Detailed proteome mapping of newly emerged honeybee worker hemolymph and comparison with the red-eye pupal stage

Abstract

-The honeybee, Apis mellifera , undergoes complete metamorphosis before transitioning to the adult stage. The newly emerged individual and the red-eye pupa stage are well defined and easily recognizable in the time life cycle honeybee and, therefore, very useful for studying physiological and developmental factors. We analyzed in detail the hemolymph proteome of newly emerged honeybee worker using 2D-E-MS/MS (pI 3-10 and 4-7). The comparison of identical hemolymph volumes (20 μL per 2D-E) for newly emerged bee and red-eye pupa revealed a dramatic decrease in the number of spots (qualitative changes) and overall protein quantity during the non-feeding stage. The results increase our knowledge about honeybee metamorphosis during the non-feeding period and clarify previous findings regarding particular proteins. The results will be useful for future comparative physiological, developmental, and host-pathogen studies on individual or population level.Apis mellifera / hemolymph / metamorphosis / hexamerin / ferritin

2D-DIGE proteomic analysis reveals changes in haemolymph proteome of 1-day-old honey bee (Apis mellifera) workers in response to infection with Varroa destructor mites

“…The three proteins, peroxiredoxin-like protein (NP_001164444.1), glutathione S-transferase S1 isoform X1 (XP_016772844.1) and T-complex protein 1 subunit zeta (XP_012349557.1), were present at increased levels in the Varroa -infected bee samples. Moreover, peroxiredoxin-like protein and glutathione S-transferase S1 are also involved in the response to oxidative stress (Du Rand et al 2015;Erban et al 2016) and Tcomplex proteins are classified as chaperone proteins (Du Rand et al 2015). In our opinion, the increased level of the latter protein suggests an increase in the amount of incorrectly folded proteins in Varroa -infected bees.…”

“…Thus, hexamerin 70a in haemolymph appears to play a different role to hexamerin 70b, 70c and 110. According to Erban et al (2016) at the time of emergence, Hex70a is not used primarily as a storage protein but has a transport function; hexamerins may transport hormones such as juvenile hormone (Braun and Wyatt 1996). In our opinion, it is interesting that the loss of haemolymph during Varroa parasitation was not accompanied by the loss of hexamerin in honey bee haemolymph.…”

“…Therefore, the level of hexamerins must be maintained at some level to play a role as storage proteins. Four hexamerin proteins, hexamerin70a, hexamerin 70b, hexamerin 70c and hexamerin 110, were identified in different stages of the honey bee (Martins et al 2010(Martins et al , 2011Erban et al 2016). At the time of emergence, 70b, 70c and 110 disappeared, and 70a appeared constant (Erban et al 2016).…”

“…Honey bee haemolymph plays a role in immune defence and is also vital for daily life functions, such as transport of nutrients and signalling molecules (Erban et al 2016). Several proteomic approaches have been used to investigate caste differences (Chan et al 2006;Ararso et al 2018) and developmental changes in honey bee haemolymph (Bogaerts et al 2009;Erban et al 2014Erban et al , 2016Feng et al 2014).…”

“…Honey bee haemolymph plays a role in immune defence and is also vital for daily life functions, such as transport of nutrients and signalling molecules (Erban et al 2016). Several proteomic approaches have been used to investigate caste differences (Chan et al 2006;Ararso et al 2018) and developmental changes in honey bee haemolymph (Bogaerts et al 2009;Erban et al 2014Erban et al , 2016Feng et al 2014). Proteome analysis of larvae as well as of the haemolymph, mushroom body, head tissues and antenna provides novel insights into honey bee resistance against Varroa infestation (Parker et al 2012;Ji et al 2015;Hu et al 2016).…”