Sex-dependent and sex-independent distribution of the beta-subunit of nerve growth factor in the central nervous and peripheral tissues of mice.
Levels of the beta-subunit of nerve growth factor (beta-NGF) were measured in the central nervous and peripheral tissues of mice using a highly sensitive, sandwich-type enzyme immunoassay system. Antiserum was raised in rabbits against the 7S form of NGF, which was purified from mouse submandibular glands. beta-NGF-specific antibody isolated on a column of Sepharose CL-4B coupled with purified beta-NGF reacted only with beta-NGF. The assay for beta-NGF was performed by incubation of F(ab')2 fragments of the antibody immobilized on a polystyrene ball with tissue extract and then with the same antibody Fab' fragments labeled with beta-D-galactosidase, followed by measurement of galactosidase activity. Our assay system was found to be highly sensitive (minimal detection limit, 0.3 pg/0.3 ml of assay mixture). Furthermore, the presence of gelatin hydrolysates and protease inhibitors during preparation of tissue extracts enabled us to determine the precise levels of beta-NGF in almost all organs of mice. The amount of beta-NGF in submandibular glands was extremely high, and its level increased rapidly until mice were 2 months of age; then, the level continued to increase slowly until mice were 1 year old (3-5 mg/g of tissue). In serum, some of the 2-month-old males, but none of the females, exhibited a fairly high level of beta-NGF (greater than 100 pg/ml).(ABSTRACT TRUNCATED AT 250 WORDS)
Katoh-Semba
R
R
Department of Perinatology, Institute for Developmental Research, Aichi, Japan.
Semba
R
R
Kashiwamata
S
S
Kato
K
K
D000311:Adrenal Glands / Q000032:analysis; D000375:Aging; D000818:Animals; D001923:Brain Chemistry; D005260:Female; D007124:Immunoenzyme Techniques; D008099:Liver / Q000032:analysis; D008297:Male; D051379:Mice; D009206:Myocardium / Q000032:analysis; D009414:Nerve Growth Factors / Q000032:analysis* / Q000097:blood; D010179:Pancreas / Q000032:analysis; D012727:Sex Characteristics; D013116:Spinal Cord / Q000032:analysis*; D013154:Spleen / Q000032:analysis; D013363:Submandibular Gland / Q000032:analysis; D014018:Tissue Distribution
D009414:Nerve Growth Factors