Studies on the function of heparan sulfate in cElegans germ line

Studies on the function of heparan sulfate in cElegans germ line

Author Cunha, Dayse Caroline Severiano da Autor UNIFESP Google Scholar
Advisor Nader, Helena Bonciani Autor UNIFESP Google Scholar
Institution Universidade Federal de São Paulo (UNIFESP)
Graduate program Ciências Biológicas (Biologia Molecular)
Abstract Germline development is a process common to all multicellular organisms and requires tight control of proliferation and stem cell maintenance. The Caenorhabditis elegans hermaphrodite germline was used in this thesis as a model to study stem cells and cellular proliferation, as well as germ cell fate specification and maintenance. In C. elegans, each germ cell is surrounded by a thickened extracellular matrix, which is comprised of a variety of macromolecules including highly modified heparan sulfate (HS). The HS chains are covalently linked to a core protein to form proteoglycans (PGs). HS chains are modified by heparan sulfate modifying enzymes (HSME). Single orthologs of all known HS-modifying enzymes are found in the C. elegans genome: hst-1/NDST, hst-2/HS2st, hse-5/GLCE, hst- 6/HS6st and two for 3-O-sulfotransferase, hst-3.1 and hst-3.2. Heparan sulfates display peculiar structural variability, which play specific roles in a diversity of cellular interactions. Yet, the understanding of the function of HS in stem cell biology in vivo remains poorly understood. In this thesis, genetic analyses of hermaphrodite germline showed that heparan sulfate is required for germline proliferation. Modifications in the HS chain by hst-2/HS2st restricts germ cell proliferation activity whereas other modifications such as, hst-1/NDST can promote mitotic activity of germ cells. On the other hand, some proteoglycans (SDN-1/syndecan, LON- 2/glypican and UNC-52/perlecan), are required for germ cell proliferation and to prevent premature entry into meiosis. Thus, these genes could be required for stem cell maintenance. However, unc-52/perlecan mutants display an increase in stem cell corpses, which may affect germline proliferation. Moreover, it was found that distal tip cell (DTC) migration defects do not correlate with changes in germline proliferation. Interestingly, no changes occur during cell cycle progression in young adult mutants of sdn-1/syndecan, lon-2/glypican and hst-2/HS2st. In addition, RNAi has the potential to shed light on the function of a gene in distinct tissues. In this thesis, it is shown that sdn-1/syndecan and lon-2/glypican are acting in the somatic gonad that surrounds the germ cells, whereas hst-2/HS2st and hst-6/HS6st act in the germline, indicating complex interactions between the extracellular matrix of the germline and the somatic gonad during normal germ cell development. In addition, it was found that crucial signaling pathways, such as GLP-1/Notch, DAF-2/IIR and DAF-7/TGF-? are genetically epistatic to hst-2/HS2ST during germline proliferation. Taken together, the results demonstrate that modification patterns in the HS chain are required for proper homeostasis of germline stem cells in the C. elegans germline.

O desenvolvimento da linhagem germinativa é um processo comum para todos os organismos multicelulares e requer controle rígido da proliferação e manutenção das células germinativas. Na presente tese, foi utilizada a linhagem germinativa de Caenorhabditis elegans (hermafrodita) como modelo para o estudo de células-tronco, proliferação celular, diferenciação e manutenção celular. Em C. elegans, cada célula germinativa está envolvida por uma matriz extracelular espessa composta por uma variedade de macromoléculas, incluindo heparam sulfato (HS). A cadeia de heparam sulfato ocorre nos tecidos como proteoglicanos (PGs), ligada de forma covalente a um "core" protéico. A cadeia de HS é sintetizada por uma série de enzimas incluindo as enzimas modificadoras da cadeia de heparam sulfato (EMHS). Genes ortólogos de todas as enzimas modificadoras de HS conhecidas são encontradas em C. elegans: hst-1/NDST (N-sulfotransferase), hst-2/HS2st (2-O- sulfotransferase), hse-5/GLCE (glucuronosil epimerase), hst-6/HS6st (6-O- sulfotransferase), e hst- 3.1 e hst-3.2 (3-O-sulfotransferases). Heparam sulfato exibe variabilidade estrutural peculiar e desempenha papéis específicos em uma diversidade de interações celulares. No entanto, a compreensão sobre a função do heparam sulfato na biologia de células-tronco in vivo é ainda pouco conhecida. Nesta tese, a análise genética da proliferação de células germinativas em hermafroditas mostrou que o heparam sulfato é necessário para este evento. Modificações na cadeia de heparam sulfato por hst-2/HS2st restringem a atividade de proliferação de células germinativas enquanto que outras modificações, como, N- sulfatação pela hst-1/NDST pode promover tal atividade. Por outro lado, alguns proteoglicanos (SDN-1/sindecam, LON-2/glipicam e UNC-52/perlecam) são necessários para a proliferação de células germinativas e podem, também, impedir a entrada prematura em meiose. Assim, estes genes parecem ser necessários para a manutenção destas células. No entanto, os mutantes unc-52/perlecam exibem aumento na expressão de CED-1
Keywords heparan sulfate
elegans
germ line
stm cell poll
proliferation
heparan sulfate
elegans
germ line
stm cell poll
proliferation
Language English
Date 2016-05-31
Published in CUNHA, Dayse Caroline Severiano da. Studies on the function of heparan sulfate in cElegans germ line. 2016. 115 f. Tese (Doutorado) - Escola Paulista de Medicina, Universidade Federal de São Paulo (UNIFESP), São Paulo, 2016.
Research area Bioquímica
Knowledge area Ciências biológicas
Publisher Universidade Federal de São Paulo (UNIFESP)
Extent 115 p.
Origin https://sucupira.capes.gov.br/sucupira/public/consultas/coleta/trabalhoConclusao/viewTrabalhoConclusao.jsf?popup=true&id_trabalho=3638029
Access rights Closed access
Type Thesis
URI http://repositorio.unifesp.br/handle/11600/46956

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