Cytological Analysis of Spermatogenesis: Live and Fixed Preparations of &lt;em&gt;Drosophila&lt;/em&gt; Testes

Sitaram, Poojitha; Hainline, Sarah G.; Lee, Laura Anne

doi:10.3791/51058-v

Cited by 3 publications

(2 citation statements)

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“…Under optimal conditions, 2‐ to 5‐day‐old males are expected to display mature sperm. By measuring SV size and the occurrence of sperm at different male ages, we can infer sperm maturation dynamics (Sitaram et al, 2014). All observations were done under a microscope (ZEISS, AxioVision Software) with an Olympus SC50 5‐megapixel color microscope camera at a 50X amplification.…”

Section: Methodsmentioning

confidence: 99%

Recovery from heat‐induced infertility—A study of reproductive tissue responses and fitness consequences in male Drosophila melanogaster

Domenech

Fricke

2022

Ecology and Evolution

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Temperature is a critical abiotic factor for many organisms and can turn into an environmental stressor, particularly for ectotherms. Elevated temperatures are known to affect an ectotherm's physiology, behavior, and, on a broader scale, its overall performance (Huey & Stevenson, 1979). When critical thermal limits are exceeded, both viability and reproductive potential are harmed (Sales et al., 2021). In recent years, most research assessing phenotypic effects under increasing temperatures has incorporated mainly

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Section: Methodsmentioning

confidence: 99%

Recovery from heat‐induced infertility—A study of reproductive tissue responses and fitness consequences in male Drosophila melanogaster

Domenech

Fricke

2022

Ecology and Evolution

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“…4a). To test whether spermatogenesis and mature sperm count were affected, testes 24,25 , and sperm numbers in the seminal vesicle 26 were evaluated (Fig. 4b).…”

Section: Fa Supplementation Rescues Mr Induced Sperm Viability and Tr...mentioning

confidence: 99%

Uncoupling of lifespan and reproductive tradeoffs by dietary methionine and one-carbon metabolism

Wei,

Liu,

Liu

et al. 2023

Preprint

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Aging has been proposed to be a consequence of reproductive ability and longevity thus occurs as a tradeoff with organismal reproduction1, 2. Lifespan extending interventions generally at the expense of fertility3. How this principle extends to nutrition and metabolism is not understood. We considered dietary methionine restriction (MR) that is linked to one carbon metabolism as well as to Mediterranean or plant-based diets4 ,5 and known to influence cancer6, metabolic health7, and longevity3, 8. Using a chemically defined diet (CDD) we developed for Drosophila melanogaster, we found that MR-mediated lifespan extension indeed occurs at the expense of reproduction. A survey of the nutritional landscape in the background of MR revealed that folic acid, a vitamin linked to one carbon metabolism, surprisingly was the lone nutrient that restored reproductive defects without compromising lifespan extension. In vivo isotope tracing, metabolomics and flux analysis identified the Tricarboxylic (TCA) cycle and redox coupling as the primary determinants of MR benefits. Interestingly, the fecundity defect occurred through altered sperm function and its restoration by folic acid supplementation also restored sperm mitochondrial metabolism. Together these findings suggest that dietary interventions connected to specific changes in metabolism can separate adverse effects that may occur by enhancing longevity.

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COX4-like, a Nuclear-Encoded Mitochondrial Gene Duplicate, Is Essential for Male Fertility in Drosophila melanogaster

Eslamieh

Mirsalehi

Markova

et al. 2022

Genes

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Recent studies on nuclear-encoded mitochondrial genes (N-mt genes) in Drosophila melanogaster have shown a unique pattern of expression for newly duplicated N-mt genes, with many duplicates having a testis-biased expression and playing an essential role in spermatogenesis. In this study, we investigated a newly duplicated N-mt gene—i.e., Cytochrome c oxidase 4-like (COX4L)—in order to understand its function and, consequently, the reason behind its retention in the D. melanogaster genome. The COX4L gene is a duplicate of the Cytochrome c oxidase 4 (COX4) gene of OXPHOS complex IV. While the parental COX4 gene has been found in all eukaryotes, including single-cell eukaryotes such as yeast, we show that COX4L is only present in the Brachycera suborder of Diptera; thus, both genes are present in all Drosophila species, but have significantly different patterns of expression: COX4 is highly expressed in all tissues, while COX4L has a testis-specific expression. To understand the function of this new gene, we first knocked down its expression in the D. melanogaster germline using two different RNAi lines driven by the bam-Gal4 driver; second, we created a knockout strain for this gene using CRISPR-Cas9 technology. Our results showed that knockdown and knockout lines of COX4L produce partial sterility and complete sterility in males, respectively, where a lack of sperm individualization was observed in both cases. Male infertility was prevented by driving COX4L-HA in the germline, but not when driving COX4-HA. In addition, ectopic expression of COX4L in the soma caused embryonic lethality, while overexpression in the germline led to a reduction in male fertility. COX4L-KO mitochondria show reduced membrane potential, providing a plausible explanation for the male sterility observed in these flies. This prominent loss-of-function phenotype, along with its testis-biased expression and its presence in the Drosophila sperm proteome, suggests that COX4L is a paralogous, specialized gene that is assembled in OXPHOS complex IV of male germline cells and/or sperm mitochondria.

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Cytological Analysis of Spermatogenesis: Live and Fixed Preparations of <em>Drosophila</em> Testes

Cited by 3 publications

References 0 publications

Recovery from heat‐induced infertility—A study of reproductive tissue responses and fitness consequences in male Drosophila melanogaster

Recovery from heat‐induced infertility—A study of reproductive tissue responses and fitness consequences in male Drosophila melanogaster

Uncoupling of lifespan and reproductive tradeoffs by dietary methionine and one-carbon metabolism

COX4-like, a Nuclear-Encoded Mitochondrial Gene Duplicate, Is Essential for Male Fertility in Drosophila melanogaster

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