Constitutive expression and enzymatic activity of Tan protein in brain and epidermis of Ceratitis capitata and of Drosophila melanogaster wild-type and tan mutants

Pérez, Martín M.; Sabio, Germán; Badaracco, Alejandra; Quesada-Allué, Luis A.

doi:10.1016/j.ibmb.2011.04.007

Cited by 9 publications

(11 citation statements)

References 29 publications

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“…Moreover, NBAD-synthase can metabolize other biogenic amines such as octopamine, tyramine, histamine and 5-HT, which are also involved in the regulation of locomotor activity and sleep in flies (Yuan et al, 2006;Crocker & Sehgal, 2008;Chen et al, 2013;Oh et al, 2013). The NBAD-hydrolase enzyme is also able to hydrolyze other β-alanyl derivatives (True et al, 2005;Pérez et al, 2011). The possibility that β-alanyl derivatives of these amines might be simultaneously synthesized and hydrolyzed by the NBAD-synthase and NBAD-hydrolase enzymes, respectively, cannot be excluded and therefore should be explored in future studies.…”

Section: Discussionmentioning

confidence: 99%

“…The ebony mutant is defective for the synthesis of N-β-alanyl derivatives (Wright, 1987;Pérez et al, 1997Pérez et al, , 2002Pérez et al, , 2004, the best known being N-β-alanyldopamine (NBAD) and N-β-alanylhistamine (carcinine). The tan mutants are unable to hydrolyze N-β-alanyl derivatives (Wright, 1987;True et al, 2005;Pérez et al, 2011). Both mutants have altered levels of neurotransmitters; the NBAD concentration is drastically reduced in homozygous ebony flies compared with wild-type, whereas DA levels are elevated approximately two-fold (Hodgetts & Konopka, 1973;Walter et al, 1996;Gruntenko et al, 2004).…”

Section: Introductionmentioning

confidence: 99%

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N‐β‐alanyldopamine metabolism, locomotor activity and sleep in Drosophila melanogaster ebony and tan mutants

et al. 2015

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Drosophila melanogaster Meigen mutants for N-β-alanyldopamine (NBAD) metabolism have altered levels of NBAD, dopamine and other neurotransmitters. The ebony 1 mutant strain has very low levels of NBAD and higher levels of dopamine, whereas the opposite situation is observed in the tan 1 mutant. Dopamine is implicated in the control of movement, memory and arousal, as well as in the regulation of sleep and wakefulness in D. melanogaster. N-β-alanyldopamine, which is best known as a cuticle cross-linking agent, is also present in nervous tissue and has been proposed to promote locomotor activity in this fly. The daily locomotor activity and the sleep patterns of ebony 1 and tan 1 mutants are analyzed, and are compared with wild-type flies. The tan 1 mutant shows reduced locomotor activity, whereas ebony 1 shows higher levels of activity than wild-type flies, suggesting that NBAD does not promote locomotor activity. Both mutants spend less time asleep than wild-type flies during night-time; ebony shows more consolidated activity during night-time and increased sleep latency, whereas tan is unable to consolidate locomotor activity and sleep in either phase of the day. The daily level of NBAD-synthase activity is measured in vitro using wild-type and tan 1 protein extracts, and the lowest NBAD synthesis is observed at the time of higher locomotor activity. The abnormalities in several parameters of the waking/sleep cycle indicate some dysfunction in the processes that regulates these behaviours in both mutants.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

N‐β‐alanyldopamine metabolism, locomotor activity and sleep in Drosophila melanogaster ebony and tan mutants

et al. 2015

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“…However, our studies in NBAD metabolism in C. capitata suggest that brownish spots in this fly would be tanned mainly by NBAD. We previously reported that the enzymatic activity responsible for the synthesis of NBAD in C. capitata epidermis is detected during narrow temporal windows, occurring during the sclerotization of larval cuticle to form the puparium, as well as during the pharate period and the first hours following imago ecdysis (Perez et al, 2004;2011).…”

Section: Introductionmentioning

confidence: 99%

Extracellular activity of NBAD-synthase is responsible for colouration of brown spots in Ceratitis capitata wings

Pérez

Bochicchio

Rabossi

et al. 2018

Journal of Insect Physiology

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After the emergence of the Ceratitis capitata imago, the pale and folded wings are expanded and sclerotized to acquire the definitive form and to stabilize the cuticle. The wings of this fly show a specific pattern of brownish and black spots. Black spots are pigmented by melanin, whereas there was scarce information about the development of the brownish spots. N-beta-alanydopamine (NBAD) is the main tanning precursor in C. capitata body cuticle, and we hypothesized that it may be responsible for the colouration of the brownish spots. We determined the topology and timing of NBAD synthesis and deposition to attain the species-specific colouration pattern. We demonstrated that during the first hours the colour of the brownish spots was principally determined by the tanning of the hairs. Haemolymph circulation through the veins is required to tan the wings. We confirmed that soon after wing spreading, most of the wing epidermal cells disappeared. Thus, the tanning of the brown spots was accomplished when the wing lamina was devoid of cells. NBAD synthase (NBAD-S; Ebony protein in D. melanogaster) activity in wings was detected in pharate adults and lasted several days after the emergence, even after the end of the tanning process. This observation is in contrast to epidermal NBAD-S activity in the body, where it was nearly undetectable 48 h post emergence. Our results indicate that NBAD-S was exported and deposited into the extracellular matrix of the brown spot areas before cell death and that tanning occurs through gradual export of NBAD precursors (dopamine and b-alanine) from veins.

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“…In Drosophila, Oncopeltus, Tribolium, Bombyx and V. cardui, e (ebony) and bl (black) KDs produce animals with dark cuticle due to loss of dopamine conversion to the tan/yellow pigment, leaving Dopamine available for the conversion to black pigment [7,13,15,46,51]. Conversely, loss of t (tan) function results in light pigmented cuticles in Drosophila, O. fasciatus, N. lugens and C. capitata [7,46,52]. The functions of the e and t genes in blood feeding insects has not been investigated yet, although a study in A. gambiae showed that a bl allele generates animals with dark cuticle and reduced fertility and vigor [53].…”

Section: An Alternative Strategy For Cuticle Pigmentation In R Prolixusmentioning

confidence: 99%

“…NBAD synthase and NBAD hydrolase, encoded by e and t, respectively, have been implicated in several different processes. They exert a role in brain neurotransmitter metabolism [52], and e is expressed in the foregut and tracheal epidermis in D. melanogaster [51]. Their effect on tyrosine detoxification, which could also explain the loss of viability, has not been explored.…”

Section: An Alternative Strategy For Cuticle Pigmentation In R Prolixusmentioning

confidence: 99%

Pigmentation loci as markers for genome editing in the Chagas disease vector Rhodnius prolixus

Berni

Bressan

Simão

et al. 2020

Preprint

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The kissing bug Rhodnius prolixus is a major vector for Chagas disease in the Americas, and also considered as the primary model for functional studies.Prospective transgenic approaches and genome editing strategies hold great promise for controlling insect populations as well as disease propagation. In this context, identifying visible genetic markers for transgenic methodologies is of paramount importance to advance the field. Here we have identified and analyzed the function of putative cuticle and eye color genes by investigating the effect of gene knockdown on fertility, viability, and the generation of visible phenotypes. Synthesis of the dark, yellow and tan pigments present in the cuticle of most insects depends on the function of key genes encoding enzymes in the tyrosine pathway. Knockdown of the R. prolixus yellow and aaNAT/pro orthologs produces striking alterations in cuticle color.Surprisingly, knockdown of ebony does not generate visible phenotypes. Since loss of ebony function results in a dark cuticle in several insect orders, we conclude that R. prolixus evolved alternative strategies for cuticle coloration, possibly including the loss of a pigmentation function for an entire branch of the tyrosine pathway. Knockdown of the scarlet and brown genes -encoding ABC transporters -alters cuticle and eye pigmentation, implying that the transport of pigment into proper organelles is an important process both for cuticle and eye coloration in this species. Therefore, this analysis identifies for the first time potential visible markers for transgenesis in a hemipteran vector for a debilitating human disease. Author SummaryThe hemipteran Rhodnius prolixus -also known as a kissing bug -is a main vector transmitting the parasite Trypanosoma cruzi, the causative agent of Chagas disease, a debilitating infection estimated to affect more than 6 million people in Central and South America. In order to limit disease spread, an important measure is insect vector control. However, kissing bugs -like other insects -develop resistance to insecticides.Alternative strategies based on transgenesis and the recently developed CRISPRbased genome edition hold great promise to control vector population or generate parasite-resistant insects. For these approaches to be feasible in R. prolixus, it is critical to identify visible phenotypic markers. Here we identify and describe several genes controlling cuticle and eye pigmentation that are well-suited putative landing sites for transformation strategies. Among these, loss-of-function mutations in the ABC transporter encoding scarlet and the tyrosine pathway enzyme encoding aaNAT/pro generate striking and easily visible phenotypes. Importantly, the knockdown of these genes does not affect insect viability and fertility under laboratory conditions. Our results suggest that R. prolixus has developed alternative strategies for cuticle coloration involving the loss of an entire branch of tanning loci, while the other branch producing cuticle patterns by generating non-pigmented a...

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Constitutive expression and enzymatic activity of Tan protein in brain and epidermis of Ceratitis capitata and of Drosophila melanogaster wild-type and tan mutants

Cited by 9 publications

References 29 publications

N‐β‐alanyldopamine metabolism, locomotor activity and sleep in Drosophila melanogaster ebony and tan mutants

N‐β‐alanyldopamine metabolism, locomotor activity and sleep in Drosophila melanogaster ebony and tan mutants

Extracellular activity of NBAD-synthase is responsible for colouration of brown spots in Ceratitis capitata wings

Pigmentation loci as markers for genome editing in the Chagas disease vector Rhodnius prolixus

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