Molecular development of the lateral geniculate nucleus in the absence of retinal waves during the time of retinal axon eye-specific segregation

BackgroundHigher mammals such as primates and carnivores have highly developed unique brain structures such as the ocular dominance columns in the visual cortex, and the gyrus and outer subventricular zone of the cerebral cortex. However, our molecular understanding of the formation, function and diseases of these structures is still limited, mainly because genetic manipulations that can be applied to higher mammals are still poorly available.ResultsHere we developed and validated a rapid and efficient technique that enables genetic manipulations in the brain of gyrencephalic carnivores using in utero electroporation. Transgene-expressing ferret babies were obtained within a few weeks after electroporation. GFP expression was detectable in the embryo and was observed at least 2 months after birth. Our technique was useful for expressing transgenes in both superficial and deep cortical neurons, and for examining the dendritic morphologies and axonal trajectories of GFP-expressing neurons in ferrets. Furthermore, multiple genes were efficiently co-expressed in the same neurons.ConclusionOur method promises to be a powerful tool for investigating the fundamental mechanisms underlying the development, function and pathophysiology of brain structures which are unique to higher mammals.

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“…Ferrets were maintained as described previously [1,2]. The day of birth was counted as postnatal day 0 (P0).…”

Section: Methodsmentioning

confidence: 99%

Rapid and efficient genetic manipulation of gyrencephalic carnivores using in utero electroporation

2012

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show abstract

“…Ferrets were maintained as described previously (Kawasaki et al, 2004; Iwai and Kawasaki, 2009; Iwai et al, 2012; Kawasaki et al, 2012). The day of birth was counted as postnatal day 0 (P0).…”

Section: Methodsmentioning

confidence: 99%

In vivo genetic manipulation of cortical progenitors in gyrencephalic carnivores using in utero electroporation

2012

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SummaryBrain structures such as the outer subventricular zone (OSVZ) and the inner fiber layer (IFL) in the developing cerebral cortex are especially prominent in higher mammals. However, the molecular mechanisms underlying the formation of the OSVZ are still largely unknown, mainly because genetic manipulations that can be applied to the OSVZ in higher mammals had been poorly available. Here we developed and validated a rapid and efficient genetic manipulation technique for germinal zones including the OSVZ using in utero electroporation in developing gyrencephalic carnivore ferrets. We also determined the optimal conditions for using in utero electroporation to express transgenes in germinal zones. Using our electroporation procedure, the morphology of GFP-positive cells in the OSVZ was clearly visible even without immunostaining, and multiple genes were efficiently co-expressed in the same cells. Furthermore, we uncovered that fibers, which seemed to correspond to those in the IFL of monkeys, also existed in ferrets, and were derived from newly generated cortical neurons. Our technique promises to be a powerful tool for investigating the fundamental mechanisms underlying the formation and abnormalities of the cerebral cortex in higher mammals.

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“…All mice were reared on a normal 12 h light/dark schedule. ME experiments were carried out as described previously with slight modifications [60]. After mice were anesthetized with isoflurane, the left eye was removed, and pieces of Gelform (Pfizer, New York, NY, USA) were inserted into the cavity.…”

Section: Methodsmentioning

confidence: 99%

“…The Genbank accession number of mouse c-fos used here was AK154418. In situ hybridization was performed as described previously with slight modifications [60]. Sections prepared from fresh-frozen tissues were treated with 4% paraformaldehyde for 10 min and 0.25% acetic anhydride for 10 min.…”

Section: Methodsmentioning

confidence: 99%

Rearrangement of Retinogeniculate Projection Patterns after Eye-Specific Segregation in Mice

Hayakawa

Kawasaki

2010

PLoS ONE

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It has been of interest whether and when the rearrangement of neuronal circuits can be induced after projection patterns are formed during development. Earlier studies using cats reported that the rearrangement of retinogeniculate projections could be induced even after eye-specific segregation has occurred, but detailed and quantitative characterization of this rearrangement has been lacking. Here we delineate the structural changes of retinogeniculate projections in the C57BL/6 mouse in response to monocular enucleation (ME) after eye-specific segregation. When ME was performed after eye-specific segregation, rearrangement of retinogeniculate axons in the dorsal lateral geniculate nucleus (dLGN) was observed within 5 days. Although this rearrangement was observed both along the dorsomedial-ventrolateral and outer-inner axes in the dLGN, it occurred more rapidly along the outer-inner axis. We also examined the critical period for this rearrangement and found that the rearrangement became almost absent by the beginning of the critical period for ocular dominance plasticity in the primary visual cortex. Taken together, our findings serve as a framework for the assessment of phenotypes of genetically altered mouse strains as well as provide insights into the mechanisms underlying the rearrangement of retinogeniculate projections.

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Molecular development of the lateral geniculate nucleus in the absence of retinal waves during the time of retinal axon eye-specific segregation

Cited by 33 publications

References 52 publications

Rapid and efficient genetic manipulation of gyrencephalic carnivores using in utero electroporation

Rapid and efficient genetic manipulation of gyrencephalic carnivores using in utero electroporation

In vivo genetic manipulation of cortical progenitors in gyrencephalic carnivores using in utero electroporation

Rearrangement of Retinogeniculate Projection Patterns after Eye-Specific Segregation in Mice

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