Proliferation and differentiation of progenitor cells in the cortex and the subventricular zone in the adult rat after focal cerebral ischemia

doi:10.1016/S0306-4522(01)00117-8

Neuroscience

Volume 105, Issue 1, 16 July 2001, Pages 33-41

https://doi.org/10.1016/S0306-4522(01)00117-8 Get rights and content

Abstract

Progenitor cells in the subventricular zone of the lateral ventricle and in the dentate gyrus of the hippocampus can proliferate throughout the life of the animal. To examine the proliferation and fate of progenitor cells in the subventricular zone and dentate gyrus after focal cerebral ischemia, we measured the temporal and spatial profiles of proliferation of cells and the phenotypic fate of proliferating cells in ischemic brain in a model of embolic middle cerebral artery occlusion in the adult rat. Proliferating cells were labeled by injection of bromodeoxyuridine (BrdU) in a pulse or a cumulative protocol. To determine the temporal profile of proliferating cells, ischemic rats were injected with BrdU every 4 h for 12 h on the day preceding death. Rats were killed 2–14 days after ischemia. We observed significant increases in numbers of proliferating cells in the ipsilateral cortex and subventricular zone 2–14 days with a peak at 7 days after ischemia compared with the control group. To maximize labeling of proliferating cells, a single daily injection of BrdU was administered over a 14-day period starting the day after ischemia. Rats were killed either 2 h or 28 days after the last injection of BrdU. A significant increase in numbers of BrdU immunoreactive cells in the subventricular zone was coincident with a significant increase in numbers of BrdU immunoreactive cells in the olfactory bulb 14 days after ischemia and numbers of BrdU immunoreactive cells did not significantly increase in the dentate gyrus. However, 28 days after the last labeling, the number of BrdU labeled cells decreased by 90% compared with number at 14 days. Clusters of BrdU labeled cells were present in the cortex distal to the infarction. Numerous cells immunostained for the polysialylated form of the neuronal cell adhesion molecule were detected in the ipsilateral subventricular zone. Only 6% of BrdU labeled cells exhibited glial fibrillary acidic protein immunoreactivity in the cortex and subcortex and no BrdU labeled cells expressed neuronal protein markers (neural nuclear protein and microtubule associated protein-2).

From these data we suggest that focal cerebral ischemia induces transient and regional specific increases in cell proliferation in the ipsilateral hemisphere and that proliferating progenitor cells may exist in the adult cortex.

Section snippets

General preparation

Adult male Wistar rats (n=44, weighing 300–350 g, obtained from Charles River, Portage, MI, USA) were used in this study. All efforts were made to minimize both the suffering and the number of animals used. All procedures were conducted in strict accordance with the NIH Guide for the Care and Use of Laboratory Animals, and have been approved by the institutional care of experimental animals committee. Animals were anesthetized with halothane (1–3.5% in a mixture of 70% N₂O and 30% O₂) using a

Results

All rats subjected to embolic MCA occlusion exhibited an ischemic lesion (for rats killed at 1–3 days after MCA occlusion) or infarction (for rats killed 7–42 days after MCA occlusion) in the ipsilateral MCA supplied territory (Fig. 2).

Discussion

Progenitor cells in the SVZ of the lateral ventricle and in the dentate gyrus of the hippocampus can proliferate throughout the life of the rodent (Gage et al., 1998, Garcia-Verdugo et al., 1998). Proliferating cells constitute an average of 10% of the cell population in the SVZ in adult mice under physiological conditions (Morshead and van der Kooy, 1992). Although cellular composition, proliferation, migration and differentiation in the SVZ and the dentate gyrus are under extensive

Acknowledgements

The authors wish to thank Cecylia Power for technical assistance. This work was supported by NINDS Grants PO1 NS23393 and RO1 NS35504.

References (32)

S. Cramer et al.
Recovery recapitulates ontogeny
Trends Neurosci.
(2000)
H. Hu et al.
The role of polysialic acid in migration of olfactory bulb interneuron precursors in the subventricular zone
Neuron
(1996)
C.B. Johansson et al.
Identification of a neural stem cell in the adult mammalian central nervous system
Cell
(1999)
N. Latov et al.
Fibrillary astrocytes proliferate in response to brain injury: a study combining immunoperoxidase technique for glial fibrillary acidic protein and radioautography of tritiated thymidine
Dev. Biol.
(1979)
S.W. Levison et al.
Both oligodendrocytes and astrocytes develop from progenitors in the subventricular zone of postnatal rat forebrain
Neuron
(1993)
T. Miyake et al.
Quantitative studies on proliferative changes of reactive astrocytes in mouse cerebral cortex
Brain Res.
(1988)
C.M. Morshead et al.
Separate blood and brain origins of proliferating cells during gliosis in adult brains
Brain Res.
(1990)
H. Tomasiewicz et al.
Genetic deletion of a neural cell adhesion molecule variant (N-CAM-180) produces distinct defects in the central nervous system
Neuron
(1993)
R.L. Zhang et al.
A rat model of embolic focal cerebral ischemia
Brain Res.
(1997)
W.M. Cheung et al.
Changes in the level of glial fibrillary acidic protein (GFAP) after mild and severe focal cerebral ischemia
Chin. J. Physiol.
(1999)

B.J. Chiasson et al.

Adult mammalian forebrain ependymal and subependymal cells demonstrate proliferative potential, but only subependymal cells have neural stem cell characteristics

J. Neurosci.

(1999)

C.M. Chuong et al.

Alterations in neural cell adhesion molecules during development of different regions of the nervous system

J. Neurosci.

(1984)

M.R. Del Bigio

The ependyma: a protective barrier between brain and cerebrospinal fluid

Glia

(1995)

F. Doetsch et al.

Cellular composition and three-dimensional organization of the subventricular germinal zone in the adult mammalian brain

J. Neurosci.

(1997)

F. Dolbeare

Bromodeoxyuridine: a diagnostic tool in biology and medicine, Part III. Proliferation in normal, injured and diseased tissue, growth factors, differentiation, DNA replication sites and in situ hybridization

Histochem. J.

(1996)

M.A. Eglitis et al.

Targeting of marrow-derived astrocytes to the ischemic brain

NeuroReport

(1999)

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Proliferation and differentiation of progenitor cells in the cortex and the subventricular zone in the adult rat after focal cerebral ischemia

Abstract

Section snippets

General preparation

Results

Discussion

Acknowledgements

Trends Neurosci.

Neuron

Cell

Dev. Biol.

Neuron

Brain Res.

Brain Res.

Neuron

Brain Res.

Changes in the level of glial fibrillary acidic protein (GFAP) after mild and severe focal cerebral ischemia

Chin. J. Physiol.

Adult mammalian forebrain ependymal and subependymal cells demonstrate proliferative potential, but only subependymal cells have neural stem cell characteristics

J. Neurosci.

Alterations in neural cell adhesion molecules during development of different regions of the nervous system

J. Neurosci.

The ependyma: a protective barrier between brain and cerebrospinal fluid

Glia

Cellular composition and three-dimensional organization of the subventricular germinal zone in the adult mammalian brain

J. Neurosci.

Bromodeoxyuridine: a diagnostic tool in biology and medicine, Part III. Proliferation in normal, injured and diseased tissue, growth factors, differentiation, DNA replication sites and in situ hybridization

Histochem. J.

Targeting of marrow-derived astrocytes to the ischemic brain

NeuroReport