Journal: International Journal of Molecular Sciences

Article Title: Hippocampal 2-Arachidonoyl Glycerol Signaling Regulates Time-of-Day- and Stress-Dependent Effects on Rat Short-Term Memory

doi: 10.3390/ijms21197316

Figure Lengend Snippet: Exploratory behavior on the training trial for vehicle- and KML29-treated rats that were subjected to no, low or high stress conditions immediately after the training trial, in the morning or afternoon.

Article Snippet: The 2-AG hydrolysis inhibitor KML29 (1,1,1,3,3,3-Hexafluoropropan-2-yl 4-[bis(1,3-benzodioxol-5-yl)-hydroxymethyl]piperidine-1-carboxylate) (2 ng or 20 ng in 0.5 μl per side; Tocris Bioscience, Bristol UK) or its vehicle were bilaterally infused into the CA1 region of the hippocampus immediately after the training trial and right before the swim stress procedure, in order to block any possible stress-induced increase in 2-AG hydrolyzation.

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Hippocampal 2-Arachidonoyl Glycerol Signaling Regulates Time-of-Day- and Stress-Dependent Effects on Rat Short-Term Memory

doi: 10.3390/ijms21197316

Figure Lengend Snippet: Exploratory behavior on the testing trial for vehicle- and KML29-treated rats that were subjected to no, low or high stress conditions immediately after the training trial, in the morning or afternoon.

Article Snippet: The 2-AG hydrolysis inhibitor KML29 (1,1,1,3,3,3-Hexafluoropropan-2-yl 4-[bis(1,3-benzodioxol-5-yl)-hydroxymethyl]piperidine-1-carboxylate) (2 ng or 20 ng in 0.5 μl per side; Tocris Bioscience, Bristol UK) or its vehicle were bilaterally infused into the CA1 region of the hippocampus immediately after the training trial and right before the swim stress procedure, in order to block any possible stress-induced increase in 2-AG hydrolyzation.

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Hippocampal 2-Arachidonoyl Glycerol Signaling Regulates Time-of-Day- and Stress-Dependent Effects on Rat Short-Term Memory

doi: 10.3390/ijms21197316

Figure Lengend Snippet: KML29 modulation of stress- and time-of-day-dependent effects on short-term memory. Discrimination index on the testing trial for rats that were administered in the CA1 region of the dorsal hippocampus with either vehicle or KML29 and then subjected to no, low or high stress conditions immediately after training, in the morning ( A ) or in the afternoon ( B ). Post hoc analysis reported significant group differences as follows: * p < 0.05, ** p < 0.01 vs the corresponding vehicle group. ♦ p < 0.05 vs the corresponding no stress group. §§ p < 0.01 vs the corresponding low stress group. # p < 0.05; ## p < 0.01, ### p < 0.0001, one-sample t-tests significantly different from zero. Data are expressed as mean ± SEM ( n = 10–12 per group).

Article Snippet: The 2-AG hydrolysis inhibitor KML29 (1,1,1,3,3,3-Hexafluoropropan-2-yl 4-[bis(1,3-benzodioxol-5-yl)-hydroxymethyl]piperidine-1-carboxylate) (2 ng or 20 ng in 0.5 μl per side; Tocris Bioscience, Bristol UK) or its vehicle were bilaterally infused into the CA1 region of the hippocampus immediately after the training trial and right before the swim stress procedure, in order to block any possible stress-induced increase in 2-AG hydrolyzation.

Techniques:

Journal: Plants

Article Title: Functional Analysis of PSRP1, the Chloroplast Homolog of a Cyanobacterial Ribosome Hibernation Factor

doi: 10.3390/plants9020209

Figure Lengend Snippet: Plastid Specific Ribosomal Protein 1 (PSRP1) abundance during a diurnal cycle in maize. Seedlings (inbred line B73) were grown in diurnal cycles, and leaves were harvested at the indicated times on the eighth day after planting. Total leaf extracts were fractionated by SDS-PAGE, and PSRP1 was detected by immunoblotting. An image of the Ponceau S-stained filter is shown to illustrate equal sample loading and the abundance of the large subunit of Rubisco (RbcL).

Article Snippet: To further examine the functions of PSRP1, we analyzed transposon insertion alleles in the maize gene encoding PSRP1 (GRMZM2G347956 or Zm00001d034897, in B73 RefGen 3 or 4, respectively).

Techniques: SDS Page, Western Blot, Staining

Journal: Plants

Article Title: Functional Analysis of PSRP1, the Chloroplast Homolog of a Cyanobacterial Ribosome Hibernation Factor

doi: 10.3390/plants9020209

Figure Lengend Snippet: Sucrose gradient sedimentation of extracts of leaves harvested at midday (8 h light) or at the end of the night (8 h dark). The extraction buffer contained 10 mM Mg ++ to maintain ribosome integrity, but lacked heparin and chloramphenicol, which are required to maintain polysome integrity (see text for rationale). Immunoblots of gradient fractions were probed to detect PSRP1, RPL4 (marker for the chloroplast 50S ribosomal subunit), or RPS1 (marker for the chloroplast 30S ribosomal subunit). In a revised nomenclature, chloroplast RPS1 and RPL4 are designated bS1c and uL4c, respectively . Excerpts of the Ponceau S-stained blots are shown to illustrate the position of Rubisco in the gradients (illustrated by its RbcL subunit). ( a ) and ( b ) The complete set of fractions from two replicate experiments were divided between two gels (demarcated by a line). The gradient in panel ( a ) was centrifuged for 105 min, whereas the gradient in panel ( b ) was centrifuged for 150 min, resulting in deeper sedimentation of PSRP1 and marker proteins. ( c ) The peak PSRP1 fractions from a third experiment (fractions 6 through 19) are displayed on a single gel. The sedimentation conditions were the same as those used in panel ( a ).

Article Snippet: To further examine the functions of PSRP1, we analyzed transposon insertion alleles in the maize gene encoding PSRP1 (GRMZM2G347956 or Zm00001d034897, in B73 RefGen 3 or 4, respectively).

Techniques: Sedimentation, Extraction, Western Blot, Marker, Staining

Journal: Plants

Article Title: Functional Analysis of PSRP1, the Chloroplast Homolog of a Cyanobacterial Ribosome Hibernation Factor

doi: 10.3390/plants9020209

Figure Lengend Snippet: Maize psrp1 mutants. ( a ) Mutant and wild-type (WT) (inbred line B73) seedlings at the developmental stage used for experiments reported here. The positions of the Mu insertions in each allele are shown below, with the target site duplications underlined. The black rectangle indicates the single exon in the gene. Additional images of mutant seedlings are shown in . ( b ) Immunoblots showing the abundance of PSRP1 and representative subunits of photosynthetic complexes in Zm- psrp1 mutants. Replicate blots were probed to detect AtpB, D2, PsaD, and PetD (subunits of the ATP synthase, Photosystem II, Photosystem I, and the cytochrome b 6 f complex, respectively). An image of one of the blots stained with Ponceau S is shown below to illustrate equal sample loading and the abundance of the large subunit of Rubisco (RbcL). An immunoblot demonstrating the absence of PSRP1 in Zm- psrp1-2 homozygotes is shown in a.

Article Snippet: To further examine the functions of PSRP1, we analyzed transposon insertion alleles in the maize gene encoding PSRP1 (GRMZM2G347956 or Zm00001d034897, in B73 RefGen 3 or 4, respectively).

Techniques: Mutagenesis, Western Blot, Staining

Journal: Plants

Article Title: Functional Analysis of PSRP1, the Chloroplast Homolog of a Cyanobacterial Ribosome Hibernation Factor

doi: 10.3390/plants9020209

Figure Lengend Snippet: Pulse-labeling analysis of Zm- psrp1-2 mutants in light and dark. Seedlings were pulse-labeled for 15 min at the end of the night (8 h dark) and after 15 min of illumination at dawn ( a ) or at midday (8 h in the light) or after one hour in the dark ( b ). Total leaf proteins were resolved by SDS-PAGE and transferred to nitrocellulose for phosphorimaging. The nitrocellulose was subsequently analyzed by immunoblotting (bottom panels) to validate the absence of PSRP1 in the mutant samples (-/-). WT siblings served as the control for the experiment performed at dawn, and the inbred line B73 served as the control for the experiment performed at midday.

Article Snippet: To further examine the functions of PSRP1, we analyzed transposon insertion alleles in the maize gene encoding PSRP1 (GRMZM2G347956 or Zm00001d034897, in B73 RefGen 3 or 4, respectively).

Techniques: Labeling, SDS Page, Western Blot, Mutagenesis, Control

Journal: Plants

Article Title: Functional Analysis of PSRP1, the Chloroplast Homolog of a Cyanobacterial Ribosome Hibernation Factor

doi: 10.3390/plants9020209

Figure Lengend Snippet: Sucrose gradient fractionation of polysomes in WT and Zm- psrp1-2 seedling leaf extract at the end of night (8 h dark) and after 15 min of illumination at dawn. These experiments differed from those in by the inclusion of heparin and chloramphenicol to maintain polysome integrity, and by the use of centrifugation conditions that resolve polysomes rather than free ribosomal subunits. An equal proportion of the RNA recovered from each fraction was analyzed by RNA gel blot hybridization, using a probe specific for the chloroplast 16S rRNA (top). Images of the same blots stained with methylene blue are shown below to illustrate the distribution of all rRNAs in the gradient. 25S and 18S are cytosolic rRNAs, and 16S and 23S* are chloroplast rRNAs. The WT and mutant tissue came from siblings on the same ear.

Article Snippet: To further examine the functions of PSRP1, we analyzed transposon insertion alleles in the maize gene encoding PSRP1 (GRMZM2G347956 or Zm00001d034897, in B73 RefGen 3 or 4, respectively).

Techniques: Fractionation, Centrifugation, Western Blot, Hybridization, Staining, Mutagenesis