76. In situ brain perfusion in a mouse model of Sanfilippo syndrome

2008 ◽  
Vol 93 (2) ◽  
pp. 33
Author(s):  
Charles Pontikis ◽  
Eva Zaccariotto
Keyword(s):  
Mouse Model ◽  
Brain Perfusion ◽  
Sanfilippo Syndrome ◽  
In Situ Brain Perfusion

An in situ brain perfusion technique to study cerebrovascular transport in the rat

1984 ◽  
Vol 247 (3) ◽  
pp. H484-H493 ◽  
Author(s):  
Y. Takasato ◽  
S. I. Rapoport ◽  
Q. R. Smith
Keyword(s):  
Brain Perfusion ◽  
External Carotid Artery ◽  
External Carotid ◽  
Perfusion Technique ◽  
Conscious Rat ◽  
Low Viscosity ◽  
Regional Flow ◽  
In Situ Brain Perfusion ◽  
The Right

The right cerebral hemisphere of the rat was perfused in situ by retrograde infusion of HCO3 saline or blood into the right external carotid artery. Infusion rate was adjusted to minimize the contribution of systemic blood to flow in the hemisphere. During perfusion with whole or artificial blood, regional cerebral blood flow and blood volume were comparable to respective values in the conscious rat, whereas perfusion with HCO3 saline increased regional flow three- to fourfold due to the low viscosity of the saline perfusate. Perfusion with whole blood for 300 S or with HCO3 saline for 60 S did not alter the permeability of the blood-brain barrier. Cerebrovascular permeability coefficients of eight nonelectrolytes ranged from 10(-8) to 10(-4) cm X S-1 and were directly proportional to the octanol-water partition coefficient of the solute. Thus the in situ brain perfusion technique is a sensitive new method to study cerebrovascular transfer in the rat and permits absolute control of perfusate composition.

scholarly journals Comparison of blood-brain barrier permeability in mice and rats using in situ brain perfusion technique

2000 ◽  
Vol 279 (3) ◽  
pp. H1022-H1028 ◽  
Author(s):  
Hideyasu Murakami ◽  
Hitomi Takanaga ◽  
Hirotami Matsuo ◽  
Hisakazu Ohtani ◽  
Yasufumi Sawada
Keyword(s):  
Blood Brain Barrier ◽  
Brain Perfusion ◽  
External Carotid Artery ◽  
Brain Barrier ◽  
External Carotid ◽  
Perfusion Technique ◽  
Perfusion Rate ◽  
In Situ Brain Perfusion ◽  
Blood Brain

Here we present a method for measuring the permeability coefficient-surface area product ( PS) values at the blood-brain barrier in mice, using the in situ brain perfusion technique originally developed for rats by Takasato et al. ( Am J Physiol Heart Circ Physiol 247: H484–H493, 1984). Retrograde infusion into the right external carotid artery increased the carotid perfusion pressure in proportion to the perfusion rate. Intravascular volume and cerebral perfusion fluid flow at a perfusion rate of 1.0 ml/min in mice were similar to those in rats. In addition, the contribution of systemic blood to total flow in the hemisphere was small (only 3.2%). These findings indicated that this perfusion rate is suitable for mice. The PS values of more than 20 different compounds were determined in mice by using the in situ brain perfusion technique, and comparisons were made with data from rats. There was a close relationship (1:1) between the PS values in mice and rats, indicating that brain capillary permeabilities are similar in mice and rats.

Brain perfusion fixation in male pigs using a safer closed system

2018 ◽  
Vol 52 (4) ◽  
pp. 413-417 ◽  
Author(s):  
Gracia U. Musigazi ◽  
Stéphanie De Vleeschauwer ◽  
Raf Sciot ◽  
Eric Verbeken ◽  
Bart Depreitere
Keyword(s):  
Closed System ◽  
Porcine Model ◽  
Brain Perfusion ◽  
External Jugular Vein ◽  
Mechanically Ventilated ◽  
In Situ Brain Perfusion ◽  
Perfusion Fixation ◽  
Large Animals ◽  
User Friendly

Tissue fixation methods are well established for rodents, but not for large animals. We present a simple technique for in situ brain perfusion fixation in a male porcine model, using cervical vessels for inflow and outflow and achieving a closed system. Thirty-four pigs, aged 4.7 ± 0.6 months and weighing 60.7 ± 10.9 kg, were anaesthetised and mechanically ventilated. The ipsilateral common carotid artery and external jugular vein were dissected and constituted the inflow and outflow access, respectively. The brains were perfused and fixed in situ with heparinised saline followed by buffered formaldehyde. Then, specimens (brain, cerebellum and brainstem) were extracted and processed for histology. Fixative fluid leakage was avoided, achieving a closed system. This technique minimises the exposure to toxic chemicals such as formaldehyde and associated hazards (inherent toxicity, eye irritation), thereby increasing operators’ safety. Perfusion was performed with a peristaltic pump for 20–30 minutes at an optimum rate of 0.20 l/min and required only 5 litres of the fixative. The specimens were sufficiently hardened to be extracted. High-quality tissues were available for histology analysis. This technique offers a user-friendly closed system for brain perfusion fixation which can be adapted for other tissues of the head, face and neck.

Blood-brain barrier tight junctions are altered during a 72-h exposure to λ-carrageenan-induced inflammatory pain

2002 ◽  
Vol 283 (4) ◽  
pp. H1531-H1537 ◽  
Author(s):  
J. D. Huber ◽  
V. S. Hau ◽  
L. Borg ◽  
C. R. Campos ◽  
R. D. Egleton ◽  
...  
Keyword(s):  
Western Blot ◽  
Tight Junctions ◽  
Blood Brain Barrier ◽  
Inflammatory Pain ◽  
Brain Perfusion ◽  
Brain Barrier ◽  
Brain Uptake ◽  
In Situ Brain Perfusion ◽  
Blood Brain

In this study, we examined the effect of λ-carrageenan-induced inflammatory pain on the functional and structural properties of the rat blood-brain barrier (BBB) over a 72-h time period. Systemic inflammation was induced by an intraplantar injection of 3% λ-carrageenan into the right hind paw of female Sprague-Dawley rats. In situ brain perfusion and Western blot analyses were performed at 1, 3, 6, 12, 24, 48, and 72 h. In situ brain perfusion showed λ-carrageenan significantly increased brain uptake of [14C]sucrose at 1, 3, 6, and 48 h (139 ± 9%, 166 ± 19%, 138 ± 13%, and 146 ± 7% compared with control, respectively). Capillary depletion analysis insured the increased brain uptake was due to increased BBB permeability and not vascular trapping. Western blot analyses for zonula occludens-1 (ZO-1) and occludin were performed on isolated cerebral microvessels. ZO-1 expression was significantly increased at 1, 3, and 6 h and returned to control expression levels by 12 h. Total occludin expression was significantly reduced at 1, 3, 6, 12, and 48 h. This investigation demonstrated that λ-carrageenan-induced inflammatory pain elicits a biphasic increase in BBB permeability with the first phase occurring from 1–6 h and the second phase occuring at 48 h. Furthermore, changes in BBB function are correlated with altered tight junctional protein expression of occludin and ZO-1. Changes in the structure of tight junctions may have important clinical ramifications concerning central nervous system homeostasis and therapeutic drug delivery.

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