Anti-SP1 (aa 68-82) polyclonal antibody

Serine racemase (SR) synthesizes 1-type serine to its enantimor, D-serine which participates in physiological processes and in pathological conditions. In the central nervous system, SR is highly expressed in neurons and astrocytes but expressed at relatively lower amount in microglia. However, the mechanism by which SR is highly expessed in neurons is hitherto unknown. We report that the SR mRNA and protein levels in Neuro-2a were increased by valproic acid (VPA), a neuron differentiation stimulator as well as a histone deacetylase inhibitor. SR proximal promoter contained nine putative Sp-binding elements and in the exon 1, three putative antioxidant elements (AREs) were conservative among human, rat, and mouse genome. The promoter constructs including 5'-, 3'-fragment, and full length fragment from mouse were individually cloned into a luciferase reporter. Using dual-luciferase assay, the promoter harboring 3'-fragment contained much lower activity than the construct containing 5'-fragment which was though resistant to VPA induction, relative to 3'-fragment. Overexpression of Sp4 or Nrf2 increased whereas knockdown of either decreased Srr mRNA and SR protein. Using site-directed mutagenesis, mutation of Sp-binding elements or AREs in the constructs significantly decreased luciferase activity of the corresponding promoter construct. With chromatin immunoprecipitation, Sp4 was demonstrated to interact directly with the Sp-binding elements whereas Nrf2 bound AREs in Srr mRNA promoter. Altogether, our study highlights that Sp4 controls constitutive expression of SR in neuron and VPA mediates SR expression in N2A cells which is associated with its effect on neuron differentiation, that is, the effect is mediated via Nrf2.

In silvopastoral (SP) systems, forage responses depend on the microenvironment in which the plants develop. Our objective was to evaluate canopy and tillering characteristics of shaded 'Marandu' palisadegrass [Brachiaria brizantha (Hochst A Rich) Stapf, syn. Urochloa brizantha] under continuous stocking in a SP system. Treatments were one full sun (FS) and three shaded systems (silvopasture, SP) corresponding to distances from tree groves: 7.5 m north (SP1), and 15 m (SP2) and 7.5 m south (SP3) studied during two rainy seasons (Year 1 and Year 2). The tree in the SP system was Eucalyptus urograndis (hybrid of Eucalyptus grandis W. Hill ex Maiden x Eucalyptus urophylla S. T. Blake). The photosynthetic active radiation was greater in FS (923 mu mol m(-2) s(-1)), followed by SP2 (811 mu mol m(-2) s(-1)), SP1 (727 mu mol m(-2) s(-1)) and SP3 (673 mu mol m(-2) s(-1)). Forage accumulation in FS was 15% greater than the mean of SP1, SP2 and SP3 (10,663 kg DM/ha). There was no difference in net accumulation of leaf, stem and dead material, averaging 3,302, 3,420 and 4,063 kg DM/ha respectively. Leaf accumulation and accumulation rate were greater in Year 2, and leaf accumulation rate was similar among treatments (19 kg DM ha(-1) day(-1)). Leaf proportion increased 14% from Year 1 to Year 2. Specific leaf area was greater for treatments SP1 and SP3 (193 cm(2)/g). Tiller population density was similar across treatments in Year 1. Shaded palisadegrass maintains leaf productivity similar to FS under continuous stocking in an SP system.