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Jan Baumgart , Nadine Baumgart
TARC (Translational Animal Research Center), University Medical Center, JGU Mainz

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In Utero Intraventricular Injection and Electroporation of E16 Rat Embryos Published 7/19/2007

In Utero Intraventricular Injection and Electroporation of E16 Rat Embryos

In Utero Intraventricular Injection and Electroporation of E15 Mouse Embryos Published 7/19/2007

In Utero Intraventricular Injection and Electroporation of E15 Mouse Embryos

Organotypic Slice Culture of E18 Rat Brains Published 7/11/2007

Organotypic Slice Culture of E18 Rat Brains

Mouse in Utero Electroporation: Controlled Spatiotemporal Gene Transfection Published 8/15/2011

Mouse in Utero Electroporation: Controlled Spatiotemporal Gene Transfection

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doi: 10.3791/53303 Published: 1/18/2016 0 Comments

This protocol describes in detail how to specifically transfect different regions in the C57BL/6 central nervous system via in utero electroporation. Included in this protocol are detailed instructions for transfections of regions that develop into the cortex, hippocampus, thalamus, hypothalamus, lateral septal nucleus and striatum.

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Figure 1.

Imbalance of dopaminergic and cholinergic modulation can result in movement impairment. () Abnormal dopamine release and enhanced FSI activity may strengthen the output D-MSN pathway and thus promote excessive locomotion. () Dopamine depletion triggers FSI circuit reorganization by reducing FSI connections to D-MSNs. In this case, the strength of the D-MSN pathway may become more dominant than that of the D-MSN pathway, resulting in reduced locomotion. (FS) fast-spiking interneuron, (Ch) cholinergic interneuron, (D1) D-MSN, (D2) D-MSN, (GPe) external globus pallidus, (GPi) internal globus pallidus, (STn) substantia nigra.

Alteration of glutamatergic transmission in the dorsal striatum by blockade of NMDARs may have a role in hyperactivity. Aberrant gamma oscillation throughout the mouse brain, including the dorsal striatum, is observed when psychotomimetics, such as ketamine or MK-801, are systemically applied ( Hakami et al. 2009 ). Hyperactivity in striatal specific NR1 subunit deficient mice is correlated with a reduction of striatal gamma oscillation and, interestingly, hyperactivity can be reduced by either applying D 1 or D 2 receptor agonists ( Ohtsuka et al. 2008 ). Such NMDAR hypofunction induced hyperactivity is thought to result from a reduced number of functional dopamine receptors ( Ohtsuka et al. 2008 ). Examining the consequences of cell-type specific NMDAR hypofunction in the dorsal striatum might be needed to explain the state of hyperactivity.

Studying the excitatory and inhibitory balance in the dorsal striatum is likely to expand our understanding of motor dysfunction (specifically, hyperactivity) that accompanies neurological and psychiatric disorders. Perhaps additional examination of FSIs at the level of channels, such as voltage-gated sodium and potassium channels ( Korotkova et al. 2010 ; Sciamanna and Wilson 2011 ; Simone Rocha Embellished Slide Sandals Discount Sneakernews Low Cost Genuine Sale Online k12iodm
), might better explain how membrane potentials of FSIs oscillate and alter under pathological states induced by an impaired balance of neuromodulators. Furthermore, manipulating specific cell types using mouse Cre-lines and optogenetic tools could provide a method for direct observation of hyperactivity.

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This work was supported by the National Honor Scientist Program, WCU, and the National Creative Research Initiative Program, Korea. K.L. was supported by the Basic Science Research Program (2011-0028240) through the National Research Foundation of Korea. J.-I.K. was supported by a BK21 fellowship.

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4 Corresponding author

The preferred, relatively low-cost alternative for many is kosher salt, which has large, pinchable flakes and typically no additives.

In the salt world, kosher has two meanings. If it's processed under rabbinical supervision, a salt can be certified kosher -- suitable for use in kosher Jewish kitchens -- irrespective of the size or shape of the grain. But kosher salt in salt-industry lingo refers to a flaky salt that adheres readily to meat and is thus well suited to the extended salting required for kosher meat.

Morton's and Diamond Crystal produce most of the boxed kosher salt on supermarket shelves. According to Dave Merriweather , technical manager for Cargill, which owns the Diamond Crystal brand, the two companies use different processes.

Morton's compacts granulated salt into thin flakes with the same equipment that cereal makers use. Cargill uses an evaporating process. Starting with brine obtained by injecting water into rock salt formations -- what's known as solution mining -- the company concentrates the brine in heated evaporating pans. It takes less than three hours for the salt to crystallize.

Because the salt forms on the surface, where the crystal can grow down but not up, the crystals are shaped like hollow pyramids, with no bottom and thin sides. Table salt, in contrast, is cubic -- a feature easily seen with magnification -- because it's made by evaporating brine quickly in a vacuum.

For some purists, the only kitchen-worthy salts are sea salts made the ancient way, by slow solar evaporation. Whether the sea water comes from the Atlantic Ocean, the Mediterranean Sea or a salt lake, the principle is the same: evaporating the water until it is concentrated enough that the salt comes out of suspension and crystallizes. For sodium chloride, saturation occurs in a solution about 10 times as salty as sea water, so there's a lot of concentrating to do.

At a traditional salt works, such as those off the coast of Brittany, source of France's most acclaimed sea salts, the ocean water is evaporated in a series of giant basins that workers have constructed in the marshy land off the coasts. According to Holly Peterson Mondavi , a Napa Valley chef and salt entrepreneur whose Sea Star sea salt comes from this region, the workers spend the winter raking their empty beds smooth and repairing breaches in the clay walls that separate them.

In March or April, when the ocean water is calm and not stirred up from a storm, they fill the first basins. Over the next few months, they move the water into a succession of other basins, leaving behind impurities that settle to the bottom.

Thanks to the work of the sun and wind, the brine becomes progressively more concentrated. By midsummer, the entire year's crop remains suspended in just a few inches of water. A bad storm at this point could dilute the brine, postponing or even ruining the harvest.


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