Investigation of MYST4 histone acetyltransferase and its involvement in mammalian gametogenesis-4

<p><b>Copyright information:</b></p><p>Taken from "Investigation of MYST4 histone acetyltransferase and its involvement in mammalian gametogenesis"</p><p>http://www.biomedcentral.com/1471-213X/7/123</p><p>BMC Developmental Biology 2007;7():123-123.</p><p>Published online 2 Nov 2007</p><p>PMCID:PMC2190771.</p><p></p> relative mRNA levels shown represent the quantity of transcript corrected for the value obtained for each pool. The highest level was attributed the relative value of 100. Shown is the relative mRNA abundance (mean ± SEM). Different letters indicate a significant difference of relative mRNA abundance (P < 0.05)

Investigation of MYST4 histone acetyltransferase and its involvement in mammalian gametogenesis-0

<p><b>Copyright information:</b></p><p>Taken from "Investigation of MYST4 histone acetyltransferase and its involvement in mammalian gametogenesis"</p><p>http://www.biomedcentral.com/1471-213X/7/123</p><p>BMC Developmental Biology 2007;7():123-123.</p><p>Published online 2 Nov 2007</p><p>PMCID:PMC2190771.</p><p></p>main zinc finger; C2HC, zinc finger; MYST HAT, conserved HAT domain characteristic of MYST family members; Acidic, glutamate/aspartate-rich region; SM-rich, serine/methionine-rich domain. MYST4 is composed of 2054 residues in which the N-terminal region and the SM-rich domain encode transcriptional repression and activation domains respectively. B) Schematic illustration of MYST4 showing the alternative MYST4 splicing variants MORF and MORFα. Conserved regions between either 2 or 3 sequences are highlighted in gray and black respectively. Regions used for primer designs are underlined in blue (forward) and in red (reverse). The number of residues in each sequence is indicated on the right. (Accession numbers: MYST4; [GenBank; ], MORF; [GenBank; ], MORFα; [GenBank; ]

Investigation of MYST4 histone acetyltransferase and its involvement in mammalian gametogenesis-6

<p><b>Copyright information:</b></p><p>Taken from "Investigation of MYST4 histone acetyltransferase and its involvement in mammalian gametogenesis"</p><p>http://www.biomedcentral.com/1471-213X/7/123</p><p>BMC Developmental Biology 2007;7():123-123.</p><p>Published online 2 Nov 2007</p><p>PMCID:PMC2190771.</p><p></p>s and round spermatids (C, D, E), round spermatids and elongating spermatids (F, G, H), round spermatids and elongated spermatids (I, J, K). Images D, G, K are enlargements of boxed sections in C, F, I respectively. In magnified sections, arrowheads indicate: round spermatocyte (D), nucleus (left) and tail (up) of an elongating spermatid (G) and nuclei of elongated spermatids located in inner (up) wall and inside (left) of the lumen (J). Positive sections were incubated with anti-MYST4 (A, C, D, F, G, I, J) and negatives were prepared by peptide-blocking assay (B, E, H, K). Original magnification 1000× (A, B)

Investigation of MYST4 histone acetyltransferase and its involvement in mammalian gametogenesis-3

<p><b>Copyright information:</b></p><p>Taken from "Investigation of MYST4 histone acetyltransferase and its involvement in mammalian gametogenesis"</p><p>http://www.biomedcentral.com/1471-213X/7/123</p><p>BMC Developmental Biology 2007;7():123-123.</p><p>Published online 2 Nov 2007</p><p>PMCID:PMC2190771.</p><p></p>oocyte nuclei (C, D); portion of large antral follicles, arrowheads indicating granulosa cells (right), theca (down) and basal lamina (left) (E, F) and blood vessels (G, H). Positive sections were incubated with anti-MYST4 (A, C, E, G,) and negatives were prepared by peptide-blocking assay (B, D, F, H). Original magnifications: 200× (C, D, E, F, G, H) and 400× (A, B)

Investigation of MYST4 histone acetyltransferase and its involvement in mammalian gametogenesis-2

<p><b>Copyright information:</b></p><p>Taken from "Investigation of MYST4 histone acetyltransferase and its involvement in mammalian gametogenesis"</p><p>http://www.biomedcentral.com/1471-213X/7/123</p><p>BMC Developmental Biology 2007;7():123-123.</p><p>Published online 2 Nov 2007</p><p>PMCID:PMC2190771.</p><p></p>with an anti-bovine MYST4. From left to right, Br, brain; Th, thymus; Mu, muscle; Lu, Lung; He, heart; Li, liver; Ki, kidney; Sp, spleen; Te, testicle; Ov, ovary; Ut, uterus; Oo, germinal vesicle stage oocytes. TUBULIN and β-ACTIN antibodies were incubated simultaneously on the same membrane and were used as control

Investigation of MYST4 histone acetyltransferase and its involvement in mammalian gametogenesis-7

<p><b>Copyright information:</b></p><p>Taken from "Investigation of MYST4 histone acetyltransferase and its involvement in mammalian gametogenesis"</p><p>http://www.biomedcentral.com/1471-213X/7/123</p><p>BMC Developmental Biology 2007;7():123-123.</p><p>Published online 2 Nov 2007</p><p>PMCID:PMC2190771.</p><p></p>main zinc finger; C2HC, zinc finger; MYST HAT, conserved HAT domain characteristic of MYST family members; Acidic, glutamate/aspartate-rich region; SM-rich, serine/methionine-rich domain. MYST4 is composed of 2054 residues in which the N-terminal region and the SM-rich domain encode transcriptional repression and activation domains respectively. B) Schematic illustration of MYST4 showing the alternative MYST4 splicing variants MORF and MORFα. Conserved regions between either 2 or 3 sequences are highlighted in gray and black respectively. Regions used for primer designs are underlined in blue (forward) and in red (reverse). The number of residues in each sequence is indicated on the right. (Accession numbers: MYST4; [GenBank; ], MORF; [GenBank; ], MORFα; [GenBank; ]

Investigation of MYST4 histone acetyltransferase and its involvement in mammalian gametogenesis-5

<p><b>Copyright information:</b></p><p>Taken from "Investigation of MYST4 histone acetyltransferase and its involvement in mammalian gametogenesis"</p><p>http://www.biomedcentral.com/1471-213X/7/123</p><p>BMC Developmental Biology 2007;7():123-123.</p><p>Published online 2 Nov 2007</p><p>PMCID:PMC2190771.</p><p></p>ly blastocyst and blastocyst) stained with an anti-MYST4 antibody (green signal) and with propidium iodide (red signal) to visualize the DNA. Original magnification 600×

Identification and Localization of the Cyclic Nucleotide Phosphodiesterase 10A in Bovine Testis and Mature Spermatozoa

<div><p>In mammals, adenosine 3’, 5’-cyclic monophosphate (cAMP) is known to play highly important roles in sperm motility and acrosomal exocytosis. It is known to act through protein phosphorylation via PRKA and through the activation of guanine nucleotide exchange factors like EPAC. Sperm intracellular cAMP levels depend on the activity of adenylyl cyclases, mostly SACY, though transmembrane-containing adenylyl cyclases are also present, and on the activity of cyclic nucleotide phosphodiesterases (PDE) whose role is to degrade cAMP into 5’-AMP. The PDE superfamily is subdivided into 11 families (PDE1 to 11), which act on either cAMP or cGMP, or on both cAMP and cGMP although with different enzymatic properties. PDE10, which is more effective on cAMP than cGMP, has been known for almost 15 years and is mostly studied in the brain where it is associated with neurological disorders. Although a high level of <i>PDE10A</i> gene expression is observed in the testis, information on the identity of the isoforms or on the cell type that express the PDE10 protein is lacking. The objective of this study was to identify the PDE10A isoforms expressed in the testis and germ cells, and to determine the presence and localization of PDE10A in mature spermatozoa. As a sub-objective, since <i>PDE10A</i> transcript variants were reported strictly through analyses of bovine genomic sequence, we also wanted to determine the nucleotide and amino acid sequences by experimental evidence. Using RT-PCR, 5’- and 3’-RACE approaches we clearly show that <i>PDE10A</i> transcript variants X3 and X5 are expressed in bovine testis as well as in primary spermatocytes and spermatids. We also reveal using a combination of immunological techniques and proteomics analytical tools that the PDE10A isoform X4 is present in the area of the developing acrosome of spermatids and of the acrosome of mature spermatozoa.</p></div

Oligonucleotide primers used for PCR amplification.

<p>Oligonucleotide primers used for PCR amplification.</p

Detection of bull sperm PDE10A at the electron microscopic level.

<p>Freshly ejaculated bull spermatozoa were washed, fixed with acrolein and pelleted in agarose. Sections of 50 μm were obtained and processed for immunodetection as described in Materials & Methods using monoclonal anti PDE10A antibody (A & B) or non-immune commercial mouse IgG (C & D). Sections were then post-fixed, embedded, and ultrathin sections were prepared and observed by transmission electron microscopy. Note that there is some non-specific signal in the midpiece region that is observed with both the PDE10A antibody and the non-immune IgG (arrowheads in A, B & C). Photographs were taken with magnification of 6800 X (A & C) and 9300 X (B & C). Acrosomal matrix (acr), plasma membrane (pm), and outer acrosomal membrane (oam; lining the acrosomal matrix) are indicated in the sperm head; mitochondria (mit), microtubules (mit) and outer dense fibers (odf) are indicated in the sperm midpiece. In all the panels, the bar scale represents 1 μm.</p