Name: microsurgical obstruction of testes fusion in spodoptera litura
Uploaded: 2021-07-16T13:00:00
Description: Watch this Scientific Journal Video about Microsurgical Obstruction of Testes Fusion in Spodoptera litura at JoVE.com

This work were supported by National Natural Science Foundation of China (Nos.:31772519, 31720103916; ) and an open grant from the State Key Laboratory of Silkworm Genome Biology, South West University (No.: sklsgb2013003).

1. Insect rearing and maintenance
<ol>
	<li>Culture the Spodoptera litura larvae in environmental simulation chambers with an artificial diet until they reach day 4 of the 6th instar (L6D4). Select male larvae when the worms enter the first day of the 6th instar (L6D0) based on the inverse triangle-shaped structure on the eighth abdomen14. 
	&#8203;NOTE: Larvae rearing and maintenance techniques were published previously4,<su...

<ol>
	<li>Zeitlinger, J., Bohmann, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Thorax+closure+in+Drosophila:+involvement+of+Fos+and+the+JNK+pathway.">Thorax closure in Drosophila: involvement of Fos and the JNK pathway.</a> Development. 126 (17), 3947-3956 (1999).</li><li>Ray, H. J., Niswander, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mechanisms+of+tissue+fusion+during+development.">Mechanisms of tissue fusion during development.</a> Development. 139 (10), 1701-1711 (2012).</li><li>Ducuing, A., Keeley, C., Mollereau, B., Vincent, S. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=DPP-mediated+feed-forward+loop+canalizes+morphogenesis+during+Drosophila+dorsal+closure.">DPP-mediated feed-forward loop canalizes morphogenesis during Drosophila dorsal closure.</a> The Journal of Cell Biology. 208 (2), 239-248 (2015).</li><li>Du, Q., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Identification+and+functional+characterization+of+doublesex+gene+in+the+testis+of+Spodoptera+litura.">Identification and functional characterization of doublesex gene in the testis of Spodoptera litura.</a> Insect Science. 26 (6), 1000-1010 (2019).</li><li>Qin, H. G., Ding, J., Ye, Z. X., Huang, S. J., Luo, R. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dynamic+analysis+of+experimental+population+of+Spodoptera+litura.">Dynamic analysis of experimental population of Spodoptera litura.</a> Journal of Biosafety. 13 (2), 45-48 (2004).</li><li>Guan, B. B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+research+in+biology+and+ecology+of+Spodoptera+litura.">The research in biology and ecology of Spodoptera litura.</a> Journal of Biosafety. 8 (1), 57-61 (1999).</li><li>Wen, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+testis+development+and+spermatogenesis+in+Spodopture+litura+(lepidoptera:+noctuidae).">The testis development and spermatogenesis in Spodopture litura (lepidoptera: noctuidae).</a> Journal of South China Normal University (Natural Science Edition. 51 (4), 47-56 (2019).</li><li>Friedl&#228;nder, M., Seth, R. K., Reynolds, S. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Eupyrene+and+apyrene+sperm:+dichotomous+spermatogenesis+in+Lepidoptera.">Eupyrene and apyrene sperm: dichotomous spermatogenesis in Lepidoptera.</a> Advances in Insect Physiology. 32, 206 (2010).</li><li>Cook, P. A., Wedell, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Non-fertile+sperm+delay+female+remating.">Non-fertile sperm delay female remating.</a> Nature. 397 (6719), 486 (1999).</li><li>Iriki, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=On+the+function+of+apyrene+spermatozoa+in+the+silk+worm.">On the function of apyrene spermatozoa in the silk worm.</a> Zoological Magazine. 53, 123-124 (1941).</li><li>Liu, L., Feng, Q. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+study+of+fusion+of+testis+in+Lepidoptera+insects.">The study of fusion of testis in Lepidoptera insects.</a> Journal of South China Normal University (Natural Science Edition). 46 (5), 1-7 (2014).</li><li>Klowden, M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Physiological systems in insects. , (2007).</li><li>Xu, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transgenic+characterization+of+two+testis-specific+promoters+in+the+silkworm,+Bombyx+mori.">Transgenic characterization of two testis-specific promoters in the silkworm, Bombyx mori.</a> Insect Molecular Biology. 24 (2), 183-190 (2015).</li><li>Guo, X. R., Zheng, S. C., Liu, L., Feng, Q. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+sterol+carrier+protein+2/3-oxoacyl-CoA+thiolase+(SCPx)+is+involved+in+cholesterol+uptake+in+the+midgut+of+Spodoptera+litura:+gene+cloning,+expression,+localization+and+functional+analyses.">The sterol carrier protein 2/3-oxoacyl-CoA thiolase (SCPx) is involved in cholesterol uptake in the midgut of Spodoptera litura: gene cloning, expression, localization and functional analyses.</a> BioMed Central Molecular Biology. 10, 102 (2009).</li><li>Kope&#263;, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Studies+on+the+necessity+of+the+brain+for+the+inception+of+insect+metamorphosis.">Studies on the necessity of the brain for the inception of insect metamorphosis.</a> Biological Bulletin. 42 (6), 323-342 (1922).</li><li>Wigglesworth, V. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Factors+controlling+moulting+and+'metamorphosis'+in+an+insect.">Factors controlling moulting and 'metamorphosis' in an insect.</a> Nature. 133 (5), 725-726 (1934).</li><li>Williams, C. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Physiology+of+insect+diapause;+the+role+of+the+brain+in+the+production+and+termination+of+pupal+dormancy+in+the+giant+silkworm,+Platysamia+cecropia.">Physiology of insect diapause; the role of the brain in the production and termination of pupal dormancy in the giant silkworm, Platysamia cecropia.</a> Biological Bulletin. 90 (3), 234-243 (1946).</li><li>Fukuda, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hormonal+control+of+molting+and+pupation+in+the+silkworm.">Hormonal control of molting and pupation in the silkworm.</a> Proceedings of the Imperial Academy Tokyo. 16 (8), 417-420 (1940).</li><li>Tian, H. J., Liu, Z. P., Bai, Y. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Common+methods+to+detect+Sperm+quality+of+mammalian.">Common methods to detect Sperm quality of mammalian.</a> Journal of Economic Zoology. 8 (4), 198-201 (2004).</li><li>Ji, X. S., Chen, S. L., Zhao, Y., Tian, Y. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Progress+in+the+quality+evaluation+of+fish+sperm.">Progress in the quality evaluation of fish sperm.</a> Chinese Fishery Science. 14 (6), 1048-1054 (2007).</li><li>Baulny, B. O. D., Vern, Y. L., Kerboeuf, D., Maisse, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Flow+cytometric+evaluation+of+mitochondrial+activity+and+membrane+integrity+in+fresh+and+cryopreserved+rainbow+trout+(Oncorhynchus+mykiss)+spermatozoa.">Flow cytometric evaluation of mitochondrial activity and membrane integrity in fresh and cryopreserved rainbow trout (Oncorhynchus mykiss) spermatozoa.</a> Cryobiology. 34 (2), 141-149 (1997).</li><li>Krasznai, Z., M&#225;ri&#225;n, T., Balkay, I., Emri, M., Tr&#243;n, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Permeabilization+and+structural+changes+in+the+membrane+of+common+carp+(Cyprinus+carpio+L.)+sperm+induced+by+hypo-osmotic+shock.">Permeabilization and structural changes in the membrane of common carp (Cyprinus carpio L.) sperm induced by hypo-osmotic shock.</a> Aquaculture. 129 (1), 134 (1995).</li><li>Kime, D. E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Computer-assisted+sperm+analysis+(CASA)+as+a+tool+for+monitoring+sperm+quality+in+fish.">Computer-assisted sperm analysis (CASA) as a tool for monitoring sperm quality in fish.</a> Comparative Biochemistry &amp; Physiology Toxicology &amp; Pharmacology Cbp. 130 (4), 425-433 (2001).</li><li>Rurangwa, E., Volckaert, F. A., Huyskens, G., Kime, D. E., Ollevier, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quality+control+of+refrigerated+and+cryopreserved+semen+using+computer-assisted+sperm+analysis+(CASA),+viable+staining+and+standardized+fertilization+in+African+catfish+(Clarias+gariepinus).">Quality control of refrigerated and cryopreserved semen using computer-assisted sperm analysis (CASA), viable staining and standardized fertilization in African catfish (Clarias gariepinus).</a> Theriogenology. 55 (3), 751-769 (2001).</li><li>Seth, R. K., Kaur, J. J., Rao, D. K., Reynolds, S. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+larval+exposure+to+sublethal+concentrations+of+the+ecdysteroid+agonists+RH-5849+and+tebufenozide+(RH-5992)+on+male+reproductive+physiology+in+Spodoptera+litura.">Effects of larval exposure to sublethal concentrations of the ecdysteroid agonists RH-5849 and tebufenozide (RH-5992) on male reproductive physiology in Spodoptera litura.</a> Journal of Insect Physiology. 50 (6), 505-517 (2004).</li><li>Sweeney, R. M., Watterson, R. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rib+development+in+chick+embryos+analyzed+by+means+of+tantalum+foil+blocks.">Rib development in chick embryos analyzed by means of tantalum foil blocks.</a> American Journal of Anatomy. 126 (2), 127-149 (1969).</li><li>Wilde, S., Logan, M. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Application+of+impermeable+barriers+combined+with+candidate+factor+soaked+beads+to+study+inductive+signals+in+the+chick.">Application of impermeable barriers combined with candidate factor soaked beads to study inductive signals in the chick.</a> Journal of Visualized Experiments. (117), e54618 (2016).</li></ol>

After microsurgically obstructing testes fusion in Spodoptera litura, the number of sperm bundles decreased, which supported the hypothesis that this fusion is beneficial to the reproductive capability. Surgical manipulation has been used to study the physiological development of insects since the early 20th century. To determine whether the cranial nerve is regulated by insect metamorphosis, some researchers performed procedures such as ligation and decapitation on different insects (including Rh...

Fusion is a common phenomenon in tissue or organ development. Examples include dorsal closure and thorax closure in Drosophila1&#160;and palate morphogenesis, neural tube morphogenesis, and heart morphogenesis in mice and chicken2. CRISPR and RNAi have been applied to investigate the roles of genes in the process of fusion2,3,4.
Spodoptera litura (S. litura, Lepidoptera: Noctuidae) is a detrimental polyphagous pest that is widely distributed in tropical and subtropical areas of Asia, including China4,5,6. The wide distribution of S. litura is partly attributed to its powerful reproductive capability, which is relevant to gonad development. Male infertility is one approach to control this pest. As shown in the schematic figure of testicular structure, the testes are enclosed by the testicular sheath, including the external sheath (peritoneal sheath) and inner basal lamina. The basal lamina extends internally to form the follicular epithelium and separates the inner area of the testis into four chambers named follicles (Figure 1).
In the follicles, spermatogonia develop into spermatozoa after mitosis and meiosis, and then the spermatozoa in the sperm sacs align in the same direction to form sperm bundles7. During spermatogenesis, the primary spermatocytes differentiate into eupyrene sperms or apyrene sperms. Spermatocytes in the larval phase develop into eupyrene sperm with a long tail connected to a head of an elongated nucleus; these can fertilize eggs. Conversely, spermatocytes in the mid-pupal phase develop into apyrene sperm with a discarded nucleus; these sperm assist the survival, motion, and fertilization of eupyrene sperm9,10. The 6th day of the pupa is the period during which the testes have abundant eupyrene and apyrene sperm bundles.
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/62524/62524fig01.jpg" /> 
Figure 1: Schematic diagram of the testicular structure of Lepidoptera insects11. <a href="https://www.jove.com/files/ftp_upload/62524/62524fig01large.jpg" target="_blank">Please click here to view a larger version of this figure.</a>
Testicular fusion occurs in most insects of the Lepidoptera order11,12, especially in those species that are agricultural pests. Testicular fusion refers to a pair of testes growing bilaterally in the larval phase, approaching and adhering to each other, eventually integrating into a single gonad11. In Spodoptera litura, it happens during metamorphosis from the larval to the pupal stage. From day 1 of the 5th instar (L5D1) to day 4 of the 6th instar (L6D4), the pair of testes grows gradually in size, and the color turns light yellow from ivory-white. It becomes faint red as it reaches the prepupal phase (L6D5 to L6D6). Two bilateral symmetrical testes approach each other during the prepupal stage, fuse into one, and twist anticlockwise (doral view) to produce a single testis in the pupal and adult phases11. This phenomenon does not occur in silkworms, which have considerable economic importance and have been domesticated for 5000 years13. Thus, it is assumed that the testes&#39; fusion improves reproductive capability.
To determine the significance of Spodoptera litura testicular fusion, it is important to investigate the effects of blocking the process. In this protocol, aluminum foil was microsurgically inserted between the testes to keep them separated, and the consequent changes in the development of the insects and their testes were studied.

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microsurgical obstruction of testes fusion in spodoptera litura

Instead of using genetic methods like RNA interference (RNAi) and clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated endonuclease Cas9, a physical barrier was microsurgically inserted between the testes of Spodoptera litura to study the impact of this microsurgery on its growth and reproduction. After inserting aluminum foil between the testes, insect molting during metamorphosis proceeded normally. Insect growth and development were not remarkably altered; however, the number of sperm bundles changed if testes fusion was stopped by the microsurgery. These findings imply that blocking testicular fusion can influence male reproduction capability. The method can be further applied to interrupt communication between organs to study the function of specific signaling pathways. Compared to conventional surgery, microsurgery only requires freezing anesthetization, which is preferable to carbon dioxide anesthetization. Microsurgery also minimizes the surgery site area and facilitates wound healing. However, the selection of materials with specific functions needs further investigation. Avoiding tissue injury is crucial when making incisions during the operation.

The effects of microsurgery on Spodoptera litura growth and development 
The microsurgery left a 2 mm-long wound on the dorsal larval epidermis that eventually stopped leaking hemolymph and healed. The larvae went through prepupal and pupal stages and eclosed, indicating that the microsurgery had no major impact on growth and development. When the larvae molted into pupae, the suture threads were discarded along with the epidermis. There were no obvious dif...

Watch this Scientific Journal Video about Microsurgical Obstruction of Testes Fusion in Spodoptera litura at JoVE.com

Microsurgical Obstruction of Testes Fusion in Spodoptera litura

Aluminum foil was microsurgically inserted between the testes of Spodoptera litura to obstruct the fusion of testis. The procedure includes freezing, fixing, disinfection, incision, placing the barrier, suturing, postoperative feeding, and inspection. This approach provides a method to interfere with tissue formation.

Microsurgical Obstruction of Testes Fusion in Spodoptera litura

Instead of using genetic methods like RNA interference (RNAi) and clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated ...

This protocol suggests that a physical method can be adopted to assist in figuring out biological problems. This technique provides a method to place material in insects with minimum incision and study the influence of microsurgery on individuals'development. This method can apply to other Lepidoptera larvae and other parts of the insects, including the head and thorax.It is very important to practice making incisions in certain material and suturing skillfully. Begin by placing Spodoptera litura male larvae at day 4 of the 6th instar, or L6D4, on ice for 10 to 30 minutes to keep them anesthetized during the operation. Place the anesthetized larva on the wax tray with the dorsal side up.Fix the head and the tail of the larva with pins and threads and disinfect the surgical area by applying 3%iodine tincture with a cotton swab to the epidermis, followed by 70%alcohol. Make a 2 millimeter-long incision on the dorsal epidermis at the 9th body segment and use a sterile cotton swab to remove leaking hemolymph and fat bodies to obtain a clear view of the surgical area. Using surgical tweezers, insert a piece of aluminum foil between the testes, and then close the epidermis with a running suture.Tie a surgical square knot with the help of a needle holder and surgical tweezers. Cut the excess suture from the loop tails with scissors, leaving a 2-millimeter thread behind. After suturing, gently lay the larva in the rearing box and maintain them in a clean environmental simulation chamber with continuous observation.The growth and development of the Spodoptera litura male larvae post-microsurgery was monitored. The larval mortality rate was slightly higher in the surgical group, whereas the percentages of pupation, adult emergence, and successful mating were slightly lower in the surgical group than in the control group. In the experimental group, the number of total sperm bundles in eupyrene and apyrene sperm bundles were significantly lower than the control and sham control group.On the other hand, the percentages of eupyrene sperm bundles had no significant differences among all three groups. After removing a unilateral testis of the larvae, the numbers of total sperm bundles and eupyrene and apyrene sperm bundles in microsurgery groups were significantly lower than the control group. The percentage of eupyrene sperm bundles on the sixth day of the pupal stage was comparable in all three groups.It is very important to notice the location of the wound and avoid damaging other organs during the incision. This technique can be used to study the biological significance of testicular fusion by applying drugs to the organs. The drugs'effect on the target organ can be explored.

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