Quantitative real-time polymerase chain reaction analyses of mRNA expression in human corneal endothelial cells at various ages

<p><b>Copyright information:</b></p><p>Taken from "Expression of senescence-related genes in human corneal endothelial cells"</p><p></p><p>Molecular Vision 2008;14():161-170.</p><p>Published online 29 Jan 2008</p><p>PMCID:PMC2254959.</p><p></p> Total RNA was isolated from HCECs of each group, and the first-strand cDNA were aynthesized from equal amounts of total RNA. Quantitative real-time PCR was performed to analyze mRNA expression. shows the expression of from the eight age groups normalized to mRNA. demonstrates fold change in expression relative to the calibrator of the 20-year-old group. is the comparison of the average level of mRNA expressed in HCECs between the young (≤ 30 years) and old (≥ 50 years). Data are presented as mean±standard deviation, and a

Immunohistochemical staining of and Representative fresh-frozen sections of human corneas from donors at various ages (: p16, 18 years; : p21, 33 years; : p27, 54 years; : p53, 18 years; : control, 68 years) are shown

<p><b>Copyright information:</b></p><p>Taken from "Expression of senescence-related genes in human corneal endothelial cells"</p><p></p><p>Molecular Vision 2008;14():161-170.</p><p>Published online 29 Jan 2008</p><p>PMCID:PMC2254959.</p><p></p> Positive staining for all four senescence-related genes are clearly visible in the endothelial nuclei. Magnification; 400×

Autophagy regulates spermatid differentiation via degradation of PDLIM1

<p>Spermiogenesis is a complex and highly ordered spermatid differentiation process that requires reorganization of cellular structures. We have previously found that <i>Atg7</i> is required for acrosome biogenesis. Here, we show that autophagy regulates the round and elongating spermatids. Specifically, we found that <i>Atg7</i> is required for spermatozoa flagella biogenesis and cytoplasm removal during spermiogenesis. Spermatozoa motility of <i>atg7</i>-null mice dropped significantly with some extra-cytoplasm retained on the mature sperm head. These defects are associated with an impairment of the cytoskeleton organization. Functional screening revealed that the negative cytoskeleton organization regulator, PDLIM1 (PDZ and LIM domain 1 [elfin]), needs to be degraded by the autophagy-lysosome-dependent pathway to facilitate the proper organization of the cytoskeleton. Our results thus provide a novel mechanism showing that autophagy regulates cytoskeleton organization mainly via degradation of PDLIM1 to facilitate the differentiation of spermatids.</p

Autophagy is required for ectoplasmic specialization assembly in sertoli cells

<p>The ectoplasmic specialization (ES) is essential for Sertoli-germ cell communication to support all phases of germ cell development and maturity. Its formation and remodeling requires rapid reorganization of the cytoskeleton. However, the molecular mechanism underlying the regulation of ES assembly is still largely unknown. Here, we show that Sertoli cell-specific disruption of autophagy influenced male mouse fertility due to the resulting disorganized seminiferous tubules and spermatozoa with malformed heads. In autophagy-deficient mouse testes, cytoskeleton structures were disordered and ES assembly was disrupted. The disorganization of the cytoskeleton structures might be caused by the accumulation of a negative cytoskeleton organization regulator, PDLIM1, and these defects could be partially rescued by <i>Pdlim1</i> knockdown in autophagy-deficient Sertoli cells. Altogether, our works reveal that the degradation of PDLIM1 by autophagy in Sertoli cells is important for the proper assembly of the ES, and these findings define a novel role for autophagy in Sertoli cell-germ cell communication.</p

The grand ERPs at contralateral electrodes.

<p>(a) at SOA 700 ms: left, enhanced N1 component in invalid-cued trials than in valid-cued trials in the healthy control group (HC); right, significantly enhanced N1 component in invalid-cued trials than in valid-cued trials in the schizophrenia group (SCZ); (b) at SOA 1200 ms: left, no significant cuing effect for N1 component in the HC group; right, significantly enhanced N1 component in invalid-cued trials than in valid-cued trials in the SCZ group.</p

The cuing effects on N1 amplitude (μv) at each SOA in patients with schizophrenia and healthy controls (Mean±S.E.).

<p>SOA: stimulus onset asynchrony = time from onset of cue to onset of target.</p><p>The cuing effects on N1 amplitude (μv) at each SOA in patients with schizophrenia and healthy controls (Mean±S.E.).</p

RTs in the covert orienting of attention task with exogenous cues in the schizophrenia group (SCZ) and the healthy control group (HC).

<p>Both the main effects of Cuing and SOA for RTs were significant (p<0.01).</p

The cuing effect and inhibition of return (IOR) at each SOA in schizophrenia patients and healthy controls (Mean±S.D.).

<p>SOA: stimulus onset asynchrony = time from onset of cue to onset of target. Cuing effect: RT_{invalid trials}-RT_{valid trials}, which indicated facilitatory effect or inhibitory effect of cuing, and the facilitatory cuing effects on RT was indicated as positive, inhibitory cuing effect on RT was indicated as negative.</p><p>The cuing effect and inhibition of return (IOR) at each SOA in schizophrenia patients and healthy controls (Mean±S.D.).</p

The procedure of modified IOR paradigm.

<p>Subjects were instructed to focus on the central cross and maintain fixation throughout the first five frames of the trial. Frame 1: the start of each trial, fixation on the central cross; Frame 2: a peripheral cue was presented randomly to the left or right of fixation; Frame 3: the cue offset for a brief inter-stimulus interval (ISI); Frame 4: then central fixation cue (the cue-back procedure); Frame 5: variable ISI₂ including 450 ms and 950 ms; Frame 6: the target appeared in the cued or uncued location with equal probability. The entire experiment consisted of 320 trials.</p