The cultural and creative function of moving image literacy in the subject of English in the Greek secondary school

Teaching media literacy as a separate school subject or as part of another school subject is lacking from the Greek educational reality, despite the international academic research and the development and application of media literacy teaching models. This thesis is an analysis of two case study research projects carried out in groups of students in two Greek secondary schools with the aim to study the students’ response to media projects, which are totally new for the Greek educational reality, realized in the English as a Foreign Language class. The data is analyzed according to Burn and Durran’s 3-Cs model of media literacy, and more precisely its Cultural and Creative functions are the aspects used that include the concepts of Cultural Taste, Identity, and Creativity. These concepts are interpreted within the framework of Cultural Studies and Psychology theories. Important theoreticians considered are Bourdieu, Bennett, Giddens, Vygotsky, Jenkins and Bakhtin. The examination of students’ participation in the media projects and their production work suggest that their cultural taste is a combination of global and local influences, a glocal result, in which the family, the peers, the media and the education play an important role. Their identity is multi-faceted, as a reflection of various aspects of their selves, and it is closely related to their cultural taste and their cultural capital. Students’ creativity is also expressed as a complex process, affected both by the guidance of the official educational context and the youth popular culture tendencies. The tensions that emerge in the expression of the students’ cultural taste, identity and creativity during moving image projects characterize the Greek adolescents’ response to the newly-learnt moving image literacy, and raise important questions for educators and researchers

<i>apkc-RA</i> and <i>orb2</i> mRNAs associate with Orb2 <i>in vivo</i>.

<p>A) The <i>apkc</i> transcription unit as annotated in <a href="http://flybase.org/reports/FBgn0261854.html" target="_blank">http://flybase.org/reports/FBgn0261854.html</a>. Transcripts expressed from four <i>apkc</i> promoters (P1–P4) plus a collection of alternatively spliced exons and UTRs are predicted to generate more than ten mRNAs. Green bar, protein coding exon; grey bar, UTR; solid black line, intron. <i>apkc-RA</i> CPE sites are labeled in red. Positions of primers used for RT-PCR experiments to identify different <i>apkc</i> mRNA species are marked in dark blue. The two dsRNA sequences used in the <i>apkc</i> RNAi knockdown experiments are marked in pink. Probes for the <i>apkc</i> Orb2 EMSAs, com-GS and RA-GS, are indicated above the gene in brown. B) <i>apkc</i> transcripts expressed in testes were detected by RT-PCR using the primer pairs as indicated. Not all of the primer pairs are specific to one of the four promoters, but some are specific to particular splice forms. C) <i>apkc-RA</i> mRNA can be immunoprecipitated with Orb2 antibodies from WT and <i>orb2^ΔQ</i> testis extracts. β-Gal antibody (a negative control) didn't immunoprecipitate <i>apkc-RA</i> mRNA. D) Orb2 antibody immunoprecipitated <i>orb2</i> mRNA from wild type testes while the control β-Gal antibody did not.</p

Spermatid Cyst Polarization in <i>Drosophila</i> Depends upon <i>apkc</i> and the CPEB Family Translational Regulator <i>orb2</i>

<div><p>Mature <i>Drosophila</i> sperm are highly polarized cells—on one side is a nearly 2 mm long flagellar tail that comprises most of the cell, while on the other is the sperm head, which carries the gamete's genetic information. The polarization of the sperm cells commences after meiosis is complete and the 64-cell spermatid cyst begins the process of differentiation. The spermatid nuclei cluster to one side of the cyst, while the flagellar axonemes grows from the other. The elongating spermatid bundles are also polarized with respect to the main axis of the testis; the sperm heads are always oriented basally, while the growing tails extend apically. This orientation within the testes is important for transferring the mature sperm into the seminal vesicles. We show here that orienting cyst polarization with respect to the main axis of the testis depends upon atypical Protein Kinase C (aPKC), a factor implicated in polarity decisions in many different biological contexts. When <i>apkc</i> activity is compromised in the male germline, the direction of cyst polarization within this organ is randomized. Significantly, the mechanisms used to spatially restrict <i>apkc</i> activity to the apical side of the spermatid cyst are different from the canonical cross-regulatory interactions between this kinase and other cell polarity proteins that normally orchestrate polarization. We show that the asymmetric accumulation of aPKC protein in the cyst depends on an mRNA localization pathway that is regulated by the <i>Drosophila</i> CPEB protein Orb2. <i>orb2</i> is required to properly localize and activate the translation of <i>apkc</i> mRNAs in polarizing spermatid cysts. We also show that <i>orb2</i> functions not only in orienting cyst polarization with respect to the apical-basal axis of the testis, but also in the process of polarization itself. One of the <i>orb2</i> targets in this process is its own mRNA. Moreover, the proper execution of this <i>orb2</i> autoregulatory pathway depends upon <i>apkc</i>.</p></div

Spermatid cysts elongate in the wrong direction in <i>apkc</i> or <i>orb2</i> mutants.

<p>A) Frequency of testes that have at least one spermatid cyst polarized in the wrong orientation so that clustered spermatid cyst nuclei are found in the apical region instead of towards the base of the testis. <i>apkc⁴⁸</i> (<i>apkc^ex48</i>), <i>06403</i> (<i>apkc⁰⁶⁴⁰³</i>) and <i>apkc⁵⁵</i> (<i>apkc^ex55</i>) are <i>apkc</i> mutant alleles. <i>34332</i> and <i>35140</i> are UAS-<i>apkc</i> RNAi lines (dsRNA targeting sites marked in <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1004380#pgen-1004380-g003" target="_blank">Fig. 3A</a>), while the Gal4 drivers are <i>nos</i> or MTD. MTD has three GAL4 drivers, <i>pCOG-Gal4</i> which has an <i>otu</i> promoter, plus two different <i>nos</i> drivers, <i>NGT-40</i> and <i>nanos</i>-<i>Gal4</i>. Error bars are standard deviation based on three experiments. Total number of testes scored for each genotype are (from left to right): Fig. A1: <i>apkc^ex48</i>: 36; <i>apkc⁰⁶⁴⁰³/+</i>:33; <i>apkc^ex55</i>:27; MTD/UAS-<i>apkc:34332</i>:43; <i>nos-GAL4/</i>UAS-<i>apkc:35140</i>: 18; MTD/UAS-<i>apkc:35140</i>:16; <i>orb³⁶/+</i>:55; <i>orb³⁶</i>: 50; <i>orb2^ΔQ</i>: 86; WT: 65. Fig. A2: <i>apkc⁰⁶⁴⁰³/+</i>: 38; <i>apkc⁰⁶⁴⁰³/</i>+; <i>orb2^ΔQ</i>/+:40; <i>apkc⁰⁶⁴⁰³/</i>+; <i>orb2³⁶</i>/+: 49; Fig. A3: <i>apkc^ex48/+</i>: 44; <i>apkc^ex48/</i>+; <i>orb2^ΔQ</i>/+: 39; <i>apkc^ex48/</i>+; <i>orb2³⁶</i>/+: 39; <i>apkc^ex55/</i>+: 27; <i>apkc^ex55/</i>+; <i>orb2^ΔQ</i>/+: 28; <i>apkc^ex55/</i>+; <i>orb2³⁶</i>/+: 37. B-D) Whole mount antibody staining of wild type or <i>apkc</i> mutant testes. Blue, DNA; red, Orb2. B) Spermatid nuclei clusters (arrow) are found in the spermatogonial region of the testis in <i>apkc</i> hypomorphic alleles. The example shown here is <i>apkc^ex55</i>. D) In the hypomorphic alleles, most spermatid nuclei clusters are located as in wild type on the basal side of testes. D) In MTD/UAS-<i>apkc:35140</i> testes, multiple spermatid nuclei clusters are found in the apical region (arrows). (Not all spermatid cysts express Orb2 and/or are in focus). E) In wild type testes spermatid cyst nuclei are never seen at the apical end of the testis. F) aPKC (green) and Orb2 (red) in a misoriented elongating <i>apkc^ex55</i> spermatid cyst. Note that the Orb2 comet head is present. G) aPKC (green) and Orb2 (red) in a misoriented elongating MTD/UAS-<i>apkc:35140</i> testis. Note that the Orb2 comet is absent. This is also the case for correctly oriented MTD/UAS-<i>apkc:35140</i> spermatid cysts (not shown). All images are orientated with apical side of the testes to the left and basal to the right. Scale bars: 50 µm.</p

<i>orb2</i> is required for polarized accumulation of <i>apkc-RA</i> mRNA and aPKC protein in spermatid cysts.

<p>A) In wild type testes, Orb2 and <i>apkc-RA</i> accumulate in a comet pattern in elongating spermatid cysts located towards the apical side of the testis. B and C) In <i>orb2^ΔQ</i> spermatid cysts that elongate in the incorrect direction, Orb2^ΔQ protein and both <i>apkc-RA</i> mRNA (detected with the <i>apkc-RA</i> probe, B) and “bulk” <i>apkc</i> mRNA (detected with the <i>apkc-com</i> probe, C) are distributed randomly instead of localizing in the comet pattern. Arrows in A indicate comet heads. In B and C the arrows indicate the expected position of the comet head. In the merged images on the far right, Red: Orb2; Green: <i>apkc</i> mRNA; Blue: DNA. D & E) aPKC protein (arrow) in <i>orb2^ΔQ</i> spermatids elongating in the correct (D) or incorrect (E) orientation. <i>orb2^ΔQ</i> spermatids elongating in the incorrect direction lack aPKC protein stripes at the tip of the elongating flagellar axonemes (compare arrows in D and E). In the merged images on the far right, Red: Tublin; Green: aPKC; Blue: DNA. F & G) <i>apkc-RA</i> (detected with the <i>apkc-RA</i> probe) and “bulk” <i>apkc</i> mRNAs (detected with the <i>apkc-com</i> probe) are expressed in <i>orb2³⁶</i> spermatids but are not localized. In the merged image on right, Red: Orb2, Green: <i>apkc-RA</i> or bulk (<i>apkc-com</i>) mRNA, and Blue: DNA. All images are orientated with apical side of the testes to the left and basal to the right. Scale bar in A–D: 20 µm; in E and F: 50 µm.</p

<i>orb2</i> is required for cyst polarization.

<p>A and B) Green, phalloidin labeled Actin; Blue, DNA; Red, Orb2. A) Clustered nuclei of spermatid cysts (arrowheads) that have completed elongation and just assembled individualization complexes (ICs) (arrows) are found in the spermatogonial/early spermatocyte region of <i>orb2^ΔQ</i> testis. B) White arrows point to two ICs in an <i>orb2^ΔQ</i> testis that are moving in opposite directions in the middle of the testis (yellow arrows indicate directions of motion). C–E) Green, Bol; blue, DNA. C) Like Orb2, the translation factor Bol is localized a comet pattern during flagellar axoneme elongation in wild type testes <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1004380#pgen.1004380-Xu1" target="_blank">[29]</a>. D) A bi-polar <i>orb2³⁶</i> spermatid cyst that has Bol concentrated at both elongating ends, while the nuclei are in the middle of the cyst (arrows). E) A partially elongated <i>orb2³⁶</i> spermatid cyst in which spermatid nuclei and Bol protein are scattered randomly. Dotted line outlines one among four spermatid cysts in the figure. Yellow arrows point out a few of the scattered nuclei. All images are orientated with apical side of the testes to the left and basal to the right. Scale bar: 50 µm.</p

<i>orb2</i> autoregulates the localization and translation of <i>orb2</i> mRNA.

<p>A) Wild type spermatid cyst showing that Orb2 protein (red) and <i>orb2</i> mRNA (green) are distributed in the characteristic comet pattern and display extensive co-localization, particularly in the comet head. B) In <i>orb2^ΔQ</i> spermatids that elongate in the incorrect direction, Orb2^ΔQ protein (red) and <i>orb2^ΔQ</i> mRNA (green) are distributed uniformly in the cyst and there is no evidence of a comet head (expected position is indicated by arrow). All images are orientated with apical to the left and basal to the right. Scale bar: 20 µm.</p

<i>orb2³⁶</i> spermatids have defects in flagellar axoneme elongation.

<p>A, B) Phase contrast images showing wild type (A) and <i>orb2³⁶</i> (B) elongated spermatid bundles. Wild type spermatid flagellar axoneme bundles (arrow) have a smooth morphology and extend in a nearly straight line. <i>orb2³⁶</i> bundles have rough and uneven morphology, are shorter, and zigzag back and forth. Individualized sperm (red arrowheads in A) are observed in wild type but not <i>orb2³⁶</i> testes. C1–C3) Wild type testes double stained with Orb2 (red) and Bol (green) antibodies showing co-localized Bol and Orb2 concentrated in a band near the tip of the elongating flagellar axonemes (arrowhead) and decreased expression level following this band. D1–D3) Bol localization at the tip is lost in <i>orb2³⁶</i> flagellar axonemes (arrow), while there is an uneven distribution along the flagellar axonemes extending behind the tip. E1–E3) Orb2 and Bol co-localization at the tip of the elongating flagellar axonemes in <i>orb2^ΔQ</i> is as in wild type. Scale bar: 20 µm.</p

Orb2 functions in meiosis and differentiation can be uncoupled in <i>orb2^ΔQ</i> allele.

<p>A) Percentage of scattered ICs is higher in <i>orb2^ΔQ</i> than wild type. A total of 34 wild type and 46 <i>orb2^ΔQ</i> testes were counted. B) Percentage of wild type or <i>orb2^ΔQ</i> testes having 0–3, 4–8, 9–13, 14–18, 19–23, 24–28, 29–33 or 34–38 ICs per testes. <i>orb2^ΔQ</i> testes have fewer ICs compared to wild type. C) <i>orb2^ΔQ</i> testes have normal spermatids. Green arrow: mature spermatocytes; yellow arrow: 64-cell spermatids cyst; arrowhead: spermatids at the beginning of elongation. D) Wild type testes stained with Orb2 (red), IC (Phalloidin, green) and DNA (blue). Yellow arrow points to where elongation usually stops in wild type testes. “d” marks the normal diameter of a testis at spermatogonia and early spermatocytes region. E–F) Flagellar axoneme bundles in <i>orb2^ΔQ</i> testes are over elongated. E) Overgrowth results in the swelling of the testis tip. Diameter of the <i>orb2^ΔQ</i> spermatogonia part of the testis is larger than that of the wild type (compare d in D and d′ in E), while the diameter of the nuclei side is relatively normal (compare d in D and d′, d″ in E). F) Another example of overly elongated flagellar axoneme bundles in <i>orb2^ΔQ</i> testis tip. The Orb2 positive axoneme bundle extended to the spermatogonial region and then changed its direction of elongation (arrows) to continue growing in the wrong direction. G, H) IC is not properly assembled in <i>orb2^ΔQ</i>. Phalloidin labeled Actin: green; DNA: blue. Arrow in G points to scattered actin cones of an <i>orb2^ΔQ</i> IC that remain relatively close together in one elongating spermatid cyst. Arrows in H are examples of widely scattered actin cones. In <i>orb2^ΔQ</i> testes with scattered IC, we can also observe what appear to be normal looking ICs, as indicated here by arrowhead in H. Scale bar: 50 µm.</p

<i>orb</i> is an Orb2 regulatory target.

<p>A) Western blots of extracts prepared from wild type, <i>orb2³⁶</i> and <i>orb2^Δ</i> testes were probed as indicated on the left. In this experiment Snf (Sans filles) was used as a loading control. Similar results were obtained using Tubulin as the loading control. B) <i>orb</i> mRNA can be immunoprecipitated with Orb2 antibodies from wild type testes extracts. <i>orb2³⁶</i> testis extract are used as a negative control for immunoprecipitation. After reverse transcription using oligo-dT primers, the <i>orb</i> cDNA was amplified using a primer set from the 3′ end of the male <i>orb</i> mRNA. L: 100 bp DNA ladder.</p