Flow-cytometric analysis of spermatogenesis in Teleogryllus commodus

doi:10.1016/0022-1910(89)90021-8

Journal of Insect Physiology

Volume 35, Issue 12, 1989, Pages 975-980

https://doi.org/10.1016/0022-1910(89)90021-8 Get rights and content

Abstract

The development of the testis of the cricket Teleogryllus commodus has been studied using flow cytometry. From a laboratory-reared population of insects, total testis tissue was prepared at approx. 48-h intervals for the entire development period extending from the fifth instar (28 days after eclosion) to the imaginal moult (about 60 days after eclosion). Cells were stained with DNA-specific fluorochromes and the temporal sequence of the development of the testis as well as the timing of specific meiotic events quantitatively determined. Testis development studied in this fashion is seen as changes in the relative frequency of cells with 1, 2, and 4C DNA content. Pre-spermiogenic processes of spermatogenesis (change in the frequency of cells with 2 and 4C DNA content) occur continuously during the fifth-eighth instars, while the appearance of 1C cells (initiation of spermiogenesis) occurs abruptly at the transition between the sixth and seventh instars.

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Cited by (3)

Parasitic castration of Pseudaletia separata by Cotesia kariyai and its association with polydnavirus gene expression
1998, Journal of Insect Physiology
Parasitization by the endoparasitoid Cotesia kariyai caused the inhibition of spermatogenesis of Pseudaletia separata. This phenomenon is called parasitic castration. The degree of castration was dependent on the host stage parasitized. Host parasitized on day 1 of the 4th stadium (the time of primary spermatocyte accumulation), had testicular cells with abnormal chromosomes appearing two days after parasitization, and spermiogenesis was completely inhibited. However, when hosts were parasitized on day 0 of the 6th (final) stadium, the degree of castration was less severe, and elongated cells appeared similar to those found in nonparasitized larvae. Results of this study involving injection of C. kariyai polydnavirus (CkPV) and venom suggested that these wasp components caused the appearance of abnormal chromosomes in specific germ cells, which were in mitotic or meiotic prophases. The amount of CkPV gene expression in host testes increased immediately after parasitization and reached a maximum 12 h later. The early-expressed CkPV gene(s) may be related to the parasitic castration phenomenon.
Chapter 5 Cell Cycle Analysis of Apoptosis Using Flow Cytometry
1995, Methods in Cell Biology
Using flow cytometry to analyze asynchronous and synchronized cell populations, this chapter describes a number of features of drug-induced apoptosis in cultured cells: (1) apoptosis can occur in any cell cycle phase; (2) a given agent can induce apoptosis in more than one cell cycle phase; (3) there is a clear temporal relationship between cell cycle “arrest” induced by different agents and the onset of apoptosis; (4) proteolysis of specific intracellular proteins represents an early event of apoptosis; ( 5 ) aberrant mitosis precedes apoptosis under a variety of conditions, including exposure to inhibitors of DNA synthesis; (6) cells with sub-G, DNA content are not always apoptotic cells; (7) DNA content can be an unreliable index of cell cycle position; and (8) under all conditions, cell cycle “stasis” precedes drug-induced apoptosis. The chapter describes specific examples of applications of standard flow cytometric methods to the analysis of drug-induced apoptosis in HeLa S3 cells.
Spermiogenesis of the testes of juvenile hormone-treated silkworm larvae, Bombyx mori
1993, Comparative Biochemistry and Physiology -- Part A: Physiology
- 1.
  1. To examine changes in the percentages of testicular cells such as 1C cells (spermatids and spermatozoa), 2C cells (G1 somatic cells, spermatogonia and secondary spermatocytes), 2–4C cells (somatic and germinal cells in S phase) and 4C cells (G2 somatic cells and primary spermatocytes), we isolated nuclei from the testes of silkworm larvae and subjected them to flow cytometric analysis.
- 2.
  2. In control testes, 1C cells appeared at day 0 of the fifth instar, increased gradually by spinning and increased steeply at 2 days later still spinning. The percentage of 2C cells decreased gradually after ecdysis to the fifth instar. The percentage of 4C cells increased from day 2 to day 5 of the fifth instar and decreased after day 11 of this instar. Cells in S phase remained constant through the fifth instar.
- 3.
  3. An injection of juvenile hormone analog, methoprene, at day 0 of the fifth instar did not inhibit spermiogenesis, but resulted in increased 2C cells and decreased 4C cells dose-dependently. In contrast, the same treatment at day 2 of the fifth instar did not change the percentages of 1C, 2C and 4C cells of the testes at all, suggesting that the testes changed sensitivity to the hormone at larval development.
- 4.
  4. Repeated injections of methoprene to induce the appearance of dauer larvae resulted in a complete block of the development of 1C cells.

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Journal of Insect Physiology

Abstract

References (20)

The premeiotic stages of spermatogenesis

Adv. Genet.

An autoradiographic study of nucleic acid synthesis during spermatogenesis in the grasshopper Melanopus differentialis

Exp. Cell Res.

The cell cycle of epidermal cells during reprogramming in the pupa of Galleria

J. Insect. Physiol.

Regulation of spatial pattern in populations of the field cricket Teleogryllus commodus

Z. Tierpsychol.

Techniques for quantitative staining of cellular DNA for flow cytometry

The regulation of spermatogenesis in insects

Ann. Rev. Ent.

Changes in accessibility of DNA to various fluorochromes during spermatogenesis

Cytometry

Ecdysteroids in vitro promote differentiation in the accessory glands of male mealworm beetles

Experientia

Physiological differences between races of Teleogryllus commodus (Walker) (Orthoptera: Gryllidae) related to a proposed genetic approach to control

Aust. J. Zool.

Cited by (3)

Parasitic castration of Pseudaletia separata by Cotesia kariyai and its association with polydnavirus gene expression

Chapter 5 Cell Cycle Analysis of Apoptosis Using Flow Cytometry

Spermiogenesis of the testes of juvenile hormone-treated silkworm larvae, Bombyx mori