Michael Crair - Academia.edu (original) (raw)

Papers by Michael Crair

Nature communications, 2015

Retinitis pigmentosa is a leading cause of inherited blindness, with no effective treatment curre... more Retinitis pigmentosa is a leading cause of inherited blindness, with no effective treatment currently available. Mutations primarily in genes expressed in rod photoreceptors lead to early rod death, followed by a slower phase of cone photoreceptor death. Rd1 mice provide an invaluable animal model to evaluate therapies for the disease. We previously reported that overexpression of histone deacetylase 4 (HDAC4) prolongs rod survival in rd1 mice. Here we report a key role of a short N-terminal domain of HDAC4 in photoreceptor protection. Expression of this domain suppresses multiple cell death pathways in photoreceptor degeneration, and preserves even more rd1 rods than the full-length HDAC4 protein. Expression of a short N-terminal domain of HDAC4 as a transgene in mice carrying the rd1 mutation also prolongs the survival of cone photoreceptors, and partially restores visual function. Our results may facilitate the design of a small protein therapy for some forms of retinitis pigment...

Developmental neurobiology, Jan 18, 2015

Spontaneous activity during early development is necessary for the formation of precise neural co... more Spontaneous activity during early development is necessary for the formation of precise neural connections, but it remains uncertain whether activity plays an instructive or permissive role in brain wiring. In the visual system, retinal ganglion cell (RGC) projections to the brain form two prominent sensory maps, one reflecting eye of origin and the other retinotopic location. Recent studies provide compelling evidence supporting an instructive role for spontaneous retinal activity in the development of eye-specific projections, but evidence for a similarly instructive role in the development of retinotopy is more equivocal. Here, we report on experiments in which we knocked down the expression of β2-containing nicotinic acetylcholine receptors (β2-nAChRs) specifically in the retina through a Cre-loxP recombination strategy. Overall levels of spontaneous retinal activity in retina-specific β2-nAChR mutant mice (Rx-β2cKO), examined in vitro and in vivo, were reduced to a degree compa...

The Journal of neuroscience : the official journal of the Society for Neuroscience, Jan 15, 2000

The mechanisms underlying changes in neural responses and connections in the visual cortex may be... more The mechanisms underlying changes in neural responses and connections in the visual cortex may be studied by occluding one eye during a critical period in early postnatal life. Under these conditions, neurons in the visual cortex rapidly lose their responses to the deprived eye and ultimately lose many of their inputs from that eye. Cats at the peak of the critical period received infusions of exogenous neurotrophin NT-4/5 into primary visual cortex beginning before a short period of monocular deprivation. Within areas affected by NT-4/5, cortical cells remained responsive to the deprived eye, and maps of ocular dominance were no longer evident using intrinsic-signal optical imaging. Cortical cells also became broadly tuned for stimulus orientation and less responsive to visual stimulation through either eye. These effects required at least 48 hr exposure to the neurotrophin and were specific for trkB, because they were not seen with the trkA or trkC ligands NGF or NT-3. Even after ...

The Journal of neuroscience : the official journal of the Society for Neuroscience, 1998

To investigate the possible anatomical basis for the functional recovery of visual cortical respo... more To investigate the possible anatomical basis for the functional recovery of visual cortical responses after reverse monocular deprivation, we have studied the morphology of single geniculocortical afferents to area 17. In kittens reverse-sutured for 10 d after an initial week of monocular deprivation, single-unit and intrinsic signal optical recordings confirmed that the effects of the initial deprivation were largely reversed. Responses through the originally nondeprived (OND) eye were drastically diminished, but remained much more selective for orientation than after an initial monocular deprivation (Crair et al., 1997). Responses through the originally deprived (OD) eye recovered completely. Geniculocortical afferent arbors in layer IV of area 17 were filled by iontophoresis of Phaseolus lectin into lamina A of the lateral geniculate nucleus (LGN) and were serially reconstructed. Arbors serving both the OD and the OND eye were analyzed. The plastic changes of both OD and OND arbo...

Development (Cambridge, England), 2002

In mice, Brn3 POU domain transcription factors play essential roles in the differentiation and su... more In mice, Brn3 POU domain transcription factors play essential roles in the differentiation and survival of projection neurons within the retina, inner ear, dorsal root and trigeminal ganglia. During retinal ganglion cell differentiation, Brn3b is expressed first, followed by Brn3a and Brn3c. Targeted deletion of Brn3b, but not Brn3a or Brn3c, leads to a loss of most retinal ganglion cells before birth. However, as a few retinal ganglion cells are still present in Brn3b(-/-) mice, Brn3a and Brn3c may partially compensate for the loss of Brn3b. To examine the role of Brn3c in retinal ganglion cell development, we generated Brn3b/Brn3c double knockout mice and analyzed their retinas and optic chiasms. Retinal ganglion cell axons from double knockout mice were more severely affected than were those from Brn3b-deficient mice, indicating that Brn3c was required for retinal ganglion cell differentiation and could partially compensate for the loss of Brn3b. Moreover, Brn3c had functions in ...

Nanosystems in Engineering and Medicine, 2012

ABSTRACT Spontaneous neural activities exist early in development and their spatiotemporal patter... more ABSTRACT Spontaneous neural activities exist early in development and their spatiotemporal patterns play important roles in the development of sensory maps such as maps of retinotopy in the visual system. We summarized different optogenetic tools, including transgenic mouse lines, viral-mediated transfection and electroporation methods to enable the expression of light-gated channelrhodopsin (ChR2) in retinal ganglion cells (RGCs) before the onset of vision. Patch-clamp and extracellular recording experiments verified that activities of ChR2-expressing cells were precisely manipulated by the patterns of optical stimuli. In chronic stimulation experiments, light-emitting diodes controlled the activity patterns of ChR2-expressing RGCs in vivo. Changes in the retinotopic map in Superior Colliculus (SC) were examined by quantifying the relative sizes of fluorescently labeled target zones. Our results revealed that various optogenetic and optical tools can manipulate retinal activities with precise temporal patterns. These techniques can be readily used in studying the development of the central nervous system of neonatal rodents.

Neuron, 2008

Cortical pyramidal neurons alter their responses to input signals depending on behavioral state. ... more Cortical pyramidal neurons alter their responses to input signals depending on behavioral state. We investigated whether changes in somatic inhibition contribute to these alterations. In layer 5 pyramidal neurons of rat visual cortex, repetitive firing from a depolarized membrane potential, which typically occurs during arousal, produced long-lasting depression of somatic inhibition. In contrast, slow membrane oscillations with firing in the depolarized phase, which typically occurs during slow-wave sleep, produced long-lasting potentiation. The depression is mediated by L-type Ca 2+ channels and GABA A receptor endocytosis, whereas potentiation is mediated by R-type Ca 2+ channels and receptor exocytosis. It is likely that the direction of modification is mainly dependent on the ratio of R-and L-type Ca 2+ channel activation. Furthermore, somatic inhibition was stronger in slices prepared from rats during slow-wave sleep than arousal. This bidirectional modification of somatic inhibition may alter pyramidal neuron responsiveness in accordance with behavioral state.

The Journal of Physiology, 2009

Gradients of molecular factors pattern the developing retina and superior colliculus (SC) and gui... more Gradients of molecular factors pattern the developing retina and superior colliculus (SC) and guide retinal ganglion cell (RGC) axons to their appropriate central target perinatally. During and subsequent to this period, spontaneous waves of action potentials sweep across the retina, providing an instructive topographic signal based on the correlations of firing patterns of neighbouring RGCs. How these activity-independent and activity-dependent factors interact during retinotopic map formation remains unclear. A typical phenotype of mutant mice lacking genes for one or more RGC axon guidance molecules is the presence of topographically inappropriate projections or 'ectopic spots'. Here, we examine mice that lack functional bone morphogenetic protein receptors (BMPRs) in the retina. Retinal BMP controls the graded expression of RGC axon guidance molecules, resulting in some dorsal RGCs projecting ectopically to locations in the SC that normally receive input from ventral retina. We examine the consequences of this anatomical phenotype in vivo by studying the receptive field (RF) properties of neurons in the superficial SC. We observe a mixture of physiological phenotypes in BMPR mutant mice; notably we find some neurons with ectopic RFs displaced in elevation, corresponding to the observed anatomical defect. However, in a result not necessarily congruent with the presence of focal ectopic projections, some neurons have split, enlarged and patchy/distorted RFs. These results are consistent with the effects of spontaneous retinal waves acting upon a disrupted molecular template, and they place significant limits on the form of an activity-dependent learning rule for the development of retinocollicular projections.

Neuroscience Research Supplements, 1993

Neuroscience Research Supplements, 1993

Neuroscience Research Supplements, 1994

[Proceedings] 1991 IEEE International Joint Conference on Neural Networks, 1991

along the center lines of ocular dominance bands (Bartfeld and Grinvald, 1992;. Deda C. Gillespie... more along the center lines of ocular dominance bands (Bartfeld and Grinvald, 1992;. Deda C. Gillespie, and Michael P. Stryker* W. M. Keck Foundation Center for Integrative

have contributed equally to this work.

Journal of Visualized Experiments, 2011

The targeting and refinement of RGC projections to the midbrain is a popular and powerful model s... more The targeting and refinement of RGC projections to the midbrain is a popular and powerful model system for studying how precise patterns of neural connectivity form during development. In mice, retinofugal projections are arranged in a topographic manner and form eye-specific layers in the Lateral Geniculate Nucleus (dLGN) of the thalamus and the Superior Colliculus (SC). The development of these precise patterns of retinofugal projections has typically been studied by labeling populations of RGCs with fluorescent dyes and tracers, such as horseradish peroxidase. However, these methods are too coarse to provide insight into developmental changes in individual RGC axonal arbor morphology that are the basis of retinotopic map formation. They also do not allow for the genetic manipulation of RGCs. Recently, electroporation has become an effective method for providing precise spatial and temporal control for delivery of charged molecules into the retina. Current retinal electroporation protocols do not allow for genetic manipulation and tracing of retinofugal projections of a single or small cluster of RGCs in postnatal mice. It has been argued that postnatal in vivo electroporation is not a viable method for transfecting RGCs since the labeling efficiency is extremely low and hence requires targeting at embryonic ages when RGC progenitors are undergoing differentiation and proliferation. In this video we describe an in vivo electroporation protocol for targeted delivery of genes, shRNA, and fluorescent dextrans to murine RGCs postnatally. This technique provides a cost effective, fast and relatively easy platform for efficient screening of candidate genes involved in several aspects of neural development including axon retraction, branching, lamination, regeneration and synapse formation at various stages of circuit development. In summary we describe here a valuable tool which will provide further insights into the molecular mechanisms underlying sensory map development.

Nature communications, 2015

Retinitis pigmentosa is a leading cause of inherited blindness, with no effective treatment curre... more Retinitis pigmentosa is a leading cause of inherited blindness, with no effective treatment currently available. Mutations primarily in genes expressed in rod photoreceptors lead to early rod death, followed by a slower phase of cone photoreceptor death. Rd1 mice provide an invaluable animal model to evaluate therapies for the disease. We previously reported that overexpression of histone deacetylase 4 (HDAC4) prolongs rod survival in rd1 mice. Here we report a key role of a short N-terminal domain of HDAC4 in photoreceptor protection. Expression of this domain suppresses multiple cell death pathways in photoreceptor degeneration, and preserves even more rd1 rods than the full-length HDAC4 protein. Expression of a short N-terminal domain of HDAC4 as a transgene in mice carrying the rd1 mutation also prolongs the survival of cone photoreceptors, and partially restores visual function. Our results may facilitate the design of a small protein therapy for some forms of retinitis pigment...

Developmental neurobiology, Jan 18, 2015

Spontaneous activity during early development is necessary for the formation of precise neural co... more Spontaneous activity during early development is necessary for the formation of precise neural connections, but it remains uncertain whether activity plays an instructive or permissive role in brain wiring. In the visual system, retinal ganglion cell (RGC) projections to the brain form two prominent sensory maps, one reflecting eye of origin and the other retinotopic location. Recent studies provide compelling evidence supporting an instructive role for spontaneous retinal activity in the development of eye-specific projections, but evidence for a similarly instructive role in the development of retinotopy is more equivocal. Here, we report on experiments in which we knocked down the expression of β2-containing nicotinic acetylcholine receptors (β2-nAChRs) specifically in the retina through a Cre-loxP recombination strategy. Overall levels of spontaneous retinal activity in retina-specific β2-nAChR mutant mice (Rx-β2cKO), examined in vitro and in vivo, were reduced to a degree compa...

The Journal of neuroscience : the official journal of the Society for Neuroscience, Jan 15, 2000

The mechanisms underlying changes in neural responses and connections in the visual cortex may be... more The mechanisms underlying changes in neural responses and connections in the visual cortex may be studied by occluding one eye during a critical period in early postnatal life. Under these conditions, neurons in the visual cortex rapidly lose their responses to the deprived eye and ultimately lose many of their inputs from that eye. Cats at the peak of the critical period received infusions of exogenous neurotrophin NT-4/5 into primary visual cortex beginning before a short period of monocular deprivation. Within areas affected by NT-4/5, cortical cells remained responsive to the deprived eye, and maps of ocular dominance were no longer evident using intrinsic-signal optical imaging. Cortical cells also became broadly tuned for stimulus orientation and less responsive to visual stimulation through either eye. These effects required at least 48 hr exposure to the neurotrophin and were specific for trkB, because they were not seen with the trkA or trkC ligands NGF or NT-3. Even after ...

The Journal of neuroscience : the official journal of the Society for Neuroscience, 1998

To investigate the possible anatomical basis for the functional recovery of visual cortical respo... more To investigate the possible anatomical basis for the functional recovery of visual cortical responses after reverse monocular deprivation, we have studied the morphology of single geniculocortical afferents to area 17. In kittens reverse-sutured for 10 d after an initial week of monocular deprivation, single-unit and intrinsic signal optical recordings confirmed that the effects of the initial deprivation were largely reversed. Responses through the originally nondeprived (OND) eye were drastically diminished, but remained much more selective for orientation than after an initial monocular deprivation (Crair et al., 1997). Responses through the originally deprived (OD) eye recovered completely. Geniculocortical afferent arbors in layer IV of area 17 were filled by iontophoresis of Phaseolus lectin into lamina A of the lateral geniculate nucleus (LGN) and were serially reconstructed. Arbors serving both the OD and the OND eye were analyzed. The plastic changes of both OD and OND arbo...

Development (Cambridge, England), 2002

In mice, Brn3 POU domain transcription factors play essential roles in the differentiation and su... more In mice, Brn3 POU domain transcription factors play essential roles in the differentiation and survival of projection neurons within the retina, inner ear, dorsal root and trigeminal ganglia. During retinal ganglion cell differentiation, Brn3b is expressed first, followed by Brn3a and Brn3c. Targeted deletion of Brn3b, but not Brn3a or Brn3c, leads to a loss of most retinal ganglion cells before birth. However, as a few retinal ganglion cells are still present in Brn3b(-/-) mice, Brn3a and Brn3c may partially compensate for the loss of Brn3b. To examine the role of Brn3c in retinal ganglion cell development, we generated Brn3b/Brn3c double knockout mice and analyzed their retinas and optic chiasms. Retinal ganglion cell axons from double knockout mice were more severely affected than were those from Brn3b-deficient mice, indicating that Brn3c was required for retinal ganglion cell differentiation and could partially compensate for the loss of Brn3b. Moreover, Brn3c had functions in ...

Nanosystems in Engineering and Medicine, 2012

ABSTRACT Spontaneous neural activities exist early in development and their spatiotemporal patter... more ABSTRACT Spontaneous neural activities exist early in development and their spatiotemporal patterns play important roles in the development of sensory maps such as maps of retinotopy in the visual system. We summarized different optogenetic tools, including transgenic mouse lines, viral-mediated transfection and electroporation methods to enable the expression of light-gated channelrhodopsin (ChR2) in retinal ganglion cells (RGCs) before the onset of vision. Patch-clamp and extracellular recording experiments verified that activities of ChR2-expressing cells were precisely manipulated by the patterns of optical stimuli. In chronic stimulation experiments, light-emitting diodes controlled the activity patterns of ChR2-expressing RGCs in vivo. Changes in the retinotopic map in Superior Colliculus (SC) were examined by quantifying the relative sizes of fluorescently labeled target zones. Our results revealed that various optogenetic and optical tools can manipulate retinal activities with precise temporal patterns. These techniques can be readily used in studying the development of the central nervous system of neonatal rodents.

Neuron, 2008

Cortical pyramidal neurons alter their responses to input signals depending on behavioral state. ... more Cortical pyramidal neurons alter their responses to input signals depending on behavioral state. We investigated whether changes in somatic inhibition contribute to these alterations. In layer 5 pyramidal neurons of rat visual cortex, repetitive firing from a depolarized membrane potential, which typically occurs during arousal, produced long-lasting depression of somatic inhibition. In contrast, slow membrane oscillations with firing in the depolarized phase, which typically occurs during slow-wave sleep, produced long-lasting potentiation. The depression is mediated by L-type Ca 2+ channels and GABA A receptor endocytosis, whereas potentiation is mediated by R-type Ca 2+ channels and receptor exocytosis. It is likely that the direction of modification is mainly dependent on the ratio of R-and L-type Ca 2+ channel activation. Furthermore, somatic inhibition was stronger in slices prepared from rats during slow-wave sleep than arousal. This bidirectional modification of somatic inhibition may alter pyramidal neuron responsiveness in accordance with behavioral state.

The Journal of Physiology, 2009

Gradients of molecular factors pattern the developing retina and superior colliculus (SC) and gui... more Gradients of molecular factors pattern the developing retina and superior colliculus (SC) and guide retinal ganglion cell (RGC) axons to their appropriate central target perinatally. During and subsequent to this period, spontaneous waves of action potentials sweep across the retina, providing an instructive topographic signal based on the correlations of firing patterns of neighbouring RGCs. How these activity-independent and activity-dependent factors interact during retinotopic map formation remains unclear. A typical phenotype of mutant mice lacking genes for one or more RGC axon guidance molecules is the presence of topographically inappropriate projections or 'ectopic spots'. Here, we examine mice that lack functional bone morphogenetic protein receptors (BMPRs) in the retina. Retinal BMP controls the graded expression of RGC axon guidance molecules, resulting in some dorsal RGCs projecting ectopically to locations in the SC that normally receive input from ventral retina. We examine the consequences of this anatomical phenotype in vivo by studying the receptive field (RF) properties of neurons in the superficial SC. We observe a mixture of physiological phenotypes in BMPR mutant mice; notably we find some neurons with ectopic RFs displaced in elevation, corresponding to the observed anatomical defect. However, in a result not necessarily congruent with the presence of focal ectopic projections, some neurons have split, enlarged and patchy/distorted RFs. These results are consistent with the effects of spontaneous retinal waves acting upon a disrupted molecular template, and they place significant limits on the form of an activity-dependent learning rule for the development of retinocollicular projections.

Neuroscience Research Supplements, 1993

Neuroscience Research Supplements, 1993

Neuroscience Research Supplements, 1994

[Proceedings] 1991 IEEE International Joint Conference on Neural Networks, 1991

along the center lines of ocular dominance bands (Bartfeld and Grinvald, 1992;. Deda C. Gillespie... more along the center lines of ocular dominance bands (Bartfeld and Grinvald, 1992;. Deda C. Gillespie, and Michael P. Stryker* W. M. Keck Foundation Center for Integrative

have contributed equally to this work.

Journal of Visualized Experiments, 2011

The targeting and refinement of RGC projections to the midbrain is a popular and powerful model s... more The targeting and refinement of RGC projections to the midbrain is a popular and powerful model system for studying how precise patterns of neural connectivity form during development. In mice, retinofugal projections are arranged in a topographic manner and form eye-specific layers in the Lateral Geniculate Nucleus (dLGN) of the thalamus and the Superior Colliculus (SC). The development of these precise patterns of retinofugal projections has typically been studied by labeling populations of RGCs with fluorescent dyes and tracers, such as horseradish peroxidase. However, these methods are too coarse to provide insight into developmental changes in individual RGC axonal arbor morphology that are the basis of retinotopic map formation. They also do not allow for the genetic manipulation of RGCs. Recently, electroporation has become an effective method for providing precise spatial and temporal control for delivery of charged molecules into the retina. Current retinal electroporation protocols do not allow for genetic manipulation and tracing of retinofugal projections of a single or small cluster of RGCs in postnatal mice. It has been argued that postnatal in vivo electroporation is not a viable method for transfecting RGCs since the labeling efficiency is extremely low and hence requires targeting at embryonic ages when RGC progenitors are undergoing differentiation and proliferation. In this video we describe an in vivo electroporation protocol for targeted delivery of genes, shRNA, and fluorescent dextrans to murine RGCs postnatally. This technique provides a cost effective, fast and relatively easy platform for efficient screening of candidate genes involved in several aspects of neural development including axon retraction, branching, lamination, regeneration and synapse formation at various stages of circuit development. In summary we describe here a valuable tool which will provide further insights into the molecular mechanisms underlying sensory map development.