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MOTS-c is a 16-amino acid mitochondrial derivative peptide reported to be involved in regulating insulin and metabolic homeostasis via the AMP activated protein kinase (AMPK). AMPK agonist AICAR has been reported to improve cognition. Previous reports also pointed out that MOTS-c may be effective as a therapeutic option toward the prevention of the aging processes. Therefore, we investigated the roles of MOTS-c in the memory recognition process. The results showed that central MOTS-c not only enhanced object and location recognition memory formation and consolidation but also ameliorated the memory deficit induced by Aβ1-42 or LPS. The memory-ameliorating effects of MOTS-c could be blocked by AMPK inhibitor dorsomorphin. Moreover, MOTS-c treatment significantly increased the phosphorylation of AMPK but not ERK, JNK, and p38 in the hippocampus. The underlying mechanism of MOTS-c neuroprotection may involve inhibiting the activation of astrocytes and microglia and production of proinflammatory cytokines. In addition, we found that peripheral administration of MOTS-c does not cross the blood-brain barrier (BBB) and plays an effect. In order to improve the brain intake of MOTS-c, we screen out (PRR)5, a cell penetrating peptides, as a carrier for MOTS-c into the brain. Then in the NOR task, intranasal or intravenous MP (cell-penetrating MOTS-c analogue) showed good memory performance on memory formation, memory consolidation, and memory impairment. Near-infrared fluorescent experiments showed the real-time biodistribution in brain after intranasal or intravenous infusion of MP. These results suggested that MOTS-c might be a new potential target for treatment of cognitive decline in AD.

The adaptation of eukaryotic cells to anaerobic conditions is reflected by substantial changes to mitochondrial metabolism and functional reduction. Hydrogenosomes belong among the most modified mitochondrial derivative and generate molecular hydrogen concomitant with ATP synthesis. The reduction of mitochondria is frequently associated with loss of peroxisomes, which compartmentalize pathways that generate reactive oxygen species (ROS) and thus protect against cellular damage. The biogenesis and function of peroxisomes are tightly coupled with mitochondria. These organelles share fission machinery components, oxidative metabolism pathways, ROS scavenging activities, and some metabolites. The loss of peroxisomes in eukaryotes with reduced mitochondria is thus not unexpected. Surprisingly, we identified peroxisomes in the anaerobic, hydrogenosome-bearing protist Mastigamoeba balamuthi . We found a conserved set of peroxin (Pex) proteins that are required for protein import, peroxisomal growth, and division. Key membrane-associated Pexs ( Mb Pex3, Mb Pex11, and Mb Pex14) were visualized in numerous vesicles distinct from hydrogenosomes, the endoplasmic reticulum (ER), and Golgi complex. Proteomic analysis of cellular fractions and prediction of peroxisomal targeting signals (PTS1/PTS2) identified 51 putative peroxisomal matrix proteins. Expression of selected proteins in Saccharomyces cerevisiae revealed specific targeting to peroxisomes. The matrix proteins identified included components of acyl-CoA and carbohydrate metabolism and pyrimidine and CoA biosynthesis, whereas no components related to either β-oxidation or catalase were present. In conclusion, we identified a subclass of peroxisomes, named “anaerobic” peroxisomes that shift the current paradigm and turn attention to the reductive evolution of peroxisomes in anaerobic organisms.

Submitted by Emi Rosane Silistino de Souza null (emi.rosane@hotmail.com) on 2020-04-27T22:06:11Z No. of bitstreams: 1 organized.pdf: 8345245 bytes, checksum: c22516224a75bd365f618cc6dfee3fb5 (MD5) Submitted by Emi Rosane Silistino de Souza null (emi.rosane@hotmail.com) on 2020-04-27T22:06:11Z No. of bitstreams: 1 organized.pdf: 8345245 bytes, checksum: c22516224a75bd365f618cc6dfee3fb5 (MD5) Rejected by Elza Mitiko Sato null (elzasato@ibilce.unesp.br), reason: Solicitamos que realize correções na submissão seguindo as orientações abaixo: 01 – No rodapé da CAPA e FOLHA DE ROSTO deve constar, cidade e ano (sem o , SP): São José do Rio Preto 2020 02 – Na ficha catalográfica solicitamos diminuir o tamanho da fonte (8) para não ultrapassar a dimensão da ficha. 03 – No rodapé da FOLHA DE APROVAÇÃO deve ser colocado a data efetiva da defesa: São José do Rio Preto 28 de fevereiro de 2020 04 - Nos agradecimentos: segundo a Portaria CAPES nº 206, de 4 de setembro de 2018, todos os trabalhos que tiveram financiamento/BOLSA CAPES deve constar a expressão EXATA: "O presente trabalho foi realizado com apoio da Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - Brasil (CAPES) - Código de Financiamento 001". 05 - Segundo a norma ABNT NBR 10520, a epígrafe deve ser referenciada. 06 - Solicitamos adequar o Sumário conforme as normas ABNT NBR 6027 e 6024. 07 - A paginação deve ser seqüencial, iniciando a contagem na folha de rosto e mostrando o número a partir da introdução, a ficha catalográfica ficará após a folha de rosto e não deverá ser contada. 08 – No SUMÁRIO e no texto, a seção REFERÊNCIAS não é numerada, e o correto é REFERÊNCIAS e não REFERÊNCIAS BIBLIOGRÁFICAS. 09 - Nas ilustrações/figuras a norma orienta que devem ter a descrição da fonte abaixo delas, mesmo que seja produção do autor. Agradecemos a compreensão on 2020-05-01T01:26:11Z (GMT) Submitted by Emi Rosane Silistino de Souza null (emi.rosane@hotmail.com) on 2020-04-27T22:06:11Z No. of bitstreams: 1 organized.pdf: 8345245 bytes, checksum: c22516224a75bd365f618cc6dfee3fb5 (MD5) Submitted by Emi Rosane Silistino de Souza null (emi.rosane@hotmail.com) on 2020-04-27T22:06:11Z No. of bitstreams: 1 organized.pdf: 8345245 bytes, checksum: c22516224a75bd365f618cc6dfee3fb5 (MD5) Submitted by Emi Rosane Silistino de Souza null (emi.rosane@hotmail.com) on 2020-05-01T21:50:58Z No. of bitstreams: 1 TESE_FINAL.pdf: 8322383 bytes, checksum: a8bfc5caf409777fe67c486bd1447eaf (MD5) Submitted by Emi Rosane Silistino de Souza null (emi.rosane@hotmail.com) on 2020-05-01T21:50:58Z No. of bitstreams: 1 TESE_FINAL.pdf: 8322383 bytes, checksum: a8bfc5caf409777fe67c486bd1447eaf (MD5) Submitted by Emi Rosane Silistino de Souza null (emi.rosane@hotmail.com) on 2020-04-27T22:06:11Z No. of bitstreams: 1 organized.pdf: 8345245 bytes, checksum: c22516224a75bd365f618cc6dfee3fb5 (MD5) Rejected by Elza Mitiko Sato null (elzasato@ibilce.unesp.br), reason: Solicitamos que realize correções na submissão seguindo as orientações abaixo: 01 – No rodapé da CAPA e FOLHA DE ROSTO deve constar, cidade e ano (sem o , SP): São José do Rio Preto 2020 02 – Na ficha catalográfica solicitamos diminuir o tamanho da fonte (8) para não ultrapassar a dimensão da ficha. 03 – No rodapé da FOLHA DE APROVAÇÃO deve ser colocado a data efetiva da defesa: São José do Rio Preto 28 de fevereiro de 2020 04 - Nos agradecimentos: segundo a Portaria CAPES nº 206, de 4 de setembro de 2018, todos os trabalhos que tiveram financiamento/BOLSA CAPES deve constar a expressão EXATA: "O presente trabalho foi realizado com apoio da Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - Brasil (CAPES) - Código de Financiamento 001". 05 - Segundo a norma ABNT NBR 10520, a epígrafe deve ser referenciada. 06 - Solicitamos adequar o Sumário conforme as normas ABNT NBR 6027 e 6024. 07 - A paginação deve ser seqüencial, iniciando a contagem na folha de rosto e mostrando o número a partir da introdução, a ficha catalográfica ficará após a folha de rosto e não deverá ser contada. 08 – No SUMÁRIO e no texto, a seção REFERÊNCIAS não é numerada, e o correto é REFERÊNCIAS e não REFERÊNCIAS BIBLIOGRÁFICAS. 09 - Nas ilustrações/figuras a norma orienta que devem ter a descrição da fonte abaixo delas, mesmo que seja produção do autor. Agradecemos a compreensão on 2020-05-01T01:26:11Z (GMT) Rejected by Elza Mitiko Sato null (elzasato@ibilce.unesp.br), reason: Solicitamos que realize correções na submissão seguindo as orientações abaixo: 01 – No rodapé da CAPA e FOLHA DE ROSTO deve constar, cidade e ano (sem o , SP): São José do Rio Preto 2020 02 – Na ficha catalográfica solicitamos diminuir o tamanho da fonte (8) para não ultrapassar a dimensão da ficha. 03 – No rodapé da FOLHA DE APROVAÇÃO deve ser colocado a data efetiva da defesa: São José do Rio Preto 28 de fevereiro de 2020 04 - Nos agradecimentos: segundo a Portaria CAPES nº 206, de 4 de setembro de 2018, todos os trabalhos que tiveram financiamento/BOLSA CAPES deve constar a expressão EXATA: "O presente trabalho foi realizado com apoio da Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - Brasil (CAPES) - Código de Financiamento 001". 05 - Segundo a norma ABNT NBR 10520, a epígrafe deve ser referenciada. 06 - Solicitamos adequar o Sumário conforme as normas ABNT NBR 6027 e 6024. 07 - A paginação deve ser seqüencial, iniciando a contagem na folha de rosto e mostrando o número a partir da introdução, a ficha catalográfica ficará após a folha de rosto e não deverá ser contada. 08 – No SUMÁRIO e no texto, a seção REFERÊNCIAS não é numerada, e o correto é REFERÊNCIAS e não REFERÊNCIAS BIBLIOGRÁFICAS. 09 - Nas ilustrações/figuras a norma orienta que devem ter a descrição da fonte abaixo delas, mesmo que seja produção do autor. Agradecemos a compreensão on 2020-05-01T01:26:11Z (GMT) Rejected by Elza Mitiko Sato null (elzasato@ibilce.unesp.br), reason: Solicitamos que realize correções na submissão seguindo as orientações abaixo: 01 - No SUMÁRIO a paginação não confere. Ex.: A Introdução no texto está na página 11 e no sumário pag. 12; a seção 1. 2 no texto está na página 14 e no sumário pag. 15. Solicitamos que faça uma revisão nas indicações. Lembramos que o arquivo depositado no Repositório deve ser igual ao impresso, o rigor com o padrão da Universidade se deve ao fato de que o seu trabalho passará a ser visível mundialmente. Agradecemos a compreensão. on 2020-05-05T20:17:48Z (GMT) Submitted by Emi Rosane Silistino de Souza null (emi.rosane@hotmail.com) on 2020-05-01T21:50:58Z No. of bitstreams: 1 TESE_FINAL.pdf: 8322383 bytes, checksum: a8bfc5caf409777fe67c486bd1447eaf (MD5) Submitted by Emi Rosane Silistino de Souza null (emi.rosane@hotmail.com) on 2020-05-06T18:32:38Z No. of bitstreams: 1 TESE_FINAL.pdf: 8296215 bytes, checksum: c098c1e8fb47b1ea91d561023e8371b1 (MD5) Submitted by Emi Rosane Silistino de Souza null (emi.rosane@hotmail.com) on 2020-05-01T21:50:58Z No. of bitstreams: 1 TESE_FINAL.pdf: 8322383 bytes, checksum: a8bfc5caf409777fe67c486bd1447eaf (MD5) Submitted by Emi Rosane Silistino de Souza null (emi.rosane@hotmail.com) on 2020-05-06T18:32:38Z No. of bitstreams: 1 TESE_FINAL.pdf: 8296215 bytes, checksum: c098c1e8fb47b1ea91d561023e8371b1 (MD5) Submitted by Emi Rosane Silistino de Souza null (emi.rosane@hotmail.com) on 2020-04-27T22:06:11Z No. of bitstreams: 1 organized.pdf: 8345245 bytes, checksum: c22516224a75bd365f618cc6dfee3fb5 (MD5) Submitted by Emi Rosane Silistino de Souza null (emi.rosane@hotmail.com) on 2020-05-01T21:50:58Z No. of bitstreams: 1 TESE_FINAL.pdf: 8322383 bytes, checksum: a8bfc5caf409777fe67c486bd1447eaf (MD5) Rejected by Elza Mitiko Sato null (elzasato@ibilce.unesp.br), reason: Solicitamos que realize correções na submissão seguindo as orientações abaixo: 01 - No SUMÁRIO a paginação não confere. Ex.: A Introdução no texto está na página 11 e no sumário pag. 12; a seção 1. 2 no texto está na página 14 e no sumário pag. 15. Solicitamos que faça uma revisão nas indicações. Lembramos que o arquivo depositado no Repositório deve ser igual ao impresso, o rigor com o padrão da Universidade se deve ao fato de que o seu trabalho passará a ser visível mundialmente. Agradecemos a compreensão. on 2020-05-05T20:17:48Z (GMT) Approved for entry into archive by Elza Mitiko Sato null (elzasato@ibilce.unesp.br) on 2020-05-08T00:42:36Z (GMT) No. of bitstreams: 1 souza_ers_dr_sjrp.pdf: 8296215 bytes, checksum: c098c1e8fb47b1ea91d561023e8371b1 (MD5) Submitted by Emi Rosane Silistino de Souza null (emi.rosane@hotmail.com) on 2020-05-01T21:50:58Z No. of bitstreams: 1 TESE_FINAL.pdf: 8322383 bytes, checksum: a8bfc5caf409777fe67c486bd1447eaf (MD5) Rejected by Elza Mitiko Sato null (elzasato@ibilce.unesp.br), reason: Solicitamos que realize correções na submissão seguindo as orientações abaixo: 01 – No rodapé da CAPA e FOLHA DE ROSTO deve constar, cidade e ano (sem o , SP): São José do Rio Preto 2020 02 – Na ficha catalográfica solicitamos diminuir o tamanho da fonte (8) para não ultrapassar a dimensão da ficha. 03 – No rodapé da FOLHA DE APROVAÇÃO deve ser colocado a data efetiva da defesa: São José do Rio Preto 28 de fevereiro de 2020 04 - Nos agradecimentos: segundo a Portaria CAPES nº 206, de 4 de setembro de 2018, todos os trabalhos que tiveram financiamento/BOLSA CAPES deve constar a expressão EXATA: "O presente trabalho foi realizado com apoio da Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - Brasil (CAPES) - Código de Financiamento 001". 05 - Segundo a norma ABNT NBR 10520, a epígrafe deve ser referenciada. 06 - Solicitamos adequar o Sumário conforme as normas ABNT NBR 6027 e 6024. 07 - A paginação deve ser seqüencial, iniciando a contagem na folha de rosto e mostrando o número a partir da introdução, a ficha catalográfica ficará após a folha de rosto e não deverá ser contada. 08 – No SUMÁRIO e no texto, a seção REFERÊNCIAS não é numerada, e o correto é REFERÊNCIAS e não REFERÊNCIAS BIBLIOGRÁFICAS. 09 - Nas ilustrações/figuras a norma orienta que devem ter a descrição da fonte abaixo delas, mesmo que seja produção do autor. Agradecemos a compreensão on 2020-05-01T01:26:11Z (GMT) Approved for entry into archive by Elza Mitiko Sato null (elzasato@ibilce.unesp.br) on 2020-05-08T00:42:36Z (GMT) No. of bitstreams: 1 souza_ers_dr_sjrp.pdf: 8296215 bytes, checksum: c098c1e8fb47b1ea91d561023e8371b1 (MD5) Submitted by Emi Rosane Silistino de Souza null (emi.rosane@hotmail.com) on 2020-05-06T18:32:38Z No. of bitstreams: 1 TESE_FINAL.pdf: 8296215 bytes, checksum: c098c1e8fb47b1ea91d561023e8371b1 (MD5) Made available in DSpace on 2020-05-08T00:42:36Z (GMT). No. of bitstreams: 1 souza_ers_dr_sjrp.pdf: 8296215 bytes, checksum: c098c1e8fb47b1ea91d561023e8371b1 (MD5) Previous issue date: 2020-02-28 Made available in DSpace on 2020-05-08T00:42:36Z (GMT). No. of bitstreams: 1 souza_ers_dr_sjrp.pdf: 8296215 bytes, checksum: c098c1e8fb47b1ea91d561023e8371b1 (MD5) Previous issue date: 2020-02-28 Submitted by Emi Rosane Silistino de Souza null (emi.rosane@hotmail.com) on 2020-05-01T21:50:58Z No. of bitstreams: 1 TESE_FINAL.pdf: 8322383 bytes, checksum: a8bfc5caf409777fe67c486bd1447eaf (MD5) Rejected by Elza Mitiko Sato null (elzasato@ibilce.unesp.br), reason: Solicitamos que realize correções na submissão seguindo as orientações abaixo: 01 – No rodapé da CAPA e FOLHA DE ROSTO deve constar, cidade e ano (sem o , SP): São José do Rio Preto 2020 02 – Na ficha catalográfica solicitamos diminuir o tamanho da fonte (8) para não ultrapassar a dimensão da ficha. 03 – No rodapé da FOLHA DE APROVAÇÃO deve ser colocado a data efetiva da defesa: São José do Rio Preto 28 de fevereiro de 2020 04 - Nos agradecimentos: segundo a Portaria CAPES nº 206, de 4 de setembro de 2018, todos os trabalhos que tiveram financiamento/BOLSA CAPES deve constar a expressão EXATA: "O presente trabalho foi realizado com apoio da Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - Brasil (CAPES) - Código de Financiamento 001". 05 - Segundo a norma ABNT NBR 10520, a epígrafe deve ser referenciada. 06 - Solicitamos adequar o Sumário conforme as normas ABNT NBR 6027 e 6024. 07 - A paginação deve ser seqüencial, iniciando a contagem na folha de rosto e mostrando o número a partir da introdução, a ficha catalográfica ficará após a folha de rosto e não deverá ser contada. 08 – No SUMÁRIO e no texto, a seção REFERÊNCIAS não é numerada, e o correto é REFERÊNCIAS e não REFERÊNCIAS BIBLIOGRÁFICAS. 09 - Nas ilustrações/figuras a norma orienta que devem ter a descrição da fonte abaixo delas, mesmo que seja produção do autor. Agradecemos a compreensão on 2020-05-01T01:26:11Z (GMT) Submitted by Emi Rosane Silistino de Souza null (emi.rosane@hotmail.com) on 2020-05-06T18:32:38Z No. of bitstreams: 1 TESE_FINAL.pdf: 8296215 bytes, checksum: c098c1e8fb47b1ea91d561023e8371b1 (MD5) Submitted by Emi Rosane Silistino de Souza null (emi.rosane@hotmail.com) on 2020-05-01T21:50:58Z No. of bitstreams: 1 TESE_FINAL.pdf: 8322383 bytes, checksum: a8bfc5caf409777fe67c486bd1447eaf (MD5) Rejected by Elza Mitiko Sato null (elzasato@ibilce.unesp.br), reason: Solicitamos que realize correções na submissão seguindo as orientações abaixo: 01 - No SUMÁRIO a paginação não confere. Ex.: A Introdução no texto está na página 11 e no sumário pag. 12; a seção 1. 2 no texto está na página 14 e no sumário pag. 15. Solicitamos que faça uma revisão nas indicações. Lembramos que o arquivo depositado no Repositório deve ser igual ao impresso, o rigor com o padrão da Universidade se deve ao fato de que o seu trabalho passará a ser visível mundialmente. Agradecemos a compreensão. on 2020-05-05T20:17:48Z (GMT) Approved for entry into archive by Elza Mitiko Sato null (elzasato@ibilce.unesp.br) on 2020-05-08T00:42:36Z (GMT) No. of bitstreams: 1 souza_ers_dr_sjrp.pdf: 8296215 bytes, checksum: c098c1e8fb47b1ea91d561023e8371b1 (MD5) Submitted by Emi Rosane Silistino de Souza null (emi.rosane@hotmail.com) on 2020-04-27T22:06:11Z No. of bitstreams: 1 organized.pdf: 8345245 bytes, checksum: c22516224a75bd365f618cc6dfee3fb5 (MD5) Submitted by Emi Rosane Silistino de Souza null (emi.rosane@hotmail.com) on 2020-05-01T21:50:58Z No. of bitstreams: 1 TESE_FINAL.pdf: 8322383 bytes, checksum: a8bfc5caf409777fe67c486bd1447eaf (MD5) Submitted by Emi Rosane Silistino de Souza null (emi.rosane@hotmail.com) on 2020-05-06T18:32:38Z No. of bitstreams: 1 TESE_FINAL.pdf: 8296215 bytes, checksum: c098c1e8fb47b1ea91d561023e8371b1 (MD5) Submitted by Emi Rosane Silistino de Souza null (emi.rosane@hotmail.com) on 2020-05-01T21:50:58Z No. of bitstreams: 1 TESE_FINAL.pdf: 8322383 bytes, checksum: a8bfc5caf409777fe67c486bd1447eaf (MD5) Approved for entry into archive by Elza Mitiko Sato null (elzasato@ibilce.unesp.br) on 2020-05-08T00:42:36Z (GMT) No. of bitstreams: 1 souza_ers_dr_sjrp.pdf: 8296215 bytes, checksum: c098c1e8fb47b1ea91d561023e8371b1 (MD5) Submitted by Emi Rosane Silistino de Souza null (emi.rosane@hotmail.com) on 2020-05-01T21:50:58Z No. of bitstreams: 1 TESE_FINAL.pdf: 8322383 bytes, checksum: a8bfc5caf409777fe67c486bd1447eaf (MD5) Made available in DSpace on 2020-05-08T00:42:36Z (GMT). No. of bitstreams: 1 souza_ers_dr_sjrp.pdf: 8296215 bytes, checksum: c098c1e8fb47b1ea91d561023e8371b1 (MD5) Previous issue date: 2020-02-28 Rejected by Elza Mitiko Sato null (elzasato@ibilce.unesp.br), reason: Solicitamos que realize correções na submissão seguindo as orientações abaixo: 01 - No SUMÁRIO a paginação não confere. Ex.: A Introdução no texto está na página 11 e no sumário pag. 12; a seção 1. 2 no texto está na página 14 e no sumário pag. 15. Solicitamos que faça uma revisão nas indicações. Lembramos que o arquivo depositado no Repositório deve ser igual ao impresso, o rigor com o padrão da Universidade se deve ao fato de que o seu trabalho passará a ser visível mundialmente. Agradecemos a compreensão. on 2020-05-05T20:17:48Z (GMT) Rejected by Elza Mitiko Sato null (elzasato@ibilce.unesp.br), reason: Solicitamos que realize correções na submissão seguindo as orientações abaixo: 01 – No rodapé da CAPA e FOLHA DE ROSTO deve constar, cidade e ano (sem o , SP): São José do Rio Preto 2020 02 – Na ficha catalográfica solicitamos diminuir o tamanho da fonte (8) para não ultrapassar a dimensão da ficha. 03 – No rodapé da FOLHA DE APROVAÇÃO deve ser colocado a data efetiva da defesa: São José do Rio Preto 28 de fevereiro de 2020 04 - Nos agradecimentos: segundo a Portaria CAPES nº 206, de 4 de setembro de 2018, todos os trabalhos que tiveram financiamento/BOLSA CAPES deve constar a expressão EXATA: "O presente trabalho foi realizado com apoio da Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - Brasil (CAPES) - Código de Financiamento 001". 05 - Segundo a norma ABNT NBR 10520, a epígrafe deve ser referenciada. 06 - Solicitamos adequar o Sumário conforme as normas ABNT NBR 6027 e 6024. 07 - A paginação deve ser seqüencial, iniciando a contagem na folha de rosto e mostrando o número a partir da introdução, a ficha catalográfica ficará após a folha de rosto e não deverá ser contada. 08 – No SUMÁRIO e no texto, a seção REFERÊNCIAS não é numerada, e o correto é REFERÊNCIAS e não REFERÊNCIAS BIBLIOGRÁFICAS. 09 - Nas ilustrações/figuras a norma orienta que devem ter a descrição da fonte abaixo delas, mesmo que seja produção do autor. Agradecemos a compreensão on 2020-05-01T01:26:11Z (GMT) Submitted by Emi Rosane Silistino de Souza null (emi.rosane@hotmail.com) on 2020-05-06T18:32:38Z No. of bitstreams: 1 TESE_FINAL.pdf: 8296215 bytes, checksum: c098c1e8fb47b1ea91d561023e8371b1 (MD5) Made available in DSpace on 2020-05-08T00:42:36Z (GMT). No. of bitstreams: 1 souza_ers_dr_sjrp.pdf: 8296215 bytes, checksum: c098c1e8fb47b1ea91d561023e8371b1 (MD5) Previous issue date: 2020-02-28 Submitted by Emi Rosane Silistino de Souza null (emi.rosane@hotmail.com) on 2020-05-01T21:50:58Z No. of bitstreams: 1 TESE_FINAL.pdf: 8322383 bytes, checksum: a8bfc5caf409777fe67c486bd1447eaf (MD5) Rejected by Elza Mitiko Sato null (elzasato@ibilce.unesp.br), reason: Solicitamos que realize correções na submissão seguindo as orientações abaixo: 01 - No SUMÁRIO a paginação não confere. Ex.: A Introdução no texto está na página 11 e no sumário pag. 12; a seção 1. 2 no texto está na página 14 e no sumário pag. 15. Solicitamos que faça uma revisão nas indicações. Lembramos que o arquivo depositado no Repositório deve ser igual ao impresso, o rigor com o padrão da Universidade se deve ao fato de que o seu trabalho passará a ser visível mundialmente. Agradecemos a compreensão. on 2020-05-05T20:17:48Z (GMT) Submitted by Emi Rosane Silistino de Souza null (emi.rosane@hotmail.com) on 2020-05-06T18:32:38Z No. of bitstreams: 1 TESE_FINAL.pdf: 8296215 bytes, checksum: c098c1e8fb47b1ea91d561023e8371b1 (MD5) Approved for entry into archive by Elza Mitiko Sato null (elzasato@ibilce.unesp.br) on 2020-05-08T00:42:36Z (GMT) No. of bitstreams: 1 souza_ers_dr_sjrp.pdf: 8296215 bytes, checksum: c098c1e8fb47b1ea91d561023e8371b1 (MD5) Approved for entry into archive by Elza Mitiko Sato null (elzasato@ibilce.unesp.br) on 2020-05-08T00:42:36Z (GMT) No. of bitstreams: 1 souza_ers_dr_sjrp.pdf: 8296215 bytes, checksum: c098c1e8fb47b1ea91d561023e8371b1 (MD5) Submitted by Emi Rosane Silistino de Souza null (emi.rosane@hotmail.com) on 2020-05-06T18:32:38Z No. of bitstreams: 1 TESE_FINAL.pdf: 8296215 bytes, checksum: c098c1e8fb47b1ea91d561023e8371b1 (MD5) Submitted by Emi Rosane Silistino de Souza null (emi.rosane@hotmail.com) on 2020-05-01T21:50:58Z No. of bitstreams: 1 TESE_FINAL.pdf: 8322383 bytes, checksum: a8bfc5caf409777fe67c486bd1447eaf (MD5) Made available in DSpace on 2020-05-08T00:42:36Z (GMT). No. of bitstreams: 1 souza_ers_dr_sjrp.pdf: 8296215 bytes, checksum: c098c1e8fb47b1ea91d561023e8371b1 (MD5) Previous issue date: 2020-02-28 Made available in DSpace on 2020-05-08T00:42:36Z (GMT). No. of bitstreams: 1 souza_ers_dr_sjrp.pdf: 8296215 bytes, checksum: c098c1e8fb47b1ea91d561023e8371b1 (MD5) Previous issue date: 2020-02-28 Approved for entry into archive by Elza Mitiko Sato null (elzasato@ibilce.unesp.br) on 2020-05-08T00:42:36Z (GMT) No. of bitstreams: 1 souza_ers_dr_sjrp.pdf: 8296215 bytes, checksum: c098c1e8fb47b1ea91d561023e8371b1 (MD5) Submitted by Emi Rosane Silistino de Souza null (emi.rosane@hotmail.com) on 2020-05-06T18:32:38Z No. of bitstreams: 1 TESE_FINAL.pdf: 8296215 bytes, checksum: c098c1e8fb47b1ea91d561023e8371b1 (MD5) Rejected by Elza Mitiko Sato null (elzasato@ibilce.unesp.br), reason: Solicitamos que realize correções na submissão seguindo as orientações abaixo: 01 – No rodapé da CAPA e FOLHA DE ROSTO deve constar, cidade e ano (sem o , SP): São José do Rio Preto 2020 02 – Na ficha catalográfica solicitamos diminuir o tamanho da fonte (8) para não ultrapassar a dimensão da ficha. 03 – No rodapé da FOLHA DE APROVAÇÃO deve ser colocado a data efetiva da defesa: São José do Rio Preto 28 de fevereiro de 2020 04 - Nos agradecimentos: segundo a Portaria CAPES nº 206, de 4 de setembro de 2018, todos os trabalhos que tiveram financiamento/BOLSA CAPES deve constar a expressão EXATA: "O presente trabalho foi realizado com apoio da Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - Brasil (CAPES) - Código de Financiamento 001". 05 - Segundo a norma ABNT NBR 10520, a epígrafe deve ser referenciada. 06 - Solicitamos adequar o Sumário conforme as normas ABNT NBR 6027 e 6024. 07 - A paginação deve ser seqüencial, iniciando a contagem na folha de rosto e mostrando o número a partir da introdução, a ficha catalográfica ficará após a folha de rosto e não deverá ser contada. 08 – No SUMÁRIO e no texto, a seção REFERÊNCIAS não é numerada, e o correto é REFERÊNCIAS e não REFERÊNCIAS BIBLIOGRÁFICAS. 09 - Nas ilustrações/figuras a norma orienta que devem ter a descrição da fonte abaixo delas, mesmo que seja produção do autor. Agradecemos a compreensão on 2020-05-01T01:26:11Z (GMT) Submitted by Emi Rosane Silistino de Souza null (emi.rosane@hotmail.com) on 2020-05-06T18:32:38Z No. of bitstreams: 1 TESE_FINAL.pdf: 8296215 bytes, checksum: c098c1e8fb47b1ea91d561023e8371b1 (MD5) Rejected by Elza Mitiko Sato null (elzasato@ibilce.unesp.br), reason: Solicitamos que realize correções na submissão seguindo as orientações abaixo: 01 - No SUMÁRIO a paginação não confere. Ex.: A Introdução no texto está na página 11 e no sumário pag. 12; a seção 1. 2 no texto está na página 14 e no sumário pag. 15. Solicitamos que faça uma revisão nas indicações. Lembramos que o arquivo depositado no Repositório deve ser igual ao impresso, o rigor com o padrão da Universidade se deve ao fato de que o seu trabalho passará a ser visível mundialmente. Agradecemos a compreensão. on 2020-05-05T20:17:48Z (GMT) Approved for entry into archive by Elza Mitiko Sato null (elzasato@ibilce.unesp.br) on 2020-05-08T00:42:36Z (GMT) No. of bitstreams: 1 souza_ers_dr_sjrp.pdf: 8296215 bytes, checksum: c098c1e8fb47b1ea91d561023e8371b1 (MD5) Approved for entry into archive by Elza Mitiko Sato null (elzasato@ibilce.unesp.br) on 2020-05-08T00:42:36Z (GMT) No. of bitstreams: 1 souza_ers_dr_sjrp.pdf: 8296215 bytes, checksum: c098c1e8fb47b1ea91d561023e8371b1 (MD5) Made available in DSpace on 2020-05-08T00:42:36Z (GMT). No. of bitstreams: 1 souza_ers_dr_sjrp.pdf: 8296215 bytes, checksum: c098c1e8fb47b1ea91d561023e8371b1 (MD5) Previous issue date: 2020-02-28 Approved for entry into archive by Elza Mitiko Sato null (elzasato@ibilce.unesp.br) on 2020-05-08T00:42:36Z (GMT) No. of bitstreams: 1 souza_ers_dr_sjrp.pdf: 8296215 bytes, checksum: c098c1e8fb47b1ea91d561023e8371b1 (MD5) Submitted by Emi Rosane Silistino de Souza null (emi.rosane@hotmail.com) on 2020-05-01T21:50:58Z No. of bitstreams: 1 TESE_FINAL.pdf: 8322383 bytes, checksum: a8bfc5caf409777fe67c486bd1447eaf (MD5) Made available in DSpace on 2020-05-08T00:42:36Z (GMT). No. of bitstreams: 1 souza_ers_dr_sjrp.pdf: 8296215 bytes, checksum: c098c1e8fb47b1ea91d561023e8371b1 (MD5) Previous issue date: 2020-02-28 Made available in DSpace on 2020-05-08T00:42:36Z (GMT). No. of bitstreams: 1 souza_ers_dr_sjrp.pdf: 8296215 bytes, checksum: c098c1e8fb47b1ea91d561023e8371b1 (MD5) Previous issue date: 2020-02-28 Submitted by Emi Rosane Silistino de Souza null (emi.rosane@hotmail.com) on 2020-05-06T18:32:38Z No. of bitstreams: 1 TESE_FINAL.pdf: 8296215 bytes, checksum: c098c1e8fb47b1ea91d561023e8371b1 (MD5) Made available in DSpace on 2020-05-08T00:42:36Z (GMT). No. of bitstreams: 1 souza_ers_dr_sjrp.pdf: 8296215 bytes, checksum: c098c1e8fb47b1ea91d561023e8371b1 (MD5) Previous issue date: 2020-02-28 Rejected by Elza Mitiko Sato null (elzasato@ibilce.unesp.br), reason: Solicitamos que realize correções na submissão seguindo as orientações abaixo: 01 - No SUMÁRIO a paginação não confere. Ex.: A Introdução no texto está na página 11 e no sumário pag. 12; a seção 1. 2 no texto está na página 14 e no sumário pag. 15. Solicitamos que faça uma revisão nas indicações. Lembramos que o arquivo depositado no Repositório deve ser igual ao impresso, o rigor com o padrão da Universidade se deve ao fato de que o seu trabalho passará a ser visível mundialmente. Agradecemos a compreensão. on 2020-05-05T20:17:48Z (GMT) Submitted by Emi Rosane Silistino de Souza null (emi.rosane@hotmail.com) on 2020-05-06T18:32:38Z No. of bitstreams: 1 TESE_FINAL.pdf: 8296215 bytes, checksum: c098c1e8fb47b1ea91d561023e8371b1 (MD5) Approved for entry into archive by Elza Mitiko Sato null (elzasato@ibilce.unesp.br) on 2020-05-08T00:42:36Z (GMT) No. of bitstreams: 1 souza_ers_dr_sjrp.pdf: 8296215 bytes, checksum: c098c1e8fb47b1ea91d561023e8371b1 (MD5) Submitted by Emi Rosane Silistino de Souza null (emi.rosane@hotmail.com) on 2020-05-01T21:50:58Z No. of bitstreams: 1 TESE_FINAL.pdf: 8322383 bytes, checksum: a8bfc5caf409777fe67c486bd1447eaf (MD5) Approved for entry into archive by Elza Mitiko Sato null (elzasato@ibilce.unesp.br) on 2020-05-08T00:42:36Z (GMT) No. of bitstreams: 1 souza_ers_dr_sjrp.pdf: 8296215 bytes, checksum: c098c1e8fb47b1ea91d561023e8371b1 (MD5) Submitted by Emi Rosane Silistino de Souza null (emi.rosane@hotmail.com) on 2020-05-06T18:32:38Z No. of bitstreams: 1 TESE_FINAL.pdf: 8296215 bytes, checksum: c098c1e8fb47b1ea91d561023e8371b1 (MD5) Submitted by Emi Rosane Silistino de Souza null (emi.rosane@hotmail.com) on 2020-05-01T21:50:58Z No. of bitstreams: 1 TESE_FINAL.pdf: 8322383 bytes, checksum: a8bfc5caf409777fe67c486bd1447eaf (MD5) Made available in DSpace on 2020-05-08T00:42:36Z (GMT). No. of bitstreams: 1 souza_ers_dr_sjrp.pdf: 8296215 bytes, checksum: c098c1e8fb47b1ea91d561023e8371b1 (MD5) Previous issue date: 2020-02-28 Made available in DSpace on 2020-05-08T00:42:36Z (GMT). No. of bitstreams: 1 souza_ers_dr_sjrp.pdf: 8296215 bytes, checksum: c098c1e8fb47b1ea91d561023e8371b1 (MD5) Previous issue date: 2020-02-28 Submitted by Emi Rosane Silistino de Souza null (emi.rosane@hotmail.com) on 2020-05-06T18:32:38Z No. of bitstreams: 1 TESE_FINAL.pdf: 8296215 bytes, checksum: c098c1e8fb47b1ea91d561023e8371b1 (MD5) Rejected by Elza Mitiko Sato null (elzasato@ibilce.unesp.br), reason: Solicitamos que realize correções na submissão seguindo as orientações abaixo: 01 - No SUMÁRIO a paginação não confere. Ex.: A Introdução no texto está na página 11 e no sumário pag. 12; a seção 1. 2 no texto está na página 14 e no sumário pag. 15. Solicitamos que faça uma revisão nas indicações. Lembramos que o arquivo depositado no Repositório deve ser igual ao impresso, o rigor com o padrão da Universidade se deve ao fato de que o seu trabalho passará a ser visível mundialmente. Agradecemos a compreensão. on 2020-05-05T20:17:48Z (GMT) Approved for entry into archive by Elza Mitiko Sato null (elzasato@ibilce.unesp.br) on 2020-05-08T00:42:36Z (GMT) No. of bitstreams: 1 souza_ers_dr_sjrp.pdf: 8296215 bytes, checksum: c098c1e8fb47b1ea91d561023e8371b1 (MD5) Approved for entry into archive by Elza Mitiko Sato null (elzasato@ibilce.unesp.br) on 2020-05-08T00:42:36Z (GMT) No. of bitstreams: 1 souza_ers_dr_sjrp.pdf: 8296215 bytes, checksum: c098c1e8fb47b1ea91d561023e8371b1 (MD5) Made available in DSpace on 2020-05-08T00:42:36Z (GMT). No. of bitstreams: 1 souza_ers_dr_sjrp.pdf: 8296215 bytes, checksum: c098c1e8fb47b1ea91d561023e8371b1 (MD5) Previous issue date: 2020-02-28 Made available in DSpace on 2020-05-08T00:42:36Z (GMT). No. of bitstreams: 1 souza_ers_dr_sjrp.pdf: 8296215 bytes, checksum: c098c1e8fb47b1ea91d561023e8371b1 (MD5) Previous issue date: 2020-02-28 Submitted by Emi Rosane Silistino de Souza null (emi.rosane@hotmail.com) on 2020-05-06T18:32:38Z No. of bitstreams: 1 TESE_FINAL.pdf: 8296215 bytes, checksum: c098c1e8fb47b1ea91d561023e8371b1 (MD5) Submitted by Emi Rosane Silistino de Souza null (emi.rosane@hotmail.com) on 2020-05-06T18:32:38Z No. of bitstreams: 1 TESE_FINAL.pdf: 8296215 bytes, checksum: c098c1e8fb47b1ea91d561023e8371b1 (MD5) Approved for entry into archive by Elza Mitiko Sato null (elzasato@ibilce.unesp.br) on 2020-05-08T00:42:36Z (GMT) No. of bitstreams: 1 souza_ers_dr_sjrp.pdf: 8296215 bytes, checksum: c098c1e8fb47b1ea91d561023e8371b1 (MD5) Approved for entry into archive by Elza Mitiko Sato null (elzasato@ibilce.unesp.br) on 2020-05-08T00:42:36Z (GMT) No. of bitstreams: 1 souza_ers_dr_sjrp.pdf: 8296215 bytes, checksum: c098c1e8fb47b1ea91d561023e8371b1 (MD5) Made available in DSpace on 2020-05-08T00:42:36Z (GMT). No. of bitstreams: 1 souza_ers_dr_sjrp.pdf: 8296215 bytes, checksum: c098c1e8fb47b1ea91d561023e8371b1 (MD5) Previous issue date: 2020-02-28 Made available in DSpace on 2020-05-08T00:42:36Z (GMT). No. of bitstreams: 1 souza_ers_dr_sjrp.pdf: 8296215 bytes, checksum: c098c1e8fb47b1ea91d561023e8371b1 (MD5) Previous issue date: 2020-02-28 Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) A subordem Heteroptera possui uma enorme diversidade de insetos distribuída em sete infraordens, dentre as quais utilizamos Gerromorpha, Nepomorpha, Cimicomorpha e Pentatomomorpha. As espécies que foram estudadas da primeira infraordem, representando os insetos semi-aquáticos, são Limnogonus aduncus e Mesovelia mulsanti, da segunda infraordem, representando os aquáticos, são Belostoma anurum, Martarega sp. e Buenoa unguis, da terceira infraordem Zelus sp. e Teleonemia sp. e da última infraordem, representando juntas os terrestres, são Largus sp., Stenocoris sp., Zicca pulchra e Dysdercus sp. para descrever as ultraestruturas de suas espermátides e posterior comparação. Durante o processo de espermiogênese em geral, ocorre uma série de modificações nas espermátides antes dessas serem transformadas em espermatozoide, inclusive nos insetos. Neste trabalho verificamos por meio da Microscopia Eletrônica de Transmissão, as modificações ocorridas nas espermátides das infraordens anteriormente mencionadas. Na maioria dos insetos com hábitat aquático foi identificada uma maior variação morfológica com relação aos derivados mitocondriais, apresentando-se tanto com simetria reniforme, com simetria e padrão diferente, quanto com assimetria. Com relação aos terrestres, foi identificado predominantemente um padrão morfológico reniforme com uma área de cada derivado mitocondrial menor que a do axonema. Somente a espécie B. unguis, de hábitat aquático, evidenciou um padrão atípico, assimétrico, de derivados mitocondriais, com relação ao comumente observado na subordem Heteroptera. Ao longo da espermiogênese foi verificada a presença de manchete ao redor do núcleo e do acrossomo nas espécies M. mulsanti, L. aduncus, Teleonemia sp. e Z. pulchra, com padrão de distribuição dos microtúbulos diferente entre os insetos de hábitat aquático e terrestre. Outra característica observada foi a presença de um forame no interior do acrossomo de M. mulsanti, L. aduncus. As variações para as características analisadas no presente trabalho podem ser utilizadas como ferramentas importantes para auxiliar em análises sistemáticas. The suborder Heteroptera has an enormous diversity of insects distributed in seven infraorders, among which Nepomorpha, Gerromorpha, Pentatomomorpha e Cimicomorpha, The species that have been studied from the first infraorder, representing semi-aquatic insects, are Limnogonus aduncus and Mesovelia mulsanti, from the second order, representing the aquatic ones, Belostoma anurum, Martarega sp. and Buenoa unguis, from the third infraorder Zelus sp. and Teleonemia sp. and the last infraorder, representing terrestrials together, are Largus sp., Stenocoris sp., Zicca pulchra and Dysdercus sp. to describe the ultrastructures of their spermatides and further comparison. During the spermiogenesis process in general, a series of changes in spermatids occur before they are transformed into sperm, including in insects In this work we verified through Transmission Electron Microscopy, the changes occurred in the spermatids of the abovementioned infraorders. In the majority of insects with aquatic habitat, a greater morphological variation was identified in relation to mitochondrial derivatives, presenting both with reniform symmetry, with different symmetry and pattern, and with asymmetry. With respect to terrestrials, a predominantly reniform morphological pattern was identified with an area of each mitochondrial derivative smaller than the axoneme. Only the species B. unguis, of aquatic habitat, showed an atypical, asymmetric pattern of mitochondrial derivatives, in relation to that commonly observed in the suborder Heteroptera. Throughout spermiogenesis it was verified the manchette around the nucleus and the acrosome in the species M. mulsanti, L. aduncus, Teleonemia sp. and Z. pulchra, with different microtubule distribution pattern between aquatic and terrestrial habitat insects. Another characteristic observed was the presence of a foramen within the acrosome of M. mulsant and L. aduncus. The variations for the characteristics analyzed in the present work can be used as important tools to assist in systematic analyzes. CAPES: 001 CNPq: 141682/2016-7

Spermatogenesis and sperm ultrastructure were investigated in Neopanorpa lui Chou & Ran and Neopanorpa lipingensis Cai & Hua by light and transmission electron microscopy. Two lateral lamellae run along the lateral grooves of the spermatid nucleus during spermiogenesis, but disappear in the mature spermatozoa. Sperm cells are very similar in ultrastructure between the two species. The spermatozoa are elongated cells, each comprising an apical bilayered acrosome inserted on the anterior nuclear region, an elongate nucleus that occupies the anterior sperm region, a neck region, and a posterior long flagellum. The nucleus has two prominent lateral grooves and assumes a helical appearance. The flagellum is helical and consists of a 9 + 2 axoneme, two mitochondrial derivatives of unequal size, one accessory body, and two extra-axonemal accessory structures. The uniqueness of spermatozoa of Neopanorpa lies in the specifically spoon-shaped larger mitochondrial derivative along most of the flagellum length. A tentative phylogenetic analysis based on sperm structure supports a possible monophyletic clade comprising Panorpidae, Panorpodidae, Bittacidae, and Boreidae of Mecoptera with two non-homoplasious characters: glycocalyx organised in longitudinal ridges and two extra-axonemal accessory structures/rods in flagellum. Closer affinities of Siphonaptera and the mecopteran clade are suggested by the characters of two asymmetric mitochondrial derivatives and the presence of mitochondrial derivative with crystalline structure. The sister group relationship of Panorpidae with Panorpodidae is supported by the presence of nuclear fiber and the irregular shape of the larger mitochondrial derivative.

Submitted by LUÍS LENIN VICENTE PEREIRA null (luislenin@gmail.com) on 2017-03-02T15:07:34Z No. of bitstreams: 1 Tese Luis Lenin Vicente Pereira.pdf: 2894913 bytes, checksum: 01d77fed0e0eb6b5cfb7979e52829667 (MD5) Approved for entry into archive by LUIZA DE MENEZES ROMANETTO (luizamenezes@reitoria.unesp.br) on 2017-03-08T13:17:27Z (GMT) No. of bitstreams: 1 pereira_llv_dr_sjrp.pdf: 2894913 bytes, checksum: 01d77fed0e0eb6b5cfb7979e52829667 (MD5) Made available in DSpace on 2017-03-08T13:17:27Z (GMT). No. of bitstreams: 1 pereira_llv_dr_sjrp.pdf: 2894913 bytes, checksum: 01d77fed0e0eb6b5cfb7979e52829667 (MD5) Previous issue date: 2017-02-17 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) A subordem Heteroptera possui sete infraordens com aproximadamente 80 famílias. A maioria ocorre em todos os continentes (exceto Antártica) e algumas ilhas. Além dos Heteroptera terrestres, há os aquáticos e semi-aquáticos que são amplamente distribuídos, e surpreendem por sua capacidade de habitar uma extraordinária variedade de ecossistemas, sendo encontrados em habitats de água doce e marinho, e variada faixa de altitude entre 0 e 4.700 m. Estudos sobre aspectos ultraestruturais da espermatogênese e, especificamente, a estrutura do espermatozoide em Heteroptera ainda são escassos, por este motivo o objetivo do presente estudo foi o de analisá-los, por meio de cortes semifinos corados com azul de toluidina ou impregnados por íons prata, e cortes ultra-finos analisados em microscopia eletrônica de transmissão, utilizando testículos de machos adultos das famílias Belostomatidae, Gelastocoridae, Gerridae, Mesoveliidae, Notonectidae e Veliidae. Após a análise ultraestrutural da espermatogênese foi possível determinar que o padrão flagelar do axonema é de 9+9+2 para todas as espécies analisadas sendo, portanto, o padrão para essa subordem, as mitocôndrias durante a espermatogênese assumem diferentes morfologias, sendo que inicialmente as mitocôndrias se unem formando o complexo mitocondrial e, posteriormente, se divide em dois derivados mitocondriais que estão posicionados bilateralmente em relação ao axonema. Os derivados mitocondriais apresentaram tamanhos diferentes para as espécies B. amnigenus (Notonectidae) e R. c. crassifemur (Gerridae) e para as demais espécies o tamanho foi semelhante. As células germinativas possuem em seu citoplasma o acúmulo de um material denominado corpo cromatóide estando localizado próximo ao núcleo. Com relação ao comportamento nucleolar da espécie Martarega brasiliensis foi observado de um a quatro corpúsculos nucleolares em células de Prófase I comprovando uma grande atividade sintética das células nessa fase da divisão celular. Células em Metáfase I apresentaram regiões organizadoras nucleolares na região telomérica de um dos autossomos. Ainda, nessa espécie, foi possível observar, em Anáfase I, vários corpúsculos nucleolares persistindo até a fase de Telófase I. Todas as ultraestruturas descritas nas espécies analisadas foram semelhantes às descritas na literatura para Heteroptera, corroborando as características sinapomórficas dessa subordem sendo elas: a) a presença de duas pontes que ligam o material intertubular do axonema flagelar às cisternas achatadas que aderem aos lados internos dos derivados mitocondriais; b) padrão flagelar do axonema de 9+9+2 e c) ausência de corpos acessórios. The suborder Heteroptera has seven infraorders with approximately 80 families. Most occur in all continents, except Antarctica and some islands. In addition to terrestrial Heteroptera, there are also widely distributed aquatic and semi-aquatic species. This suborder have adapted to live in an extraordinary variety of ecosystems as freshwater and marine habitats and at altitudes ranging from 0 m to 4,700 m. The research concerning the ultrastructural aspects of spermatogenesis is a large and growing field of study, however, in the case of Heteroptera, research is still scarce. For this reason, the aim of this study was to analyze the ultrastructures and spermatogenesis through semi-thin sections stained with toluidine blue or silver ions (Ag-NOR) and ultrathin sections examined in transmission electron microscopy, using testes of adult males ofthe following families: Belostomatidae, Gelastocoridae, Gerridae, Mesoveliidae, Notonectidae and Veliidae. After ultrastructural analysis of spermatogenesis, it was possible to determine that the flagellar pattern of the axoneme is 9+9+2 for all species, being therefore, the pattern for this suborder. As spermatogenesis progresses, the mitochondria begins to cluster and concentrate on only one side of the cell. Then, the mitochondria combine to form a single mitochondrial complex, which subsequently divides into two mitochondrial derivatives. They are positioned on opposite sides of the axoneme. The mitochondrial derivatives presented different sizes for the species B. amnigenus (Notonectidae) and R. c. crassifemur (Gerridae) and for the other species the size was similar. The germ cells have in their cytoplasm the accumulation of a material denominated the chromatoid body, being located near the nucleus. Regarding the nucleolar behavior, M. brasiliensis showed nucleus in prophase I composed by the nucleolus and nucleolar corpuscles that varied from one to four, emphasizing that this insect has great synthetic activity during meiosis. The analysis of cells in metaphase I, showed that M. brasiliensis presents nucleolar organizing region (NOR) in at least one autosome. Furthermore, was not observed the phenomenon of nucleolar persistence. All the ultrastructures described in the analyzed species were similar to those described in the literature for Heteroptera, corroborating the synapomorphic characteristics of this suborder, being them: a) two opposite bridges in the axoneme connect the flattened cisterns adherent to the internal side of each mitochondrial derivative to the intertubular material; b) flagellar pattern of the axoneme of 9+9+2; c) accessory bodies are absent all along the flagellum.

SummaryAlmost all animals contain mitochondria of maternal origin only, but the exact mechanisms underlying this phenomenon are still vague. We investigated the fate of Drosophila paternal mitochondria after fertilization. We demonstrate that the sperm mitochondrial derivative (MD) is rapidly eliminated in a stereotypical process dubbed paternal mitochondrial destruction (PMD). PMD is initiated by a network of vesicles resembling multivesicular bodies and displaying common features of the endocytic and autophagic pathways. These vesicles associate with the sperm tail and mediate the disintegration of its plasma membrane. Subsequently, the MD separates from the axoneme and breaks into smaller fragments, which are then sequestered by autophagosomes for degradation in lysosomes. We further provide evidence for the involvement of the ubiquitin pathway and the autophagy receptor p62 in this process. Finally, we show that the ubiquitin ligase Parkin is not involved in PMD, implying a divergence from the autophagic pathway of damaged mitochondria.

In this study, the sperm morphology of the parasitoids Trichospilus diatraeae and Palmistichus elaeisis (Eulophidae) was investigated using light and transmission electron microscopy. In the two species, the sperm are spiral along their entire length and measure about 130 μm and 195 μm in length, respectively. The head region consists of the acrosome and nucleus. The acrosome is composed of an acrosomal vesicle and, in P. elaeisis, a perforatorium. In both species, an extracellular layer in which several filaments are radiated covers the acrosome and the anterior nuclear region. The nuclei are filled with homogeneous and compact chromatin and measure about 50 μm in length in P. elaeisis and 20 μm in T. diatraeae. The flagellum consists of an axoneme with the 9 + 9 + 2 microtubule arrangement spiraled in a long helix, two mitochondrial derivatives coiling around the axoneme and, in P. elaeisis, two accessory bodies. In T. diatraeae were observed transverse striations throughout the central region of the axoneme, whereas the central pair of microtubules was rarely observed. In the final flagellar region in T. diatraeae, different from P. elaeisis, one mitochondrial derivative ends well before the other and both end before the axoneme. The sperm of these two species exhibit features that discriminate one species from each other, as well as characteristics suggest that Eulophidae is closely related to Trichogrammatidae and both of these families are more similar to Eurytomidae than Agaonidae.

This study represents the first sperm description of a Vespinae species (Vespa crabro). The acrosome consists of an acrosomal vesicle and a perforatorium. The nucleus has compact chromatin and shows lenticular structures on the nuclear envelope. These structures, which have never been observed in a hymenopteran sperm, could be clusters of nuclear pores. The centriolar adjunct has an asymmetric pattern and shows a structured periphery. The centriole consists of 9 accessory microtubules and 9 doublet microtubules devoid of arms and spokes. The axoneme has a 9 + 9 + 2 microtubule pattern and the accessory microtubules have 16 protofilaments. The mitochondrial derivatives differ in length and diameter. The larger one is adjacent to the nuclear base, while the smaller one begins below the centriolar adjunct. They possess three distinct areas and a large paracrystalline region, which occurs only in the large one. The large mitochondrial derivative ends first, followed by the small one. The axoneme gradually disorganizes: first the central microtubules disappear, then the doublets, which show opened B-tubules, and finally the accessory microtubules. The sperm morphology of V. crabro is very similar to that of the polistine wasp, Agelaia vicina. This can indicate that, in Vespidae, sperm morphology is maintained without important variations among subfamilies and/or that this similarity indicates close phylogenetic relationship between these two subfamilies. Although Vespidae phylogenetically related to Formicidae, these data suggest that the former more closely related to Apoidea than to Formicidae.

The sperm ultrastructure of a few species of Limoniidae (Limonia nigropunctata; L. nubeculosa; Chionea n. sp.; C. alpina; C. lutescens) was studied. The two species of Limonia have a monolayered acrosome with crystallized material, a three-lobed nucleus in cross section, a ring of centriole adjunct material and a flagellum which consists of a 9 + 9 + 1 axoneme and a single mitochondrial derivative. The central axonemal tubule is provided with 15 protofilaments in its tubular wall, while the accessory tubules have 13 protofilaments and are flanked by the electron-dense intertubular material. The three species of Chionea share a monolayered acrosome, a nucleus with two longitudinal grooves, a centriole adjunct material which surrounds the centriole and the initial part of the axoneme. The axoneme is of conventional type, with 9 + 9 + 2 microtubular pattern, with accessory tubules provided with 13 protofilaments and intertubular material. However, in C. lutescens the accessory tubules start with 15 protofilaments and transform into a tubule with 13 protofilaments. These data are discussed in the light of the phylogenetic relationship between Limoniidae and Tipulidae. For this purpose, the sperm ultrastructure of Nephrotoma appendiculata was also considered comparatively.

Translational control is crucial for proper timing of developmental events that take place in the absence of transcription, as in meiotic activation in oocytes, early embryogenesis in many organisms, and spermatogenesis. Here we show that a novel form of the translation initiation complex component eIF4G in Drosophila, eIF4G2, is required specifically for male germ cells to undergo meiotic division and proper spermatid differentiation. Flies mutant for eIF4G2 are viable and female fertile but male sterile. Spermatocytes form, but the germ cells in mutant males skip the major events of the meiotic divisions and form aberrant spermatids with large nuclei. Consistent with the failure to undergo the meiotic divisions, function of eIF4G2 is required post-transcriptionally for normal accumulation of the core cell cycle regulatory proteins Twine and CycB in mature spermatocytes. Loss of eIF4G2 function also causes widespread defects in spermatid differentiation. Although differentiation markers Dj and Fzo are expressed in late-stage eIF4G2 mutant germ cells, several key steps of spermatid differentiation fail, including formation of a compact mitochondrial derivative and full elongation. Our results suggest that an alternate form of the translation initiation machinery may be required for regulation and execution of key steps in male germ cell differentiation.

Studies on the ultrastructural aspects of spermatogenesis and, specifically, the structure of sperm in aquatic Heteroptera are scarce. Therefore, the objective of this study was to analyse of the histology and ultrastructure of spermatogenesis. Semi-fine sections of the testicles of adult male Gelastocoris flavus flavus were stained with toluidine blue or impregnated with silver ions, and ultra-fine sections were analysed by transmission electron microscopy. The ultrastructural features observed during spermatogenesis of the species showed the presence of several small mitochondria uniformly distributed in the cytoplasm of cells in prophase I. These mitochondria then came together to form fewer, larger structures, which converged and formed the mitochondrial complex. Later, this mitochondrial complex was divided into two structures, termed mitochondrial derivatives, which were arranged bilaterally to the axoneme. The axoneme showed a flagellar pattern of 9+9+2. A vesicle was observed that originated in the early stages of spermiogenesis and was composed of many argyrophilic granules that united to form a single structure. This vesicle contained some highly stained structures in its interior. Thus, this paper describes histological and ultrastructural characteristics during spermatogenesis, contributing to the reproductive knowledge of these aquatic Heteroptera

To verify the questionable description of sperm structure of Trichogramma dendrolimi, in relation to Chalcidoids, a reinvestigation was undertaken. The spermatozoa appear wavy along their entire length. A small acrosome, together with the anterior nuclear region, is surrounded by an extracellular sheath, from which filaments radiate. The nucleus is helicoidal and attached to the flagellum by a centriolar adjunct. The axoneme has the 9 + 9 + 2 microtubule arrangement pitched in a long helix, with the spiralling mitochondrial derivatives coiling around it. Therefore, the spermatozoa of T. dendrolimi are very similar to those of other chalcidoids and different from the first description. It is now possible to affirm that the helical sperm structures are an apomorphic homology for Trichogrammatidae as well as Eulophidae, Pteromalidae, Eurytomidae, Torymidae, Mymaridae and for some other chalcidoids, if not all.

In the present study, spermatozoon ultrastructure was documented in two species of hangingflies, Bittacus strigosus Hagen (Mecoptera: Bittacidae) and B. stigmaterus Say. Structures considered important to phylogenetic assessment that were observed in B. strigosus and B. stigmaterus included a short bilayered acrosome, elongated nucleus, tube-like glycocalyx, centriole adjunct material, accessory bodies, two mitochondrial derivatives, extra axonemal rods, globular units, and 9+2 arrangement of microtubules in the axoneme. Comparisons were made to Bittacus planus Cheng, which was previously examined by electron microscopy ( Xie and Hua 2010 ). Similarities among the ultrastructural characteristics of the three Bittacus species support the monophyly of this genus. Displacement of a mitochondrial derivative by an accessory body was documented for the first time. This paper includes clarifications on differences between accessory bodies and extra axonemal rods, which are issues important to phylogenetic placement.

The Scelionidae sperm are distinguished from those of all hymenopterans already studied at least by the presence of a single mitochondrial derivative and the absence of a centriolar adjunct. The absence of an acrosome, in Telenomus podisi, is also unique. The helical nucleus and mitochondrial derivative spiralling around a twisted axoneme can be considered as synapomorphies shared with the Chalcidoidea, and the mitochondrial derivative running together with the nucleus for a long distance can be considered as a synapomorphy shared with the Diapriidae. Therefore, from a consideration of these features, it is possible to suppose that the Scelionidae, Chalcidoidea and Diapriidae are more closely related between themselves than are any of them to the Cynipoidea, since the latter does not share any of the above-mentioned features. This supposition agrees with phylogenetic analyses that supported the inclusion of Platygastroidea (Scelionidae and Platygastridae) and Chalcidoidea within the Proctotrupomorpha lineage, as well as the close relationship of these to the Diapriidae, and the exclusion of the Cynipoidea from this lineage.

Results from a transmission electron microscope study of the spermiogenesis and spermatozoon of Meara stichopi (Nemertodermatida, Platyhelminthes) indicate that the sperm type of the Nemertodermatida has evolved from the primitive metazoan sperm type rather than from an aberrant biflagellar sperm type as found in many other flatworms. The spirally coiled mitochondrial derivative in the mature spermatozoon develops from two large oval mitochondria in the early spermatid stages. A single flagellum grows out from a peripheral basal body adjacent to a perpendicularly placed accessory centriole. The basal body moves to a distal depression of the nucleus, and becomes equipped with an anchoring fibre apparatus. Most of the flagellum becomes axially incorporated into the developing spermatid. No trace of a second flagellum was found in any stage of the spermiogenesis. Rounded vesicles appear around the proximal, tapering end of the elongating nucleus. Most probably these vesicles form a thin acrosomal structure in the mature spermatozoon. No dense bodies, characteristic of many other ‘turbellarian’ flatworm sperm types, were found.

The spermatogenesis of Allostoma sp. and the organization of mature spermatozoa are described by means of transmission electron microscopy. All stages of sperm development were observed in mature testes, which are bordered by a layer of fibrous connective tissue. The primary spermatogonium produces 64 spermatids, grouped as a cytophore. The process of spermiogenesis occurs at the periphery of the cytophore, when the sperm body is formed and released at complete maturation. The mature spermatozoon is a flattened, filiform structure. It is aciliary. Its anterior end is not distinguishable from the posterior one in the testis. The nucleus, showing an approximately rectangular cross section, is situated centrally and the chromatin is densely packed. A membranous cylinder, located on each side of the nucleus, contains conspicuous stacks of clear thick-envelope vesicles, and a long mitochondrial derivative placed peripherally. The vesicles show contacts with the cylinder membrane and the outer sperm membrane. Their supposed Golgi-derived content may thus be released into the external medium. A hypothesis is proposed about the role of these vesicles in sperm motility.

The Y chromosome of Drosophila melanogaster, which is required only for male fertility, contains six loci that are essential for spermatogenesis. In primary spermatocytes, three of these loci form large lampbrush loops containing RNA transcripts and associated proteins. The identities and functions of these Y chromosome loop-binding proteins are largely unknown. This report demonstrates that the RB97D protein, which is essential for spermatogenesis, bound to a specific lampbrush loop. RB97D contains two copies of a well-characterized RNA binding domain, the RNA recognition motif, followed by a proline-glutamine rich domain. Immunohistochemical and immunofluorescence experiments showed that in the testis, RB97D was found only in primary spermatocyte nuclei and associated with the C loop from the ks-1 fertility locus in an RNAse-sensitive manner. The anti-RB97D antibodies also bound a single Y chromosome loop in D. hydei, suggesting that the protein and its loop-binding function have been evolutionarily conserved. These results demonstrate that the proteins that bind lampbrush loops can be essential for fertility. Since RB97D was present only premeiotically, its function is likely to be directly related to the metabolism of the C loop transcripts.

In this study, the morphology of spermatozoa of Bethylidae Dissomphalus connubialis (Pristocerinae) was analyzed using light and transmission electron microscopy. Spermatozoa of this species are thin, measure approximately 130 μm in length and comprise a head region and flagellum region. The head is formed by the acrosome and nucleus. The acrosome consists of the acrosome vesicle and the perforatorium, the posterior portion of which is inserted into a cavity at the anterior extremity of the nucleus. The nucleus is compact, electron-dense and measures 15 μm in length. The flagellum possesses two mitochondrial derivatives, two accessory bodies and one axoneme with a 9+9+2 microtubular pattern. The nucleus is connected to the flagellum by the centriole adjunct. Mitochondrial derivatives are compact, apparently without paracrystalline material and with rare mitochondrial cristae. They are asymmetric in length, such that the larger mitochondrial derivative begins parallel to the posterior region of the nucleus and the smaller mitochondrial derivative begins just below the centriole adjunct. The basic structure of spermatozoa of D. connubialis is similar to that of other Aculeata studied. However, this species shows characteristics not seen in other Hymenoptera, such as the wide electron-lucid region that separates the acrosomal vesicle from the perforatorium and the depth of the cavity in the anterior extremity of the nucleus, into which the base of the perforatorium is inserted. There are also characteristics that distinguish this species from Bethylidae Prorops nasuta, including the fact that one of the mitochondrial derivatives lies to parallel to the nucleus over a long distance, the small quantity of cristae, the absence of paracrystalline material in these organelles, and the fact that the accessory microtubules are the first to terminate in the final portion of the flagellum.

Spermatozoa of the triatomideo Rhodnius prolixus possess an axoneme with a 9 + 9 + 2 microtubule pattern and two mitochondrial derivatives. Bridges occur between axoneme and mitochondrial derivatives. Two paracrystalline structures embedded in amorphous regions were observed in the mitochondrial derivative. The use of the negative staining technique shows a zig-zag profile in the mitochondrial derivatives due to infolding to the cristae, regularly spaced with approximately 50 nm. This spacing is also observed in the distribution of the strands of particles in the mitochondrial membrana as seen in freeze-fracture replicas. In the P-fracture face of the flagellar plasma membrane, a regular array of the intramembranous particles was observed. This array consists of two rows, with 12-15 particles, and occurs in the space between the mitochondrial derivatives. Thus R. prolixus spermatozoon present a membrane domain, localized in the flagellar region, and bridges between mitochondrial membrane derivatives and the plasma membrane are probably attached to the flagellar components. These membrane specializations may be related to the production of co-ordinated flagellar movement, and can contribute significantly to further phylogenetic studies.

Made available in DSpace on 2014-06-11T19:26:05Z (GMT). No. of bitstreams: 0 Previous issue date: 2011-07-29Bitstream added on 2014-06-13T20:14:37Z : No. of bitstreams: 1 pereira_llv_me_sjrp.pdf: 1182871 bytes, checksum: 6053df49fe569dc60c6513fafdffaa9d (MD5) No presente trabalho verificamos que os testículos possuem morfologias diferentes podendo ser arredondados, arredondados/espiralados ou alongados/espiralados. Com relação à morfometria das células em prófase I, B. micantulum e R. zela foram as que apresentaram as menores células, G. f. flavus foi a que apresentou maior tamanho e R. c. crassifemur e M. brasiliensis apresentaram tamanho intermediário. A avaliação da espermatogênese nos permitiu concluir que as características observadas são semelhantes às das outras espécies de Heteroptera, descritas na literatura, diferindo apenas com relação à morfologia dos testículos, o número de cromossomos e o sistema cromossômico do sexo. A análise das ultraestruturas observadas durante a espermatogênese de Gelastocoris flavus flavus e Martarega uruguayensis mostraram a presença de várias mitocôndrias pequenas e uniformemente distribuidas pelo citoplasma em células em profase I, de ambas espécies, que foram se unindo formando o complexo mitocondrial, que possui no seu interior as mitocôndrias enoveladas, posteriormente este complexo mitocondrial se divide em duas estruturas denominadas derivados mitocondriais, que se dispõem bilateralmente ao axonema. O axonema dessas espécies possui o padrão de 9+9+2. A formação do acrossomo inicia-se nos primeiros estágios da espermiogênese sendo composto de muitas vesículas acrossomais que se unem formando uma única estrutura, sendo observada regiões e algumas estruturas mais coradas em seu interior. Basicamente o processo de espermiogênese não diferiu entre as duas espécies analisadas In this study, we found different morphologies for testes of the Heteroptera species Belostoma anurum, B. micantulum, Gelastocoris angulatus, G. flavus flavus, Rheumatobates crassifemur crassifemur, Buenoa amnigenus, B. unguis, Martarega brasiliensis, M. membranacea, M. uruguayensis, Rhagovelia tenuipes and R. zela. They can by round, round/spiral and elongated/spiral. The size of prophase I cells also varied, being the smallest ones detected in B. micantulum and R. zela, the largest in G. f. flavus, and the intermediate in R. c. crassifemur and M. brasiliensis. The analyses of spermatogenesis allowed us to conclude that, in the studied species, the features are similar to those of other previously described Heteroptera species, differing only as to the testicular morphology, the chromosome number, and the sex chromosome system. Ultrastructural analysis of the spermatogenesis showed several small mitochondrias evenly distributed throughout the cytoplasm, in cells at prophase I of G. f. flavus and M. uruguayensis. The small mitochondrias joined to form the mitochondrial complex. Later, this mitochondrial complex divided into two structures called mitochondrial derivatives, located bilaterally to the axoneme. The axoneme of these species showed the flagellar pattern 9+9+2. The acrosome started to be formed in the early stages of spermiogenesis, being composed of many acrosome vesicles that join to form a single structure. Some regions within this structure were more strongly stained. Basically the process of spermiogenesis did not differ between the species G. f. flavus and M. uruguayensis

Background Mammals and Drosophila melanogaster share some striking similarities in spermatogenesis. Mitochondria in spermatids undergo dramatic morphological changes and syncytial spermatids are stripped from their cytoplasm and then individually wrapped by single membranes in an individualization process. In mammalian and fruit fly testis, components of the mitochondrial iron metabolism are expressed, but so far their function during spermatogenesis is unknown. Here we investigate the role of Drosophila mitoferrin (dmfrn), which is a mitochondrial carrier protein with an established role in the mitochondrial iron metabolism, during spermatogenesis. Results We found that P-element insertions into the 5'-untranslated region of the dmfrn gene cause recessive male sterility, which was rescued by a fluorescently tagged transgenic dmfrn genomic construct (dmfrn venus ). Testes of mutant homozygous dmfrn SH115 flies were either small with unorganized content or contained some partially elongated spermatids, or testes were of normal size but lacked mature sperm. Testis squashes indicated that spermatid elongation was defective and electron micrographs showed mitochondrial defects in elongated spermatids and indicated failed individualization. Using a LacZ reporter and the dmfrn venus transgene, we found that dmfrn expression in testes was highest in spermatids, coinciding with the stages that showed defects in the mutants. Dmfrn-venus protein accumulated in mitochondrial derivatives of spermatids, where it remained until most of it was stripped off during individualization and disposed of in waste bags. Male sterility in flies with the hypomorph alleles dmfrn BG00456 and dmfrn EY01302 over the deletion Df(3R)ED6277 was increased by dietary iron chelation and suppressed by iron supplementation of the food, while male sterility of dmfrn SH115 /Df(3R)ED6277 flies was not affected by food iron levels. Conclusions In this work, we show that mutations in the Drosophila mitoferrin gene result in male sterility caused by developmental defects. From the sensitivity of the hypomorph mutants to low food iron levels we conclude that mitochondrial iron is essential for spermatogenesis. This is the first time that a link between the mitochondrial iron metabolism and spermatogenesis has been shown. Furthermore, due to the similar expression patterns of some mitochondrial iron metabolism genes in Drosophila and mammals, it is likely that our results are applicable for mammals as well.

Spermatozoa of Sergentomyia minuta were examined by light, scanning, and transmission electron microscopy. They were seen to be formed by a head and a tail. The head was characterized by an acrosomal complex with a thin space containing granular material organized as in a perforatorium. The tail contained a “9 + 9 + 0”; axoneme and one mitochondrial derivative. A comparison with the spermatozoon ultrastructure of other Diptera, particularly Psychodidae, is discussed.

We have tested the functional capacity of different beta tubulin isoforms in vivo by expressing beta 3-tubulin either in place of or in addition to beta 2-tubulin in the male germ line of Drosophila melanogaster. The testes-specific isoform, beta 2, is conserved relative to major metazoan beta tubulins, while the developmentally regulated isoform, beta 3, is considerably divergent in sequence. beta 3-tubulin is normally expressed in discrete subsets of cells at specific times during development, but is not expressed in the male germ line. beta 2-Tubulin is normally expressed only in the postmitotic germ cells of the testis, and is required for all microtubule-based functions in these cells. The normal functions of beta 2-tubulin include assembly of meiotic spindles, axonemes, and at least two classes of cytoplasmic microtubules, including those associated with the differentiating mitochondrial derivatives. A hybrid gene was constructed in which 5' sequences from the beta 2 gene were joined to protein coding and 3' sequences of the beta 3 gene. Drosophila transformed with the hybrid gene express beta 3-tubulin in the postmitotic male germ cells. When expressed in the absence of the normal testis isoform, beta 3-tubulin supports assembly of one class of functional cytoplasmic microtubules. In such males the microtubules associated with the membranes of the mitochondrial derivatives are assembled and normal mitochondrial derivative elongation occurs, but axoneme assembly and other microtubule-mediated processes, including meiosis and nuclear shaping, do not occur. These data show that beta 3 tubulin can support only a subset of the multiple functions normally performed by beta 2, and also suggest that the microtubules associated with the mitochondrial derivatives mediate their elongation. When beta 3 is coexpressed in the male germ line with beta 2, at any level, spindles and all classes of cytoplasmic microtubules are assembled and function normally. However, when beta 3-tubulin exceeds 20% of the total testis beta tubulin pool, it acts in a dominant way to disrupt normal axoneme assembly. In the axonemes assembled in such males, the doublet tubules acquire some of the morphological characteristics of the singlet microtubules of the central pair and accessory tubules. These data therefore unambiguously demonstrate that the Drosophila beta tubulin isoforms beta 2 and beta 3 are not equivalent in intrinsic functional capacity, and furthermore show that assembly of the doublet tubules of the axoneme imposes different constraints on beta tubulin function than does assembly of singlet microtubules.

In the seminal vesicle of the ‘symphyta’ Arge pagana the spermatozoa are stored in motile spermatodesm bundles, maintained by an anterior cap of extracellular material. This cap consists of a denser cortex and of an internal matrix, where part of the sperm heads are embedded. The number of spermatozoa per bundle is variable. The spermatozoa are short, only 30 μm long, with a head region of about 23 μm, and a very short flagellum of about 7 μm. The head includes the acrosome, with a perforatorium, and the nucleus. The flagellum consists of an axoneme, with a 9 + 9 + 2 microtubule pattern, a centriolar adjunct, two mitochondrial derivatives and two accessory bodies. The mitochondrial derivatives are very slender and of different lengths. The longer begins at the base of the nucleus, while the shorter one starts just below the base of the centriolar adjunct. This latter is asymmetric and appears at the nuclear base, extending parallel to the axoneme up to the anterior end of the smaller mitochondrial derivative. The short spermatodesmata and the small mitochondrial derivatives characterize the A. pagana sperm. In addition, the centriolar adjunct asymmetry and the occurrence of spermatodesm bundles might be considered plesiomorphic states present in the basal Tenthredinoidea.

Background Drosophila Merlin, the homolog of the human Neurofibromatosis 2 (NF2) gene, is important for the regulation of cell proliferation and receptor endocytosis. Male flies carrying a Mer 3 allele, a missense mutation (Met177→Ile) in the Merlin gene, are viable but sterile; however, the cause of sterility is unknown. Results Testis examination reveals that hemizygous Mer 3 mutant males have small seminal vesicles that contain only a few immotile sperm. By cytological and electron microscopy analyses of the Mer 3, Mer 4 (Gln170→stop), and control testes at various stages of spermatogenesis, we show that Merlin mutations affect meiotic cytokinesis of spermatocytes, cyst polarization and nuclear shaping during spermatid elongation, and spermatid individualization. We also demonstrate that the lethality and sterility phenotype of the Mer 4 mutant is rescued by the introduction of a wild-type Merlin gene. Immunostaining demonstrates that the Merlin protein is redistributed to the area associated with the microtubules of the central spindle in telophase and its staining is less in the region of the contractile ring during meiotic cytokinesis. At the onion stage, Merlin is concentrated in the Nebenkern of spermatids, and this mitochondrial localization is maintained throughout sperm formation. Also, Merlin exhibits punctate staining in the acrosomal region of mature sperm. Conclusion Merlin mutations affect spermatogenesis at multiple stages. The Merlin protein is dynamically redistributed during meiosis of spermatocytes and is concentrated in the Nebenkern of spermatids. Our results demonstrated for the first time the mitochondrial localization of Merlin and suggest that Merlin may play a role in mitochondria formation and function during spermatogenesis.

Sperm ultrastructure has been studied in three species of the taxa Mecoptera and Siphonaptera. The spermatozoon of the scorpion fly Panorpa germanica shows an apical bilayered acrosome, a helicoidal nucleus, a centriolar region and a 9+2 flagellar axoneme helicoidally arranged around a long mitochondrial derivative. A second mitochondrial derivative is very short and present only in the centriolar region. A single accessory body is present and it is clearly formed as a prolongation of the centriole adjunct material. Two lateral lamellae run parallel to the nucleus. The snow fly Boreus hyemalis has a conventional sperm structure and shows a bilayered acrosome, a long nucleus, a centriolar region, two mitochondrial derivatives and two accessory bodies. The axoneme is of the 9+2 type and is flattened at the tail tip. Both P. germanica and B. hyemalis have two longitudinal extra-axonemal rods and have a glycocalyx consisting of longitudinal parallel ridges or filaments. The spermatozoon of the flea Ctenocephalides canis has a long apical bilayered acrosome, a nucleus, a centriolar region, a 9+2 axoneme wound around two unequally sized mitochondrial derivatives, and two triangular accessory bodies. In the posterior tail end the flagellar axoneme disorganises and a few microtubular doublets run helicoidally around the remnant mitochondrial derivative. The glycocalyx consists of fine transverse striations. In all three species, the posterior tail tip is characterised by a dense matrix embedding the disorganised axoneme. From this comparative analysis of the sperm structure it is concluded that Mecoptera, as traditionally defined, is monophyletic and that B. hyemalis is a member of Mecoptera rather than of Siphonaptera.

Sperm ultrastructure is examined in representatives of five genera of the nudibranch gastropod family Chromodorididae: (Chromodoris, Hypselodoris, Glossodoris, Risbecia and Pectenodoris) and the results compared with previous work on other gastropods, especially other nudibranchs. As chromodoridid phylogeny is still incompletely understood, this study partly focuses on the search for new and as yet untapped sources of informative characters. Like spermatozoa of most other heterobranch gastropods, those of the Chromodorididae are elongate, complex cells composed of an acrosomal complex (small, rounded acrosomal vesicle, and columnar acrosomal pedestal), a condensed nucleus, sub-nuclear ring, a highly modified mid-piece (axoneme + coarse fibres surrounded by a glycogen-containing, helically-coiled mitochondrial derivative) and terminally a glycogen piece (or homologue thereof). The finely striated acrosomal pedestal is a synapomorphy of all genera examined here, but interestingly also occurs in at least one dorid (Rostanga arbutus). Substantial and potentially taxonomically informative differences were also observed between genera in the morphology of the nucleus, the neck region of the mid-piece, and also the terminal glycogen piece. The subnuclear ring is shown for the first time to be a segmented, rather than a continuous structure; similarly, the annular complex is shown to consist of two structures, the annulus proper and the herein-termed annular accessory body.

The spermatozoa of Solenopsis invicta are long and slender, measuring about 70 μm in length, of which the head region measures approximately 13 μm. The head consists of an acrosome formed by an acrosomal vesicle covering a perforatorium and a nucleus. This latter measures about 12 μm, has compact chromatin, and many translucent areas. The flagellum consists of an axoneme of 9+9+2 microtubules that begins just below the nuclear base. The two mitochondrial derivatives are of almost equal diameter, and there are two accessory bodies. The nucleus is attached to the flagellum by a centriolar adjunct located between the baso-lateral region of the nucleus and the anterior mitochondrial derivative extremities. Therefore, the nucleus–flagellum transition region of the spermatozoa of S. invicta is very similar to that of other ants and differs from the first description of this species [Ann. Entomol. Soc. Am. 67 (1967) 632]. In relation to other aculeates, it is possible that three characteristics (type of centriolar adjunct, presence of paracrystalline areas in each mitochondrial derivative, and the position of these areas within each derivative) may be autopomorphies for the Formicidae.

The flagellum of Apis mellifera (Hymenoptera, Apidae) consists of two mitochondrial derivatives, an axoneme and two accessory bodies. The mitochondrial derivatives are of unequal size and lie parallel to the axoneme. In the larger derivative four regions can be distinguished while in the smaller, only three. The region occurring only in the larger derivative consists of paracystalline material. The smaller mitochondrial derivative terminates anterior to the larger one. An extremely long centriolar adjunct is observed between the nucleus and the smaller mitochondrial derivative. This adjunct is compact, very electron dense and gradually tapers from base toward apex, finishing at the anterior extremity of the axonemal microtubules. In this flagellar region, there is only one accessory body present between the larger mitochondrial derivative and the axoneme. Anteriorly, the tips of the axonemal microtubules are inserted in a well developed mass of granular appearance. This material surrounds the nuclear base, separating it from the anterior end of the larger mitochondrial derivative. We believe that the structure identified here as a centriolar adjunct is homologous to that observed in Formicidae, Ichneumonoidea and Symphyta. Therefore, very probably, it is common to most Hymenoptera.

The spermatozoon of Onchidella celtica shows many peculiar features. The acrosome is formed by an apical vesicle and a pedestal consisting of a principal body which is flanked by two electron-dense elliptical elements. The mitochondrial derivative consists of three different components: peripheral layers of paracrystalline material, concentric cisterns, and two cylindrical compartments. The 9 + 9 + 2 axoneme shows 9 small deposits of glycogen granules in correspondence with the axonemal “spokeheads,” and a small assembly of possibly proteic material lying close to the two central tubules. Another intraaxonemal deposit of glycogen granules is present at the level of the “glycogen piece.” Most of the observed features are in substantial agreement with those of another species of the Onchidiidae. This confirms the constancy of sperm structure among the members of a single family of the Gastropoda.

The giant aflagellate spermatozoa of P. quadrioculatum are composed of two different parts: a thicker head piece and a more slender tail piece. In the head there exist a large elongated nucleus and an elongated mitochondrial derivative situated in a groove-like cavity of the nucleus. In mature spermatozoa the nuclear material is arranged in many small membrane bounded areas. Both structures, nucleus and mitochondrial derivative, are spirally coiled. The outer part of the membrane in the mitochondrial derivative forms many loop-like foldings. Both organelles continue to the tail in form of two small, helically coiled ribbons; the nucleus is anchored within the mitochondrial derivative by an electron-opaque process. A sheath of spirally-orientated cortical microtubules starting from the tip of the head runs to the tip of the tail under the cell membrane. In addition, a second sheath of tubules occurs in the tail region, these tubules also run parallel to each other, but in the opposite direction to the microtubules of the outer sheath.

Ultrastructural observations on spermiogenesis and spermatozoa of selected pyramidellid gastropods (species ofTurbonilla, Pyrgulina, Cingulina andHinemoa) are presented. During spermatid developement, the condensing nucleus becomes initially anterio-posteriorly compressed or sometimes cup-shaped. Concurrently, the acrosomal complex attaches to an electrondense layer at the presumptive anterior pole of the nucleus, while at the opposite (posterior) pole of the nucleus a shallow invagination is formed to accommodate the centriolar derivative. Midpiece formation begins soon after these events have taken place, and involves the following processes: (1) the wrapping of individual mitochondria around the axoneme/coarse fibre complex; (2) later internal metamorphosis resulting in replacement of cristae by paracrystalline layers which envelope the matrix material; and (3) formation of a glycogen-filled helix within the mitochondrial derivative (via a secondary wrapping of mitochondria). Advanced stages of nuclear condensation (elongation, transformation of fibres into lamellae, subsequent compaction) and midpiece formation proceed within a microtubular sheath (‘manchette’). Pyramidellid spermatozoa consist of an acrosomal complex (round to ovoid apical vesicle; column-shaped acrosomal pedestal), helically-keeled nucleus (short, 7–10 μm long, shallow basal invagination for axoneme/coarse fibre attachment), elongate helical midpiece (composed of axoneme, coarse fibres, paracrystalline and matrix materials, glycogen-filled helix), glycogen piece (length variable, preceeded by a dense ring structure at junction with midpiece). The features of developing and mature spermatozoa observed in the Pyramidellidae are as observed in opisthobranch and pulmonate gastropods indicating that the Pyramidelloidea should be placed within the Euthyneura/Heterobranchia, most appropriately as a member group of the Opisthobranchia.

In this paper the spermatozoon of a plecopteran and of a trichopteran is described. The former is an “orthopteroid type” spermatozoon, with the following peculiarities: acrosome lying in lateral position with respect to the nucleus; proteinaceous rodlet arising from the adjunct centriole and flanking the two mitochondrial derivatives in the tail; accessory fibers of the axial-filament complex hollow even at maturation. This spermatozoon is therefore quite different from Odonata and Ephemeroptera sperms previously described, and it represents the simplest case of structure of the outer accessory fibers known in insects. The spermatozoon of the trichopteran exhibits the following characteristics: absence of an acrosome, a third supernumerary proteinaceous slender fibril between the two principal central ones, only one mitochondrial derivative entirely occupied by parallel transverse cristae and lacking a crystalline axis. The outer accessory fibers of the flagellum are 300 A in diameter and, in this respect, closely recall those of Lepidoptera, comparatively reexamined in this paper. Like the principal central units, these fibers are filled with glycogen-like polysaccharides and proteins, and they are longer than the 9 principal peripheral doublets. This model of Trichoptera and Lepidoptera outer fibers is quite different from the pattern hitherto described in other insects.

Epoxy embedding methods of Glauert and Kushida have been modified so as to yield rapid, reproducible, and convenient embedding methods for electron microscopy. The sections are robust and tissue damage is less than with methacrylate embedding.

The mitochondrial derivative of the sperm cell of the Gastropod Helix is endowed with a compartment loaded with glycogen (Personne et André, 1964). The aim of this work is to establish whether this mitochondrial glycogen is synthesized in the mitochondrial derivative itself or elsewhere in the cell. For this purpose, living sperm were first incubated in a medium containing glucose 1-phosphate, then fixed, sectioned, and stained specifically for polysaccharides by the phosphotungstic acid technique or the periodic acid-thiosemicarbazide-silver proteinate technique. Comparison with controls shows that a synthesis of glycogen occurred during the time of incubation. It can be inferred from this result that an amylophosphorylase system controlling the metabolism of polysaccharides is present in the mitochondrial derivative itself. Results obtained with the iodine technique indicate that the original glycogen molecules are elongated during the experiment. It seems probable that the amylophosphorylase system demonstrated here accounts for at least a part of the proteinaceous coat morphologically detectable around each individual glycogen particle.

Although 9-membered centrioles are found in somatic tissues of Sciara, the centriole which lies at the spindle pole of the second meiotic division in male Sciara is composed of a row of approximately 70 short tubules in an oval array. Shortly after telophase of this unequal division, in the daughter cell destined to undergo spermiogenesis, microtubules become confluent with the tubules of the centriole. These tubules have the same density as other cytoplasmic microtubules after glutaraldehyde-OsO4 fixation and, like them, are not preserved with OsO4 fixation. As the centriole, now with approximately 70 attached, posteriorly directed, doublet tubules, migrates from the polar to the apolar end of the nucleus to take a final position in an oval groove which forms in the nuclear envelope, the tubules lengthen and become demonstrable after OsO4 fixation and more electron opaque than other cytoplasmic microtubules following glutaraldehyde-OsO4 fixation. Later, a singlet tubule appears peripherally to each doublet of the oval and 4 "arms" develop at specific sites on the tubules. Posteriorly, where the oval of tubules becomes discontinuous and forms a spiral, the arrangement of arms is different and the singlet tubules are lacking. Dense solid bodies develop inside this odd flagellum and become enclosed by a smooth double membrane. A single mitochondrial derivative has three components: a central area of homogeneous, moderately electron-opaque, proteinaceous material; a peripheral ring of cristae; and a crystalloid which is specifically oriented with respect to the flagellar tubules.

Though the fagellum of Sciara sperm arises from a blepharoplast and is characterized by doublet tubules with arms, it differs markedly from the familiar type of flagella in the number and arrangement of its tubules. The axial filament complex in sperm from the testis of Sciara consists of approximately 70 doublet tubules, each with an associated singlet tubule. Near the nucleus these tubules are displaced in an oval array. Posteriorly the oval breaks and coils from one free end so that the axial filament complex at posterior levels has the form of a spiral. The singlet tubules do not extend the full length of the sperm but terminate in order from inside the spiral. Farther posteriorly the axial filament complex reverses the direction of coiling, and the doublets terminate from outside the spiral. Four arms are specifically positioned on the singlet and doublet tubules. A single mitochondrial derivative extends most of the length of the sperm; it consists of a large mass of proteinacious material, a crystalloid located adjacent to the axial filament complex, and peripheral cristae. In the female genital tract, sperm undergo gross morphological changes which include sloughing of practically all the mitochondrial material except the crystalloid, repositioning of the crystalloid, and uncoiling and subsequent recoiling of the axial filament complex into a different configuration. From analysis of serial sections it was determined that the orientation of arms, when the axial filament is viewed from base to tip, is the same as in conventional flagella.

Objective To investigate the effect of human contraceptive (HC) as ability to suppress the reproductive success of blow fly, Chrysomya megacephala (Fabricius) ( C. megacephala ) and offspring under controlled laboratory conditions. Methods Adult C. megacephala were fed with low (0.036 mg/mL) and high dose (0.072 mg/mL) HC (Microgest®, Thailand), containing levonorgestrel and ethinyl estradiol, in their drinking water for 7 days. Three experiments were set; experiment I with fed only in parental males, experiment II with fed only in parental females and experiment III with fed in both males and females. All experiments were then maintained for 3 generations after crossing and inbreeding. Results A lower ovariole production and less fully mature ovarioles were evident in F1, F2 and F3 than control when parent males, females and both had been fed with high dose HC. Cellular changes during spermatogenesis in F1, F2 and F3 testes was confirmed using transmission electron microscopy (TEM), showing the low level of condensed chromatin, necrotic chromatin, irregularities and degenerated nuclear envelope in the nucleus. In the cytoplasm, mitochondrial swelling, rough endoplasmic reticulum swelling as well as vacuolated cytoplasm were noticed. As for the sperm per se , we found the degenerated nuclei and/or incomplete mitochondrial derivative, axoneme and vacuolated flagella. Regarding deformity in F1, F2 and F3 ovariole, ultrastructural alteration observed by scanning electron microscopy (SEM) included malformations involving fragile enveloping peritoneal sheath, cracked ovarioles, peel away chorion, crumbled eggshell and incomplete development; whereas TEM presented malformed and disorganized mass of cells, proteic yolk granules and vacuolated vesicles. Conclusions Administer of HC to adult C. megacephala caused ovariole reduction, less matured ovariole and affected cellular changes in testes and ovariole of offspring up to F3.

Drosophila parkin, the ortholog of the human parkin gene, responsible for a familiar form of autosomal recessive juvenile parkinsonism, has been shown previously to be involved in Drosophila male fertility. Loss-of-function mutations in the parkin gene cause failure of spermatid individualization by affecting the proper progression of the actin-based investment cones that assemble in the nuclear region, but fail to translocate in synchrony down the cyst. In parkin mutants, the investment cones are scattered along the post-elongated spermatid bundles and fail to act properly in the process of sperm individualization. Using phase-contrast and electron microscopy analysis, we demonstrate that the parkin spermatids assemble a seemingly normal onion-stage nebenkern, but when the axoneme elongates only one mitochondrial derivative unfurls from the nebenkern. This unique mitochondrial derivative undergoes abnormal shaping and condensation during spermatid elongation. Our results indicate that parkin gene function is necessary for mitochondrial morphogenesis during earlier and later phases of spermiogenesis. The failure of cyst individualization may be due to the sensitivity of investment cone movement to the perturbation of mitochondrial morphology during spermatid elongation.

The mitochondrial derivative of the sperm of the gastropod pulmonate Biomphalaria glabrata was studies to ascertain succinic dehydrogenase localization cytochemically. Two techniques were compared. One technique depends on a tetrazolium salt that yields an osmiophilic formazan upon reduction. The other technique is dependent on the reduction of copper ferricyanide. The effects of several electron transport inhibitors were studied. The reaction product observed in the matrix of the mitochondrial derivative using the former technique is sensitive to rotenone and is believed to be nicotinamide adenine dinucleotide-dependent. The reaction product observed in the intracristal spaces using the copper ferricyanide method is insensitive to rotenone and is believed to cytochemically demonstrate succinic dehydrogenase in this material.

1. The study of certain structures, like the acrosome and Nebenkern, in the developing sperms of Arvelius is facilitated by the fact that they are present in greatly enlarged dimensions in certain testicular lobes while retaining normal size in adjoining lobes. 2. The dictyosomes or Golgi bodies in the spermatocytes are stained red by the PAS treatment. They are assembled to form the similarly stained acroblast in the spermatid, and the acrosome that is finally evolved is also PAS-positive in its color reaction. All the evidence indicates that in the process of acrosome formation there is a direct utilization of dictyosomal material which is already present two cell generations earlier. 3. The substance responsible for the red staining of the dictyosomes and acrosome is a polysaccharide with a 1,2 glycol grouping. There is also direct evidence for the presence of small quantities of proteins which probably form a complex with the polysaccharides. This complex is similar or perhaps identical with a mucopolysaccharide. 4. The Nebenkern, which is a mitochondrial derivative, does not contain polysaccharides with a 1,2 glycol grouping. On the other hand, it contains considerable amounts of proteins which, in contrast to those of the acrosome, are easily digested by pepsin. The evidence does not support the suggestion, frequently made, that the acrosome and Nebenkern are closely interrelated. 5. The enzymatic nature of the acrosome and its possible role in fertilization and sterility are discussed.