The use of parthenogenetic silkworm (Bombyx mori) strains, which eliminate the problem of recombination, is a useful tool for maintaining transgenic clonal lines. The generation of genetically identical individuals is becoming an important tool in genetic engineering, allowing replication of an existing advantageous trait combination without the mixing that occurs during sexual reproduction. Thus, an animal with a particular genetic modification, such as the ability to produce transgenic proteins, can reproduce more rapidly than by natural mating. One obstacle to the widespread use of parthenogenesis in silkworm genetic engineering is the relatively low efficiency of downstream transgenesis techniques. In this work, we seek to optimize the use of transgenesis in conjunction with the production of parthenogenetic individuals. We found that a very important parameter for the introduction of foreign genes into a parthenogenetic strain is the precise timing of embryo microinjection. Our modification of the original method increased the efficiency of transgene injection as well as the survival rate of injected embryos. We also provide a detailed description of the methodological procedure including a graphical overview of the entire protocol.

• Klíčová slova
• Bombyx mori, embryonic development, genetic engineering, ovary transplantation, overcoming diapause, parthenogenesis, transgenesis,
• Publikační typ
• časopisecké články MeSH

Insect adipokinetic hormones (AKHs) are short peptides produced in the corpora cardiaca and are responsible for mobilizing energy stores from the fat body to the hemolymph. Three related peptides, AKH1, AKH2, and AKH/corazonin-related peptide (ACP) as well as three AKH receptors have been reported in Bombyx mori. AKH1 and AKH2 are specific for the AKHR1 receptor, whereas ACP interacts with the other two AKHRs. To assess the effect of the two silkworm AKHs and ACP in the regulation of energy homeostasis we examined the expression pattern of the three peptides and their receptors as well as their effect on the level of carbohydrates and lipids in the hemolymph. Our results support the hypothesis that only AKH1 and AKH2 peptides together with the AKHR1 receptor are involved in the maintenance of energy homeostasis. Because Bombyx AKHR1 (BmAKHR1) seems to be a true AKHR we generated its mutation. The BmAKHR1 mutant larvae display significantly lower carbohydrate and lipid levels in the hemolymph and reduced sensitivity to starvation. Our study clarifies the role of BmAKHR1 in energy homeostasis.

• Klíčová slova
• BMSK0010951, Bommo-AKH1, Bommo-AKH2, NM_001043584, TALEN, silkworm, targeted mutagenesis,
• MeSH
• bourec růst a vývoj   metabolismus   MeSH
• energetický metabolismus MeSH
• hemolymfa metabolismus   MeSH
• hmyzí hormony genetika   metabolismus   MeSH
• hmyzí proteiny genetika   metabolismus   MeSH
• kyselina pyrrolidonkarboxylová analogy a deriváty   metabolismus   MeSH
• larva metabolismus   MeSH
• lipidy analýza   MeSH
• mutageneze MeSH
• neuropeptidy genetika   metabolismus   MeSH
• oligopeptidy genetika   metabolismus   MeSH
• protein - isoformy genetika   metabolismus   MeSH
• receptory glukagonu genetika   metabolismus   MeSH
• regulace genové exprese MeSH
• sacharidy analýza   MeSH
• signální transdukce * MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• adipokinetic hormone MeSH Prohlížeč
• corazonin protein, insect MeSH Prohlížeč
• hmyzí hormony MeSH
• hmyzí proteiny MeSH
• kyselina pyrrolidonkarboxylová MeSH
• lipidy MeSH
• neuropeptidy MeSH
• oligopeptidy MeSH
• protein - isoformy MeSH
• receptory glukagonu MeSH
• sacharidy MeSH