Thesis submitted for the degree of Doctor of Philosophy in the University of the Witwatersrand, Johannesburg. / Biology, having outgrown its purely descriptive phase, has, for nearly a century, been in an era of causal analysis. It has been a period of extreme compartmentalisation of the general field into many disciplines, each endowed with a defined range of problems, with peculiar materials of study and with special approaches and techniques.From the nature of things, it is inevitable that each biologist should have been a specialist. The field of living things ramified so vastly that the species of scientist known as the biologist or naturalist became largely extinct: instead, there were geneticists, systematists, physiologists, embryologists, biochemists, cytologlsts and others. Specialisation did not stop even at that point for the systematists split into mammalogists, ornithologists, helminthologists, etc.; the geneticists into experimental geneticists, cytogeneticists, phenogenetlclsts, and so on. Good and bad consequences flowed from this tendency. The advantage of specialisation was a great increase in the store of factual information; the disadvantage lay in the isolation between representatives of the various disciplines and in the absence of cross-pollination in the development and evaluation of concepts. / WHSLYP2017

Submitted by Fabíola Silva (fabiola.silva@famerp.br) on 2016-06-22T19:13:02Z No. of bitstreams: 1 luizrobertofalleirosjunior_dissert.pdf: 2855726 bytes, checksum: 19672bd7d4c60e301a52bec076ff8240 (MD5) / Made available in DSpace on 2016-06-22T19:13:02Z (GMT). No. of bitstreams: 1 luizrobertofalleirosjunior_dissert.pdf: 2855726 bytes, checksum: 19672bd7d4c60e301a52bec076ff8240 (MD5) Previous issue date: 2015-09-28 / Fundação de Amparo à Pesquisa do Estado de São Paulo - FAPESP / The prostate is an accessory gland of genital system responsible for producing an alkaline liquid which ensures sperm survival and capacitation. Its development occurs under the influence of an androgen and estrogen regulated and precise control, so sensible interferences may predispose this gland to developing diseases such as benign prostatic hyperplasia and cancer during adult and senile life. Inappropriate exposure to exogenous estrogens in embryonic and neonatal development period has been related to permanent disturbances in morphophysiology of various organs of the male reproductive system. Epidemiological studies indicate that men who were exposed to estrogenic compounds during intrauterine and neonatal development had higher probability of infertility and testicular and prostate cancer. Until now, it is not possible to say whether there are intrinsic factors of development that may predispose prostatic gland to the development of cancer with age. However, as early exposure to steroid hormones can permanently alter various reproductive organs, it is questionable whether such agents may interfere in the prostate structure and physiology. Based on these reports, the aim of this study was to analyze morphologically the ventral prostate of adult gerbils exposed to ethynylestradiol (EE) during the first week of prenatal development. To this, we employed morphological, stereological-morphometrical, immunohistochemical and ultrastructural methods. The results showed that the postnatal exposure to EE duplicated the prostatic complex weight, increasing the relative frequency of epithelial and stromal compartments, besides the secretory activity of the ventral lobe of the prostate. All glands exposed to EE showed strong stromal reshuffling and some foci of epithelial hyperplasia and inflammatory infiltrated in both luminal and epithelial or stromal compartments. Cells positive for AR and PCNA increased into the epithelial and stromal tissues. ERα-positive cells, which are normally found into stromal compartment of intact prostates, were frequently observed in the prostatic epithelial of treated animals. This study demonstrated that the exposure to EE during the postnatal development causes histophysiological alterations of this gland, predisposing to the development of prostatic lesions during life. These results are important taking account public health, considering the EE has been largely used by women worldwide. Moreover, the bioaccumulation of this chemical has been increased in different ecosystems. / A próstata é uma glândula acessória do sistema genital responsável pela produção de um líquido alcalino que garante a capacitação e sobrevivência dos espermatozóides. O seu desenvolvimento ocorre sob influência de um controle androgênico e estrogênico regulado e preciso, de forma que sensíveis interferências podem predispor esta glândula a desenvolver doenças como hiperplasia prostática benigna e câncer durante a vida adulta e senil. A exposição inapropriada a estrógenos exógenos no período de desenvolvimento embrionário e neonatal tem sido relacionada a distúrbios permanentes na morfofisiologia de vários órgãos do sistema reprodutor masculino. Estudos epidemiológicos indicam que homens que foram expostos a componentes estrogênicos durante o desenvolvimento intra-uterino e neonatal apresentaram maior probabilidade de ocorrência de infertilidade e de câncer de testículo e próstata. Até o momento, não é possível afirmar se existem fatores intrínsecos do desenvolvimento que podem predispor a glândula prostática ao desenvolvimento de câncer com a idade. Porém, como a exposição precoce a hormônios esteróides é capaz de alterar permanentemente vários órgãos reprodutivos, torna-se questionável se esses agentes também podem interferir na estrutura e fisiologia prostática. Com base nesses relatos, o objetivo deste estudo foi analisar morfologicamente a próstata ventral de gerbilos adultos expostos ao ethynylestradiol (EE) durante a primeira semana de desenvolvimento pós-natal. Para isso, foram empregados métodos morfológicos, morfométricos-estereológicos, imunohistoquímicos e ultraestruturais. Os resultados obtidos demonstraram que a exposição pós-natal ao EE duplicou o peso do complexo prostático, aumentando a frequência relativa dos compartimentos epitelial e estromal e a atividade secretora no lobo ventral da próstata. Todas as glândulas expostas ao EE apresentaram intensa remodelação estromal, focos de hiperplasia epitelial e de infiltrados inflamatórios intersticiais e intraluminais. Células AR e PCNA-positivas tornaram-se significativamente mais frequentes no epitélio e no estroma. Células ERα-positivas, que foram restritas ao compartimento estromal em próstatas controle, foram frequentemente observadas no epitélio prostático dos animais tratados. O presente estudo demonstrou que a exposição ao EE durante o desenvolvimento pós-natal da próstata ventral pode causar alterações organizacionais na histofisiologia deste órgão, predispondo os indivíduos precocemente a lesões prostáticas. Estes resultados são de grande importância para a saúde pública, visto que o EE tem sido cada vez mais utilizado por mulheres em todo o mundo, e que a bioacumulação deste químico ambiental tem atingido níveis cada vez mais altos nos diferentes ecossistemas.

Introdução: Muitos estudos veem sendo realizados com a finalidade de identificar agentes que possam ter efeito benéfico no tratamento ou prevenção das lesões causadas nos neurônios devido à isquemia. A hipotermia já demonstrou resultados consistentes em estudos experimentais e a Estimulação Magnética Transcraniana (EMTr) já foi usada visando reduzir danos em neurônios hipocampais de animais submetidos a isquemia cerebral. Com a propriedade de aumentar ou diminuir a excitabilidade cortical a partir do estímulo magnético, estima-se que ocorra uma interferência na produção de alguns neurotransmissores e receptores de membrana, que promoveriam efeito protetor a estas células. Neste estudo avaliamos a capacidade da EMTr de proteger os neurônios de uma lesão por hipóxia, e sua possível interferência no efeito protetor da hipotermia, tentando identificar alguns mecanismos que possivelmente estariam envolvidos neste fenômeno. Métodos: Como modelo de isquemia, foram utilizados Gerbils previamente submetidos a uma avaliação de comportamento e memória por meio do teste de esquiva. O protocolo de EMTr foi a partir de sessões diárias com 25 séries de 5 segundo a 25Hz, com um intervalo de 45 segundos entre as séries, por sete dias consecutivos, com um total de 21 875 pulsos com uma intensidade de 100% do limiar motor, e sendo realizada a indução da isquemia logo após o término da última sessão, ou na isquemia após a EMTr, em sessões diárias com 25 séries de 5 segundos a 25Hz, com um intervalo de 45 segundos entre as séries, durante 3 dias consecutivos, começando imediatamente após a cirurgia. Foi mantida a temperatura de 36 °C durante o período de oclusão do vaso e os 30 minutos consecutivos, ou 31 a 32 °C quando em hipotermia. O preparo das lâminas teve cortes envolvendo a região do hipocampo, corados com hematoxilina e eosina, além de outros preparos, a marcação de TUNEL e Caspase, que visam evidenciar a ocorrência de apoptose. Resultados: Embora sem significância estatística, os animais que receberam EMTr aparentemente tiveram uma melhor performance no teste da esquiva, principalmente se aplicado após a indução da isquemia. A hipotermia demonstrou uma eficiência significativa, tanto na análise histológica quanto no teste da esquiva, associado ou não à EMTr, e nestes animais submetidos a isquemia durante a hipotermia, os que receberam EMTr tiveram área de sobrevida no hipocampo significativamente maior na análise histológica com hematoxilina e eosina. Nos animais submetidos à isquemia durante a temperatura normal, a EMTr não demonstrou aumentar a área de sobrevida das células do hipocampo. Conclusões: A EMTr (ativa ou placebo, prévia ou posterior à isquemia) pareceu ter um efeito positivo no teste de esquiva. O procedimento de estimulação pareceu bastante traumático e estressante para os animais, tendo ocorrido alguns óbitos durante a imobilização, provavelmente por asfixia. A EMTr apresentou efeito protetor significativo apenas nos animais submetidos a isquemia durante hipotermia / Introduction: Over the time many researches have been conducted with the aim of identifying agents that may have beneficial effects in the treatment or prevention of cerebral ischemia, hypothermia has shown consistent results in experimental trials and Repetitive Trans Cranial Magnetic Stimulation (rTMS) has been used in a study attempting to reduce damage in hippocampal neurons. With the property to increase or decrease cortical excitability from the repetitive magnetic stimulus, it is estimated that an interference occurs in the production of some neurotransmitters and receptors of neuronal membrane, which therefore protects these cells from hypoxia. In this study we evaluated the ability of rTMS to protect neurons from injury due to hypoxia, and its possible interference in the protective effect of hypothermia and we tried to identify some mechanisms that possibly are involved in this phenomenon. Methods: Ischemia model was performed using Gerbil that was subsequently submitted to an evaluation of behavior and memory through passive avoidance task. The rTMS protocol was daily sessions with 25 series of 5 seconds at 25Hz with an interval of 45 seconds between series, for 7 consecutive days, with a total of 21 875 pulses with an intensity of 100% of motor threshold, and being carried through the induction of ischemia soon after the end of the last session, or rTMS after ischemia, in daily sessions with 25 series of 5 seconds at 25Hz with an interval of 45 seconds between series, for 3 consecutive days, starting immediately after surgery. The temperature of 36 °C was maintained during the period of vessel occlusion and subsequent 30 minutes, or 31 °C to 32 °C when in hypothermia. The preparation of the slices had sections of the region involving the hippocampus, stained with hematoxylin and eosin in addition to other preparations, TUNEL and caspase, which aim to evidence the occurrence of apoptosis. Results: Although not statistically significant, animals that received rTMS, apparently had better performance in passive avoidance task especially when applied after ischemia. The hypothermia demonstrated a significant efficiency, both in the histological analysis and in the passive avoidance task, associated or not to applications of rTMS and, in these animals undergoing ischemia during hypothermia, the ones who received rTMS had survival area in hippocampus significantly higher in histological analysis with hematoxylin and eosin. In animals undergone to ischemia during normal temperature, the rTMS has not shown to increase the area of hippocampal cell survival. Conclusions: rTMS (placebo or active, after or before the ischemia) seems to have a positive effect on passive avoidance task. The stimulation procedure appeared to be very traumatic and stressful for the animal, in which a few deaths occurred during the procedure, probably from asphyxiation due to restraint. The rTMS had a significant protective effect only in animals undergoing ischemia during hypothermia, as demonstrated in the histological analysis with hematoxylin and eosin

Introdução: Muitos estudos veem sendo realizados com a finalidade de identificar agentes que possam ter efeito benéfico no tratamento ou prevenção das lesões causadas nos neurônios devido à isquemia. A hipotermia já demonstrou resultados consistentes em estudos experimentais e a Estimulação Magnética Transcraniana (EMTr) já foi usada visando reduzir danos em neurônios hipocampais de animais submetidos a isquemia cerebral. Com a propriedade de aumentar ou diminuir a excitabilidade cortical a partir do estímulo magnético, estima-se que ocorra uma interferência na produção de alguns neurotransmissores e receptores de membrana, que promoveriam efeito protetor a estas células. Neste estudo avaliamos a capacidade da EMTr de proteger os neurônios de uma lesão por hipóxia, e sua possível interferência no efeito protetor da hipotermia, tentando identificar alguns mecanismos que possivelmente estariam envolvidos neste fenômeno. Métodos: Como modelo de isquemia, foram utilizados Gerbils previamente submetidos a uma avaliação de comportamento e memória por meio do teste de esquiva. O protocolo de EMTr foi a partir de sessões diárias com 25 séries de 5 segundo a 25Hz, com um intervalo de 45 segundos entre as séries, por sete dias consecutivos, com um total de 21 875 pulsos com uma intensidade de 100% do limiar motor, e sendo realizada a indução da isquemia logo após o término da última sessão, ou na isquemia após a EMTr, em sessões diárias com 25 séries de 5 segundos a 25Hz, com um intervalo de 45 segundos entre as séries, durante 3 dias consecutivos, começando imediatamente após a cirurgia. Foi mantida a temperatura de 36 °C durante o período de oclusão do vaso e os 30 minutos consecutivos, ou 31 a 32 °C quando em hipotermia. O preparo das lâminas teve cortes envolvendo a região do hipocampo, corados com hematoxilina e eosina, além de outros preparos, a marcação de TUNEL e Caspase, que visam evidenciar a ocorrência de apoptose. Resultados: Embora sem significância estatística, os animais que receberam EMTr aparentemente tiveram uma melhor performance no teste da esquiva, principalmente se aplicado após a indução da isquemia. A hipotermia demonstrou uma eficiência significativa, tanto na análise histológica quanto no teste da esquiva, associado ou não à EMTr, e nestes animais submetidos a isquemia durante a hipotermia, os que receberam EMTr tiveram área de sobrevida no hipocampo significativamente maior na análise histológica com hematoxilina e eosina. Nos animais submetidos à isquemia durante a temperatura normal, a EMTr não demonstrou aumentar a área de sobrevida das células do hipocampo. Conclusões: A EMTr (ativa ou placebo, prévia ou posterior à isquemia) pareceu ter um efeito positivo no teste de esquiva. O procedimento de estimulação pareceu bastante traumático e estressante para os animais, tendo ocorrido alguns óbitos durante a imobilização, provavelmente por asfixia. A EMTr apresentou efeito protetor significativo apenas nos animais submetidos a isquemia durante hipotermia / Introduction: Over the time many researches have been conducted with the aim of identifying agents that may have beneficial effects in the treatment or prevention of cerebral ischemia, hypothermia has shown consistent results in experimental trials and Repetitive Trans Cranial Magnetic Stimulation (rTMS) has been used in a study attempting to reduce damage in hippocampal neurons. With the property to increase or decrease cortical excitability from the repetitive magnetic stimulus, it is estimated that an interference occurs in the production of some neurotransmitters and receptors of neuronal membrane, which therefore protects these cells from hypoxia. In this study we evaluated the ability of rTMS to protect neurons from injury due to hypoxia, and its possible interference in the protective effect of hypothermia and we tried to identify some mechanisms that possibly are involved in this phenomenon. Methods: Ischemia model was performed using Gerbil that was subsequently submitted to an evaluation of behavior and memory through passive avoidance task. The rTMS protocol was daily sessions with 25 series of 5 seconds at 25Hz with an interval of 45 seconds between series, for 7 consecutive days, with a total of 21 875 pulses with an intensity of 100% of motor threshold, and being carried through the induction of ischemia soon after the end of the last session, or rTMS after ischemia, in daily sessions with 25 series of 5 seconds at 25Hz with an interval of 45 seconds between series, for 3 consecutive days, starting immediately after surgery. The temperature of 36 °C was maintained during the period of vessel occlusion and subsequent 30 minutes, or 31 °C to 32 °C when in hypothermia. The preparation of the slices had sections of the region involving the hippocampus, stained with hematoxylin and eosin in addition to other preparations, TUNEL and caspase, which aim to evidence the occurrence of apoptosis. Results: Although not statistically significant, animals that received rTMS, apparently had better performance in passive avoidance task especially when applied after ischemia. The hypothermia demonstrated a significant efficiency, both in the histological analysis and in the passive avoidance task, associated or not to applications of rTMS and, in these animals undergoing ischemia during hypothermia, the ones who received rTMS had survival area in hippocampus significantly higher in histological analysis with hematoxylin and eosin. In animals undergone to ischemia during normal temperature, the rTMS has not shown to increase the area of hippocampal cell survival. Conclusions: rTMS (placebo or active, after or before the ischemia) seems to have a positive effect on passive avoidance task. The stimulation procedure appeared to be very traumatic and stressful for the animal, in which a few deaths occurred during the procedure, probably from asphyxiation due to restraint. The rTMS had a significant protective effect only in animals undergoing ischemia during hypothermia, as demonstrated in the histological analysis with hematoxylin and eosin

Thesis (MSc)--Stellenbosch University, 2013. / ENGLISH ABSTRACT: The objectives of this research were to: (1) accurately identify chromosome homology, (2) identify chromosome rearrangements leading to diploid number variation and chromosome evolution to formulate the ancestral gerbil karyotype and distinguish homoplasy from hemiplasy, and (3) construct a phylogeny using chromosomal characters based on G-banding and fluorescence in situ hybridization (FISH) of ten gerbil species representing five genera (Gerbilliscus, Desmodillus, Psammomys, Meriones, Taterillus), with probes derived from flow-sorted chromosomes of G. paeba (GPA, 2n = 36), and polarised with the murid outgroup, Micaelamys namaquensis. All paints successfully hybridized to all ingroup and outgroup taxa. Three of the 19 G. paeba painting probes (GPA 7, 9 and X [including the X; autosome translocation in T. pygargus]) were conserved as whole chromosomes, and 16 were rearranged (GPA 1-6, 8, 10-17). Chromosome painting correctly identified the homology of the heterochromatic GPA 7, which was conserved as whole chromosomes in all gerbils. Thirteen previously misidentified G-band homologies were correctly identified with FISH; one in D. auricularis, and six each in G. kempi and G. gambianus. Homology maps identified 57 syntenic associations and that 19 rearrangements are responsible for diploid number differentiation among species. Parsimony analysis of the two matrices (syntenic association and rearrangements) retrieved a sister-species relationship between G. gambianus and G. kempi, and P. obesus and M. persicus (syntenic associations), an unresolved clade that included D. auricularis, G. gambianus, P. obesus and M. persicus (chromosome rearrangements) and a basal position for T. pygargus. Phylogenies derived from chromosomal data failed to resolve the deeper nodes. Consequently, characters were subsequently mapped on a molecular consensus tree (including a chronogram). This allowed inferences on the rate of chromosome evolution, which indicates that the basal D. auricularis is separated from Gerbilliscus by nine rearrangements (four Robertsonian fusions, five inversions), at a rate of 1.25/Myr. Gerbilliscus species evolved with an average of 10 Robertsonian rearrangements involving GPA 1–6, 8, 10 – 12, of which four are homoplasies (GPA 1-3, 5), one a potential hemiplasy (GPA 5) to southern African taxa, one a synapomorphy to G. paeba and G. tytonis (GPA 6), two synapomorphies in G. kempi and G. gambianus (GPA 11, 12), and three are synapomorphic to Gerbilliscus (GPA 4, 8, 10). Homoplasic characters across the two clades include GPA 3 (T. pygargus, G. paeba and G. tytonis) and GPA 5p-q prox (D. auricularis, P. obesus and M. persicus). Gerbilliscus (excluding G. paeba and G. tytonis) had the slowest chromosome evolutionary rate of < 1/Myr; G. paeba and G. tytonis were slightly faster at 2/Myr. The clade comprised of M. persicus, P. obesus and T. pygargus evolved faster, at a rate of 4/Myr (seven fissions, five fusions, two inversions), 2.3/Myr (seven fissions, two fusions, four inversions) and 16/Myr (eight fusions), respectively, indicating heterogeneity among Gerbillinae: A slow rate in Desmodillus and Gerbilliscus, and a fast evolutionary rate in Psammomys, Meriones and Taterillus. The putative ancestral karyotype was postulated to be 2n = 56, and included five biarmed autosomes and X chromosome, and 22 acrocentrics. This is provisional, since Brachiones, Desmodilliscus, Pachyuromys, Sekeetamys, Gerbillus and Rhombomys were not analysed. / AFRIKAANSE OPSOMMING: Die objektief van hierdie navorsing was om: (1) akkurate chromosoom homologie te identifiseer, (2) chromosoom herranskikkings te identifiseer wat mag lei tot diploide chromosoom getal variasie en chromosoom evolusie ten einde te formuleer die voorouer karyotiepe van “gerbils” sowel as om te onderskei tussen homoplasie en hemiplasie, en (3) die konstruksie van 'n filogenetiese boomstam gebasseer op chromosoom karakters verkry vanaf G-banding en FISH (fluoressensie in situ hibridisasie) van tien “gerbil” spesies wat vyf genera verteenwoordig (Gerbilliscus, Desmodillus, Psammomys, Meriones, Taterillus), deur van geskikte sondes gebruik te maak wat verkry in deur floei-sorteerde chromosome van G. peaba (GPA, 2n = 36), wat gepolariseerd was met die Murid buitegroep, Micealamys namaquensis. Alle chromosoom verwe het suksesvolgehibridiseer aan al die ingroep en buitegroep taxa. Drie van die 19 G.peaba verfwe (GPA 7, 9 and X (including the X; autosome translocation in T. pygargus) was bewaar as heel chromosome, en 16 herrangskik (GPA 1-6, 8, 10-17). Chromosoom verfwing kon suksesvol die homologie van die heterochromatise GPA7 identifiseer wat gekonserveerd was as heel chromosome in al die “gerbils”, wat moontlik aandui die teenwoordigheid van funksionele gene. Dertien voorheen mis geidentifiseerde G-band homologieë was gekorregeer deurmiddel van FISH, een in D. auricularis, en ses elk in G. kempi en G. gambianus. Homologie kaarte het 57 sintesiese assosiasies geidentifiseer en dat 19 herrangskikings verantwoordelik was vir diploied nommer differensiasies tussen spesies. Parsimonie analises van die twee matrikse (sinteniese assosiasies en herrangskikings) wys 'n suster-spesie verwantskap tussen G. gambianus en G. kempi, en P. obesus en M .persicus (sinteniese assosiasies), 'n unopgeloste klade wat D. auricularis, G. gambianus, P. obesus en M. persicus (chromosoom herrangskikkings) opmaak vorm die basale posisie vir Taterillus pygargus. Filogenetise boomstamme verkry vanaf die chromosomale data misluk egter om die dieper nodes op te los. Karakters was daarna geplot op 'n konsensus boom (insluitend 'n chronogram). Dit het dieper insigte toegelaat soos die tempo van chromosoom evolusie, wat aandui dat die basale D. auricularis geskei is vanaf Gerbilliscus met nege herrangskikkings (vier Robertsonian, vyf inversies) teen 'n tempo van 1.25/Mja. Gerbilliscus spesies het verander met 13 herranskikinge (11 saamsmeltings en twee inversies), waarvan vier potensiele homoplasies/hemiplasies (GPA 1-3, 5). Met die uitsluitsel van G. paeba en G. tytonis, het Gerbilliscus die laagste chromosoom evolutionêre tempo van al die “gerbils” < 1 /Mja, G. paeba en G. tytonis was ietwat vinniger met 'n tempo van 2/Mja. The klade wat bestaan uit M. persicus, P. obesus en T. pygargus verander vinniger as Desmodillus en Gerbilliscus, met 'n evolutionêre tempo van 4/Mja (sewe fissies, vyf samesmeltings, twee inversies) en 2.3 Mja (sewe fissies, twee samesmeltings, vier inversies) onderskeidelik, wat grootendeels tandem was. Die karyotiepe van Taterillus pygarus het agt samesmeltings gehad wat predominant tandem was, teen 'n tempo van 16/Mja. Terwyl meeste van die herrangskikinge synapomorfies was, was sommige homoplasties of hemiplasties. Homoplastiese karakters wat gedeel was tussen die twee klades sluit in GPA 3 (in T pygargus en G. paeba en G. tytonis) en GPA 5p-q prox (D. auricularis, P. obesus en M. persicus). GPA 5 was hemiplasties aan alle suider Afrikaanse taxa. Die analise van sinteniese assosiasies en chromosoom herrangskikings was geanaliseer in PAUP, en gepolariseer met die murid Micealamys namaquensis. Taterillus pygarus het 'n basale posisie in beide filogenetiese boomstamme. Die data stel voor dat FISH meer akkurate resultate lewer op chromosoom homologie as die streng gebruik making van banding patrone. Verder het die tempo van chromosoom evolusie gevarieër vanaf stadig (Desmodillus en Gerbilliscus) tot vinnig (Psammomys, Meriones en Taterillus), chromosoom karakters egter was nie sterk genoeg om dieper filogenetiese verwantskappe te ondersoek nie. Die voornemende voorouerlike karytiepe van “gerbils” was hier gehipotiseer as 2n = 56. Drie bevindinge resoneer uit hierdie studie. Eerstens, chromosoom verwing kon chromosoom homologieë wat voorheen deur banding studies mis ge-identifiseer was korrek identifiseer: hierdie sluit in een konflik in D. auricularis, en ses elk vir G. kempi en G. gambianus. Tweedens, die homologie van die heterochromatiese of C-positiewe autosome, GPA 7, was gedemonstreer as bewaar as 'n heel chromosoom as beide heterochromaties en euchromatiese chromosome in alle “gerbils”, wat aandui dat dit functionele gene dra. Derdens gebasseer op simpleisiomorfe wat geidentifiseer was vanaf die homologie kaarte en vergelykbare opleidings, hipotiseer ek dat die voorgestelde voorouer karyotiepe bestaan uit ses autosome (GPA 7, 9, 13, 15, 16, 17) en die X chromosoom, wat onveranderd gebly het tussen alle suider Afrikaanse taxa. Met die uitsluitsel van GPA 7p/7q, was almal behoue as twee-armige chromosome in die voor ouer karyotiepe. In lyn met hierdie is, 21 akrosentries GPA1p,1q, 2p, 2q, 3p, 3q, 4p, 4q, 5p, 5q, 6p, 6q, 8p, 8q, 10q, 10p, 11p,11q, 12p, 12q en 14, wat lei tot die voorgestelde voor ouerlike diploiede chromosoom getal van 2n = 56. Diè karyotiepe word voorgestel as 'n werkende hipotese deels omdat Brachiones, Desmodilliscus, Pachyuromys, Sekeetamys, Gerbillus en Rhombomys nie geanaliseer was nie, en dat die idiale buite groep vir “gerbils” van die Acomyinae nie gebruik was om die karakters te polariseer was nie.

Orientador: Sebastião Roberto Taboga / Tese (doutorado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-09T15:30:54Z (GMT). No. of bitstreams: 1 Souza_CristianiZanetoniIsraelde_D.pdf: 8114014 bytes, checksum: 02df40337836deefc11fdc588dbdccb0 (MD5) Previous issue date: 2007 / Resumo: O câncer de próstata atualmente é o tumor mais comum em homens com mais de 50 anos de idade. Entre os fatores de risco que contribuem para o aumento dessa doença destaca-se o envelhecimento, período em que ocorrem acentuados desequiliorios hormonais. Há muitas dificuldades na obtenção de material humano para estudos do desenvolvimento de tumores prostáticos, pois esta questão esbarra na ética médica. Vários grupos de pesquisa vêm tentando desenvolver, caracterizar e validar modelos roedores para análise do câncer de próstata. Modelos autóctones, nos quais são estudadas as lesões prostáticas espontâneas, têm desempenhado papel relevante nas pesquisas dessa neoplasia. Além disso, inúmeras investigações têm sido feitas sobre a indução experimental de tumores na próstata de roedores de laboratório: ratos, camundongos e cobaias. Em uma primeira etapa deste trabalho, foram realizadas análises morfológicas (estruturais e ultra-estruturais), quantitativas e funcionais dos componentes celulares dos compartimentos epitelial e estromal do lobo ventral da próstata do gerbilo" velho (Meliones unguiculatus). A morfologia prostática nesses animais revelou que em uma mesma glândula puderam ser observadas regiões funcionais com epitélio secretor normal e outras áreas com alterações histopatológicas atípicas. Nesses, o declínio de testosterona esteve associado a alterações proliferativas na glândula, levando ao entendimento da importância desse andrógeno na homeostase e funcionalidade prostática. Devido a essas constatações, em uma segunda etapa, foi feita a indução experimental de tumores na próstata do gerbilo adulto, após tratamento conjugado de N-metil-N-rutrosouréia com propionato de testosterona. Depois de estabelecidos os tumores, as próstatas foram processadas para estudos rustológico, imunocitoquímico e ultra-estrutural. Os resultados mostraram que em gerbilos o surgimento de lesões prostáticas ocorreu em períodos experimentais de até 9 meses e que tanto o cancerígeno como a testosterona, associados ou não, foram indutores de neoplasias. Sugere-se que as células atípicas possam apresentar potencial invasivo pela observação da ruptura da membrana basal pelos métodos de imunocitoquímica para laminina e análise ultra-estrutural. Detectou-se também no gerbilo a expressão da proteína citoplasmática Alfa-Metilacil-CoARacemase (PS04S) em células prostáticas neoplásicas, bem como é observado no câncer prostático do homem. Assim, este trabalho é pioneiro na demonstração da expressão de P504S em roedores. A partir dos resultados apresentados constatou-se que o gerbilo é um modelo animal para estudos de carcinogênese química prostática, que somados aos dados da literatura, levarão ao melhor entendimento da biologia de lesões da próstata / Abstract: The prostate cancer is the most common tumor that attacks men from the 50's decade. Among the risk factors that contribute to this disease's increase stand out the aging process, when hormonal unbalances happen frequendy. Developmental studies of prostatic tumors have been complicated because of difficult in obtaining human material, once this question lies on medical ethics. Researchers group have had tried to develop, characterize and validate some rodent models to analyze the prostate cancer. Autoctone models used to study spontaneous prostatic lesions have performed important role to these kind of neoplasia. ln addition, lots of innstigations have been done about experimental induction of prostatic tumors in rats, mice and guinea pig. ln a first phase of the present work, it was realized in the ventral lobe of old gerbil's prostate veriones unguiculatllm) structural and ultra-structural morphological, quantitative and functional analyzes of the cellular compounds of epithelial and stromal compartments. The prostatic features of theses animaIs revealed that in a same gland could be noted functional regions containing normal secretof)' epithelium and regions completely altered, showing histopathological lesions. A testosterone concentration decrease associated to these prolifera tive sites confmns the importam role of this androgen to the prostatic homeostasis. Because of these data, in the second phase of this. work, it was done a prostatic tumors experimental induction in the adult gerbil, after conjugate treatment of N-metil-N-nitrosourea with testosterone propionate. Once the tumors were established, prostate fragments were processed for histological, immunocytocl)emical and ultra-structural studies. The results showed that in gerbils, the emergence of prostatic lesions happened in experimental periods of until 9 months and both the carcinogen and testosterone, associated or not, was able to induce these adenocarcinomas. The invase potencial of anomalus cells could be proved by ultra-structural analyz~s and by the immunocytochemical test for laminima, noted in the basal membrane disruption. On the other hand, it was observed that gerbil is one of the pioneers in expression of the citoplasmatic protein Alfa-lvletilacilCoARacemase (P504S), which is found in the neoplastic cells, likely in prostatic human cancer. Concluding, this study is pioneer in the demonstration of the P504S expression in rodents. Based on these presented data, it was verified that gerbil is a good experimental model to chemical carcinogenesis research of the prostate, which taken together with previous literature will give a better and profounder understanding of prostatic lesions / Doutorado / Biologia Celular / Doutor em Biologia Celular e Estrutural