Sexual differences in the total content of ribosomal RNA, established by cytophotometric measurements in neurones from selected brain regions, were studied in rats of the Wistar strain. In females of reproductive age, cyclic changes of RNA were synchronized with their oestrous cycle, the values being higher in the oestrous phase than in dioestrus. These changes were observed in pyramidal cells of the hippocampus and of the frontal cortex, in cells of anterior thalamus, of ventromedial and lateral hypothalamus and of tuberculi olfactorii. However, cycling cells were not disclosed in septum and thalamus posterior. A dependence upon the actual level of ovarial hormones was found in ventromedial hypothalamic cells only. In general, the RNA values in males of the same age corresponded to values of dioestrous females. The differences between newborn and 7-day-old pups were not marked enough and did not allow to define the critical period responsible for initiation of this sexual difference. In senescent rats, this difference persisted. During the stable phase of long-lasting dioestrus, the total RNA content in cells of the frontal cortex, hippocampus and hypothalamus was higher in females than in males of the same age which may suggest a faster reduction of this substance in aged males. The prolonged influence of oestrogens in the oestrous phase of the climacteric period (preceding the permanent dioestrus) decreased the RNA values in hippocampal and hypothalamic neurones even below the level established during the permanent dioestrus (and thus reached male values). On the contrary, in frontal cortical neurones, the female values remained higher in the permanent dioestrus as well as during long-lasting oestrus. A discussion concerns the possible participation of genetic determination and of the actual state of ovarial hormones in the manifestation of sexual differences in brain cells of the rat.
An allogeneic reaction among brain cortex cells (mixed reaction) was demonstrated previously by H-2 alloantigen- induced uncoupling of oxidative metabolism (Kovářů Med. Biol. 58: 273, 1980). In the present study we have demonstrated that alloantigen already increased cell surface Na + ,K+-ATPase activity after 100 min when the enzyme activation was highest at Mg2+/ATP ratio 4: 1. The allogeneic cell reaction was accompanied by an elevation of membrane lipid fluidity and probably also by a thermotropic lipid phase transition which might influence the membrane lipid-dependent Na+,K+-ATPase activity, while Mg2+-ATPase remained unaffected. Furthermore, the effects of proteins and peptides released into the supernatant during the allogeneic reaction were analyzed in brain cortex cells. One of the isolated active peptide fractions, Fa (m.w. lower than 2.5 kD), was able to enhance Na+,K+-ATPase activity as well as to block K+-evoked O2 uptake by brain cortex cells. Thus the Fa fraction simulated primary allorecognition events. The data indicate that various brain cell surface domains were influenced by a regulatory peptide fraction of the cytokine type during the early phase of allogeneic reaction. Allorecognition among brain cortex cells is directed against functionally important metabolic reactions.
This study describes the effects of cytokine peptides released into the supernatant during an early allogeneic reaction (AR) of mouse spleen lymphocytes or brain cortex cells which differ in their major histocompatibility complex (MHC). The peptides were isolated by ultrafiltration, liquid chromatography and HPLC. We found that both peptides stimulated the cell surface Na+,K+-ATPase and Ca2+-ATPase activities of quiescent spleen lymphocytes in vitro and mimicked early allogeneic cell interactions. Both brain and spleen AR peptides inhibited Concanavalin A-stimulated spleen lymphocyte proliferation, whereas 3H-TdR incorporation into DNA of the E7 neuroblastoma cell Une was stimulated by these peptides. The peptide isolated from the supernatant of the allogeneic brain ceU reaction inhibited phagocytosis in phorbol myristate-stimulated LA5-9/8 mouse macrophage cell line. Immunosuppressive activity of spleen AR peptide is supported by inhibition of spontaneous E rosette formation by lymphocytes. The immunosuppressive effect of isolated peptide cytokines on lectin-activated lymphocytes was comparable with the serum thymic factor (FTS, Lenfant et al. 1983). These changes demonstrate the pleiotropic cytokine actions mediated by plasma membrane of immune system and brain cells.