• Osgin1
• Osgin2
• flavin-dependent monooxygenase
• oxidative stress
• tumorigenesis

• Oxidative Stress/genetics
• Humans
• Animals
• Multigene Family
• Autophagy/genetics
• Apoptosis/genetics
• Phylogeny
• Mixed Function Oxygenases/genetics
• Mixed Function Oxygenases/metabolism

• 1R15HD114135-01 NIH HHS

• NKCC cotransporter
• annelid
• invertebrate
• osmoconformer organisms
• physiology

• Doctorate fellowship to S.M. Coordenação de Aperfeicoamento de Pessoal de Nível Superior
• #302829/2015-6 National Council for Scientific and Technological Development

• ANO2
• Ca2+-activated Cl¯ currents
• olfactory sensory neurons
• olfactory transduction

• Animals
• Mice
• Anoctamins/genetics
• Calcium/metabolism
• Mice, Knockout
• Olfactory Mucosa/metabolism
• Olfactory Receptor Neurons/metabolism

• R01 DC009613 NIDCD NIH HHS
• R01 DC016647 NIDCD NIH HHS
• R01DC016647 NIH HHS
• National Institutes of Health

• artificial olfactory systems
• biological olfactory systems
• odorants
• olfaction
• sensors

The prevalence of metabolic diseases is growing worldwide. Accumulating evidence suggests that solute carrier (SLC) transporters contribute to the etiology of various metabolic diseases. Consistent with metabolic characteristics, the top five organs in which SLC transporters are highly expressed are the kidney, brain, liver, gut, and heart. We aim to understand the molecular mechanisms of important SLC transporter-mediated physiological processes and their potentials as drug targets. SLC transporters serve as ‘metabolic gate’ of cells and mediate the transport of a wide range of essential nutrients and metabolites such as glucose, amino acids, vitamins, neurotransmitters, and inorganic/metal ions. Gene-modified animal models have demonstrated that SLC transporters participate in many important physiological functions including nutrient supply, metabolic transformation, energy homeostasis, tissue development, oxidative stress, host defense, and neurological regulation. Furthermore, the human genomic studies have identified that SLC transporters are susceptible or causative genes in various diseases like cancer, metabolic disease, cardiovascular disease, immunological disorders, and neurological dysfunction. Importantly, a number of SLC transporters have been successfully targeted for drug developments. This review will focus on the current understanding of SLCs in regulating physiology, nutrient sensing and uptake, and risk of diseases.

• Slc12a1/Nkcc2a
• Slc12a5/Kcc2
• glucagon
• glucose homeostasis
• insulin

• Cynopterus sphinx
• MiR-132
• NMDA receptors
• Olfactory learning
• PRTFDC1
• SERT

• Bumetanide
• Cognitive decline
• Hypoxia-ischemia brain damage
• NKCC1
• Neurogenesis

AIM: To investigate the intestinal functions of the NKCC1 Na⁺-K⁺-2Cl cotransporter (SLC12a2 gene), differential mRNA expression changes in NKCC1-null intestine were analyzed.

METHODS: Microarray analysis of mRNA from intestines of adult wild-type mice and gene-targeted NKCC1-null mice (n = 6 of each genotype) was performed to identify patterns of differential gene expression changes. Differential expression patterns were further examined by Gene Ontology analysis using the online Gorilla program, and expression changes of selected genes were verified using northern blot analysis and quantitative real time-polymerase chain reaction. Histological staining and immunofluorescence were performed to identify cell types in which upregulated pancreatic digestive enzymes were expressed.

RESULTS: Genes typically associated with pancreatic function were upregulated. These included lipase, amylase, elastase, and serine proteases indicative of pancreatic exocrine function, as well as insulin and regenerating islet genes, representative of endocrine function. Northern blot analysis and immunohistochemistry showed that differential expression of exocrine pancreas mRNAs was specific to the duodenum and localized to a subset of goblet cells. In addition, a major pattern of changes involving differential expression of olfactory receptors that function in chemical sensing, as well as other chemosensing G-protein coupled receptors, was observed. These changes in chemosensory receptor expression may be related to the failure of intestinal function and dependency on parenteral nutrition observed in humans with SLC12a2 mutations.

CONCLUSION: The results suggest that loss of NKCC1 affects not only secretion, but also goblet cell function and chemosensing of intestinal contents via G-protein coupled chemosensory receptors.

• SLC12a2
• Chemosensory
• Chemosensitivity
• Gastrointestinal
• Dyspepsia

Traumatic brain injury (TBI) results in significant impairments in hippocampal synaptic plasticity. A molecule critically involved in hippocampal synaptic plasticity, 3′,5′-cyclic adenosine monophosphate, is downregulated in the hippocampus after TBI, but the mechanism that underlies this decrease is unknown. To address this question, we determined whether phosphodiesterase (PDE) expression in the hippocampus is altered by TBI. Young adult male Sprague Dawley rats received sham surgery or moderate parasagittal fluid-percussion brain injury. Animals were analyzed by western blotting for changes in PDE expression levels in the hippocampus. We found that PDE1A levels were significantly increased at 30 min, 1 h and 6 h after TBI. PDE4B2 and 4D2 were also significantly increased at 1, 6, and 24 h after TBI. Additionally, phosphorylation of PDE4A was significantly increased at 6 and 24 h after TBI. No significant changes were observed in levels of PDE1B, 1C, 3A, 8A, or 8B between 30 min to 7 days after TBI. To determine the spatial profile of these increases, we used immunohistochemistry and flow cytometry at 24 h after TBI. PDE1A and phospho-PDE4A localized to neuronal cell bodies. PDE4B2 was expressed in neuronal dendrites, microglia and infiltrating CD11b⁺ immune cells. PDE4D was predominantly found in microglia and infiltrating CD11b⁺ immune cells. To determine if inhibition of PDE4 would improve hippocampal synaptic plasticity deficits after TBI, we treated hippocampal slices with rolipram, a pan-PDE4 inhibitor. Rolipram partially rescued the depression in basal synaptic transmission and converted a decaying form of long-term potentiation (LTP) into long-lasting LTP. Overall, these results identify several possible PDE targets for reducing hippocampal synaptic plasticity deficits and improving cognitive function acutely after TBI.

• CREB
• cAMP
• fluid-percussion
• hippocampus
• inflammation
• phosphodiesterase
• traumatic brain injury

• NKCC1
• NKCC2
• glucose homeostasis
• insulin secretion
• β-cell

• Animals
• Bumetanide/pharmacology
• Down-Regulation/drug effects
• Gene Expression/drug effects
• Glucose/metabolism
• Glucose Tolerance Test
• Heterozygote
• Insulin/metabolism
• Insulin-Secreting Cells/drug effects
• Insulin-Secreting Cells/metabolism
• Mice
• Mice, Transgenic
• Sodium Potassium Chloride Symporter Inhibitors/pharmacology
• Solute Carrier Family 12, Member 1/genetics
• Solute Carrier Family 12, Member 1/metabolism
• Solute Carrier Family 12, Member 2/genetics
• Solute Carrier Family 12, Member 2/metabolism

• P30 DK020593 NIDDK NIH HHS
• R01 DK097829 NIDDK NIH HHS

Adenylyl Cyclase 3 (AC3) plays an important role in the olfactory sensation-signaling pathway in mice. AC3 deficiency leads to defects in olfaction. However, it is still unknown whether AC3 deficiency affects gene expression or olfactory signal transduction pathways within the main olfactory epithelium (MOE). In this study, gene microarrays were used to screen differentially expressed genes in MOE from AC3 knockout (AC3^−/−) and wild-type (AC3^+/+) mice. The differentially expressed genes identified were subjected to bioinformatic analysis and verified by qRT-PCR. Gene expression in the MOE from AC3^−/− mice was significantly altered, compared to AC3^+/+ mice. Of the 41266 gene probes, 3379 had greater than 2-fold fold change in expression levels between AC3^−/− and AC3^+/+ mice, accounting for 8% of the total gene probes. Of these genes, 1391 were up regulated, and 1988 were down regulated, including 425 olfactory receptor genes, 99 genes that are specifically expressed in the immature olfactory neurons, 305 genes that are specifically expressed in the mature olfactory neurons, and 155 genes that are involved in epigenetic regulation. Quantitative RT-PCR verification of the differentially expressed epigenetic regulation related genes, olfactory receptors, ion transporter related genes, neuron development and differentiation related genes, lipid metabolism and membrane protein transport etc. related genes showed that P75NTR, Hinfp, Gadd45b, and Tet3 were significantly up-regulated, while Olfr370, Olfr1414, Olfr1208, Golf, Faim2, Tsg101, Mapk10, Actl6b, H2BE, ATF5, Kirrrel2, OMP, Drd2etc. were significantly down-regulated. In summary, AC3 may play a role in proximal olfactory signaling and play a role in the regulation of differentially expressed genes in mouse MOE.

• adenylyl cyclase 3
• differentially expressed genes
• main olfactory epithelium
• microarray analysis

A developmental “switch” in chloride transporters occurs in most neurons resulting in GABA_A mediated hyperpolarization in the adult. However, several neuronal cell subtypes maintain primarily depolarizing responses to GABA_A receptor activation. Among this group are gonadotropin-releasing hormone-1 (GnRH) neurons, which control puberty and reproduction. NKCC1 is the primary chloride accumulator in neurons, expressed at high levels early in development and contributes to depolarization after GABA_A receptor activation. In contrast, KCC2 is the primary chloride extruder in neurons, expressed at high levels in the adult and contributes to hyperpolarization after GABA_A receptor activation. Anion exchangers (AEs) are also potential modulators of responses to GABA_A activation since they accumulate chloride and extrude bicarbonate. To evaluate the mechanism(s) underlying GABA_A mediated depolarization, GnRH neurons were analyzed for 1) expression of chloride transporters and AEs in embryonic, pre-pubertal, and adult mice 2) responses to GABA_A receptor activation in NKCC1^-/- mice and 3) function of AEs in these responses. At all ages, GnRH neurons were immunopositive for NKCC1 and AE2 but not KCC2 or AE3. Using explants, calcium imaging and gramicidin perforated patch clamp techniques we found that GnRH neurons from NKCC1^-/- mice retained relatively normal responses to the GABA_A agonist muscimol. However, acute pharmacological inhibition of NKCC1 with bumetanide eliminated the depolarization/calcium response to muscimol in 40% of GnRH neurons from WT mice. In the remaining GnRH neurons, HCO₃^- mediated mechanisms accounted for the remaining calcium responses to muscimol. Collectively these data reveal mechanisms responsible for maintaining depolarizing GABA_A mediated transmission in GnRH neurons.