Metabolic effects of manganese in the nematode Caenorhabditis elegans through DAergic pathway and transcription factors activation.

Manganese (Mn) is an essential trace element for physiological functions since it acts as an enzymatic co-factor. Nevertheless, overexposure to Mn has been associated with a pathologic condition called manganism. Furthermore, Mn has been reported to affect lipid metabolism by mechanisms which have yet to be established. Herein, we used the nematode Caenorhabditis elegans to examine Mn's effects on the dopaminergic (DAergic) system and determine which transcription factors that regulate with lipid metabolism are affected by it. Worms were exposed to Mn for four hours in the presence of bacteria and in a liquid medium (85 mM NaCl). Mn increased fat storage as evidenced both by Oil Red O accumulation and triglyceride levels. In addition, metabolic activity was reduced as a reflection of decreased oxygen consumption caused by Mn. Mn also affected feeding behavior as evidenced by decreased pharyngeal pumping rate. DAergic neurons viability were not altered by Mn, however the dopamine levels were significantly reduced following Mn exposure. Furthermore, the expression of sbp-1 transcription factor and let-363 protein kinase responsible for lipid accumulation control was increased and decreased, respectively, by Mn. Altogether, our data suggest that Mn increases the fat storage in C. elegans, secondary to DAergic system alterations, under the control of SBP-1 and LET-363 proteins.

Human bone marrow mesenchymal progenitors: perspectives on an optimized in vitro manipulation.

When it comes to regenerative medicine, mesenchymal stem cells (MSCs) are considered one of the most promising cell types for use in many cell therapies and bioengineering protocols. The International Society of Cellular Therapy recommended minimal criteria for defining multipotential MSC is based on adhesion and multipotency in vitro, and the presence or absence of select surface markers. Though these criteria help minimize discrepancies and allow some comparisons of data generated in different laboratories, the conditions in which cells are isolated and expanded are often not considered. Herein, we propose and recommend a few procedures to be followed to facilitate the establishment of quality control standards when working with mesenchymal progenitors isolation and expansion. Following these procedures, the classic Colony-Forming Unit-Fibroblast (CFU-f) assay is revisited and three major topics are considered to define conditions and to assist on protocol optimization and data interpretation. We envision that the creation of a guideline will help in the identification and isolation of long-term stem cells and short-term progenitors to better explore their regenerative potential for multiple therapeutic purposes.

Senemorphism: a novel perspective on aging patterns and its implication for diet-related biology.

Aging can be described as the accumulation of changes in organisms over time. Aging in organisms undergoing caloric restriction (CR) is widely considered as a slowed version of aging under ad libitum (AL) conditions. However, here we argue that aging under optimized CR is fundamentally different from aging under AL based on the following facts: (1) Comparing the two dietary groups, several age-related changes run in the opposite direction over time; (2) Switching from an AL to a CR diet clearly reverts (not only delays) several "normal" accumulated changes; (3) major causes of death are as different between both groups as they are between species. These observations support the idea that CR and AL initially modulate different metabolic and physiological programs, which exclusively over time generate two biologically different organisms. Such distinct diet-related senescence is analogous to the divergent aging processes and causes of death observed between castes of social insects, such as queens versus workers ("caste-related-senescence") and also between breeding versus non-breeding semelparous animals ("reproduction-related-senescence"). All these aging phenotypes are different not because they accumulate changes at a different rate, but because they accumulate different changes over time. Thus, the environment does not simply affect the individual aging rate through stochastic effects (e.g. U.V.) but also modulates the activation of a particular program/strategy that influences lifespan (e.g. caste, calorie intake). We refer to the environment-dependent aging patterns encoded by the genome as "senemorphism". Based on this idea we propose experimental schemes for aging, evolution and biomedical research.

Divergent regulation of adipose tissue metabolism by calorie restriction and inhibition of growth hormone signaling.

Calorie restriction (CR) and a reduced growth hormone (GH) signal affect insulin sensitivity and lifespan in mammals in a similar manner. We investigated the effects of CR and moderate inhibition of GH on glucose-stimulated activation of insulin signaling and the expression of genes related to fat metabolism in white adipose tissue (WAT) in rats. We used 10-month-old male, wild-type (W) Wistar rats, fed ad libitum (AL) or a 30% CR diet from 6weeks of age, and transgenic (Tg) rats with moderately suppressed GH signaling. Rats were killed 15min after an intraperitoneal injection of glucose or saline. In control W-AL rats, the levels of serum insulin, phosphorylated (p) insulin receptor (pY-IR), p-Akt, and the expression of glucose transporter (Glut) 4 in the membrane fraction were greater in the glucose-injected group than in the saline-injected group, indicating significant activation of insulin signaling in response to glucose loading. In the W-CR and Tg-AL rats, the serum insulin and pY-IR levels were lower than those in the W-AL rats. The Akt-Glut pathway was up-regulated even after saline-injection. Expression levels of adipogenic and lipogenic genes including PPARgamma, adiponectin, and its receptors, were higher in the W-CR rats than in the W-AL and Tg-AL rats. The present findings indicate adipose tissue metabolic profiles specific to CR.

Identification and characterization of an insulin receptor substrate 4-interacting protein in rat brain: implications for longevity.

The hypothalamus is organized as a collection of distinct, autonomously active nuclei that regulate discrete functions, such as feeding activity and metabolism. We used suppression subtractive hybridization (SSH) to identify genes that are enriched in the hypothalamus of the rat brain. We screened a subtractive library of 160 clones, and 4 genes that were predominantly expressed in the hypothalamus, compared to other brain regions. The mRNA for a member of the WD-repeat family of proteins, WDR6, was abundantly expressed in the hypothalamus, and we found that WDR6 interacted with insulin receptor substrate 4 (IRS-4) in the rat brain. Interestingly, WDR6 gene expression in the hypothalamic arcuate nucleus was decreased by caloric restriction, and in growth hormone (GH)-antisense transgenic rats, both of which are associated with an increased life span. Insulin-like growth factor (IGF)-I and insulin treatment increased WDR6 gene expression in mouse hypothalamus-derived GT1-7 cells. Our results might suggest that WDR6 participates in insulin/IGF-I signaling and the regulation of feeding behavior and longevity in the brain.