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Visualisation of the copepod female reproductive system using confocal laser scanning microscopy and two-photon microscopy

Lookup NU author(s): Susan Fitzer, Dr Gary Caldwell, Professor Tony Clare, Professor Robert Upstill-Goddard, Professor Matt Bentley

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Abstract

We examined mating behaviour in the harpacticoid copepod Tisbe battagliai, in particular the process of delivering seminal contents to the female urosome. Labelling using 4’6’ diamidino-2-phenylindole (DAPI) coupled with two-photon confocal laser scanning microscopy successfully visualised the spermatophore and female internal reproductive system. Sections of the female urosome were imaged to examine seminal fluid stores. The female tissues were found to auto-fluoresce as red emission under green excitation, requiring no additional tissue labelling. DAPI-labelled seminal fluid stores were identified within the female reproductive system. The details observed agreed with previous descriptions of copepod reproductive anatomy and of spermatophores. Specimens cultured under pH 8.10 and a simulated ocean acidification scenario (pH 7.67) were compared for changes in reproductive anatomy and spermatophore size and site attachment. No differences were observed in spermatophore attachment or the female reproductive system but spermatophore size was reduced significantly at pH 7.67 compared with pH 8.10. This size reduction was, however, in proportion to an overall reduction in female body size at reduced pH. Confocal microscopy is shown here to be a valuable tool to investigate detailed reproductive processes in copepods.


Publication metadata

Author(s): Fitzer SC, Bishop JDD, Caldwell GS, Clare AS, Upstill-Goddard RC, Bentley MG

Publication type: Article

Journal: Journal of Crustacean Biology

Year: 2012

Volume: 32

Issue: 5

Pages: 685-692

Print publication date: 01/08/2012

ISSN (print): 0278-0372

ISSN (electronic): 1937-240X

Publisher: BioOne

URL: http://dx.doi.org/10.1163/193724012X639788

DOI: 10.1163/193724012X639788


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