Effects of methyl-beta-cyclodextrin on blood-brain barrier permeability in angiotensin II-induced hypertensive rats.

The blood-brain barrier (BBB) permeability primarily increases in cerebral venules during acute hypertension. Methyl-ß-cyclodextrin (MßCD), a cholesterol-depleting agent, decreases the expression of caveolins disrupting caveolar structures. We aimed to determine the effects of MßCD on the BBB permeability of angiotensin (ANG) II-induced hypertensive rats. Three minutes after MßCD administration (5 mg/kg), acute hypertension was induced by ANG II (60 µg/kg). Evans blue (EB) and horseradish peroxidase (HRP) tracers were used to assess BBB permeability. Immunohistochemistry for caveolin (Cav)-1 and tight junction protein claudin-5 was performed. EB dye content significantly increased in both cerebral cortices and left hippocampus in MßCD (P < 0.05), right cerebral cortex and both hippocampi in ANG II (P < 0.05; P < 0.01), and both cerebral cortices and hippocampi in MßCD plus ANG II groups compared with controls (P < 0.05; P < 0.01). A significant decrease in claudin-5 immunostaining intensity was observed in animals treated with MßCD compared with controls (P < 0.05). Cav-1 immunostaining intensity increased in ANG II group (P < 0.05). Ultrastructurally, HRP reaction products were observed in endothelial cells of the microvessels in the hippocampus region in MßCD group while the tracer was mainly localized in astrocytes and neurons in ANG II, and MßCD plus ANG II groups. The endothelial cells of the venules in the cerebral cortex of the animals in the latter experimental groups also showed an abundance of caveolar vesicles devoid of HRP reaction products. Our results revealed that MßCD did not provide overall protective effects on BBB integrity in acute hypertension and even led to BBB disruption in normotensive animals.

Recombinant activin A enhances the growth and survival of isolated preantral follicles cultured three-dimensionally in extracellular basement matrix protein (matrigel) under serum-free conditions.

Development of in vitro technologies that will allow the culture of early stage follicles before antral stage is an essential part of research in reproductive biology in order to understand the ovarian folliculogenesis better. Current evidence suggests that oocyte and somatic cells-derived growth factors interacting with each other and extracellular matrix proteins at paracrine level are involved in this early, gonadotrophin-independent phase of follicle growth. Basement membrane matrix protein (Matrigel™) is a soluble gel rich in extracellular matrix proteins and growth factors. Activin A promotes preantral follicle growth in vivo by inducing the proliferation of granulosa cells and by upregulating the expression of FSH receptor and aromatase enzyme. We hypothesized that activin A and matrigel may provide a better in vitro environment for early stage preantral follicles. Preantral follicles isolated from 14-21 day old BALB/c mice were cultured in matrigel ± activin A for four days. The growth (119.4% versus 45.4%, p < 0.05; respectively) and survival rates (76.3% versus 43.7%, p < 0.05; respectively) of the follicles treated with activin A were significantly higher compared to those without activin A. These results suggest that Activin A and matrigel provide a better in vitro milieu for the growth of isolated ovarian follicles.