Pineal glands were grafted under the kidney capsule of mature male rats for periods of 20, 40, 60 and 100 days. Each grafted gland was then excised and divided into two halves. One half was processed for conventional electron microscopy and the other was fixed in aldehydes and then incubated in a zinc-iodide-osmium tetroxide mixture at pH 4.4 (A-ZIO-4.4). During the forty days following the operation pinealocytes showed the typical ultrastructural features associated with cells with a high protein and/or peptide secretory activity. On the other hand, during this period, the number of granular vesicles decreased progressively. From day 40 on, the grafted pinealocytes lacked granular vesicles. During the second half of the experimental period the ultrastructure of the pinealocytes indicated that their secretory activity was considerably decreased. During the acute phase of the experimental period numerous structures regarded as the tip of growing axons as well as typical nerve fibres appeared around blood vessels and within the paren chyma of the grafted gland. In the transplanted tissue obtained 60 and 100 days after the operation the growth cones were scarce, whereas typical nerve endings became numerous. These endings contained small clear vesicles which reacted positively when the tissue was treated with A-ZIO-4.4. The secretory activity of the grafted pineal gland and the nature of the nerve fibres which innervate the graft are discussed.

The aim of this study was to characterize the methylation system of phosphatidylethanolamine (PE) in the ovarian membranes and identify their possible role on the ovarian physiology. The presence of two methylating activities was found, by using S-adenosyl-L-[methyl-3H] methionine (SAM) as methyl donor. One of them (Met I) uses PE as substrate, and the other one, (Met II), catalyses the two successive methylations of phosphatidyl-N-monomethylethanolamine (PME) to phosphatidylcholine (PC). Met I shows a Km value of 5.71 microM and a Vm of 1.53 pmol/mg protein x min, working at pH 6.5, which seems to be the optimum. Met II exhibits low affinity for SAM, a Km of 50 microM, a Vm of 1.2 pmol/mg protein x min and it works an optimum pH of 9.5. Those enzymatic properties are in agreement with the amount of monomethylated and trimethylated products found working at pH 6.5 and 9.5, respectively. In order to investigate whether this enzyme system is affected or not by the hormonal environment influencing the ovary during the estrous cycle, the methylating activity was measured at its different stages. Both methylating activities were induced on proestrus but only Met I on diestrus. The total methylating activity increases when the ovarian membranes were obtained from rats injected with pregnant mare serum gonadotrophin (PMSG). Those results suggest that phospholipid methylation could be regulated by the hormonal environment during the estrous cycle.