Impaired cortical mitochondrial function following TBI precedes behavioral changes.

Traumatic brain injury (TBI) pathophysiology can be attributed to either the immediate, primary physical injury, or the delayed, secondary injury which begins minutes to hours after the initial injury and can persist for several months or longer. Because these secondary cascades are delayed and last for a significant time period post-TBI, they are primary research targets for new therapeutics. To investigate changes in mitochondrial function after a brain injury, both the cortical impact site and ipsilateral hippocampus of adult male rats 7 and 17 days after a controlled cortical impact (CCI) injury were examined.

A microplate technique to simultaneously assay calcium accumulation in endoplasmic reticulum and SERCA release of inorganic phosphate.

Traditional analyses of calcium homeostasis have separately quantified either calcium accumulation or release mechanisms. To define the system as a whole, however, requires multiple experimental techniques to examine both accumulation and release. Here we describe a technique that couples the simultaneous quantification of radio-labeled calcium accumulation in endoplasmic reticulum (ER) microsomes with the release of inorganic phosphate (Pi) by the hydrolytic activity of sarco-endoplasmic reticulum calcium ATPase (SERCA) all in the convenience of a 96-well format.

The cytokine temporal profile in rat cortex after controlled cortical impact.

Cerebral inflammatory responses may initiate secondary cascades following traumatic brain injury (TBI). Changes in the expression of both cytokines and chemokines may activate, regulate, and recruit innate and adaptive immune cells associated with secondary degeneration, as well as alter a host of other cellular processes. In this study, we quantified the temporal expression of a large set of inflammatory mediators in rat cortical tissue after brain injury.

Craniotomy: true sham for traumatic brain injury, or a sham of a sham?

Abstract Neurological dysfunction after traumatic brain injury (TBI) is caused by both the primary injury and a secondary cascade of biochemical and metabolic events. Since TBI can be caused by a variety of mechanisms, numerous models have been developed to facilitate its study. The most prevalent models are controlled cortical impact and fluid percussion injury.

In cold-hardy insects, seasonal, temperature, and reversible phosphorylation controls regulate sarco/endoplasmic reticulum Ca2+-ATPase (SERCA).

Winter cold hardiness of insects typically involves one of two major strategies for survival below 0 degrees C: freeze avoidance and freeze tolerance. The two strategies have some common features, including the accumulation of high concentrations of cryoprotectant polyols and the frequent occurrence of diapause. Entry into the hypometabolic state of diapause requires coordinated suppression of major ATP-consuming metabolic processes, and ion motive ATPases are important targets for regulation.

Regulation of Akt during torpor in the hibernating ground squirrel, Ictidomys tridecemlineatus.

The 13-lined ground squirrel (Ictidomys tridecemlineatus) is capable of entering into extended periods of torpor during winter hibernation. The state of torpor represents a hypometabolic shift wherein the rate of oxygen consuming processes are strongly repressed in an effort to maintain cellular homeostasis as the availability of food energy becomes limited. We are interested in studying hibernation/torpor because of the robust state of tolerance to constrained oxygen delivery, oligemia, and hypothermia achieved by the tissues of hibernating mammals.

The regulation of AMPK signaling in a natural state of profound metabolic rate depression.

In response to energy stress (and elevated AMP), the AMP-activated protein kinase (AMPK) coordinates the restoration of energy homeostasis. We determined that AMPK is activated in a model system (desert snail Otala lactea) during a physiological state of profound metabolic rate depression (estivation) in the absence of a rise in AMP. Kinetic characterization indicated a strong increase in AMPK activity and phosphorylation in estivation, consistent with an increase in P-Ser428 LKB, an established regulator of AMPK.

Suppression of Na+K+ -ATPase activity by reversible phosphorylation over the winter in a freeze-tolerant insect.

Larvae of the gall fly, Eurosta solidaginis, use the cold hardiness strategy of freeze tolerance as well as entry into a hypometabolic state (diapause) to survive the winter. Cold hardiness strategies have been extensively explored in this species, but the metabolic features of winter hypometabolism have received little attention. A primary consumer of energy in cells is the ATP-dependent sodium-potassium ion pump (Na(+)K(+)-ATPase) so inhibitory controls over transmembrane ion movements could contribute substantially to energy savings over the winter months.

Expression of Nrf2 and its downstream gene targets in hibernating 13-lined ground squirrels, Spermophilus tridecemlineatus.

Mammalian hibernation is associated with wide variation in heart rate, blood flow, and oxygen delivery to tissues and is used as a model of natural ischemia/reperfusion. In non-hibernators, ischemia/reperfusion is typically associated with oxidative stress but hibernators seem to deal with potential oxidative damage by enhancing antioxidant defenses in an anticipatory manner. The present study assesses the role of the Nrf2 transcription factor in the regulation of antioxidant defenses during hibernation.

Mitochondria of cold hardy insects: responses to cold and hypoxia assessed at enzymatic, mRNA and DNA levels.

Winter survival for larvae of goldenrod gall insects, the freeze-avoiding Epiblema scudderiana, and the freeze tolerant, Eurosta solidaginis, includes entry into diapause (a torpid state of arrested development) and expression of a variety of cryoprotective adaptations. Diapause and cold winter temperatures, as well as freezing in E. solidaginis, all strongly reduce the need for mitochondrial activity.

Protein SUMOylation is massively increased in hibernation torpor and is critical for the cytoprotection provided by ischemic preconditioning and hypothermia in SHSY5Y cells.

Hibernation torpor provides an excellent natural model of tolerance to profound reductions in blood flow to the brain and other organs. Here, we report that during torpor of 13-lined ground squirrels, massive SUMOylation occurs in the brain, liver, and kidney. The level of small ubiquitin-related modifier (SUMO) conjugation coincides with the expression level of Ubc9, the SUMO specific E2-conjugating enzyme.

HIF-1alpha involvement in low temperature and anoxia survival by a freeze tolerant insect.

Winter survival for many insect species relies on the ability to endure the freezing of extracellular body fluids. Because freezing impedes oxygen delivery to tissues, one component of natural freeze tolerance is a well-developed anoxia/ischemia resistance. The present study explores the responses of the hypoxia-inducible factor-1alpha (HIF-1alpha) to cold, freezing and anoxia exposures in the freeze tolerant goldenrod gall fly larva, Eurosta solidaginis.