Role In Insect Reproduction Research Articles

The first vitellogenin receptor from a Lepidopteran insect: molecular characterization, expression patterns and RNA interference analysis

The vitellogenin receptor (VgR) belongs to the low-density lipoprotein receptor (LDLR) superfamily, and is an important carrier for the uptake of vitellogenin (Vg) into developing oocytes of all oviparous species. The first full-length message for a VgR from a Lepidopteran insect was cloned and sequenced from the ovary of Spodoptera litura Fabricius (GenBank accession no. GU983858). The coding region consisted of 5370 bp flanked by a 49 bp 5'-untranslated region (UTR) and a 177 bp 3'-UTR, which encoded a 1798-residue protein with a predicted molecular weight (MW) of 201.69 kDa. S. litura VgR (SlVgR)comprised two ligand binding sites with four LDLR class A repeats in the first domain and seven in the second domain, an epidermal growth factor-like domain containing an LDLR class B repeat and a YWXD motif, a transmembrane domain and a cytoplasmic domain. A phylogenetic relationship placed SlVgR as a separate group from the other insects. SlVgR messenger RNA (mRNA) was specifically expressed in the ovarian tissues. The developmental expression patterns showed that VgR mRNA was first transcribed in 6(th) day female pupae and the maximum level of VgR mRNA appeared in 36-h-old adults. Immunoblot analysis detected an ovary-specific VgR protein with a MW of ∼200 kDa, whose development profiles were consistent with VgR mRNA expression patterns. RNA inteference (RNAi) specifically disrupted the VgR gene by injection of 3 or 5 µg VgR double-stranded RNA per insect in 4(th) or 6(th) day pupae. RNAi of SlVgR led to a phenotype characterized by high Vg accumulation in the haemolymph, low Vg deposition in the ovary and the failure of insect spawning. These results mean that VgR is critical for binding Vg and transporting it into the oocytes of the insect ovary, thus playing an important role in insect reproduction.

Effects of the NMDA receptor antagonist MK-801 on female reproduction and juvenile hormone biosynthesis in the cricketGryllus bimaculatusand the butterflyBicyclus anynana

Apart from regulating insect development, juvenile hormones (JHs) play an important role in insect reproduction, where they initiate vitellogenin synthesis and regulate the uptake of yolk by the ovary. JH synthesis is a tightly regulated process controlled by neurons and peptidergic neurosecretory cells. One of the known stimulatory regulators of JH biosynthesis is glutamate, and its N-methyl-d-aspartate (NMDA) receptor has been recently found in the cockroach Diploptera punctata. In this study we demonstrate a strong reduction in reproductive output in the tropical butterfly Bicyclus anynana and the Mediterranean field cricket Gryllus bimaculatus caused by the NMDA receptor antagonist MK-801. Such inhibiting effects on reproduction could be overruled by the application of JH mimics. In G. bimaculatus, MK-801 inhibits in vitro JH biosynthesis in the corpora allata and reduces in vivo JH haemolymph titres in a dose-dependent manner. These results suggest that JH biosynthesis in the corpora allata is at least in part controlled by an NMDA receptor with Ca2+ as a second level messenger. Based on our findings we consider NMDA receptor antagonists as important tools for manipulating juvenile hormone biosynthesis and therefore for gaining a better understanding of the mechanistic basis of reproduction.

Immunocytochemical distribution of angiotensin I-converting enzyme-like immunoreactivity in the brain and testis of insects

Angiotensin converting enzyme (ACE) is Zn 2+ metallopeptidase which plays an important role in blood pressure homeostasis in mammals and other vertebrates. Homologues of ACE involved in the biosynthesis of mammalian peptide hormones have also been identified in the insects, Musca domestica, Drosophila melanogaster and Haematobia irritans exigua. In the pursuit of the biological role of insect ACE, this work focused on the tissue and cellular distribution of ACE in several insect species. The localisation of ACE in the central nervous system and reproductive tissues from a number of insect species suggests that ACE is of physiological importance in these tissues. By means of an antiserum to housefly ACE, we found that ACE-like immunoreactivity was abundantly present in the neuropil areas of the brain of all insects investigated, suggesting a role for ACE in the metabolic inactivation of peptide neurotransmitters. Especially in the fleshfly, Neobellieria bullata neuropile staining is abundant. In the cockroach Leucophaea maderae, immunoreactive staining was abundant in the neuronal perikarya as well as in the neuropilar regions. Staining in neurosecretory cells was also observed in the brains of the lepidopteran species, Bombyx mori and Mamestra brassica. The localisation of ACE in neurosecretory cells is consistent with the role as a processing hormone, involved in the generation of active peptide hormones. ACE was found to be co-localised with peptides of the FXPRLamide family in M. brassica and in B. mori, suggesting a role for the biosynthesis of these hormones. Finally, we found ACE-like immunoreactivity in the testis of Locusta migratoria, N. bullata and Leptinotarsa decemlineata, providing additional evidence for its important role in insect reproduction.

Nutrient modulation of hormone production: Dietary essential fatty acids, tissue prostaglandins and their probable role in insect reproduction

The essential polyunsaturated fatty acids required by insects in their food appear to be needed for prostaglandinogenesis. Prostaglandins themselves are likely to be widely distributed in insects playing perhaps an hitherto unsuspected important role as in reproduction.

Extracellular localization of cyclic GMP in the house cricket male accessory reproductive gland and its fate in mating.

The male accessory reproductive gland (ARG) of the house cricket, Acheta domesticus (L.), contains an exceedingly high concentration of cyclic GMP, about 1,000 pmol/mg protein. Immunofluorescent localization and radioimmunoassay measurements show that cyclic GMP is concentrated in a small number of tubules. It accumulates in the tubule lumina where it is protected from degradation by phosphodiesterases. Cyclic GMP is secreted by the ARG and is incorporated into spermatophores. Over 80% of spermatophore cyclic GMP is found in the handle-capillary tube, a thin conduit through which sperm pass during transfer to the female. The concentration of cyclic GMP in the insemination fluid is about 20 microM but does not appear to be specifically associated with the sperm. Cyclic GMP enters the female spermatheca during insemination but disappears rapidly. Physiological effects of cyclic GMP on sperm were not observed nor was an effect of cyclic GMP observed on egg laying by mated females. Cyclic AMP was localized on sperm flagella in the spermatophore and in the spermatheca. These studies indicate that cyclic nucleotides have important roles in insect reproduction and that the house cricket is a good model for elucidating these functions.