<p>Leydig-ove&nbsp; ćelije&nbsp; testisa&nbsp; su&nbsp; primarno&nbsp; mesto&nbsp; sinteze mu&scaron;kih polnih hormona. Ovi hormoni su neophodani za reproduktivno,&nbsp; ali&nbsp; i&nbsp; za&nbsp; op&scaron;te&nbsp; zdravlje&nbsp; budući&nbsp; da&nbsp; su<br />ozbiljni zdravstveni problemi često povezani sa njihovom smanjenom produkcijom.&nbsp; Insulin i insulinu sličan faktor rasta&nbsp; 1,&nbsp; IGF1&nbsp; <em>(engl.</em>&nbsp; insulin&nbsp; like&nbsp; growth&nbsp; factor&nbsp; 1),&nbsp; i<br />signalizacija koju pokreću preko svojih receptora&nbsp; (INSR i IGF1R),&nbsp; su&nbsp; jedan&nbsp; od&nbsp; ključnih&nbsp; faktora&nbsp; koji&nbsp; reguli&scaron;u specifični razvoj tkiva, pa i samih gonada. Ipak,&nbsp; uloga&nbsp; i<br />mehanizmi&nbsp; delovanja&nbsp; ovih&nbsp; receptora&nbsp; u&nbsp; steroidogenim tkivima nisu&nbsp; u potpunosti&nbsp; poznati.&nbsp; Stoga je&nbsp; istraživanje&nbsp; uokviru ove&nbsp; doktorske&nbsp; disertacije&nbsp; koncipirano sa ciljem da se,&nbsp; na&nbsp; modelu&nbsp; prepubertalnih&nbsp; (P21)&nbsp; i&nbsp; adultnih&nbsp; (P80) mužjaka mi&scaron;eva sa kondicionalnom delecijom<em> Insr </em>i <em>Igf1</em>r gena&nbsp; u&nbsp; steroidogenim&nbsp; ćelijama&nbsp; (Insr/Igf1r-DKO), defini&scaron;e uloga INSR i IGF1R u regulisanju diferencijacije i&nbsp; steroidogene&nbsp; funkcije&nbsp; Leydig-ovih&nbsp; ćelija.&nbsp; Pored&nbsp; toga, mužjaci&nbsp; i&nbsp; ženke&nbsp; P21&nbsp; mi&scaron;eva&nbsp; sa&nbsp; istom&nbsp; delecijom&nbsp; su kori&scaron;ćeni&nbsp; za&nbsp; praćenje&nbsp; ekspresije&nbsp; glavnih&nbsp; markera mitohondrijalne&nbsp; biogeneze&nbsp; i&nbsp; fuzije/arhitekture&nbsp; u&nbsp; Leydigovim&nbsp; ćelijama,&nbsp; ovarijumima&nbsp; i&nbsp;&nbsp; nadbubrežnim&nbsp; žlezdama. Rezultati&nbsp; su&nbsp; potvrdili&nbsp; da&nbsp; delecija&nbsp; Insr&nbsp; i&nbsp; Igf1r&nbsp; u<br />steroidogenim&nbsp; tkivima&nbsp; utiče&nbsp; na&nbsp; diferencijaciju&nbsp; i funkcionalne karakteristike Leydig-ovih ćelija P21 i P80 mi&scaron;eva,&nbsp; upućujući&nbsp; na&nbsp; pojavu&nbsp; tzv.&nbsp; &bdquo;feminizacije&ldquo;.&nbsp; Broj<br />Leydig-ovih&nbsp; ćelija&nbsp; izolovanih&nbsp; iz&nbsp; P21&nbsp; i&nbsp; P80&nbsp; Insr/Igf1rDKO&nbsp; mi&scaron;eva&nbsp; bio&nbsp; je&nbsp; smanjen,&nbsp; a&nbsp; morfologija&nbsp; i ultrastruktura&nbsp; ovih&nbsp; ćelija&nbsp; izmenjene&nbsp; kod&nbsp; P21&nbsp; Insr/Igf1rDKO&nbsp; mi&scaron;eva.&nbsp; Steroidogeni&nbsp; kapacitet&nbsp; i&nbsp; aktivnost,&nbsp; kao&nbsp; i ekspresija&nbsp; glavnih&nbsp; elemenata&nbsp; steroidogene&nbsp; ma&scaron;inerije <em>(Lhcgr, Star, Cyp11a1, Cyp17a1, Hsd3b1&nbsp; i&nbsp; 6, Hsd17b3,</em><br /><em>Sf</em>1)&nbsp; bili su&nbsp; smanjeni&nbsp; u Leydig-ovim ćelijama P21 i P80 <em>Insr/Igf1</em>r-DKO mi&scaron;eva,&nbsp; dok je ekspresija transkripcionih represora&nbsp; steroidogeneze&nbsp; (Arr19&nbsp; i&nbsp; Dax1)&nbsp; bila&nbsp; povećana specifično&nbsp; u&nbsp; istim&nbsp; ćelijama,&nbsp; ali&nbsp; ne&nbsp; i&nbsp; u&nbsp; ostatku&nbsp; testisa.<br />Transkripcioni&nbsp; profil&nbsp; markera&nbsp; mu&scaron;kog&nbsp; pola&nbsp; (<em>Sry,&nbsp; Sox9, Amh</em>)&nbsp; bio&nbsp; je&nbsp; izmenjen&nbsp; u Leydig-ovim ćelijama P21 i P80 <em>Insr/Igf1r</em>-DKO&nbsp; mi&scaron;eva.&nbsp; Transkripcija&nbsp; markera&nbsp; ženskog pola (<em>Rspo1, Wnt4</em>) u testisima,&nbsp; kao i ekspresija&nbsp; Cyp19a1 i&nbsp; produkcija estradiola (E2) u Leydig-ovim ćelijama,&nbsp; P21 i&nbsp; P80&nbsp;<em> Insr/Igf1r</em>-DKO&nbsp; mi&scaron;eva&nbsp; bile&nbsp; su&nbsp; povećane. Transkripcija&nbsp; markera&nbsp; mitohondrijalne&nbsp; biogenze (<em>Ppargc1a,&nbsp; Tfam</em>,&nbsp; <em>Mtnd1</em>)&nbsp; bila&nbsp; je&nbsp; smanjena&nbsp; u&nbsp; Leydigovim&nbsp; ćelijama&nbsp; P21&nbsp; <em>Insr/Igf1r</em>-DKO&nbsp; mi&scaron;eva,&nbsp; dok&nbsp; supromene&nbsp; ekspresije&nbsp; izostale&nbsp; u&nbsp; ovarijumima&nbsp; ženki&nbsp; istog&nbsp; genotipa.&nbsp; Isti&nbsp; markeri&nbsp; su&nbsp; bili&nbsp; povećani&nbsp; u&nbsp; nabdubrežnim&nbsp; žlezdama&nbsp; oba&nbsp; pola.&nbsp; Markeri&nbsp; mitohondrijalne fuzije/arhitekture&nbsp; (<em>Mfn1&nbsp; i&nbsp; Mfn2)</em>&nbsp; bili&nbsp; su&nbsp; povećani&nbsp; u Leydig-ovim ćelijama P21 <em>Insr/Igf1r</em>-DKO mi&scaron;eva, &scaron;to je&nbsp; praćeno&nbsp; i&nbsp; naru&scaron;enom&nbsp; mitohondrijalnom&nbsp; fazom steroidogeneze (produkcija progesterona), kao i brojem i&nbsp; morfologijom ovim organela.&nbsp; Ekspresija istih markera u ovarijumima&nbsp; bila&nbsp; je&nbsp; nepromenjena.&nbsp; Sumirano,&nbsp; rezultati ovog istraživanja&nbsp; su&nbsp; pokazali&nbsp; da su&nbsp; INSR i IGF1R&nbsp; važni za&nbsp; diferencijaciju&nbsp; i&nbsp; steroidogenu&nbsp; funkciju&nbsp; Leydig-ovih&nbsp; ćelija&nbsp; P21&nbsp; i&nbsp; P80&nbsp; mi&scaron;eva.&nbsp; Takođe,&nbsp; ovi&nbsp; receptori&nbsp; su&nbsp; važni regulatori&nbsp; markera&nbsp; mitohondrijalne&nbsp; biogeneze&nbsp; i fuzije/arhiteture u steroidogenim ćelijama mu&scaron;kih gonada&nbsp; P21 mi&scaron;eva, ali ne i u steroidogenim ćelijama ovarijuma.&nbsp;</p> / <p>Leydig cells of testes are the primary site of the male sex hormones&nbsp; synthesis.&nbsp; These&nbsp; hormones&nbsp; are&nbsp; indispensable for&nbsp; both&nbsp; reproductive&nbsp; and&nbsp; general&nbsp; health&nbsp; since&nbsp; serious health&nbsp; problems&nbsp; are&nbsp; often&nbsp; associated&nbsp; with&nbsp; their&nbsp; reduced production.&nbsp; Insulin&nbsp; and&nbsp; insulin-like&nbsp; growth&nbsp; factor&nbsp; 1, IGF1&nbsp; (insulin&nbsp; like&nbsp; growth&nbsp; factor&nbsp; 1),&nbsp; and&nbsp; signaling triggered through&nbsp; their receptors (INSR and IGF1R), are&nbsp; one of the key&nbsp; factors&nbsp; that regulate specific development of&nbsp; tissue&nbsp; including&nbsp; gonads.&nbsp; However,&nbsp; the&nbsp; role&nbsp; and mechanisms&nbsp; of&nbsp; these&nbsp; receptors&nbsp; action&nbsp; in&nbsp; steroidogenic tissues are not known enough. This study was designed to&nbsp; observe &nbsp; the role of INSR and IGF1R in regulating the differentiation and steroidogenic function of Leydig cells by using the model of prepubertal (P21) and adult (P80) male mice with the conditional deletion of the&nbsp; Insr&nbsp; and Igf1r&nbsp; genes&nbsp; in&nbsp; steroidogenic&nbsp; cells&nbsp; (<em>Insr/Igf1r-</em>DKO).&nbsp; In addition,&nbsp; male&nbsp; and&nbsp; female&nbsp; P21&nbsp; mice&nbsp; with&nbsp; the&nbsp; samedeletion were used to monitor the expression of the main markers&nbsp; of&nbsp; mitochondrial&nbsp; biogenesis&nbsp; and fusion/architecture&nbsp; in&nbsp; Leydig&nbsp; cells,&nbsp; ovaries&nbsp; and&nbsp; adrenal glands.&nbsp; The&nbsp; results&nbsp; confirmed&nbsp; that&nbsp; deletion&nbsp; of&nbsp;<em> Insr</em>&nbsp; and<em> Igf1r&nbsp;</em> in&nbsp; steroidogenic&nbsp; tissues&nbsp; influences&nbsp; differentiation and&nbsp; functional&nbsp; characteristics&nbsp; of&nbsp; Leydig&nbsp; cells&nbsp; isolated from&nbsp; P21&nbsp; and&nbsp; P80&nbsp; mice,&nbsp; suggesting&nbsp; an&nbsp; appearance&nbsp; of &quot;feminization&quot;.&nbsp; The&nbsp; number&nbsp; of&nbsp; Leydig&nbsp; cells&nbsp; isolated from&nbsp; both&nbsp; P21&nbsp; and&nbsp; P80&nbsp; <em>Insr/Igf1</em>r-DKO&nbsp; mice&nbsp; was reduced.&nbsp; Morphology&nbsp; and&nbsp; ultrastructure&nbsp; of&nbsp; Leydig&nbsp; cells were&nbsp; disturbed&nbsp; in&nbsp; P21&nbsp; <em>Insr/Igf1r-</em>DKO&nbsp; mice. Steroidogenic capacity and activity, as well as expression of the main elements of&nbsp; steroidogenic machinery (<em>Lhcgr, Star, Cyp11a1, Cyp17a1, Hsd3b1&nbsp; and&nbsp; 6, Hsd17b3, Sf1) </em>were&nbsp; decreased&nbsp; in&nbsp; Leydig&nbsp; cells&nbsp; from&nbsp; P21&nbsp; and&nbsp; P80 I<em>nsr/Igf1</em>r-DKO&nbsp; mice,&nbsp; while&nbsp; the&nbsp; expression&nbsp; of transcriptional&nbsp; repressors&nbsp; of&nbsp; steroidogenesis&nbsp; (<em>Arr19</em>&nbsp; and <em>Dax1) </em>was increased&nbsp; in the same cells, but not in the rest of&nbsp; the&nbsp; testes.&nbsp; Transcription&nbsp; profile&nbsp; of&nbsp; the&nbsp; male&nbsp; sex markers&nbsp; (<em>Sry,&nbsp; Sox9</em>,&nbsp; <em>Amh</em>)&nbsp; was&nbsp; altered&nbsp; in&nbsp; Leydig&nbsp; cells from&nbsp; P21&nbsp; and&nbsp; P80&nbsp; <em>Insr/Igf1</em>r-DKO&nbsp; mice.&nbsp; Transcription of the female sex markers (<em>Rspo1, Wnt4</em>) in the testes, as well&nbsp; as&nbsp; <em>Cyp19a1&nbsp; </em>expression&nbsp; and&nbsp; estradiol&nbsp; (E2) production in Leydig cells,&nbsp; from P21 and P80&nbsp; I<em>nsr/Igf1</em>rDKO&nbsp; mice&nbsp; were&nbsp; increased.&nbsp; Transcription&nbsp; of mitochondrial&nbsp; biogenesis&nbsp; markers&nbsp; (<em>Ppargc1a,&nbsp; Tfam, Mtnd1</em>)&nbsp; was&nbsp; declined&nbsp; in&nbsp; Leydig&nbsp; cells&nbsp; from&nbsp; P21<em> Insr/Igf1r-</em>DKO mice, while changes were absent in&nbsp; the ovaries of the same genotype.&nbsp; Transcription of the&nbsp; same markers&nbsp; was&nbsp; increased&nbsp; in&nbsp; the&nbsp; adrenal&nbsp; glands&nbsp; of&nbsp; both sexes.&nbsp; The&nbsp; mitochondrial&nbsp; fusion/architecture&nbsp; markers (<em>Mfn1</em>&nbsp; and&nbsp; <em>Mfn2</em>)&nbsp; were&nbsp; increased&nbsp; in&nbsp; Leydig&nbsp; cells&nbsp; from<em> Insr/Igf1r</em>-DKO&nbsp; mice&nbsp; and&nbsp; followed&nbsp; by&nbsp; disturbedmitochondrial&nbsp; phase&nbsp; of&nbsp; steroidogenesis&nbsp; (progesterone production), as well as&nbsp; decreased&nbsp; number and&nbsp; disturbed morphology&nbsp; of&nbsp; mitochondria.&nbsp;&nbsp; Expression&nbsp; of&nbsp; the&nbsp; same markers&nbsp; in&nbsp; the&nbsp; ovaries&nbsp; was&nbsp; unchanged.&nbsp; In&nbsp; summary, results&nbsp; of&nbsp; this&nbsp; study&nbsp; showed&nbsp; that&nbsp; INSR&nbsp; and&nbsp; IGF1R&nbsp; are important in differentiation and steroidogenic function of Leydig&nbsp; cells&nbsp; from&nbsp; P21&nbsp; and&nbsp; P80&nbsp; mice.&nbsp; Also,&nbsp; these receptors&nbsp; are&nbsp; important&nbsp; regulators&nbsp; of&nbsp; mitochondrial biogenesis&nbsp; and&nbsp;&nbsp; fusion/architecture&nbsp; markers&nbsp; in steroidogenic&nbsp; cells&nbsp; of&nbsp; P21&nbsp; male&nbsp; mice,&nbsp; but&nbsp; not&nbsp; in steroidogenic cells of ovaries.</p>

The location of Černá louka is situated between the city-centre of Moravská Ostrava and an area of Slezská Ostrava next to the confluence of two rivers – Ostravice and Lučina. The development at the building site of Černá louka is a result from an international competition 2010, which winner was the project Culture Meadow from studio Maxwan. The main idea of the project is to concentrate cultural and educational buildings around a culture meadow. This design has been used as a default situation for the master´s thesis. The complex of schools is situated at the parcel of existing Miniuni-site between cultural cluster Černá louka and the residential quarter Nová Karolina. The volume of the school with its front-spaces reacts to the surrounding structures. In the middle of the complex arises a half-public open space, where are located the internal entrances. Because of the placing of the school next to the city-centre the multifunctional pitch is situated on the roof of the gym. The design of the school respects principles of the age differentiation just as the horizontal integration. That´s why the substance makes up a symbiosis of 5 parts - common space, gym, first and second level, gymnasium. These parts are connected together and at the same time enable to use individual parts separately. The building of kindergarten with its garden is separated from the school because of differential needs of the smallest children.