The eupyrene-apyrene dichotomous spermatogenesis of Lepidoptera. The relationship with postembryonic development and the role of the decline in juvenile hormone titer toward pupation.

The relationships between the stages of postembryonic development and the occurrence of eupyrene and apyrene spermatogenesis, and the effects of the decline of the juvenile hormone (J.H.) titer toward pupation in these processes, were studied in the carob moth, Ectomyelois ceratoniae. The accurate timing of the spermatogenetic events was determined daily from the 2nd instar larva to the imago in squashes and electron microscope preparations of testes. Eupyrene spermatids elongate in two phases. In the first, beginning in late 4th instar larva, only flagella elongate, while in the second, beginning in the mid 5th instar larva, both flagella and nuclei elongate. Apyrene spermatogenesis starts just after the beginning of the nuclear elongation of eupyrene spermatids, in the mid 5th instar larva and not in the pupa, as is commonly believed. Using ligatures, topical applications of a J.H. mimic, and testes transplantation, it was found that the nuclear elongation begins in the 5-day-old eupyrene spermatid and cannot be induced earlier; the elongation is inhibited by high titer of the J.H. mimic. Elongation of the flagella, however, is unaffected by fluctuations of the J.H. titer. The onset of the apyrene spermatogenesis, which occurs in the very early 5th instar larva or before, was found to be unrelated to the decline in the J.H. titer toward pupation.     

Immune responses to weakly immunogenic virally induced tumors. III. Genetically unrestricted cytolysis of allogeneic tumor target cells.

Splenocytes from A mice injected with YAC-1 or RBL5 could generate, after in vitro culture with or without stimulation, a genetically nonrestricted cytotoxic response against the allogenic tumor RBL5. YAC-1 tumor is an in vitro carried tumor induced in A mice (H-2a) by Moloney virus. RBL5 tumor is a Rauscher virus-induced tumor of C57BL/6 mice (H-2b). These tumors cross-react serologically. The effector cells that were generated after the in vitro cultivation recognized tumor-associated antigens on the target cells. H-2 alloantigens were not recognized by the effector cells. The effector cells that killed RBL5 tumor in a genetically nonrestricted manner were identified as T cells. The in vivo carried tumor YAC, in contrast to the in vitro carried tumor YAC-1, could not induce anti-RBL5 reactive cells in A mice. Instead, YAC tumor induced suppressor cells in A mice, which could abrogate the anti-RBL5 cytotoxic response of RBL5-primed splenocytes, but not that of YAC-1 primed splenocytes.     

Dissociation between release and gene expression of gonadotropin alpha-subunit in gonadotropin-releasing hormone-stimulated alpha T3-1 cell line.

The alpha T3-1 cell line which was derived by targeted tumorigenesis in transgenic mice [Windle et al. (1990) Mol. Endocrinol. 4, 597-603] possesses high-affinity binding sites for GnRH analogs coupled to enhanced phosphoinositide turnover and phospholipase D activity. Incubation of alpha T3-1 cells with [D-Trp6]-GnRH analog (GnRH-A) resulted in a rapid increase in gonadotropin alpha-subunit mRNA levels which was detected already at 30 min of incubation (0.1 nM GnRH-A, 3-fold, p < 0.01). The effect diminished with time to reach basal levels at about 12 h of incubation, with a secondary rise in alpha mRNA levels between 12 and 24 h of incubation. Addition of the protein kinase C activator 12-O-tetradecanoylphorbol 13-acetate (TPA, 100 ng/mL) or the Ca2+ ionophore ionomycin (1 microM) to alpha T3-1 cells also resulted in a rapid increase in alpha-subunit mRNA levels. Surprisingly, GnRH-induced alpha-subunit release was detected only after a lag of 4 h of incubation. Thus, dissociation between exocytosis and gene expression can be demonstrated in GnRH-stimulated alpha T3-1 cell line.