissues in vivo without stopping the blood supply, which was referred to as “in vivo cryotechnique”.This method has been also suitable for obtaining informations about dynamic morphological changes.Seven female Balb/c mice were anesthetized peritoneally with sodium pentobarbital (100μg/g body weight), and their abdomen was opened through a pararectus incision. For artificial cardiac arrest, some mice were anesthetized with an excessive dose of the anesthetic (500μg/g body weight), their respiration and heart-beat were completely stopped, and the following procedures were done within one minute. A liver was put on a plastic plate without disturbance of blood circulation, and the “in vivo cryotechnique” was performed. Briefly, a cryoknife was pushed into the liver as fast as possible and the tissue was immediately poured with liquid isopentane-propane mixture (-193°C) (Fig.la,b).

Visualization of microvascular blood flow in mouse kidney and spleen by quantum dot injection with “in vivo cryotechnique”

2010 ◽  
Vol 80 (3) ◽  
pp. 491-498 ◽  
Author(s):  
Nobuo Terada ◽  
Yurika Saitoh ◽  
Sei Saitoh ◽  
Nobuhiko Ohno ◽  
Takashi Jin ◽  
...  
Keyword(s):  
Blood Flow ◽  
Quantum Dot ◽  
Mouse Kidney ◽  
Microvascular Blood Flow ◽  
In Vivo Cryotechnique
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