Many male arthropods package their sperm in a spermatophore for transfer to the female. We propose to analyze the structure, composition, and formation of the spermatophore of a model insect, Tenebrio molitor. We have previously described the development of the glands which make the wall of the spermatophore. These glands export a heterogenous structured product which is molded into the multi-layered wall of the spermatophore within the male reproductive tract. After its ejection from the male aedeagus, the spermatophore goes through a programmed sequence of expansions and bursts to release its seminal contents. The structure of the spermatophore will be investigated by differential interference contrast, histochemistry, and electron microscopy, with particular attention to the changes in the layers and their properties as the expansions take place. In some ways, the spermatophore is like cuticle without chitin. There are over/15 structural proteins in the wall of the spermatophore. In order to follow the formation and expansion of the spermatophore, we need specific probes for some of the proteins of the wall. The protein constituents of the spermatophore can be studied best with probes for specific proteins of the wall. We will prepare monoclonal antibodies to wall proteins by fusion of spaenic lymphocytes from BALB/c mice with myeloma X63/Ag 8.653. Clones will be screened by ELISA and then characterized further by immunoprecipitation and electrophoresis. Specific antibodies will be coupled with enzymes or fluorochromes for immunocytochemical localization of wall components. In the future, we can use these antibodies to isolate the structural proteins of the spermatophore, and we can extend our model study to other species such as Simulium, Culicoides, or Glossina, the vectors of filariasis, blue tongue virus, and trypanosomes, respectively. Increased knowledge of the reproductive biology of these vectors may aid in the development and evaluation of programs for their control.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI015662-08
Application #
3126334
Study Section
Tropical Medicine and Parasitology Study Section (TMP)
Project Start
1978-07-01
Project End
1986-06-30
Budget Start
1985-07-01
Budget End
1986-06-30
Support Year
8
Fiscal Year
1985
Total Cost
Indirect Cost
Name
University of Vermont & St Agric College
Department
Type
Schools of Arts and Sciences
DUNS #
066811191
City
Burlington
State
VT
Country
United States
Zip Code
Paesen, G C; Feng, X; Happ, G M (1996) Structure of a D-protein gene and amino-acid sequences of the highly repetitive D-proteins secreted by the accessory glands of the mealworm beetle. Biochim Biophys Acta 1293:171-6
Feng, X; Happ, G M (1996) Isolation and sequencing of the gene encoding Sp23, a structural protein of spermatophore of the mealworm beetle, Tenebrio molitor. Gene 179:257-62
Yaginuma, T; Mizuno, T; Mizuno, C et al. (1996) Trehalase in the spermatophore from the bean-shaped accessory gland of the male mealworm beetle, Tenebrio molitor: purification, kinetic properties and localization of the enzyme. J Comp Physiol B 166:1-10
Paesen, G C; Happ, G M (1995) The B proteins secreted by the tubular accessory sex glands of the male mealworm beetle, Tenebrio molitor, have sequence similarity to moth pheromone-binding proteins. Insect Biochem Mol Biol 25:401-8
Paesen, G C; Happ, G M (1994) cDNA-inferred amino-acid sequence of a C protein, a heparin-binding, basic secretion product of the tubular accessory sex glands of the mealworm beetle, Tenebrio molitor. Insect Biochem Mol Biol 24:21-7
Paesen, G C; Schwartz, M B; Peferoen, M et al. (1992) Amino acid sequence of Sp23, a structural protein of the spermatophore of the mealworm beetle, Tenebrio molitor. J Biol Chem 267:18852-7
Happ, G M (1992) Maturation of the male reproductive system and its endocrine regulation. Annu Rev Entomol 37:303-20
Weyda, F (1991) Postembedding immunostaining of ultrathin sections realized after their staining with heavy metals. J Electron Microsc Tech 19:269-70
Weyda, F (1991) Possibility of use of primary antibody drop for repeated immunostaining. J Electron Microsc Tech 18:450-1
Weyda, F (1990) Notes on the use of Nanoplast FB 101 for transmission electron microscopy. J Electron Microsc Tech 16:356-7

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