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    Nucleologenesis of Dipetalogaster maxima (AD). Note the nucleolar persistence phenomenon (C, D, arrow). Bar = 10 μm.

  • 1.

    World Health Organization, 2015. Chagas disease (American trypanosomiasis). Wkly Epidemiol Rec 90: 3344.

  • 2.

    Almeida CE, Duarte R, Guerra do Nascimento R, Pacheco RS, Costa J, 2002. Triatoma rubrovaria (Blanchard, 1843) (Hemiptera, Reduviidae, Triatominae) II: trophic resources and ecological observations of five populations collected in the state of Rio Grande do Sul, Brazil. Mem Inst Oswaldo Cruz 97: 11271131.

    • Search Google Scholar
    • Export Citation
  • 3.

    Almeida CE, Folly-Ramos E, Agapito-Souza R, Magno-Esperança G, Pacheco RS, Costa J, 2005. Triatoma rubrovaria (Blanchart, 1843) (Hemiptera-Reduviidae-Triatominae) IV: bionomic aspects on the vector capacity of nymphs. Mem Inst Oswaldo Cruz 100: 231235.

    • Search Google Scholar
    • Export Citation
  • 4.

    Alevi KCC, Rosa JA, Azeredo-Oliveira MTV, 2014. Spermatogenesis in Triatoma melanica Neiva and Lenti, 1941 (Hemiptera, Triatominae). J Vector Ecol 39: 231233.

    • Search Google Scholar
    • Export Citation
  • 5.

    Alevi KCC, Castro NFC, Oliveira J, Rosa JA, Azeredo-Oliveira MTV, 2015. Cystic spermatogenesis in three species of the prolixus complex (Hemiptera: Triatominae). Ital J Zool (Modena) 82: 172178.

    • Search Google Scholar
    • Export Citation
  • 6.

    Morielle-Souza A, Taboga SR, Azeredo-Oliveira MTV, 2010. Ultrastructural analysis of the nucleolar aspects at spermiogenesis in triatomines (Heteroptera, Triatominae). Micron 41: 791796.

    • Search Google Scholar
    • Export Citation
  • 7.

    Alevi KCC, Guerra AL, Imperador CHL, Rosa JA, Azeredo-Oliveira MTV, 2016. Reproductive biology of Triatoma brasiliensis (Hemiptera, Triatominae) during the imaginal molt. Am J Trop Med Hyg 94: 689690.

    • Search Google Scholar
    • Export Citation
  • 8.

    Tartarotti E, Azeredo-Oliveira MTV, 1999. Patterns of nucleolar activity during spermatogenesis of two triatomines, Panstrongylus megistus and P. herreri. Caryologia 52: 177184.

    • Search Google Scholar
    • Export Citation
  • 9.

    Alevi KCC, Castro NFC, Lima ACC, Ravazi A, Morielle-Souza A, Oliveira J, Rosa JA, Azeredo-Oliveira MTV, 2014. Nucleolar persistence during spermatogenesis of the genus Rhodnius (Hemiptera, Triatominae). Cell Biol Int 38: 977980.

    • Search Google Scholar
    • Export Citation
  • 10.

    Howell WM, Black DA, 1980. Controlled silver-staining of nucleolus organizer regions with a protective colloidal developer: a 1-step method. Experientia 36: 10141015.

    • Search Google Scholar
    • Export Citation
  • 11.

    Costa LC, Azeredo-Oliveira MTV, Tartarotti E, 2008. Spermatogenesis and nucleolar activity in Triatoma klugi (Triatomine, Heteroptera). Genet Mol Biol 31: 438444.

    • Search Google Scholar
    • Export Citation
  • 12.

    Alevi KCC, Mendonça PP, Pereira NP, Rosa JA, Azeredo-Oliveira MTV, 2013. Análise das possíveis regiões organizadoras nucleolares e da atividade nucleolar em Triatoma melanocephala e T. lenti, importantes vetores da doença de Chagas. Rev Cienc Farm Básica e Apl 34: 417421.

    • Search Google Scholar
    • Export Citation
  • 13.

    Bardella VB, Azeredo-Oliveira MTV, Tartarotti E, 2008. Cytogenetic analysis in the spermatogenesis of Triatoma melanosoma (Reduviidae; Heteroptera). Genet Mol Res 7: 326335.

    • Search Google Scholar
    • Export Citation
  • 14.

    Severi-Aguiar GDC, Azeredo-Oliveira MTV, 2005. Cytogenetic study on three species of the genus Triatoma (Heteroptera: Reduviidae) with emphasis on nucleolar organizer regions. Caryologia 58: 293299.

    • Search Google Scholar
    • Export Citation
  • 15.

    Severi-Aguiar GDC, Lourenço LB, Bicudo HEMC, Azeredo-Oliveira MTV, 2006. Meiosis aspects and nucleolar activity in Triatoma vitticeps (Triatominae, Heteroptera). Genetica 126: 141151.

    • Search Google Scholar
    • Export Citation
  • 16.

    Pereira NP, Alevi KCC, Mendonça PP, Azeredo-Oliveira MTV, 2015. Spermatogenesis and nucleolar behavior in Triatoma vandae and Triatoma williami (Hemiptera, Triatominae). Genet Mol Res 14: 1214512151.

    • Search Google Scholar
    • Export Citation
  • 17.

    Borgueti AO, Alevi KCC, Silistino-Souza R, Rosa JA, Azeredo-Oliveira MTV, 2015. Immunofluorescence and ultrastructural analysis of the chromatoid body during spermatogenesis of Triatoma platensis and T. rubrovaria (Hemiptera, Triatominae). Micron 74: 4446.

    • Search Google Scholar
    • Export Citation
  • 18.

    Madeira FF, Lima ACC, Rosa JA, Azeredo-Oliveira MTV, Alevi KCC, 2016. Nucleolar persistence phenomenon during spermatogenesis in genus Meccus (Hemiptera, Triatominae). Genet Mol Res 15. doi:10.4238/gmr.15017427.

    • Search Google Scholar
    • Export Citation
  • 19.

    Morielle A, Azeredo-Oliveira MTV, 2004. Description of the nucleolar activity and karyotype in germinative cell lines of Rhodnius domesticus (Triatominae, Heteroptera). Caryologia 57: 3137.

    • Search Google Scholar
    • Export Citation
  • 20.

    Alevi KCC, Mendonça PP, Pereira NP, Rosa JA, Azeredo-Oliveira MTV, 2014. Is there post-meiotic transcriptional activity during hemipteran spermiogenesis? Invertebr Reprod Dev 58: 193198.

    • Search Google Scholar
    • Export Citation

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Nucleolar Persistence: Peculiar Characteristic of Spermatogenesis of the Vectors of Chagas Disease (Hemiptera, Triatominae)

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  • 1 Departamento de Biologia, Instituto de Biociências, Letras e Ciências Exatas, Universidade Estadual Paulista–São José do Rio Preto, São Paulo, Brazil.
  • 2 Departamento de Ciências Biológicas, Faculdade de Ciências Farmacêuticas, Universidade Estadual Paulista–Araraquara, São Paulo, Brazil.

All species of triatomines are considered potential vectors of Chagas disease and the reproductive biology of these bugs has been studied by different approaches. In 1999, nucleolar persistence during meiosis was observed in the subfamily for the first time. Recently, it has been observed that all species within the genus Rhodnius exhibit the same phenomenon, suggesting that it may be a synapomorphy of the triatomines. Thus, this article aims to analyze the nucleolar behavior during spermatogenesis of 59 triatomine species. All analyzed species exhibited nucleolar persistence during meiosis. Recently, it has been suggested that nucleolar persistence may be fundamental for the spermatogenesis of these vectors, since it is related to the formation of the chromatoid body. Therefore, we emphasize that this phenomenon is a peculiarity of the Triatominae subfamily and that further studies are required to analyze whether the nucleolar material that persists is active.

The Triatominae subfamily is composed of 150 species (148 living species and two fossils), grouped into 18 genera and five or six tribes. All triatomine species are considered potential vectors of Trypanosoma cruzi, the etiologic agent of Chagas disease.1

Chagas disease is a neglected disease that has no cure. The main way to minimize the incidence of this disease in human populations is through vector control.1 It is estimated that more than 5 million people are infected by T. cruzi, the parasite that causes Chagas disease, and that 70 million still live at risk, which places this illness among the most serious parasitic diseases in the world.1 Thus, improving the knowledge on several fields on triatomine vector potentiality (such as ecology2 and biology3) may provide important information for control measures.

The reproductive biology of these bugs has been studied by different approaches, such as cytogenetic,4 structural,5 and ultrastructural6 analysis. Furthermore, the spermatogenesis of the triatomines is characterized as cystic,5,6 and it has been suggested that, during imaginal molt (transition from the fifth instar nymph to adult), the cell division is disrupted, aiming to reduce energy costs, and the differentiation into sperm is stimulated to ensure the paternity of the adult male.7

In 1999, Tartarotti and Azeredo-Oliveira,8 while studying the spermatogenesis of Panstrongylus megistus and Panstrongylus herreri (= Panstrongylus lignarius), noted that these triatomines exhibited a different nucleolar behavior than the one described for other eukaryotes: the nucleolus persisted during all stages of meiosis. The authors characterized this phenomenon as nucleolar persistence.

Recently, Alevi and others9 have observed that all species within the genus Rhodnius feature nucleolar persistence during meiosis as well, and they suggested that analyses of the nucleolar behavior should be carried out in a large range of species of triatomines to examine whether that characteristic is a synapomorphy of the Triatominae subfamily.

Thus, this article aims to analyze the nucleolar behavior during spermatogenesis of 59 triatomine species to determine whether nucleolar persistence occurs in all bugs of the Triatominae subfamily.

We analyzed at least two adult male specimens of each species (Cavernicola pilosa, Psammolestes tertius, Rhodnius brethesi, Rhodnius colombiensis, Rhodnius domesticus, Rhodnius ecuadoriensis, Rhodnius milesi, Rhodnius montenegrensis, Rhodnius nasutus, Rhodnius neglectus, Rhodnius neivai, Rhodnius pallescens, Rhodnius pictipes, Rhodnius prolixus, Rhodnius robustus, Rhodnius stali, Dipetalogaster maxima, Eratyrus cuspidatus, Meccus pallidipennis, Meccus longipennis, Mepraia spinolai, Nesotriatoma bruneri (= Nesotriatoma flavida), Panstrongylus lignarius, Panstrongylus megistus, Triatoma arthurneivai, Triatoma baratai, Triatoma brasiliensis, Triatoma brasiliensis macromelasoma, Triatoma carcavalloi, Triatoma circummaculata, Triatoma costalimai, Triatoma delpontei, Triatoma dimidiata, Triatoma garciabesi, Triatoma guasayana, Triatoma guazu, Triatoma infestans, Triatoma juazeirensis, Triatoma jurbergi, Triatoma klugi, Triatoma lectularia, Triatoma lenti, Triatoma maculata, Triatoma matogrossensis, Triatoma melanica, Triatoma melanocephala, Triatoma petrochiae, Triatoma platensis, Triatoma protracta, Triatoma pseudomaculata, Triatoma pintodiasi, Triatoma rubrovaria, Triatoma sherlocki, Triatoma sordida, Triatoma tibiamaculata, Triatoma vandae, Triatoma vitticeps, Triatoma williami, Triatoma wygodzinskyi) that were provided by the “Insetário de Triatominae,” from Faculdade de Ciências Farmacêuticas, Universidade Estadual Paulista, Araraquara, São Paulo, Brazil, and by the “Insetário do Laboratório Nacional e Internacional de Referência em Taxonomia de Triatomíneos,” from Fundação Oswaldo Cruz, Rio de Janeiro, Brazil. The bugs were dissected, the testicles were removed, the slides were mounted through cell squashing, and stained through silver impregnation.10

The analysis of the nucleolar behavior during the meiosis of these bugs has highlighted that all species feature the nucleolar persistence phenomenon. Therefore, as the observed results were the same for all triatomines, we represent the nucleologenesis (Figure 1AD) and the nucleolar persistence phenomenon (Figure 1C and D, arrow) with D. maxima. Our results are in agreement with the initial works carried out for the Triatoma,1117 Meccus,18 Rhodnius,9,19 and Panstrongylus9 genera, which likewise have exhibited this phenomenon during spermatogenesis of these vectors.

Figure 1.
Figure 1.

Nucleologenesis of Dipetalogaster maxima (AD). Note the nucleolar persistence phenomenon (C, D, arrow). Bar = 10 μm.

Citation: The American Society of Tropical Medicine and Hygiene 95, 5; 10.4269/ajtmh.16-0149

Recently, it has been suggested that the nucleolar persistence can be essential for the spermiogenesis of these vectors, since during this differentiation stage, the nucleolus—although present—is inactive by epigenetic factors, and the chromatoid body (CB) is an essential organelle to supply all the transcriptional activity required for spermiogenesis.20 Thus, it has been suggested that the nucleolus that persists during meiosis can be a structure responsible for forming the CB, because if it is active, all the transcribed RNA is directed to the formation of the CB.17

Therefore, we describe the nucleolar persistence as a synapomorphy of the triatomines and suggest that further analysis should be conducted to assess whether the persistent nucleolus is really active, thus clarifying the importance of the persistence of the nucleolus for spermatogenesis of these vectors of Chagas disease.

  • 1.

    World Health Organization, 2015. Chagas disease (American trypanosomiasis). Wkly Epidemiol Rec 90: 3344.

  • 2.

    Almeida CE, Duarte R, Guerra do Nascimento R, Pacheco RS, Costa J, 2002. Triatoma rubrovaria (Blanchard, 1843) (Hemiptera, Reduviidae, Triatominae) II: trophic resources and ecological observations of five populations collected in the state of Rio Grande do Sul, Brazil. Mem Inst Oswaldo Cruz 97: 11271131.

    • Search Google Scholar
    • Export Citation
  • 3.

    Almeida CE, Folly-Ramos E, Agapito-Souza R, Magno-Esperança G, Pacheco RS, Costa J, 2005. Triatoma rubrovaria (Blanchart, 1843) (Hemiptera-Reduviidae-Triatominae) IV: bionomic aspects on the vector capacity of nymphs. Mem Inst Oswaldo Cruz 100: 231235.

    • Search Google Scholar
    • Export Citation
  • 4.

    Alevi KCC, Rosa JA, Azeredo-Oliveira MTV, 2014. Spermatogenesis in Triatoma melanica Neiva and Lenti, 1941 (Hemiptera, Triatominae). J Vector Ecol 39: 231233.

    • Search Google Scholar
    • Export Citation
  • 5.

    Alevi KCC, Castro NFC, Oliveira J, Rosa JA, Azeredo-Oliveira MTV, 2015. Cystic spermatogenesis in three species of the prolixus complex (Hemiptera: Triatominae). Ital J Zool (Modena) 82: 172178.

    • Search Google Scholar
    • Export Citation
  • 6.

    Morielle-Souza A, Taboga SR, Azeredo-Oliveira MTV, 2010. Ultrastructural analysis of the nucleolar aspects at spermiogenesis in triatomines (Heteroptera, Triatominae). Micron 41: 791796.

    • Search Google Scholar
    • Export Citation
  • 7.

    Alevi KCC, Guerra AL, Imperador CHL, Rosa JA, Azeredo-Oliveira MTV, 2016. Reproductive biology of Triatoma brasiliensis (Hemiptera, Triatominae) during the imaginal molt. Am J Trop Med Hyg 94: 689690.

    • Search Google Scholar
    • Export Citation
  • 8.

    Tartarotti E, Azeredo-Oliveira MTV, 1999. Patterns of nucleolar activity during spermatogenesis of two triatomines, Panstrongylus megistus and P. herreri. Caryologia 52: 177184.

    • Search Google Scholar
    • Export Citation
  • 9.

    Alevi KCC, Castro NFC, Lima ACC, Ravazi A, Morielle-Souza A, Oliveira J, Rosa JA, Azeredo-Oliveira MTV, 2014. Nucleolar persistence during spermatogenesis of the genus Rhodnius (Hemiptera, Triatominae). Cell Biol Int 38: 977980.

    • Search Google Scholar
    • Export Citation
  • 10.

    Howell WM, Black DA, 1980. Controlled silver-staining of nucleolus organizer regions with a protective colloidal developer: a 1-step method. Experientia 36: 10141015.

    • Search Google Scholar
    • Export Citation
  • 11.

    Costa LC, Azeredo-Oliveira MTV, Tartarotti E, 2008. Spermatogenesis and nucleolar activity in Triatoma klugi (Triatomine, Heteroptera). Genet Mol Biol 31: 438444.

    • Search Google Scholar
    • Export Citation
  • 12.

    Alevi KCC, Mendonça PP, Pereira NP, Rosa JA, Azeredo-Oliveira MTV, 2013. Análise das possíveis regiões organizadoras nucleolares e da atividade nucleolar em Triatoma melanocephala e T. lenti, importantes vetores da doença de Chagas. Rev Cienc Farm Básica e Apl 34: 417421.

    • Search Google Scholar
    • Export Citation
  • 13.

    Bardella VB, Azeredo-Oliveira MTV, Tartarotti E, 2008. Cytogenetic analysis in the spermatogenesis of Triatoma melanosoma (Reduviidae; Heteroptera). Genet Mol Res 7: 326335.

    • Search Google Scholar
    • Export Citation
  • 14.

    Severi-Aguiar GDC, Azeredo-Oliveira MTV, 2005. Cytogenetic study on three species of the genus Triatoma (Heteroptera: Reduviidae) with emphasis on nucleolar organizer regions. Caryologia 58: 293299.

    • Search Google Scholar
    • Export Citation
  • 15.

    Severi-Aguiar GDC, Lourenço LB, Bicudo HEMC, Azeredo-Oliveira MTV, 2006. Meiosis aspects and nucleolar activity in Triatoma vitticeps (Triatominae, Heteroptera). Genetica 126: 141151.

    • Search Google Scholar
    • Export Citation
  • 16.

    Pereira NP, Alevi KCC, Mendonça PP, Azeredo-Oliveira MTV, 2015. Spermatogenesis and nucleolar behavior in Triatoma vandae and Triatoma williami (Hemiptera, Triatominae). Genet Mol Res 14: 1214512151.

    • Search Google Scholar
    • Export Citation
  • 17.

    Borgueti AO, Alevi KCC, Silistino-Souza R, Rosa JA, Azeredo-Oliveira MTV, 2015. Immunofluorescence and ultrastructural analysis of the chromatoid body during spermatogenesis of Triatoma platensis and T. rubrovaria (Hemiptera, Triatominae). Micron 74: 4446.

    • Search Google Scholar
    • Export Citation
  • 18.

    Madeira FF, Lima ACC, Rosa JA, Azeredo-Oliveira MTV, Alevi KCC, 2016. Nucleolar persistence phenomenon during spermatogenesis in genus Meccus (Hemiptera, Triatominae). Genet Mol Res 15. doi:10.4238/gmr.15017427.

    • Search Google Scholar
    • Export Citation
  • 19.

    Morielle A, Azeredo-Oliveira MTV, 2004. Description of the nucleolar activity and karyotype in germinative cell lines of Rhodnius domesticus (Triatominae, Heteroptera). Caryologia 57: 3137.

    • Search Google Scholar
    • Export Citation
  • 20.

    Alevi KCC, Mendonça PP, Pereira NP, Rosa JA, Azeredo-Oliveira MTV, 2014. Is there post-meiotic transcriptional activity during hemipteran spermiogenesis? Invertebr Reprod Dev 58: 193198.

    • Search Google Scholar
    • Export Citation

Author Notes

* Address correspondence to Kaio Cesar Chaboli Alevi, Instituto de Biociências, Letras e Ciências Exatas, Universidade Estadual Paulista (IBILCE–UNESP), Rua Cristovão Colombo, 2265, Jardim Nazareth, CEP 15054-000, São José do Rio Preto, São Paulo, Brazil. E-mail: kaiochaboli@hotmail.com

Financial support: The study was supported by Fundação de Amparo à Pesquisa do Estado de São Paulo (process numbers 2013/19764-0 and 2015/14762-5-FAPESP, Brazil) and Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq, Brazil).

Authors' addresses: Fernanda Fernandez Madeira, Kelly Cristine Borsatto, Anna Claudia Campaner Lima, Amanda Ravazi, Maria Tercília Vilela Azeredo-Oliveira, and Kaio Cesar Chaboli Alevi, Departamento de Biologia, Instituto de Biociencias, Letras e Ciencias Exatas, Universidade Estadual Paulista Julio de Mesquita Filho–Campus de Sao Jose do Rio Preto, São Paulo, Brazil, E-mails: fernanda.bio56@hotmail.com, kellyborsatto@gmail.com, anna_campaner@hotmail.com, amandaravazi95@gmail.com, tercilia@ibilce.unesp.br, and kaiochaboli@hotmail.com. Jader de Oliveira and João Aristeu da Rosa, Departamento de Ciências Biológicas, Faculdade de Ciencias Farmaceuticas, Universidade Estadual Paulista Julio de Mesquita Filho–Campus de Araraquara, São Paulo, Brazil, E-mails: jdr.oliveira@hotmail.com and joaoaristeu@gmail.com.

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