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Article ; Online: Visualization of Apoptotic Ovarian Follicles during

Isoe, Jun / Miesfeld, Roger L / Riehle, Michael A

Cold Spring Harbor protocols

2023

Abstract: In insects, oocyte resorption (oosorption) or follicular atresia is one of the key physiological processes and evolutionary strategies used to optimize reproductive fitness. Mosquitoes are ideal model organisms for studying egg maturation in arthropods, ... ...

Abstract	In insects, oocyte resorption (oosorption) or follicular atresia is one of the key physiological processes and evolutionary strategies used to optimize reproductive fitness. Mosquitoes are ideal model organisms for studying egg maturation in arthropods, as their follicle development is initiated only following the ingestion of a blood meal, followed by a carefully orchestrated series of hormonally regulated events leading to egg maturation. A cohort of approximately 100 follicles per mosquito ovary begin developing synchronously. However, a significant fraction of follicles ultimately undergo apoptosis and oosorption, especially when available resources from the blood meal are limited. Therefore, simple, rapid, and reliable techniques to accurately evaluate follicular atresia are necessary to understand mechanisms underlying follicle development in insects. This protocol describes how to detect apoptotic follicle cells within the
Language	English
Publishing date	2023-12-13
Publishing country	United States
Document type	Journal Article
ISSN	1559-6095
ISSN (online)	1559-6095
DOI	10.1101/pdb.prot108226
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Mosquito Egg Development and Eggshell Formation.

Isoe, Jun / Riehle, Michael A / Miesfeld, Roger L

Cold Spring Harbor protocols

2023

Abstract: Anautogenous female mosquitoes, which ingest a blood meal from warm-blooded vertebrates to produce eggs, have become a valuable model organism for investigating signaling pathways and physiological processes that occur during egg development. Different ... ...

Abstract	Anautogenous female mosquitoes, which ingest a blood meal from warm-blooded vertebrates to produce eggs, have become a valuable model organism for investigating signaling pathways and physiological processes that occur during egg development. Different molecular pathways tightly regulate the initiation of egg development and are governed by a balance among different insect hormones. Gravid (mature egg-carrying) females deposit fully developed eggs at the end of each gonotrophic cycle, which is defined as the time interval between the ingestion of a blood meal to oviposition. An intact eggshell protects the oocyte and embryo inside from external factors such as desiccation, physical damage, etc., and the various eggshell proteins are spatially and temporary deposited during oogenesis. Additionally, follicle resorption (oosorption) during blood meal-induced mosquito ovarian follicle development is an adapted physiological process that optimizes reproductive fitness. Mosquito oocytes grow and mature synchronously throughout oogenesis; however, during the later stages of oogenesis, some oocytes may undergo oosorption if sufficient nutrients are unavailable. This introduction highlights how mosquito egg development can be used to investigate follicular resorption and identify proteins involved in eggshell formation in
Language	English
Publishing date	2023-12-13
Publishing country	United States
Document type	Journal Article
ISSN	1559-6095
ISSN (online)	1559-6095
DOI	10.1101/pdb.top107669
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Article ; Online: Starving the Beast: Limiting Coenzyme A Biosynthesis to Prevent Disease and Transmission in Malaria.

Riske, Brendan F / Luckhart, Shirley / Riehle, Michael A

International journal of molecular sciences

2023 Volume 24, Issue 18

Abstract: Malaria parasites must acquire all necessary nutrients from the vertebrate and mosquito hosts to successfully complete their life cycle. Failure to acquire these nutrients can limit or even block parasite development and presents a novel target for ... ...

Abstract	Malaria parasites must acquire all necessary nutrients from the vertebrate and mosquito hosts to successfully complete their life cycle. Failure to acquire these nutrients can limit or even block parasite development and presents a novel target for malaria control. One such essential nutrient is pantothenate, also known as vitamin B5, which the parasite cannot synthesize de novo and is required for the synthesis of coenzyme A (CoA) in the parasite. This review examines pantothenate and the CoA biosynthesis pathway in the human-mosquito-malaria parasite triad and explores possible approaches to leverage the CoA biosynthesis pathway to limit malaria parasite development in both human and mosquito hosts. This includes a discussion of sources for pantothenate for the mosquito, human, and parasite, examining the diverse strategies used by the parasite to acquire substrates for CoA synthesis across life stages and host resource pools and a discussion of drugs and alternative approaches being studied to disrupt CoA biosynthesis in the parasite. The latter includes antimalarial pantothenate analogs, known as pantothenamides, that have been developed to target this pathway during the human erythrocytic stages. In addition to these parasite-targeted drugs, we review studies of mosquito-targeted allosteric enzymatic regulators known as pantazines as an approach to limit pantothenate availability in the mosquito and subsequently deprive the parasite of this essential nutrient.
Language	English
Publishing date	2023-09-10
Publishing country	Switzerland
Document type	Journal Article ; Review
ZDB-ID	2019364-6
ISSN	1422-0067 ; 1422-0067 ; 1661-6596
ISSN (online)	1422-0067
ISSN	1422-0067 ; 1661-6596
DOI	10.3390/ijms241813915
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Article ; Online: Midgut Mitochondrial Function as a Gatekeeper for Malaria Parasite Infection and Development in the Mosquito Host.

Luckhart, Shirley / Riehle, Michael A

Frontiers in cellular and infection microbiology

2020 Volume 10, Page(s) 593159

Abstract: Across diverse organisms, various physiologies are profoundly regulated by mitochondrial function, which is defined by mitochondrial fusion, biogenesis, oxidative phosphorylation (OXPHOS), and mitophagy. Based on our data and significant published ... ...

Abstract	Across diverse organisms, various physiologies are profoundly regulated by mitochondrial function, which is defined by mitochondrial fusion, biogenesis, oxidative phosphorylation (OXPHOS), and mitophagy. Based on our data and significant published studies from
MeSH term(s)	Animals ; Anopheles ; Drosophila melanogaster ; Host-Parasite Interactions ; Malaria ; Mitochondria ; Mosquito Vectors
Language	English
Publishing date	2020-12-11
Publishing country	Switzerland
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Review
ZDB-ID	2619676-1
ISSN	2235-2988 ; 2235-2988
ISSN (online)	2235-2988
ISSN	2235-2988
DOI	10.3389/fcimb.2020.593159
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Article ; Online: Chemical Probes to Interrogate the Extreme Environment of Mosquito Larval Guts.

Guzmán, Lindsay E / Wijetunge, Anjalee N / Riske, Brendan F / Massani, Brooke B / Riehle, Michael A / Jewett, John C

Journal of the American Chemical Society

2024 Volume 146, Issue 12, Page(s) 8480–8485

Abstract: Mosquito control methods are vital to curtail the spread of life-threatening illnesses, such as dengue fever, malaria, and yellow fever. Vector control technologies must be selective to minimize deleterious effects on our ecosystem. Successful methods ... ...

Abstract	Mosquito control methods are vital to curtail the spread of life-threatening illnesses, such as dengue fever, malaria, and yellow fever. Vector control technologies must be selective to minimize deleterious effects on our ecosystem. Successful methods that control mosquito larva populations utilize the uniquely high alkaline nature of the midgut. Here, we present novel protected triazabutadienes (pTBD) that are deprotected under basic conditions of the larval midgut, releasing an aryl diazonium ion (ADI) that results in protein modification. The probes contain a bioorthogonal terminal alkyne handle, enabling a selective Cu-click reaction with an azidofluorophore for quantification by SDS PAGE and visualization using fluorescence microscopy. A control TBD, unable to release an ADI, did not label the midgut. We envision our chemical probes will aid in the development of new selective mosquito control methods, thus preventing the spread of mosquito-borne illnesses with minimal impact on other organisms in the ecosystem.
MeSH term(s)	Animals ; Ecosystem ; Larva ; Extreme Environments ; Mosquito Control/methods ; Malaria/prevention & control
Language	English
Publishing date	2024-03-14
Publishing country	United States
Document type	Journal Article
ZDB-ID	3155-0
ISSN	1520-5126 ; 0002-7863
ISSN (online)	1520-5126
ISSN	0002-7863
DOI	10.1021/jacs.3c14598
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Article ; Online: Identification of Mosquito Eggshell Proteins from

Isoe, Jun / Parsawar, Krishna / David, Cynthia L / Riehle, Michael A / Miesfeld, Roger L

Cold Spring Harbor protocols

2023

Abstract: The insect eggshell is a multifunctional structure with several important roles, including generating an entry point for sperm via the micropyle before oviposition, serving as an oviposition substrate attachment surface, and functioning as a protective ... ...

Abstract	The insect eggshell is a multifunctional structure with several important roles, including generating an entry point for sperm via the micropyle before oviposition, serving as an oviposition substrate attachment surface, and functioning as a protective layer during embryo development. Eggshell proteins play major roles in eggshell tanning and hardening following oviposition and provide molecular cues that define dorsal-ventral axis formation. Precise eggshell formation during ovarian follicle maturation is critical for normal embryo development and the synthesis of a defective eggshell often gives rise to inviable embryos. Therefore, simple and accurate methods for identifying eggshell proteins will facilitate our understanding of the molecular pathways regulating eggshell formation and the mechanisms underlying normal embryo development. This protocol describes how to isolate and enrich eggshells from mature oocytes of
Language	English
Publishing date	2023-12-13
Publishing country	United States
Document type	Journal Article
ISSN	1559-6095
ISSN (online)	1559-6095
DOI	10.1101/pdb.prot108227
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Genetic Variation in

Brophy, Maureen / Riehle, Michael A / Mastrud, Nikki / Ravenscraft, Alison / Adamson, Johnathan E / Walker, Kathleen R

International journal of environmental research and public health

2022 Volume 19, Issue 7

Abstract: Rhipicephalus ... ...

Abstract	Rhipicephalus sanguineus
MeSH term(s)	Animals ; Arizona ; Dog Diseases/epidemiology ; Dog Diseases/genetics ; Dogs ; Genetic Variation ; Phylogeny ; Rhipicephalus sanguineus/genetics ; Southwestern United States
Language	English
Publishing date	2022-04-01
Publishing country	Switzerland
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2175195-X
ISSN	1660-4601 ; 1661-7827
ISSN (online)	1660-4601
ISSN	1661-7827
DOI	10.3390/ijerph19074223
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Article ; Online: Characterization of essential eggshell proteins from Aedes aegypti mosquitoes.

Isoe, Jun / Simington, Carter J / Oscherwitz, Max E / Peterson, Alyssa J / Rascón, Alberto A / Massani, Brooke B / Miesfeld, Roger L / Riehle, Michael A

BMC biology

2023 Volume 21, Issue 1, Page(s) 214

Abstract: Background: Up to 40% of the world population live in areas where mosquitoes capable of transmitting the dengue virus, including Aedes aegypti, coexist with humans. Understanding how mosquito egg development and oviposition are regulated at the ... ...

Abstract	Background: Up to 40% of the world population live in areas where mosquitoes capable of transmitting the dengue virus, including Aedes aegypti, coexist with humans. Understanding how mosquito egg development and oviposition are regulated at the molecular level may provide new insights into novel mosquito control strategies. Previously, we identified a protein named eggshell organizing factor 1 (EOF1) that when knocked down with RNA interference (RNAi) resulted in non-melanized and fragile eggs that did not contain viable embryos. Results: In this current study, we performed a comprehensive RNAi screen of putative A. aegypti eggshell proteins to identify additional proteins that interact with intracellular EOF1. We identified several proteins essential for eggshell formation in A. aegypti and characterized their phenotypes through a combination of molecular and biochemical approaches. We found that Nasrat, Closca, and Polehole structural proteins, together with the Nudel serine protease, are indispensable for eggshell melanization and egg viability. While all four proteins are predominantly expressed in ovaries of adult females, Nudel messenger RNA (mRNA) expression is highly upregulated in response to blood feeding. Furthermore, we identified four additional secreted eggshell enzymes that regulated mosquito eggshell formation and melanization. These enzymes included three dopachrome-converting enzymes (DCEs) and one cysteine protease. All eight of these eggshell proteins were essential for proper eggshell formation. Interestingly, their eggshell surface topologies in response to RNAi did not phenocopy the effect of RNAi-EOF1, suggesting that additional mechanisms may influence how EOF1 regulates eggshell formation and melanization. Conclusions: While our studies did not identify a definitive regulator of EOF1, we did identify eight additional proteins involved in mosquito eggshell formation that may be leveraged for future control strategies.
MeSH term(s)	Animals ; Humans ; Female ; Aedes/genetics ; Egg Proteins/genetics ; Egg Proteins/metabolism ; RNA Interference ; Ovary/metabolism
Chemical Substances	chorion proteins ; Egg Proteins
Language	English
Publishing date	2023-10-13
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2133020-7
ISSN	1741-7007 ; 1741-7007
ISSN (online)	1741-7007
ISSN	1741-7007
DOI	10.1186/s12915-023-01721-z
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Article: Chemical probes to interrogate the extreme environment of mosquito larval guts.

Guzmán, Lindsay E / Wijetunge, Anjalee N / Riske, Brendan F / Massani, Brooke B / Riehle, Michael A / Jewett, John C

bioRxiv : the preprint server for biology

2023

Abstract: Mosquito control methods are vital for the spread of life-threatening illnesses such as dengue fever, malaria, and yellow fever. Vector control technologies must be selective to minimize deleterious effects to our ecosystem. Successful methods that ... ...

Abstract	Mosquito control methods are vital for the spread of life-threatening illnesses such as dengue fever, malaria, and yellow fever. Vector control technologies must be selective to minimize deleterious effects to our ecosystem. Successful methods that control mosquito larva populations utilize the uniquely high alkaline nature of the midgut. Here, we present novel protected triazabutadienes (pTBD) which are deprotected under basic conditions of the larval midgut, releasing an aryl diazonium ion (ADI) that results in protein modification. The probes contain a bioorthogonal terminal alkyne handle, enabling a selective Cu-click reaction with an azido-fluorophore for quantification by SDS PAGE and visualization using fluorescence microscopy. A control TBD, unable to release an ADI, did not label the midgut. We envision our chemical probes will aid in the development of new selective mosquito control methods thus preventing the spread of mosquito-borne illnesses with minimal impact on other organisms in the ecosystem.
Language	English
Publishing date	2023-12-27
Publishing country	United States
Document type	Preprint
DOI	10.1101/2023.12.27.573438
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Article ; Online: Increased Akt signaling in the fat body of Anopheles stephensi extends lifespan and increases lifetime fecundity through modulation of insulin-like peptides.

Hun, Lewis V / Luckhart, Shirley / Riehle, Michael A

Journal of insect physiology

2019 Volume 118, Page(s) 103932

Abstract: Insulin-like peptides (ILPs) and the insulin/insulin-like growth factor 1 signaling (IIS) cascade regulate numerous physiological functions, including lifespan, reproduction, immunity, and metabolism, in diverse eukaryotes. We previously demonstrated ... ...

Abstract	Insulin-like peptides (ILPs) and the insulin/insulin-like growth factor 1 signaling (IIS) cascade regulate numerous physiological functions, including lifespan, reproduction, immunity, and metabolism, in diverse eukaryotes. We previously demonstrated that in female Anopheles stephensi and Aedes aegypti mosquitoes, activation of the IIS cascade in the fat body led to a significant increase in lifespan. In this work, we elucidated two putative mechanisms in A. stephensi behind the observed lifespan extension and assessed whether this lifespan extension confers an overall fitness advantage to the mosquito. Specifically, we demonstrated that increased Akt signaling in the mosquito fat body following a blood meal significantly suppressed the expression of ILP2 in the head. Moreover, overexpression of active Akt in the fat body altered the expression of a putative insulin binding protein ortholog, Imaginal morphogenesis protein-Late 2 (Imp-L2), in response to transgene expression. Combined, these two factors may act to reduce overall levels of circulating ILP2 or other ILPs in the mosquito, in turn conferring increased survival. We also examined the impact increased fat body IIS had on lifetime fecundity and demonstrated that transgenic female mosquito populations had higher lifetime fecundity relative to non-transgenic sibling controls. These studies provide new insights into the complex hormonal and molecular mechanisms regulating the interplay between IIS, aging, and reproduction in this important vector of human malaria parasites.
MeSH term(s)	Aging/genetics ; Aging/metabolism ; Animals ; Animals, Genetically Modified ; Anopheles/genetics ; Anopheles/growth & development ; Anopheles/metabolism ; Blood ; Fat Body/metabolism ; Female ; Fertility/genetics ; Fertility/physiology ; Humans ; Insect Proteins/genetics ; Insect Proteins/metabolism ; Insulin-Like Growth Factor I/metabolism ; Longevity/genetics ; Longevity/physiology ; Proto-Oncogene Proteins c-akt/metabolism ; Signal Transduction
Chemical Substances	Insect Proteins ; Insulin-Like Growth Factor I (67763-96-6) ; Proto-Oncogene Proteins c-akt (EC 2.7.11.1)
Language	English
Publishing date	2019-08-22
Publishing country	England
Document type	Journal Article ; Research Support, N.I.H., Extramural
ISSN	1879-1611
ISSN (online)	1879-1611
DOI	10.1016/j.jinsphys.2019.103932
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