Proteins in spermathecal gland secretion and spermathecal fluid and the properties of a 29 kDa protein in queens of Apis mellifera

Abstract

-One and two dimensional SDS-PAGE were used to characterize the protein pattern of the spermathecal gland secretion and spermathecal fluid in Apis mellifera queen pupae and emerged queens of different ages. The concentration of protein varied from 8.5 and 15.3 mg/mL in the spermathecal fluid, and from 5 to 8.5 mg/mL in the secretion. Development of the protein pattern of the gland secretion and spermathecal fluid was identical from pupae until the age of 3 days. In sexually mature queens (10 days or older) the gland secretion and spermathecal fluid each had one additional band at 79 kDa and at 29 kDa respectively. The 29 kDa protein was N-terminal blocked but several peptide fragments were sequenced after digestion with LysC protease. Only 2 of the sequences showed a distinct homology to the N-terminal half of the glycolytic enzyme triosephosphate isomerase (TPI). TPI antibody reacted with the 29 kDa protein, but the enzymatic activity was only 1/100 compared to TPI of hemolymph. The possible function of the protein is discussed. spermatheca / sperm storage / protein pattern / 29 kDa protein / Apis mellifera

Instrumental insemination of Apis mellifera queens with hetero- and conspecific spermatozoa results in different sperm survival

“…The results can be interpreted as differences in the physiological conditions between the two multiplecomb cavity and the two single-comb open-air nesting Apis species. This supports the idea of the significance of spermathecal fluid for sperm storage, as suggested by Klenk et al (2004). Spermathecal fluid and spermathecal gland secretion should be compared in the species.…”

“…Further the spermathecal compartment contains several sugars and soluble proteins . The concentration of proteins varies from 5 to 15.3 mg/mL according to age and season (Klenk et al, 2004). A 29 kDa protein only occurred in the spermathecal fluid of sexually mature queens.…”

“…A 29 kDa protein only occurred in the spermathecal fluid of sexually mature queens. It may have a function for longterm sperm storage, but further studies are required (Klenk et al, 2004). Antioxidases (CAT, SOD and GST) found in the spermathecae of mated queens may be also involved in the storage process by protecting the spermatozoa from oxidative stress (Weirich et al, 2002).…”

“…In some insects, such as Drosophila melanogaster, male-derived proteins play a major role for the physiology of the spermatozoa (Chapman, 2001). For the long term storage of 3 up to 5 years in A. mellifera, secretions of the queen into the spermatheca seem to be of significance (Klenk et al, 2004). Further the spermathecal fluid contains several sugars and its pH value is 8.6 (Gessner and Gessner, 1976).…”

Morphological and morphometrical assessment of spermathecae of Aedes aegypti females

“…Depending on the insect species, the clustered cells have either individual ducts that transport secreted substances directly into the reservoir (Curtin & Jones 1961) or a common glandular duct (Lawson & Thomas 1970, Martins et al 2008. Secreted substances are thought to include nutrients that keep the spermatozoa viable during the entire reproductive life of the adult female (Schoeters & Billen 2000, Klenk et al 2004.…”