Journal of Biosciences

, Volume 32, Issue 2, pp 309–328 | Cite as

Programmed cell death in the larval salivary glands of Apis mellifera (Hymenoptera, Apidae)

  • E C M Silva-Zacarin
  • G A Tomaino
  • M R Brocheto-Braga
  • S R Taboga
  • R L M Silva de Moraes
Article
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Accepted:

Abstract

The morphological and histochemical features of degeneration in honeybee (Apis mellifera) salivary glands were investigated in 5th instar larvae and in the pre-pupal period. The distribution and activity patterns of acid phosphatase enzyme were also analysed. As a routine, the larval salivary glands were fixed and processed for light microscopy and transmission electron microscopy. Tissue sections were subsequently stained with haematoxylin-eosin, bromophenol blue, silver, or a variant of the critical electrolyte concentration (CEC) method. Ultrathin sections were contrasted with uranyl acetate and lead citrate. Glands were processed for the histochemical and cytochemical localization of acid phosphatase, as well as biochemical assay to detect its activity pattern. Acid phosphatase activity was histochemically detected in all the salivary glands analysed. The cytochemical results showed acid phosphatase in vesicles, Golgi apparatus and lysosomes during the secretory phase and, additionally, in autophagic structures and luminal secretion during the degenerative phase. These findings were in agreement with the biochemical assay. At the end of the 5th instar, the glandular cells had a vacuolated cytoplasm and pyknotic nuclei, and epithelial cells were shed into the glandular lumen. The transition phase from the 5th instar to the pre-pupal period was characterized by intense vacuolation of the basal cytoplasm and release of parts of the cytoplasm into the lumen by apical blebbing; these blebs contained cytoplasmic RNA, rough endoplasmic reticule and, occasionally, nuclear material. In the pre-pupal phase, the glandular epithelium showed progressive degeneration so that at the end of this phase only nuclei and remnants of the cytoplasm were observed. The nuclei were pyknotic, with peripheral chromatin and blebs. The gland remained in the haemolymph and was recycled during metamorphosis. The programmed cell death in this gland represented a morphological form intermediate between apoptosis and autophagy.

Keywords

Degeneration honeybee larva prepupa silk gland 

Abbreviations used

CEC

Critical electrolyte concentration

HE

haematoxylin-eosin

IRS

insect Ringer solution

p-NPP

p-nitrophenyl phosphate

PP

pre-pupal phase

TEM

transmission electron microscope

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Copyright information

© Indian Academy of Sciences 2007

Authors and Affiliations

  • E C M Silva-Zacarin
    • 1
  • G A Tomaino
    • 2
  • M R Brocheto-Braga
    • 2
  • S R Taboga
    • 3
  • R L M Silva de Moraes
    • 2
  1. 1.Avenida Darci Carvalho DaffenerUniversidade Federal de São Carlos-UFSCarSorocaba/SPBrasil
  2. 2.Departamento de Biologia, Instituto de BiociênciasUniversidade Estadual Paulista-UNESPRio Claro/SPBrasil
  3. 3.Departamento de Biologia, Instituto de Biociências, Letras e Ciências ExatasUniversidade Estadual Paulista-UNESPSão José do Rio PretoBrazil

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