Cortical barrelfields in organotypic slice cultures from rat somatosensory cortex

Behan, Mary; Kroker, Axel; Bolz, Jürgen

doi:10.1016/0304-3940(91)90567-d

Cited by 12 publications

(5 citation statements)

References 25 publications

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“…Unlike the laminae already formed by the time of culture preparation, layers II-IV were not clearly distinguishable in these cultures. These findings are consistent with other reports suggesting that cortical cytoarchitectonic features formed by the time of slice preparation are maintained in culture (De Jong et al, 1988;Behan et al, 1991;Distler and Robertson, 1992). Neurons that would normally give rise to layers 11,111, and IV are generated later and are either contained within the undifferentiated compact cortical plate or are still in the process of migration to the cortical plate at PO ( Angevine and Sidman, 196 1 ;Berry and Rogers, 1965;Walsh and Cepko, 1992).…”

Section: Laminae In Vifrosupporting

confidence: 93%

Aspects of early postnatal development of cortical neurons that proceed independently of normally present extrinsic influences

1993

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To examine the contribution of local versus extrinsic influences on postnatal development of cortical neurons, we compared the maturation of deep (infragranular) layer neurons in isolated slices of neocortex grown in organotypic culture to a similar population of neurons developing in vivo. All slice cultures were prepared from sensorimotor cortices of newborn mice (P0) and neurons in these cultures were examined at daily intervals during the first 9 days in vitro (DIV). The maturational state of neurons developing in vivo over this same time period was assessed in acute slices prepared from animals of equivalent postnatal age, P1-P9. Electrophysiological recordings were obtained from neurons in both cultured and acute slices, using Lucifer yellow filled whole-cell recording electrodes, enabling subsequent morphometric analysis of the labeled cells. We report significant changes in both cellular morphology and electrical membrane properties of these deep layer cortical neurons during the first week in culture. Morphological maturation over this time period was characterized by a two- to three-fold increase in cell body size and total process length, and an increase in dendritic complexity. In this same population of cells a three-fold decrease in input resistance and changes in the action potential waveform, including a two-fold decrease in the AP duration, also occur. The degree of morphological and electrophysiological differentiation of individual neurons was highly correlated across developmental ages, suggesting that the maturational state of a cell is reflected in both cellular morphology and intrinsic membrane properties. A remarkably similar pattern of neuronal maturation was observed in neurons in layers V, VI/SP examined in acute slices prepared from animals between P1-P9. Because our culture system preserves many aspects of the local cortical environment while eliminating normal extrinsic influences (including thalamic, brainstem, and callosal connections), our findings argue that this early phase of neuronal differentiation, including the rate and extent of dendritic growth and development of AP waveform, results from instructive and/or permissive local influences, and appears to proceed independently of the many normally present extrinsic factors.

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Section: Laminae In Vifrosupporting

confidence: 93%

Aspects of early postnatal development of cortical neurons that proceed independently of normally present extrinsic influences

1993

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“…In these preparations not only was it found that PNs in explants from P2 animals developed cytotypically in contrast to the organotypic appearance of P6 explants, but also that the PN dendritic tree was normal in the absence of excitatory neurotransmission and BDNF signaling (Adcock et al, 2004;Fenili and De Boni, 2003). Annis et al, 1990Annis et al, , 1993Baker et al, 1992;Behan et al, 1991;Brussaard and Baker, 1995;Caeser et al, 1989;Chattopadhyaya et al, 2004;Cheema et al, 2000;Corthesy-Theulaz et al, 1991;Dayer et al, 2008;Del Rio et al, 1996;Gotz and Bolz, 1989;Humpel and Weis, 2002;Ide et al, 1996;Jacobsen and Miller, 2003;Kierstein et al, 1996;Leiman and Seil, 1986;Molnar and Blakemore, 1999;Neely et al, 2007;Patz et al, 2003;Plenz and Kitai, 1996b;Prewitt and Wilson, 2007;Robertson et al, 1997;Romijn et al, 1988;Ross and Porter, 2002;Soares and Sotelo, 2004;Toran-Allerand, 1991;Vogt Weisenhorn et al, 1998;Wang and Johnson, 2007;Wirth et al, 1998;Yamamoto et al, 1989;Zhou et al, 1993;Zink et al, 2004Striatum Aleksandrova et al, 2001Gutierrez et al, 2004;Humpel et al, 1996;Liu et al, 1995; These exemplificative observations s...…”

Section: Organotypic Culturesmentioning

confidence: 99%

Cell death and proliferation in acute slices and organotypic cultures of mammalian CNS

Lossi

Alasia

Salio

et al. 2009

Progress in Neurobiology

127

125

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“…This implies that the neurons in our sample developed, to a large degree, without sensory input, and may therefore have abnormal properties. Previous studies have documented similarities in many of the morphological and electrophysiological properties of neurons which developed in organotypic slice cultures to those of neurons which developed in vivo, suggesting that proper development of these properties does not require intact sensory input (Bolz et al, 1990;Behan et al, 1991;Bolz et al, 1992;Novak and Bolz, 1993;Sieg et al, 1998). Similar conclusions were reached in a recent study that focused specifically on morphological and physiological properties of inhibitory cortical interneurons (Klostermann and Wahle, 1999).…”

Section: Potential Caveatssupporting

confidence: 68%

Dendritic development of GABAergic cortical interneurons revealed by biolistic transfection with GFP

Jin¹

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Dendritic Development of GABAergic Cortical Interneurons Revealed by Biolistic Transfection with GFP Xiaoming Jin Neurons using γ-aminobutyric acid (GABA) as a neurotransmitter are critically important as inhibitory neurons in the CNS. A major hindrance to further studies of this important and highly diverse class of neurons is their relative scarcity, which makes it difficult to identify and target them for morphological and physiological studies. To overcome this difficulty, we developed a novel protocol for visualizing detailed morphology of GABAergic neurons in living mouse brain slice cultures, using biolistic transfection with GAD67-GFP. The morphological, immunocytochemical and physiological characteristics of GAD67-GFP expressing neurons indicated that they were functional GABAergic neurons. Additionally, our data demonstrate that this technique allows patch-clamp recording of firing patterns and postsynaptic currents from GABAergic cortical interneurons, and unitary inhibitory postsynaptic currents from neurons postsynaptic to them. We imaged and reconstructed dendritic morphology of 150 GAD67-GFP expressing cortical interneurons in organotypic slice cultures. Using a novel computational metric of dendritic polarization, we demonstrated that dendrites of about two thirds of these GAD67-GFP expressing neurons were not randomly oriented, but instead showed a highly significant tendency to extend along the vertical axis of the cortex, indicating that GABAergic interneurons make an important contribution to the vertical bias of the neocortical neuropil. We further tested the hypothesis that brain derived neurotrophic factor (BDNF) is involved in activity-dependent dendritic growth of cortical GABAergic interneurons. By repeated confocal imaging of the same living GFPexpressing neurons at 5-day intervals, we demonstrated that both high KCl medium and BDNF enhance dendritic growth of GABAergic neurons, and that the enhancing effect of depolarization on dendritic growth is mediated by BDNF, while the BDNF effect is not dependent on neuronal activity. These results suggest that enhanced neuronal activity acts to promote dendritic growth of GABAergic interneurons by increasing expression and release of BDNF from neighboring neurons. Altogether, we demonstrated two modes of dendritic development in GABAergic cortical interneurons: vertically-oriented growth and activity-dependent growth, with each probably serving different functional requirements of the cerebral cortex. Our results also demonstrate that biolistic transfection with GAD67-GFP is an efficient, flexible and reliable technique for electrophysiological and morphological studies of living GABAergic interneurons.

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Cortical barrelfields in organotypic slice cultures from rat somatosensory cortex

Cited by 12 publications

References 25 publications

Aspects of early postnatal development of cortical neurons that proceed independently of normally present extrinsic influences

Aspects of early postnatal development of cortical neurons that proceed independently of normally present extrinsic influences

Cell death and proliferation in acute slices and organotypic cultures of mammalian CNS

Dendritic development of GABAergic cortical interneurons revealed by biolistic transfection with GFP

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