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Highlights the sixth International Botanical Microscopy meeting in Saint Andrews, Scotland. Coverage of the meeting on research and technology in plant cell biology; Participation of speakers and audiences; Organization of the meeting through sessions.

We have fused the signal anchor sequences of a rat sialyl transferase and a human galactosyl transferase along with the Arabidopsis homologue of the yeast HDEL receptor (AtERD2) to the jellyfish green fluorescent protein (GFP) and transiently expressed the chimeric genes in tobacco leaves. All constructs targeted the Golgi apparatus and co-expression with DsRed fusions along with immunolabelling of stably transformed BY2 cells indicated that the fusion proteins located all Golgi stacks. Exposure of tissue to brefeldin A (BFA) resulted in the reversible redistribution of ST-GFP into the endoplasmic reticulum. This effect occurred in the presence of a protein synthesis inhibitor and also in the absence of microtubules or actin filaments. Likewise, reformation of Golgi stacks on removal of BFA was not dependent on either protein synthesis or the cytoskeleton. These data suggest that ER to Golgi transport in the cell types observed does not require cytoskeletal-based mechanochemical motor systems. However, expression of an inhibitory mutant of Arabidopsis Rab 1b (AtRab1b(N121I) significantly slowed down the recovery of Golgi fluorescence in BFA treated cells indicating a role for Rab1 in regulating ER to Golgi anterograde transport.

Hemicastration of rams at 1 week of age resulted in compensatory growth of the remaining testis. This was associated with an increase in the concentration of FSH in peripheral plasma which was maintained until the rams were approximately 10 weeks old. Plasma concentrations of LH were similar in both entire and hemicastrated lambs during this period. Pulsatile release of LH was observed in all rams from approximately 6 weeks of age onwards. The frequency of these pulses increased and the size of each pulse declined as the animals grew older. At 16 weeks pulsatile release was almost undetectable and the basal levels of LH had increased. The increased concentration of FSH in plasma was related to the size of the remaining testis but not to its development. The return of the plasma concentration of FSH to values similar to those found in entire rams preceded the appearance of a lumen or of any primary spermatocytes in the seminiferous tubules. Restricting the development of the remaining testis in hemicastrated rams, by trapping it in the inguinal region, did not alter the FSH response to hemicastration. The administration of gonadotrophin releasing hormone (10 microgram) at 10, 13 and 18 weeks of age provoked release of similar quantities of LH in both entire and hemicastrated rams. There was also a substantial release of FSH at 10 weeks of age but this response declined with age and by 18 weeks there was little FSH released in any of the animals. This loss of response occurred earlier in the hemicastrated rams. The data suggest that in prepubertal rams the release of FSH but not of LH is restricted by a substance produced by the developing Sertoli cells.

The concentrations of prolactin in plasma were measured before and after the injection of TRH in pregnant ewes and in non-pregnant ewes during the breeding season and seasonal anoestrus. During the first 80 days of pregnancy basal concentrations or prolactin were low and stable but thereafter increased progressively to reach maximum values at Day 140. During a comparable time of year (December-April) basal concentrations of prolactin in non-pregnant ewes were not significantly different from those found between Days 20 and 120 of pregnancy. At other times of the year basal concentrations of prolactin in non-pregnant ewes were elevated during anoestrus and declined markedly at the onset of the breeding season. In pregnant and non-pregnant ewes the responsiveness of the pituitary gland to stimulation with TRH was significantly correlated with the basal concentration of prolactin in plasma: the raised concentration of prolactin at Day 140 of pregnancy and in July in non-pregnant ewes was associated with the greatest release of prolactin from the pituitary gland. A change in the responsiveness of the pituitary gland may play an important role in the overall control of prolactin secretion in the ewe.

The levels of FSH, LH and testosterone in the plasma were measured in cross-bred male lambs which were bilaterally castrated, hemicastrated or left entire at 1 week of age. Removal of one testis caused the level of FSH in the circulation to increase to a value midway between the levels found in entire and castrated lambs up to 12 weeks of age. The concentrations of LH and testosterone in the plasma were similar in the control and hemicastrated lambs. The concentration of FSH in the plasma of the hemicastrated lambs fell at 12--14 weeks of age, the expected time of puberty, to the level found in control animals of the same age. Although the possibility of increased release of LH cannot be excluded, the increase in the secretion of FSH is believed to account for the compensatory hypertrophy of the remaining testis observed in the hemicastrated lambs. Removal of one testis from mature Soay rams in sping (May) caused a small selective increase in the concentration of RSH in the plasma, compared with entire animals. This difference was maintained until July when there was a marked increase in the levels of FSH and LH in the hemicastrated rams such that an exaggeration of the normal seasonal changes in these hormones was observed. Compensatory hypertrophy of the remaining testis was observed in the hemicastrated animals.

The plasma concentrations of FSH, LH and prolactin in ewes were measured at frequent intervals during 24-h periods in anoestrus at different times from the onset of the breeding season. Ewes kept under natural daylength conditions were compared with ewes in which the onset of the breeding season was advanced by exposure to constant short days (8L : 16D). The concentrations of FSH during mid-anoestrus did not vary during the day and there was no effect of short days or any changes which could be associated with the onset of ovulation. In all of the ewes pulsatile releases of LH were observed on each sampling occasion. During mid-anoestrus the occurrence of LH pulses varied between 1 and 3/day and one of the pulses appeared to be synchronized with dawn. Exposure to short days did not affect the frequency of LH release. In both groups of ewes an increased frequency of LH pulses was observed in the period 12-14 days before the first ovulation but this was associated with a decrease in the magnitude of each pulse. Prolactin concentrations were raised during anoestrus and tended to be higher during the hours of darkness and in the early morning. Exposure to short days for 3 weeks abolished these diurnal changes and reduced the concentrations to non-detectable amounts.

In the rat a single i.v. injection of donor whole blood (0.5 ml) 7 days before a renal allograft leads to complete suppression of rejection and permanent acceptance of the graft in DA(RT1a)-to-LEW(RT1(1], LEW-to-DA, and (DA X LEW)F1-to-DA combinations. This effect is donor-specific. Purified erythrocyte preparations were used for pretreatment (8 X 10(9) erythrocytes) 7 days before transplantation in the same strain combinations. The contamination by leukocytes was 1300 per 8 X 10(9) erythrocytes after one method of preparation, but only 1.3 per 8 X 10(9) erythrocytes after further purification by affinity chromatography. Both preparations of purified erythrocytes were able to suppress rejection and induce permanent acceptance of an allograft. The role of contaminating leukocytes in the first preparation was further excluded by pretreating rats with 1300 lymphocytes, but in these animals allografts were rejected in the normal fashion. The effect of the erythrocyte pretreatment was donor-specific, dose-dependent, only effective if given by the i.v. route, and not effective if given less than 7 days before transplantation. Furthermore neither lymphocytotoxic nor erythrocyte binding antibody could be detected in rats pretreated with purified erythrocytes. These experiments show convincingly that, in the rat--in the strain combinations tested--pretreatment with erythrocytes, that express cell surface class I MHC antigens, but not class II products, can induce specific suppression of rejection of a renal allograft.