A simple stereologic method for analysis of cerebral cortical microvessels using image analysis

Manoonkitiwongsa, Panya S.; McMillan, Paul J.; Schultz, Robert L.; Jackson-Friedman, Catherine; Lyden, Patrick D.

doi:10.1016/s1385-299x(01)00087-3

Cited by 14 publications

(13 citation statements)

References 20 publications

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“…1). Surface density (S v ), is defined as the total surface of microvessels per unit volume of tissue (mm 2 /mm 3 ) (Manoonkitiwongsa et al, 2001). We determined the surface density of RECA-1-positive capillaries in the SGZ in the same randomly selected serial sections of the hippocampus that were used for quantification for capillary number.…”

Section: Vascular Niches Of the Subgranular Zone: Measurement Of Surfmentioning

confidence: 99%

Quantitative changes in hippocampal microvasculature of chronically stressed rats: No effect of fluoxetine treatment

et al. 2009

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Exposure to chronic stress alters the number and morphology of neurons and glia in the hippocampal formation; however, little is known about possible changes in vasculature. Here, we examined the effect of chronic social defeat stress on hippocampal vascular supply in rats. Recent reports document that antidepressant treatment can influence angiogenesis in the hippocampus; therefore, we also studied the effect of antidepressant drug treatment on hippocampal capillarization. Animals were subjected to 5 weeks of daily social defeat by an aggressive conspecific and received concomitant, daily, oral fluoxetine (10 mg/kg) treatment during the last 4 weeks. Rat endothelial cell antigen-1 (RECA-1)-labeling of capillaries and quantitative stereological techniques were used to evaluate the treatment effects on capillary number. Special attention was paid to analysis of the vascular supply of the subgranular zone, which is regarded as an important component of the neurogenic niche for adult hippocampal neurogenesis. Chronic stress significantly decreased the number of microvessels by 30% in all hippocampal subregions, whereas fluoxetine treatment had no influence on capillary number. Furthermore, chronic stress decreased the capillarization of the subgranular zone to a similar extent, indicating that chronic stress affects the vascular niche for adult hippocampal neurogenesis. However, fluoxetine treatment had no impact on capillarization in the subgranular zone. We also detected a decrease in hippocampal volume in the animals as a result of stress, which was mildly altered by fluoxetine treatment. These pronounced changes in vascular supply may explain why the hippocampus is more vulnerable to insults when chronic stress precedes or coincides with other harmful conditions. Reduced microvasculature may also contribute to hippocampal volume decrease in stress-related disorders.

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Section: Vascular Niches Of the Subgranular Zone: Measurement Of Surfmentioning

confidence: 99%

Quantitative changes in hippocampal microvasculature of chronically stressed rats: No effect of fluoxetine treatment

et al. 2009

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“…[18] with the expression (6) ⌬R* 2 Ϸ 4 3 f␥B 0 ⌬ , [19] then gives the criterion 3⌬R* 2 4 f␥B 0 0 Ͼ Ͼ1. [20] Here B 0 is the magnitude of the main field, and ␥ ϭ 2.675 ϫ 10 8 s -1 /T is the proton gyromagnetic ratio. Adopting the estimate 0 ϭ 4 ϫ 10 -8 (6), Eq.…”

Section: Validity Testmentioning

confidence: 99%

Microvessel density estimation in the human brain by means of dynamic contrast‐enhanced echo‐planar imaging

Jensen

Inglese

2006

Magnetic Resonance in Med

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Animal studies have shown that in vivo estimates of microvessel density in the brain may be obtained from an MRI-measurable index (Q) provided that a sufficiently high dose of an intravascular paramagnetic contrast agent is employed. Q is determined from the shifts in the transverse relaxation rates induced by the contrast agent, and a high dose is required for the validity of analytic expressions relating Q to the microvessel density. However, the steady-state imaging techniques used in these prior investigations are not appropriate for humans, as the required contrast agent dose is too large. Here results of a pilot study with three subjects are reported. The results suggest that reliable Q measurements can be performed in the human brain at 1.5 T by using an interleaved spin-echo ( The microvessel density (MVD) is a useful prognostic indicator in many tumors (1,2), and is also of interest in the study of ischemia and aging (3-5). It was recently proposed that an MRI-measurable index (referred to as Q) may allow for in vivo quantitative estimates of the MVD (6,7). In particular, a good correspondence between MVD estimates based on Q and histologic values in the mouse brain was demonstrated in a study using superparamagnetic iron oxide particles (7).However, in order to obtain useful MVD values from Q measurements, a high dose of an intravascular paramagnetic or superparamagnetic contrast agent is required. Indeed, for a steady-state experiment, it has been argued that the necessary dose of gadopentetate dimeglumine (Gd-DTPA) at a field level of 1.5 T should be much greater than the standard clinical dose of 0.1 mmol/kg (6). In this article we show that one can overcome this limitation by measuring Q during the first passage of a contrast agent bolus, which makes Q-based MVD estimation feasible for clinical studies.In addition to the need for a high dose, a limitation of the previously proposed method is that it yields, within the model assumptions, a lower bound for the MVD. In regions where the distribution of vessel radii is narrow, this lower bound is expected to be a good MVD predictor (6,7). However, the presence of larger venules and arterioles can broaden the vessel distribution to the extent that the MVD will be significantly underestimated. For this reason, we introduce here a second Q-based quantity that is formally an MVD upper bound. Indeed, our results suggest that this upper bound actually provides a better estimator of the MVD than does the lower bound.Our method utilizes an interleaved spin-echo (SE)/gradient-echo (GE) echo-planar imaging (EPI) sequence and a bolus injection of triple the standard dose of Gd-DTPA administered at a rate of 5 ml/s, in order to achieve a high first-pass contrast agent concentration. Since triple-dose Gd-DTPA injections are safe and well tolerated (8), the method is suitable for human subjects. The Q-values are measured at the concentration peak during the first passage of the bolus through the brain's microvascular network, to ensure sufficient contrast-agent conc...

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“…Vessels have been identified using standard histological stains such as silver (Gallyas, 1970) or toluidine blue (Manoonkitiwongsa et al, 2001), histochemical assays for enzymes enriched in blood vessels (Lee et al, 2005;Bell et al, 1984) as well as immunohistochemistry for vascular associated antigens. In animals, the cerebrovasculature has also been labeled in vivo by perfusion with photographic emulsions (Boero et al, 1999;D'Andrea, 2004), India ink (Shimada et al, 1992) or lectins .…”

Section: Introductionmentioning

confidence: 99%

Pepsin pretreatment allows collagen IV immunostaining of blood vessels in adult mouse brain

Franciosi¹,

Gasperi²,

Dickstein³

et al. 2007

Journal of Neuroscience Methods

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While the brain vasculature can be imaged with many methods, immunohistochemistry has distinct advantages due to its simplicity and applicability to archival tissue. However, immunohistochemical staining of the murine brain vasculature in aldehyde fixed tissue has proven elusive and inconsistent using current protocols. Here we investigated whether antigen retrieval methods could improve vascular staining in the adult mouse brain. We found that pepsin digestion prior to immunostaining unmasked widespread collagen IV staining of the cerebrovasculature in the adult mouse brain. Pepsin treatment also unmasked widespread vascular staining with laminin, but only marginally improved

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A simple stereologic method for analysis of cerebral cortical microvessels using image analysis

Cited by 14 publications

References 20 publications

Quantitative changes in hippocampal microvasculature of chronically stressed rats: No effect of fluoxetine treatment

Quantitative changes in hippocampal microvasculature of chronically stressed rats: No effect of fluoxetine treatment

Microvessel density estimation in the human brain by means of dynamic contrast‐enhanced echo‐planar imaging

Pepsin pretreatment allows collagen IV immunostaining of blood vessels in adult mouse brain

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